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Kurd, Soleiman, Sara Hosseini, Fardin Fathi, Vahid Jajarmi, and Mohammad Salehi. "Dimethyl sulphoxide and electrolyte-free medium improve exogenous DNA uptake in mouse sperm and subsequently gene expression in the embryo." Zygote 26, no. 5 (October 2018): 403–7. http://dx.doi.org/10.1017/s0967199418000436.
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SummaryOne of the methods to generate transgenic animals is called sperm-mediated gene transfer (SMGT). Mature sperm cells can take up exogenous DNA molecules intrinsically and transfer them into the oocyte during fertilization. This study assessed the effect of dimethyl sulfoxide (DMSO) and electrolyte-free medium (EFM) on DNA uptake (EGFP–N1plasmid) in mouse sperm. Sperms cells cultured in human tubular fluid (HTF) without any treatment were considered as the control group. Sperms cells that were incubated in EFM and HTF with DNA/DMSO at 4°C were classified into EFM and HTF groups. Sperm motility and viability were assessed following treatment. In vitro fertilization (IVF) with sperm in all groups was performed. Fertilization, embryo development and GFP-positive blastocyst rates were analyzed and compared. The result showed that sperm motility and viability in EFM were better than those in the HTF group. The rate of development to reach the blastocyst stage and GFP-positive blastocysts was significantly higher in the EFM group compared with the HTF group (P<0.05). Our data demonstrate that sperm stored in the EFM group can improve the efficiency of SMGT for the generation of GFP-positive blastocysts.
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Belayeva, N. S. "Relation of the male gametes with embryo sec cells. The hypothesis of double fertilization." Acta Societatis Botanicorum Poloniae 50, no. 1-2 (2014): 173–75. http://dx.doi.org/10.5586/asbp.1981.026.
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The sperms one after another get out of synergids. The front sperm gets the first into the egg cell attraction zone and then the sperm comes into contact with egg membrane. At this moment Attraction ceases and the second sperm is led by a current of cytoplasma to the central nucleus. In the egg cell the sperm nucleus is led to the nucleus by cytoplasmic current too. After fertilization the character of cytoplasmic motion changes, because of a cell membrane damage. The presence of the sperm in the female nuclei may also serve as a regulating factor.
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Xiong, Hui, Xiangchen Li, Qingyun Hu, Pengfei Hu, and Weijun Guan. "Chicken Model for Inducing Primordial Germ Cells Differentiating into Motile Sperms in vitro." Avian Biology Research 10, no. 1 (February 2017): 12–18. http://dx.doi.org/10.3184/175815617x14799886572986.
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Stem cell technologies have been widely used in the study of spermatogenesis. However, deriving motile sperm from stem cells in vitro is still rarely achieved. We found that chicken primordial germ cells could directly differentiate into sperm by using retinoic acid in a non-testicular culture system. The induced sperms were characterised by RT-PCR, immunofluorescence and flow cytometry techniques. Results suggested that chicken primordial germ cells could produce motile sperm in vitro. Our work has provided a novel animal model of spermatogenesis in vitro, which might be used for male reproductive mechanism research.
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Bukowska, D., B. Kempisty, J. Sikora, M. Jackowska, M. Wozna, P. Antosik, H. Piotrowska, J. Budna, and JM Jaskowski. "The effect of swim-up purification and incubation of cells on sperm viability in dogs of different ages." Veterinární Medicína 56, No. 5 (June 10, 2011): 248–54. http://dx.doi.org/10.17221/1560-vetmed.
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The influence of selected semen extenders on the motility of frozen-thawed dog spermatozoa has been clearly demonstrated in several studies, although there are no reports indicating the effect of swim-up purification on sperm viability in this species of mammals. Therefore, this study was aimed at investigating phosphatidylserine (PS) externalization and necrosis in sperm after variable lengths of time of in vitro incubation after swim-up purification. Dog semen samples were collected from (i) ten dogs aged six months to 1.5 year, (ii) ten dogs aged six to eight years, and (iii) ten dogs aged 11 to 13 years. A flow cytometric method was employed to evaluate dog sperm viability in animals of different age groups after employment of a swim-up (SU) purification technique. After SU spermatozoa were incubated for 15, 30, 45 and 60 min in Sperm-TALP medium. We observed an increase in the number of viable sperm (double negatives) after 15 min of incubation compared to sperm undergoing PS externalization and late necrotic sperms (P < 0.001) in each group of dogs. We also found a higher number of early necrotic sperm after 60 min of in vitro incubation (P < 0.001). The amounts of late necrotic sperm and cells with PS externalization were similar among animals of different age groups. We show for the first time that most viable sperm are recovered after an in vitro incubation step of 15 min (control samples in this study) because as the time of incubation increases so does the number of degenerated or damaged cells. The higher number of early necrotic cells after 60 min of in vitro incubation may be a special feature of this species and may result from the induction of necrosis in the sperm. This knowledge may be used in future experiments for the preparation of spermatozoa following in vitro fertilization in dogs.
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Xie, Lan, Rui Ma, Chao Han, Kai Su, Qiufang Zhang, Tian Qiu, Lei Wang, et al. "Integration of Sperm Motility and Chemotaxis Screening with a Microchannel-Based Device." Clinical Chemistry 56, no. 8 (August 1, 2010): 1270–78. http://dx.doi.org/10.1373/clinchem.2010.146902.
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BACKGROUND Sperm screening is an essential step in in vitro fertilization (IVF) procedures. The swim-up method, an assay for sperm motility, is used clinically to select the ideal sperm for subsequent manipulation. However, additional parameters, including acrosome reaction capability, chemotaxis, and thermotaxis, are also important indicators of mammalian sperm health. To monitor both sperm motility and chemotaxis simultaneously during sperm screening, we designed and constructed a microdevice comprising a straight channel connected with a bibranch channel that mimics the mammalian female reproductive tract. METHODS The width and length of the straight channel were optimized to select the motile sperms. We selectively cultured cumulus cells in the bibranch channel to generate a chemoattractant-forming chemical gradient. Sperm chemotaxis was represented by the ratio of the sperm swimming toward different branches. RESULTS The percentage of motile sperms improved from 58.5% (3.8%) to 82.6% (2.9%) by a straight channel 7 mm in length and 1 mm in width. About 10% of sperms were found to be chemotactically responsive in our experiment, which is consistent with previous studies. CONCLUSIONS For the first time, we achieved the combined evaluation of both sperm motility and chemotaxis. The motile and chemotactically responsive sperms can easily be enriched on a lab-on-a-chip device to improve IVF outcome.
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L Zanella, Eraldo. "Melatonin Effect on Cryopreserved Sperm Cells of Crioulo Stallions." Open Access Journal of Veterinary Science & Research 5, no. 2 (2020): 1–8. http://dx.doi.org/10.23880/oajvsr-16000202.
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The freezing/thawing process of spermatozoa can cause cellular damage to the male gamete, decreasing the fertilization potential due to the increase in the production of reactive oxygen species (ROS). Melatonin is a potent endogenous antioxidant that protects the body against the damage caused by ROS. This study has evaluated different melatonin concentrations on the sperm viability of cryopreserved semen of Crioulo stallions. For that, three ejaculates were collected from five stallions diluted in a commercial extender followed by centrifugation and resuspension in a commercial freezing extender supplemented with 0; 1.25; 2.5. 5mM of Melatonin before the cryopreservation process. After thawing, the evaluation was performed assessing motility and flow cytometry evaluations: the plasma membrane integrity (PI), the integrity of the acrosomal membrane (FITC-PNA), mitochondrial membrane potential (JC1), and ROS generation (DCF-DA). Our results showed that sperm motility in the group without Melatonin and the 1.25mM group did not show the difference; however, the groups 2.5mM and 5mM presented a reduction in sperm motility. The 1.25 mM concentration was able to protect the plasma membrane during the cryopreservation process, in addition to showing a significant reduction in the production of ROS and increasing the percentage of sperm with integral acrosome. It can also be seen that high concentrations of Melatonin did not show beneficial effects. In conclusion, the addition of 1.25 mM of the Melatonin in Crioulo sperm cells showed to have a protective effect on the sperm cell during cryopreservation.
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Ding, Qiang, Xiuhu Ding, Shuwen Xia, Fang Zhao, Kunlin Chen, Yong Qian, Shaoxian Cao, et al. "Bta-miR-6531 Regulates Calcium Influx in Bovine Leydig Cells and Is Associated with Sperm Motility." Genes 13, no. 10 (October 3, 2022): 1788. http://dx.doi.org/10.3390/genes13101788.
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MicroRNAs (miRNAs) play key roles in sperm as the regulatory factors involved in fertility and subsequent early embryonic development. Bta-miR-6531 is specifically a highly enriched miRNA in low-motility sperms in previous study. To investigate the mechanism of bta-miR-6531, 508 shared target genes of bta-miR-6531 were predicted using two miRNA target databases (TargetScan7 and miRWalk). According to the Kyoto Encyclopedia of Genes and Genomes (KEGG), the calcium and cAMP signaling pathways were the most enriched of the target genes. A dual-luciferase assay indicated that bta-miR-6531 targeted ATP2A2 mRNA by binding to the coding sequence region. In bovine Leydig cells, bta-miR-6531 overexpression affected the intracellular calcium concentration by restraining ATP2A2 expression. Moreover, we observed high calcium concentrations and high ATP2A2 protein levels in high-motility sperm compared with those in low-motility sperms. Furthermore, high-linkage single-nucleotide polymorphisms (SNPs) of the pre-bta-miR-6531 sequence were identified that related to sperm traits. Genotype TCTC of bta-miR-6531 showed high sperm motility and density and low deformity rate in Holstein bulls. However, the mutation in pre-miR-6531 did not significantly affect mature bta-miR-6531 expression in the sperm or cell models. Our results demonstrate that bta-miR-6531 might involve in sperm motility regulation by targeting ATP2A2 of the calcium signaling pathway in bovine spermatozoa.
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Kondracki, Stanisław, Anna Wysokińska, Magdalena Kania, and Krzysztof Górski. "Application of two staining methods for sperm morphometric evaluation in domestic pigs." Journal of Veterinary Research 61, no. 3 (September 26, 2017): 345–49. http://dx.doi.org/10.1515/jvetres-2017-0045.
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Abstract Introduction: The effect of two smear staining methods on the dimensions and shape of sperm cells in the semen of domestic pigs was evaluated. Material and Methods: The studies were carried out on 30 ejaculates collected from 15 boars, which included five Duroc boars, five Pietrain boars, and five hybrid Duroc × Pietrain boars. Each ejaculate was next sampled to make two microscopic slides, of which one was stained with eosin-nigrosin and the other with eosin-gentian dye. In total, 600 measurements of sperm cells were made. Each sperm was measured for the following morphometric parameters: head length, head width, head area, head perimeter, tail length, and the total sperm length. Results: Sperms measured on slides stained with eosin-nigrosin showed lower dimensions as compared with those stained with the eosin-gentian dye method. Sperm stained with eosin-nigrosin had shorter and narrower heads than sperm stained with eosin-gentian dye. The method of staining, therefore, affected not only the dimensions of the sperm, but also the proportions of the dimensions defining the shape of the sperm. Conclusions: The size and shape parameters in porcine sperm may take on different values depending on the method of semen staining. Sperm cells stained with eosin-nigrosin are smaller than the sperm stained with eosin-gentian dye. The sensitivity of the sperm to the type of dye used for the fixation may be associated with genetic factors.
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Khodamoradi, Maedeh, Saeed Rafizadeh Tafti, Seyed Ali Mousavi Shaegh, Behrouz Aflatoonian, Mostafa Azimzadeh, and Patricia Khashayar. "Recent Microfluidic Innovations for Sperm Sorting." Chemosensors 9, no. 6 (June 1, 2021): 126. http://dx.doi.org/10.3390/chemosensors9060126.
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Sperm selection is a clinical need for guided fertilization in men with low-quality semen. In this regard, microfluidics can provide an enabling platform for the precise manipulation and separation of high-quality sperm cells through applying various stimuli, including chemical agents, mechanical forces, and thermal gradients. In addition, microfluidic platforms can help to guide sperms and oocytes for controlled in vitro fertilization or sperm sorting using both passive and active methods. Herein, we present a detailed review of the use of various microfluidic methods for sorting and categorizing sperms for different applications. The advantages and disadvantages of each method are further discussed and future perspectives in the field are given.
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Bhat, Ghulam Rasool, Farooz Ahmad Lone, and Nafis Ibni Assad. "Spermbots: A Promising Futuristic Innovation in Assisted Reproductive Technology." Indian Journal of Animal Reproduction 44, no. 2 (December 29, 2023): 14–17. http://dx.doi.org/10.48165/ijar.2023.44.02.3.
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Spermbots are robotic sperms formed out of sperm cells conjugated to artificial microstructures, having potential applications ranging from biomedical processes, drug delivery systems, in situ real time imagery and assisted reproduction. The robotic sperm can act as an exploratory microdevice in biological networks. Incorporation of a biological entity like sperm into microstructures under the environment of magnetic or electric fields helps in shape templating and carrying chemotherapeutic agents to target sites. Besides its role in drug delivery systems, spermbots can potentially contribute in combating infertility, especially in oligo-zoospermia and necro-zoospermia in males. Numerous checkpoints may impede sperm cells to reach the oocyte in vivo. Spermbots bypass these sites and carry sperm to oocyte. Targeted delivery always requires interventions of natural functional aspects of living systems. Sperm flagellum, being a biological motor in nature, can be harnessed as a driving force in spermbots to ensure delivery of fertile sperm cells with impaired motile machinery to the target. Moreover, the technology has a potential to unravel sperm migration patterns and understanding the processes in vivo. After a review of documented literature on possible use of spermbot technology in assisted reproduction, we hereby discuss the application of this new and innovative technology in humans and animals. The paper also highlights certain shortfalls in the widespread application of this cargo delivery technology in assisted reproduction.
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HENRY, CELIA. "SORTING OUT SPERM CELLS." Chemical & Engineering News Archive 83, no. 3 (January 17, 2005): 15. http://dx.doi.org/10.1021/cen-v083n003.p015.
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Friedrich, B. M., and F. Julicher. "Chemotaxis of sperm cells." Proceedings of the National Academy of Sciences 104, no. 33 (August 8, 2007): 13256–61. http://dx.doi.org/10.1073/pnas.0703530104.
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Brugger, Peter, Ervin Macas, and Jürgen Ihlemann. "Do sperm cells remember?" Behavioural Brain Research 136, no. 1 (October 2002): 325–28. http://dx.doi.org/10.1016/s0166-4328(02)00127-4.
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Weber, Martina. "Unusual sperm cells inApiaceae." Plant Systematics and Evolution 159, no. 3-4 (1988): 273–76. http://dx.doi.org/10.1007/bf00935978.
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Gupta, Sachin, Ruchi Yadav, and Mayank Thakur. "In-vitro sperm immobilization activity and biocompatibility study of NVD terpolymer." International Journal of Basic & Clinical Pharmacology 6, no. 5 (April 24, 2017): 1221. http://dx.doi.org/10.18203/2319-2003.ijbcp20171680.
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Background: Boronate derivatives have been used in affinity chromatography for separation of cells based on their glycoprotein content. Boronate containing polymers when used intravaginally before sexual intercourse could bind to the glycoproteins present on sperm cell surface and render them immobile, which potentially may work as female-controlled contraceptive. To study this hypothesis NVD terpolymer which contains boronic acid was studied on goat sperm and its biocompatibility was accessed on NIH3T3 fibroblast.Methods: Sperm motility study was carried out on goat sperm cells. The study was divided into two groups, test group (NVD terpolymer in simulated vaginal fluid) and negative control (simulated vaginal fluid only) performed using Sander-crammer assay. In the test group, the study was started from 0.1 % of the polymer solution, until the half of the sperms became non-motile as compared to normal control. The biocompatibility study was performed by culturing the NIH3T3 fibroblast with different concentrations of NVD polymer, followed by cell viability assay by performing 3-(4, 5-dimethylthiazol-2-Yl)-2, 5-diphenyltetrazolium bromide (MTT) assay at 6, 12 and 24 hour spectrophotometrically.Results: Sander-crammer assay resulted in significant (P-value < 0.05) decrease in motile sperm count in test group when compared to control. At 7.5 % concentration, the half of the sperms rendered immobile, and this was termed as effective concentration 50 (EC50). In-vitro biocompatibility study using NIH3T3 fibroblasts culture and MTT assay with cultured cells at 6, 12 and 24 hour, revealed that the polymer is biologically compatible as there were no significant change (P-value < 0.05) in the absorbance.Conclusions: Boronate containing polymer, such as NVD terpolymer has in-vitro sperm immobilizing activity in goat sperm model, with further research in this area could yield a potential female control contraceptive agent.
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Krishnamurthy, Thejasvi, and Yashica Gowda R. "Correlation of Total Sperm Count with Presence of Round Cells and Abnormal Sperm Morphology in Semen Analysis." Annals of Pathology and Laboratory Medicine 6, no. 5 (May 24, 2019): A271–276. http://dx.doi.org/10.21276/apalm.2372.
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Amiri, Banafshe, and Bahare Amiri. "The effect of human papilloma virus on women reproductive system, pregnancy outcome of mothers: a review." Nursing & Care Open Access Journal 8, no. 3 (December 1, 2022): 88–90. http://dx.doi.org/10.15406/ncoaj.2022.08.00245.
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Human papillomavirus is a type of sexually transmitted virus that is common in people with factors such as multiple sexual partners, a weakened immune system caused by anxiety, or alcohol and tobacco users. This virus (human papilloma) causes negative effects on vaginal and cervical epithelial cells, sperm quality and other fertility parameters. Human papilloma virus can exist in the seminal fluid caused by ejaculation and play a role in the fertility quality of men. Different viruses may directly or indirectly infect male gametes, sperms. Decreased fertility quality is caused by changes in sperm structure, increased sperm DNA fragmentation, and increased sperm aneuploidy. As a result, it causes a decrease in the quality of fertility in women and men and failure of assisted reproductive methods, and pregnancy complications such as frequent miscarriages, premature births and weight loss at birth. HPV causes precancerous cells, low-grade and high-grade cancer (LSIL-HSIL).
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Cass, David D., and George C. Fabi. "Structure and properties of sperm cells isolated from the pollen of Zea mays." Canadian Journal of Botany 66, no. 5 (May 1, 1988): 819–25. http://dx.doi.org/10.1139/b88-119.
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Sperm cells can be isolated from maize pollen grains by gentle osmotic treatment. After the initial osmotic burst, the slurry is sieved through nylon mesh and then subjected to sucrose density gradient centrifugation to remove as much starch as possible. The isolated cells are spherical and approximately 7.2 μm in diameter, exclude Evans blue stain up to several hours at room temperature, and can be frozen and stored with reasonable retention of viability. Little alteration of the appearance of the living cells is seen after preparation for transmission electron microscopy. There is ultrastructural confirmation that maize sperm cells are protoplasts. Furthermore, there is no evidence of vegetative cell inner membrane on the outside of the sperms. Internally, there is a normal organelle complement including mitochondria, dictyosomes, protein bodies, and endoplasmic reticulum. The latter component is extensive and has associated ribosomes. Some coated vesicles occur in maize sperm protoplasts. Nuclear morphology is quite variable, including spherical, lobed, and irregular shapes.
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Solomon, I. P., E. F. Istifanus, and E. J. Idang. "Effect of frequency of ejaculation and graded dietary levels of Roselle (Hibiscussabdariffa) calyx meal supplementation on sperm profile of mongrel rabbit bucks." Nigerian Journal of Animal Production 49, no. 5 (October 20, 2022): 162–73. http://dx.doi.org/10.51791/njap.v49i5.3780.
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Changes in semen quality occasioned by ejaculation frequency is one of the important yet neglected factor considered in artificial insemination. Several factors have been shown to influence semen parameters, one of which is sexual abstinence. Rabbits have a very short interval between successive ejaculations hence, semen can easily be collected for quality examination. This experiment was carried out to evaluate the effect of frequency of ejaculation and graded dietary levels of Roselle calyx meal supplementation on sperm profile of mongrel rabbit bucks. Twenty healthy mongrel rabbit bucks were used for the study. The rabbits were managed intensively and were provided with feed, water and forages ad libitum. Four experimental diets were formulated to contain dietary levels of Roselle calyx meal at 0.00% (control), 2.00%, 4.00% and 6.00% and coded as T1 , T2 , T3 and T4 respectively.The four treatment groups were assigned to four experimental diets in a completely randomized design (CRD) each replicated three times with two (2) rabbits per replicate. Each replicate received an assigned diet for 8 weeks. The experimental data obtained were subjected to analysis of variance (ANOVA) procedure. Semen volume, colour, motility, concentration, live sperm percentage, abnormal sperm percentage, total sperm cells per ejaculate and reaction time were evaluated in this study. From the results obtained, semen volume, sperm concentration, live sperm cells, reaction time and total cells per ejaculate were significantly (p<0.05) influenced by frequency of ejaculation while sperm motility and abnormal cells did not show (p>0.05) statistical variation. Roselle calyx meal supplementation in mongrel rabbit bucks' diet influenced all sperm indices except sperm motility and reaction time. The highest semen volume was observed in once per week frequency of ejaculation with a value of 1.01ml while twice per week frequency of ejaculation recorded a lower value of 0.85ml. There was interaction (p<0.05) between frequency of ejaculation and graded dietary levels of Roselle calyx meal on semen volume of bucks in this study. Live sperm cells were higher (82.51%) in once per week frequency of ejaculation than in twice per week (77.08%). From the findings of this study, once a week frequency of ejaculation and Roselle calyx meal supplementation at 2.0% improved sperm characteristics of mongrel rabbit bucks.
Les changements de qualité de sperme occasionnés par la fréquence d'éjaculation sont l'un des facteurs importants mais négligés considérés dans l'insémination artificielle. Il a été démontré que plusieurs facteurs influencent les paramètres de sperme, dont l'une abstinence sexuelle. Les lapins ont un très court intervalle entre éjaculations successives donc, le sperme peut facilement être collecté pour un examen de qualité. Cette expérience a été réalisée pour évaluer l'effet de la fréquence de l'éjaculation et des niveaux diététiques de RoselleCalyx Supplémentation des repas sur le profil de sperme de Daims de lapin de Mongrel. Vingt daims de lapin de Son ont été utilisés pour l'étude. Les lapins ont été gérés de manière intensive et ont été fournis avec des aliments pour animaux, de l'eau et des forages ad-libitum. Quatre régimes expérimentaux ont été formulés pour contenir des niveaux diététiques de farine de CalyxRoselle à 0,00% (contrôle), de 2,00%, 4,00% et 6,00% et codés comme T , T , T et T respectivement. Les quatre groupes de traitement ont été attribués à quatre régimes expérimentaux dans une conception entièrement randomisée (CER) répliquée à trois reprises avec deux (2) lapins par réplication. Chaque réplique a reçu une alimentation assignée pendant 8 semaines. Les données expérimentales obtenues ont été soumises à une analyse de la procédure de variance (ANOVA). Volume de sperme, couleur, motilité, concentration, pourcentage de sperme en direct, pourcentage de sperme anormal, cellules de sperme total par éjaculation et temps de réaction ont été évalués dans cette étude. Des résultats obtenus, le volume de sperme, la concentration de sperme, les spermatozoïdes en direct, le temps de réaction et les cellules totales par éjaculation ont été significativement (p <0,05) influencés par la fréquence d'éjaculation tandis que la motilité des spermatozoïdes et les cellules anormales n'ont pas montré (p> 0,05) variation statistique. RoselleCalyx Supplément repas dans le régime de Daims de lapin de Mongrel influencé tous les indices de sperme sauf la motilité du sperme et le temps de réaction. Le volume de sperme le plus élevé a été observé en fréquence d'éjaculation une fois par semaine avec une valeur de 1,01 ml, tandis que la fréquence d'éjaculation de deux fois par semaine enregistrait une valeur inférieure de 0,85 ml. Il y avait une interaction (p <0,05) entre la fréquence d'éjaculation et les niveaux diététiques gradués du repas RoselleCalyx sur le volume de sperme de dollars dans cette étude. Les spermatozoïdes en direct étaient plus élevés (82,51%) en fréquence d'éjaculation une fois par semaine qu'à deux fois par semaine (77,08%). Des conclusions de cette étude, une fois par semaine, une fréquence d'éjaculation et une supplémentation de repas RoselleCalyx à 2,0% des caractéristiques de sperme améliorées de Daims de lapin de Mongrel.
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Kavaldzhieva, K. K., D. K. Dimitrova-Dikanarova, K. S. Mladenova, V. V. Lazarov, and N. Y. Mladenov. "The Death of Sperm Cells." Acta Medica Bulgarica 50, no. 4 (December 1, 2023): 69–72. http://dx.doi.org/10.2478/amb-2023-0046.
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Abstract A major factor affecting male fertility is excessive death of germ cells, both immature germ cells and mature spermatozoa. It can be due to various factors causing testicular and/or post-testicular damage, such as infections, obstructive conditions, toxins, oxidative stress, hormonal imbalance, hyperthermia, and anti-sperm antibodies. Massive death of spermatozoa leads to a high proportion of dead sperm cells in the ejaculate (necrozoospermia or necrospermia) while death of immature germ cells can lead to low sperm count (oligozoospermia or oligospermia). Cell death can occur both by necrosis and by apoptosis; in recent decades, it has been found that apoptosis of mature spermatozoa is not only possible but quite common, and can contribute to infertility. Treatment approaches are primarily directed to the underlying condition, i.e. removing the cause(s) of sperm cell death whenever possible, but include also attempts to bypass the cell death event by intracytoplasmic sperm injection with testicular spermatozoa.
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Feng, Y. L., L. Ma, J. L. McKeeby, B. Y. Chen, and J. L. Hall. "Sexing Individual Sperm Cells During Intracytoplasmic Sperm Injection (ICSI)." Fertility and Sterility 74, no. 3 (September 2000): S18—S19. http://dx.doi.org/10.1016/s0015-0282(00)00774-3.
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Shamsadini, Fatemeh, Hadi Zare, Hossein Eslami, Mojtaba Ansari, Farzaneh Fesahat, and Esmat Mangoli. "Sambucus Nigra: black fruits that makes sperms come out ahead in in vitro fertilization." Acta Bioclínica 14, no. 27 (2024): 98–112. http://dx.doi.org/10.53766/acbio/2024.14.27.11.
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Freezing is a non-physiological method that includes a series of processes such as dilution, incubation, cooling, freezing and thawing, which affect the structure, biochemistry and function of sperm. Human sperm freezing has overcome many spatial and temporal limitations and is now an integral part of assisted reproductive technologies (ART). Freezing causes biochemical, functional and structural changes in sperm. These changes include plasma and sperm acrosome membranes, which increase their permeability and leads to disruption of morphology, motility and chromatin structure. Therefore, freezing causes irreversible damage to sperm, which leads to the loss of movement and biological function of most sperm. Sperm health is one of the important factors in the fertility of couples. Therefore, reducing the harmful effects caused by freezing and properly performing the in vitro Fertilization (IVF) process, the development of useful solutions is inevitable. The extract from black fruits of Elderberry (Sambucus nigra) has potent in vitro effect on the sperm motility, vitality and oxidative profile. This extract can largely improve the sperms parameters and stereological parameters (spermatogonia, primary spermatocyte, round spermatid, and Leydig cells), decrease of TNF-a and caspase-3 expression, increase of the serum testosterone, increase of mitochondrial activity, and reduction of reactive oxygen species. So, it can also be used as in vivo status. We suggested that black fruits of Sambucus nigra makes sperms come out ahead in IVF techniques.
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Schoeller, Simon F., William V. Holt, and Eric E. Keaveny. "Collective dynamics of sperm cells." Philosophical Transactions of the Royal Society B: Biological Sciences 375, no. 1807 (July 27, 2020): 20190384. http://dx.doi.org/10.1098/rstb.2019.0384.
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While only a single sperm may fertilize the egg, getting to the egg can be facilitated, and possibly enhanced, by sperm group dynamics. Examples range from the trains formed by wood mouse sperm to the bundles exhibited by echidna sperm. In addition, observations of wave-like patterns exhibited by ram semen are used to score prospective sample fertility for artificial insemination in agriculture. In this review, we discuss these experimental observations of collective dynamics, as well as describe recent mechanistic models that link the motion of individual sperm cells and their flagella to observed collective dynamics. Establishing this link in models involves negotiating the disparate time- and length scales involved, typically separated by a factor of 1000, to capture the dynamics at the greatest length scales affected by mechanisms at the shortest time scales. Finally, we provide some outlook on the subject, in particular, the open questions regarding how collective dynamics impacts fertility. This article is part of the theme issue ‘Multi-scale analysis and modelling of collective migration in biological systems’.
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Hansen, Jan, Sebastian Rassmann, Jan Jikeli, and Dagmar Wachten. "SpermQ–A Simple Analysis Software to Comprehensively Study Flagellar Beating and Sperm Steering." Cells 8, no. 1 (December 26, 2018): 10. http://dx.doi.org/10.3390/cells8010010.
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Motile cilia, also called flagella, are found across a broad range of species; some cilia propel prokaryotes and eukaryotic cells like sperm, while cilia on epithelial surfaces create complex fluid patterns e.g., in the brain or lung. For sperm, the picture has emerged that the flagellum is not only a motor but also a sensor that detects stimuli from the environment, computing the beat pattern according to the sensory input. Thereby, the flagellum navigates sperm through the complex environment in the female genital tract. However, we know very little about how environmental signals change the flagellar beat and, thereby, the swimming behavior of sperm. It has been proposed that distinct signaling domains in the flagellum control the flagellar beat. However, a detailed analysis has been mainly hampered by the fact that current comprehensive analysis approaches rely on complex microscopy and analysis systems. Thus, knowledge on sperm signaling regulating the flagellar beat is based on custom quantification approaches that are limited to only a few aspects of the beat pattern, do not resolve the kinetics of the entire flagellum, rely on manual, qualitative descriptions, and are only a little comparable among each other. Here, we present SpermQ, a ready-to-use and comprehensive analysis software to quantify sperm motility. SpermQ provides a detailed quantification of the flagellar beat based on common time-lapse images acquired by dark-field or epi-fluorescence microscopy, making SpermQ widely applicable. We envision SpermQ becoming a standard tool in flagellar and motile cilia research that allows to readily link studies on individual signaling components in sperm and distinct flagellar beat patterns.
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Berg, B. "Microscopic analysis of MTT stained boar sperm cells." Open Veterinary Journal 5, no. 2 (2015): 58. http://dx.doi.org/10.5455/ovj.2015.v5.i1.p58.
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The ability of sperm cells to develop colored formazan by reduction of MTT was used earlier to develop a spectrophotometric assay to determine the viability of sperm cells for several mammalian species. It was the objective of the present study to visualize microscopically the location of the formazan in boar sperm cells. The MTT staining process of boar sperm cells can be divided into a series of morphological events. Incubation of the sperm cells in the presence of MTT resulted after a few min in a diffuse staining of the midpiece of the sperm cells. Upon further incubation the staining of the midpiece became more intense, and gradually the formation of packed formazan granules became more visible. At the same time, a small formazan stained granule appeared medially on the sperm head, which increased in size during further incubation. After incubation for about 1 h the midpiece granules were intensely stained and more clearly distinct as granules, while aggregation of sperm cells occurred. Around 90% of the sperm cells showed these staining events. At the end of the staining the formazan granules have disappeared from both the sperm cells and medium, whereas formazan crystals appeared as thin crystal threads, that became heavily aggregated in the incubation medium. It was concluded that formazan is taken up by lipid droplets in the cytoplasm. Further, the use of the MTT assay to test for sperm viability should be regarded as a qualitative assay, whereas its practical use at artificial insemination (AI) Stations is limited.
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Mapeka, M. H., K. C. Lehloenya, M. L. Mphaphathi, and T. L. Nedambale. "219 IN VITRO FERTILIZATION RATE OF MATURED PIG OOCYTES BY FROZEN - THAWED KOLBROEK PIG SPERM CELLS." Reproduction, Fertility and Development 23, no. 1 (2011): 208. http://dx.doi.org/10.1071/rdv23n1ab219.
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No studies have investigated the IVF rate of South African indigenous Kolbroek sperm cells following cryopreservation. The objective of this study was to test if frozen–thawed Kolbroek pig sperm cells could penetrate pig oocytes matured in vitro. Pig ovaries were collected from a local abattoir and cumulus–oocytes complexes were obtained by aspiration and were then in vitro matured in TCM-199 supplemented with 10% pig follicular fluid, 10% fetal bovine serum, and 1 μg mL–1 of FSH and LH. Following 44 h of incubation, 200 matured pig oocytes were randomly assigned to 2 treatments with frozen–thawed and fresh (control) Kolbroek pig sperm cells. For IVF, Kolbroek sperm cells were in vitro capacitated using Brackett and Oliphant’s sperm wash medium. Matured pig oocytes and sperm cells were co-incubated for 24 h in Brackett and Oliphant’s IVF medium. Following fertilization, presumptive zygotes were in vitro cultured at 39°C in 5% CO2, 5% O2, and 90% N2. Rate of fertilization was identified by the number of cleaved zygotes. Data were analysed by ANOVA. The total motility of Kolboek pig sperm cells used for IVF was 40% for frozen–thawed sperm cells and 80% for fresh sperm cells. The results showed that Kolbroek pig sperm cells were able to penetrate pig oocytes in vitro. However, no significant (P < 0.05) difference was observed in the percentage of cleavage of pig oocytes fertilized with either frozen–thawed (13.25%) or fresh (13.0%) Kolbroek pig sperm cells. The percentage of embryos that developed to the morulae stage was 2% in frozen–thawed sperm cells and was 0% in fresh Kolbroek sperm cells. Furthermore, oocytes fertilized with Kolboek sperm cells did not develop to the blastocyst stage in either treatment. In conclusion, this study demonstrated that frozen–thawed Kolbroek sperm cells are able to fertilize matured pig oocytes in vitro. This study was funded by the Department of Agriculture Forestry and Fishery, ARC, DST-PDP (RT19000), and National Research Foundation (NRF, Grant No. RT21 and 24000).
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Nassir, Mayssam, Mattan Levi, and Natan T. Shaked. "Dynamic 3D Modeling for Human Sperm Motility through the Female Cervical Canal and Uterine Cavity to Predict Sperm Chance of Reaching the Oocyte." Cells 12, no. 1 (January 3, 2023): 203. http://dx.doi.org/10.3390/cells12010203.
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Sperm motility in the female genital tract is a key factor in the natural selection of competent cells that will produce a healthy offspring. We created a dynamic three-dimensional (3D) mechanical model of human sperm cells swimming inside cervical canal and uterine cavity dynamic 3D models, all generated based on experimental studies. Using these simulations, we described the sperm cells’ behaviors during swimming inside the 3D tract model as a function of 3D displacement and time. We evaluated normal- and abnormal-morphology sperm cells according to their chances of reaching the oocyte site. As expected, we verified that the number of normal sperm cells that succeeded in reaching the fallopian tube sites is greater than the number of abnormal sperm cells. However, interestingly, after inspecting various abnormal sperm cells, we found out that their scores changed compared to swimming in an infinite medium, as is the case with in vitro fertilization. Thus, the interactions of abnormal sperm cells and the complicated geometry and dynamics of the uterus are significant factors in the filtering of abnormal sperm cells until they reach the oocyte site. Our study provides an advanced tool for sperm analysis and selection criteria for fertility treatments.
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Manko, B. V., N. M. Kozopas, ,. H. M. Mazur, A. М. Voityk, B. O. Manko, and V. V. Manko. "Bioenergetic functions of mitochondria in liver, pancreatic acinar cells, and sperm cells of rats fed short-term high-fat or high-fat high-sugar diets." Ukrainian Biochemical Journal 95, no. 5 (November 6, 2023): 51–60. http://dx.doi.org/10.15407/ubj95.05.051.
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An unhealthy diet often is a cause of obesity, chronic inflammation, and metabolic disruption in multiple organs. However, the direct influence of elevated lipid or sugar consumption on liver, pancreatic, and sperm mitochondria is not well understood. The aim of the study was to investigate the functional activity of mitochondria of liver, pancreatic acinar cells, and sperm cells in rats on a short-term (7 weeks) diet with high fat or high fat and high sugar content. Male Wistar rats were on a basic, high-fat or high-fat high-sugar diet for 7 weeks. At the end of the experiment, visceral fat mass, blood glucose and lipids were measured. Mitochondrial functional activity was evaluated with oxygen consumption assay. In isolated pancreatic acinar cells, NAD(P)H autofluorescence and mitochondrial membrane potential were also studied. No difference in body mass was observed between the 3 groups at the end of the experiment. Visceral fat mass was slightly but significantly elevated in rats on a high-fat high-sugar diet. Both diets did not affect plasma glucose or triglyceride levels but caused a modest elevation of total plasma cholesterol. Respiration and oxidative phosphorylation of isolated liver mitochondria were not affected by any experimental diet. In pancreatic acinar cells, a high-fat diet caused a significant decrease of basal respiration by ~15%, but no effects were observed on the maximal rate of uncoupled respiration, mitochondrial membrane potential, or NAD(P)H autofluorescence. In these cells, a ketone body 3-hydroxybutyrate caused elevation of uncoupled respiration and NAD(P)H level irrespectively of the diet. Diets did not cause any change in sperm concentration, viability or motility. Surprisingly, in animals on a high-fat high-sugar diet, a significant increase in both basal and maximal respiration of sperm cells was observed. Collectively, these data show that while the elevated fat and sugar content in the diet does not cause significant obesity, no detrimental effects on mitochondria of the liver, pancreas, and sperm cells are observed. Keywords: diet, liver, mitochondria, pancreatic acinar cells, sperm
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Nassir, Mayssam, Mattan Levi, Gili Dardikman-Yoffe, Simcha K. Mirsky, and Natan T. Shaked. "Prediction of Sperm Progression in Three Dimensions Using Rapid Optical Imaging and Dynamic Mechanical Modeling." Cells 11, no. 8 (April 13, 2022): 1319. http://dx.doi.org/10.3390/cells11081319.
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We present a multidisciplinary approach for predicting how sperm cells with various morphologies swim in three-dimensions (3D), from milliseconds to much longer time scales at spatial resolutions of less than half a micron. We created the sperm 3D geometry and built a numerical mechanical model using the experimentally acquired dynamic 3D refractive-index profiles of sperm cells swimming in vitro as imaged by high-resolution optical diffraction tomography. By controlling parameters in the model, such as the size and shape of the sperm head and tail, we can then predict how different sperm cells, normal or abnormal, would swim in 3D, in the short or long term. We quantified various 3D structural factor effects on the sperm long-term motility. We found that some abnormal sperm cells swim faster than normal sperm cells, in contrast to the commonly used sperm selection assumption during in vitro fertilization (IVF), according to which sperm cells should mainly be chosen based on their progressive motion. We thus establish a new tool for sperm analysis and male-infertility diagnosis, as well as sperm selection criteria for fertility treatments.
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Bonyadi, F., S. Hasanzadeh, H. Malekinejad, and G. Najafi. "Cyclopiazonic acid decreases sperm quality and in vitro fertilisation rate in mice." World Mycotoxin Journal 11, no. 4 (December 7, 2018): 599–610. http://dx.doi.org/10.3920/wmj2018.2337.
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The presence of cyclopiazonic acid (CPA) as a mycotoxin has been reported in feed and foodstuffs. The aim of this investigation was to determine the effects of CPA on reproductive functions of male mice. In this experiment, 40 mature male mice were randomly assigned into five groups (n=8): control, control-sham, CPA (0.03 mg/kg, body weight (BW)), CPA (0.06 mg/kg, BW) and CPA (0.12 mg/kg, BW). Following 28 days exposure to CPA, sperm quality parameters, in vitro fertilisation (IVF) capacity of sperms, serum testosterone level, Leydig cells number and serum total antioxidant capacity (TAC) were analysed. The results revealed a significant (P<0.05) reduction in sperm count, sperm viability, sperm motility, chromatin quality of sperm, sperms with intact DNA, IVF rate, testosterone level, Leydig cell distribution and TAC in comparison to the control group. The most prominent detrimental effects of CPA were found at the highest given dose level. Our results suggest that CPA at higher dose levels exerts detrimental effects on the male reproductive system. Moreover, these descriptive warrant further investigations into the specific mechanisms of action and the effects of CPA on spermatogenesis.
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MELENDREZ, CARMAN S., JACK L. RUTTLE, DENNIS M. HALLFORD, PREM S. CHAUDHRY, and EDMUND R. CASILLAS. "Polyamines in Ejaculated Ram Spermatozoa and Their Relationship with Sperm Motility." Journal of Andrology 13, no. 4 (July 8, 1992): 293–96. http://dx.doi.org/10.1002/j.1939-4640.1992.tb00318.x.
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ABSTRACT: Intracellular polyamine levels in ejaculated spermatozoa and seminal fluid from rams were determined by fluorescent spectroscopy of their dansyl derivatives. Relationships between the sperm polyamine content and sperm motility of six mature and eight pubescent rams were studied. Samples were collected from both groups once a month from August through October. Mature rams had a greater percentage of motile sperm cells than lambs (94% versus 73% in September and 92% versus 78% in October); higher spermidine content (36 versus 9 pmol/108 cells in September and 162 versus 55 pmol/108 cells in October); higher spermine content (984 versus 205 pmol/108 cells in September and 1,229 versus 414 pmol/108 cells in October); and higher total sperm polyamine content (1,021 versus 216 pmol/108 cells in September and 2,258 versus 973 pmol/108 cells in October). In the lambs, spermidine content increased (55 versus 9 pmol/108 cells); spermine content increased (414 versus 205 pmol/108 cells); and total sperm polyamine content increased (973 versus 215 pmol/108 cells) in October compared to September. Ejaculates with sperm motility higher than 85% had greater spermine (848 versus 234 pmol/108 cells in September and 1064 versus 449 pmol/108 cells in October), and total sperm polyamine content (882 versus 244 pmol/108 cells in September and 2,015 versus 1,008 pmol/108 cells in October) than ejaculates with less than 85% motility. Sperm motility in ejaculates with less than 450 pmol total sperm polyamines/108 cells was 68% ± 6% compared to 90% ± 4% in cells with greater than 450 pmol (average for all ejaculates) total sperm polyamines/108 cells. These data suggest a positive relationship between sperm polyamine content and sperm motility. Sperm‐free seminal plasma from mature rams contained 7.5 and 0.28 nmol/ml of spermine and spermidine, respectively. Seminal fluids from pre‐pubertal rams also contained similar amounts of polyamines.
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Zhang, Zhaojie, Huiling Xu, Mohan B. Singh, and Scott D. Russell. "Isolation and collection of two populations of viable sperm cells from the pollen of Plumbago zeylanica." Zygote 6, no. 4 (November 1998): 295–98. http://dx.doi.org/10.1017/s0967199498000240.
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A protocol is described for individually collecting two populations of sperm cells, Svn and Sua, from pollen of Plumbago zeylanica. Pollen grains were burst in 10 mM MOPS buffer containing 0.8 M mannitol (pH 4.6). Paired sperm cells released from pollen were separated using a microinjector. Svn and Sua were then collected individually with a microinjector, based upon known size differences. Collected sperm cells were washed with isolation medium and transferred to liquid nitrogen until use. Fluorochromatic reaction (FCR) test of isolated sperm cells showed a positive reaction, indicating that the isolated sperm cells are viable; most of the sperm cells retain viability for at least 2 h.
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Ambrosini, Annarina, Rosamaria Fiorini, and Giovanna Zolese. "Endocannabinoids and Human Sperm Cells." Pharmaceuticals 3, no. 10 (October 12, 2010): 3200–3211. http://dx.doi.org/10.3390/ph3103200.
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Harrison, Sarah E. "Somatic cells raise sperm barriers." Science 354, no. 6310 (October 20, 2016): 298.1–298. http://dx.doi.org/10.1126/science.354.6310.298-a.
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Reynaud, Karine, Zeev Schuss, Nathalie Rouach, and David Holcman. "Why so many sperm cells?" Communicative & Integrative Biology 8, no. 3 (May 4, 2015): e1017156. http://dx.doi.org/10.1080/19420889.2015.1017156.
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Bains, Rani, Jude Adeghe, and Ray J. Carson. "Human sperm cells express CD44." Fertility and Sterility 78, no. 2 (August 2002): 307–12. http://dx.doi.org/10.1016/s0015-0282(02)03230-2.
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Rosales-Cruzaley, E., P. A. Cota-Elizondo, D. Sánchez, and Blanca H. Lapizco-Encinas. "Sperm cells manipulation employing dielectrophoresis." Bioprocess and Biosystems Engineering 36, no. 10 (October 20, 2012): 1353–62. http://dx.doi.org/10.1007/s00449-012-0838-6.
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Muchlisin, Zainal A., Dian Afriani, Kartini Eriani, Iwan Hasri, Firman M. Nur, Siti Maulida, Luvi S. Handayani, et al. "Improvement of Sperm Quality of the Depik Fish, Rasbora tawarensis, After Cryopreservation Using Antioxidant." Cryoletters 44, no. 1 (January 1, 2023): 13–19. http://dx.doi.org/10.54680/fr23110110312.
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BACKGROUND: The cryopreservation of the sperm of the depik fish, Rasbora tawarensis, has previously been developed. However, the quality of the sperm post cryopreservation was not satisfactory and might be improved through the application of antioxidants. OBJECTIVE: To determine the most suitable antioxidant for the cryopreservation of the depik fish spermatozoa. MATERIALS AND METHODS: A completely randomized design with a non-factorial experiment was used and the tested antioxidants were glutathione, β-carotene, ascorbic acid, and butylated hydroxytoluene (BHT) at 6% concentrations. All treatments had three replications. The sperms were collected from 10 male fishes and diluted with Ringer solution in a ratio of 1: 20 (v/v, sperm: Ringer solution). Then 5% DMSO and 5% egg yolk were added to the diluted sperms. Furthermore, 6% of the tested antioxidants were added to the diluents, and then, cryopreservation was carried out in liquid nitrogen for 14 days. RESULTS: The ANOVA test showed that the application of antioxidants significantly affected the sperm motility, fertility, and hatching rates of the eggs (P> 0.05). Furthermore, the antioxidants also protected the sperm cells during cryopreservation, with glutathione being the best antioxidant. CONCLUSION: The application of antioxidants during the cryopreservation of depik fish sperm had a significant effect on motility, fertility and hatchability of eggs post-cryo. Furthermore, glutathione was the most suitable antioxidant.
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Sgueglia, Joanne B., Hailey Holt, Erin Hanson, Justina Nichols, Tim Kalafut, Mah-ro Khan, Thomas Walsh, et al. "Interlaboratory Comparison of SpermX™ and Conventional Differential Extractions Indicated High Male DNA Recovery by the SpermX Method." Forensic Sciences 3, no. 4 (December 1, 2023): 592–610. http://dx.doi.org/10.3390/forensicsci3040043.
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The National Institute of Justice reported that current methods for processing sexual assault samples have a high failure rate, with 60 to 80 percent of tested kits unable to produce usable DNA profiles. Even when samples test positive for male DNA, 34 percent of sexual assault kits (SAKs) do not yield recovered male DNA after differential extraction. Less than 30% recovery of available sperm DNA contributes to this low success rate. The SpermX™ method (SX) has been shown to recover 80 percent or more of sperm DNA from sexual assault samples. An interlaboratory evaluation compared SX to standard differential extraction (DE) protocols. Mock samples with known ratios of female epithelial cells and sperm cells were processed using both methods. Results revealed that SX consistently provided CODIS up-loadable DNA profiles, even with as few as 25 sperm cells, whereas DE failed to produce usable results. On average, SX yielded a seven-fold increase in the recovery of unshared male alleles compared to DE. In conclusion, SX outperformed DE in recovering higher quantities of male DNA with minimal female carryover in sexual assault-type samples. This improved success rate in obtaining usable DNA profiles can significantly aid in solving sexual assault cases.
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Kon, Y., H. Iwata, H. Shiono, K. Matsubara, T. Kuwayama, and Y. Monji. "302 EFFECT OF MONOSACCHARIDES ON SPERM ABILITY TO BIND TO OVIDUCTAL EPITHELIAL CELLS." Reproduction, Fertility and Development 19, no. 1 (2007): 266. http://dx.doi.org/10.1071/rdv19n1ab302.
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The attachment of sperm to the oviductal epithelium is one of the major factors responsible for successful in vivo fertilization. Carbohydrates are reported to be involved in this attachment. In the present study, sperm were separated into 2 fractions based on their ability to bind to the oviductal epithelial cells (OECs; sperm that do not bind to OECs, UB-sperm; sperm that bind to OECs, B-sperm). Subsequently, the fertilization competence of these sperm fractions was examined. Furthermore, the effect of various monosaccharides on the ability of sperm to bind to OECs was investigated. Oviducts of cows whose ipsilateral ovaries demonstrated new clot formation (ovulation fossa) were collected from a slaughterhouse, and those that contained ovulated oocytes were further selected for experimentation. OECs were collected from these oviducts. OECs and frozen-thawed semen were each suspended in a medium (synthetic oviduct fluid, SOF, supplemented with 5 mg mL-1 of BSA and heparin), at a final concentration of 1 � 106/mL. These suspensions were mixed and centrifuged to separate the 2 abovementioned fractions. Immediately after the suspensions of sperm and OECs were mixed, d-mannose, d-fucose, or n-acetyl-d-glucosamine (GlcNAc) was added (final concentration of each monosaccharide, 5 mM). Next, the number of live sperm in each sperm population was estimated at 0 and 3 h. In addition, the oocytes collected from the ovaries were fertilized with the UB-sperm or non-separated (control) sperm, and the percentage of fertilization was examined. The percentage of fertilization and the number of sperm binding to the zona pellucida (ZP) were lower (P < 0.05) for the UB-sperm than for the control sperm. When sperm (106) were separated in the presence of GlcNAc, the number of motile sperm attached to OECs was significantly higher than that among sperm that were separated in the absence of monosaccharides (0.49 � 106 cells vs. 0.16 � 106 cells). The motility of B-sperm that were separated in the absence of monosaccharides was maintained for a longer time than that of the control sperm (percentage of motile sperm: 74.6% vs. 38.4%, P < 0.05). The B-sperm separated in the presence of GlcNAc or d-fucose lost their motility to a greater extent than the B-sperm separated in the absence of monosaccharaides (27.3% vs. 49.9%, P < 0.05). Further, when GlcNAc (5 mM) was added to the suspension for sperm cryopreservation, the number of sperm that bound to the OECs also increased. In conclusion, it was shown that sperm with low ability to bind to OECs have low fertilization competence, and GlcNAc increased the number of sperm binding to the OECs.
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Mirihagalle, Supipi, Jennifer Rose Hughes, and David Joel Miller. "Progesterone-Induced Sperm Release from the Oviduct Sperm Reservoir." Cells 11, no. 10 (May 12, 2022): 1622. http://dx.doi.org/10.3390/cells11101622.
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In mammalian females, after sperm are deposited in the reproductive tract, a fraction of sperm migrates to the lower oviduct (isthmus) and forms a sperm storage site known as the functional sperm reservoir. The interactions between sperm membrane proteins and oviduct epithelial cells facilitate sperm binding to the oviductal epithelium and retention in the reservoir. Sperm are bound by glycans that contain specific motifs present on isthmic epithelial cells. Capacitated sperm are released from the reservoir and travel further in the oviduct to the ampulla where fertilization occurs. For decades, researchers have been studying the molecules and mechanisms of sperm release from the oviductal sperm reservoir. However, it is still not clear if the release of sperm is triggered by changes in sperm, oviduct cells, oviduct fluid, or a combination of these. While there is a possibility that more than one of these events are involved in the release of sperm from the reservoir, one activator of sperm release has the largest accumulation of supporting evidence. This mechanism involves the steroid hormone, progesterone, as a signal that induces the release of sperm from the reservoir. This review gathers and synthesizes evidence for the role of progesterone in inducing sperm release from the oviduct functional sperm reservoir.
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Woo, Sun Hee, Taiji Adachi, Seung Keun Jong, and Clayton G. Campbell. "Isolation of protoplasts from viable sperm cells of common buckwheat (Fagopyrum esculentum Moench.)." Canadian Journal of Plant Science 80, no. 3 (July 1, 2000): 583–85. http://dx.doi.org/10.4141/p99-118.
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Protoplasts from sperm cells were isolated. Sperm cells from germinated pollen placed in solution were released, sieved and removed after sucrose density gradient centrifugation. The isolated cells were spherical and approximately 5.0–6.0 µm in diameter. Cytological observations confirmed that sperm cells of common buckwheat are true protoplasts. Key words: Common buckwheat (F. esculentum Moench.), sperm cell isolation, osmotic shock
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Pessoa, G. A., J. M. Trentin, A. P. Martini, D. R. Dotto, L. A. M. Centeno, M. L. Jardim, K. V. Aires, and M. I. B. Rubin. "180 SPERM SELECTION OF STALLION PONIES THROUGH GLASS WOOL." Reproduction, Fertility and Development 26, no. 1 (2014): 204. http://dx.doi.org/10.1071/rdv26n1ab180.
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Two techniques of sperm concentration (centrifugation or filtering) and sperm separation technique with glass wool were applied to the sperm samples collected from 3 pony stallions (6 ejaculates; 2 from each stallion). Ejaculates were extended to a final concentration of 50 × 106 spermatozoa mL–1 using a nonfat dry milk-based extender and evaluations occurred at 24, 48, and 72 h after immediate ejaculate dilution and cooling. Each stallion was considered as a block, and semen from each stallion was assigned to 4 treatments: Group A (control): extended semen alone; Group B: extended-centrifuged semen; Group C: extended-sperm filtered semen; Group D: extended-glass wool-separated semen. All groups were tested for pH, osmolarity, motility, morphology, membrane functionality (hyposmotic), and cell viability (MTT assay). The experimental design was performed using a split-plot model. Data analysis at the level of 5% was performed using ANOVA and Bonferroni as post-hoc test. Data are presented as mean ± standard error. Group D had the highest rate of viable cells (P < 0.05) after the separation procedure (Table 1). Group B had a higher percentage of cells with tail defects after processing compared with the controls and Groups A, C, and D (P < 0.05). More than 60% of the cells retained on the filter showed defects (P < 0.001). Progressive motility was greater in group D at 0, 24, and 48 h (P < 0.05). Seventy-two hours after cooling, motility in groups A and B was lower than in Group D (P < 0.01). Group D showed a higher number of cells with mitochondrial activity during the cooling period. In conclusion, the technique of sperm selection by gravity using a glass wool filter resulted in an increased number of viable sperms after cooling pony semen for 24, 48, and 72 h. Table 1.Effect of sperm concentration and separation techniques on mean ± standard error percent of intact sperm from 3 stallions ponies (2 ejaculates/pony) stained with eosin-nigrosin
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Saha, Amit, Mohammad Asaduzzaman, and Farida Yeasmin Bari. "Cryopreservation Techniques for Ram Sperm." Veterinary Medicine International 2022 (April 30, 2022): 1–16. http://dx.doi.org/10.1155/2022/7378379.
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Germplasm storage and transportation in artificial insemination (AI) and other advanced technologies are facilitated by cryopreservation. In reproduction, the cryopreservation of sperm allows it to be transported across vast distances and used even after the sire’s death. However, the technique of cryopreservation might damage sperm and limit their activity. Several cryobiological investigations have reported that the integrity of the sperm membrane is frequently involved in the physical and biological elements that affect sperm survival at low temperatures during the cryopreservation process. However, successful cryopreservation of ram sperm is still a work in progress because a considerable percentage of sperm do not survive the freezing and thawing process. Sperms are destroyed during cryopreservation of semen due to varying concentrations of cryoprotective chemicals and if semen is not cooled at optimal cooling rates. Hence, it is crucial to know the optimum cooling rates with freezing and thawing protocols for maximum recovery of viable and functional sperm cells for a successful cryo-freezing of ram spermatozoa. Therefore, the current study compiled and compared the research on the impact of different cryopreservation procedures, cooling rates, equilibration time, and thawing protocols on post-thaw ram semen quality.
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Dwi Juniatiningrum, Ratna, Taufiqurrachman Nasihun, and Israhnanto Isradji. "HIGH DOSE VITAMIN C ADMINISTRATION EFFECT IN LEYDIG CELLS, SERTOLI CELLS NUMBER, AND SPERM QUALITY ON MALE WISTAR RATS." Sains Medika : Jurnal Kedokteran dan Kesehatan 9, no. 1 (November 11, 2018): 18. http://dx.doi.org/10.30659/sainsmed.v9i1.2468.
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Introduction: Most infertile male are associated with poor spermatogenesis due to oxidative stress, and can be prevented with vitamin C. However, excessive amount of high dose of vitamin C can hindered and lower the sperm quality. Objective: To prove that high dose vitamin C is capable to decrease the number of leydig cells, sertoli cells, and sperm quality on male wistar rats.Methods: This research was using experimental method with Post Test Only Controlled Group Design. Of 24 male Wistar rats, divided randomly to 4 groups. Normal groups (Nor -G), only given 2 ml/day distilled water; vitamin C group (VC18-G, VC36-G, and VC72-G) given 18 mg/day, 36 mg/day and 72 mg/day vitamin C respectively, dissolved in 2 ml of distilled water. Sperm, the number of Leydig cells and Sertoli cells were taken from the epididymis and left right testicle on day 21. Sperm analysis using WHO standard, while the number of Leydig cells and Sertoli cells with HE staining.Results: Mann Whitney analysis indicated that the number of sperm in VC36-G and VC72-G are lower compared to that of Nor-G and VC18-G groups, p <0.05. Post-Hoc LSD analysis showed that the lowest number of Leydig and Sertoli Cell, and the weakest sperm motility and morphology in VC36-G and VC72-G groups, compared to that of Nor-G and VC18-G groups, p <0.05 .Conclusion: Vitamin C 36 and 72 mg/day were capable of reducing Leydig and Sertoli cells number, and worsen sperm quality, characterized by decreased in sperm concentration, motility and morphology in Wistar male- rats.
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Naz, R. K., K. Ahmad, and P. Kaplan. "Expression and function of ras proto-oncogene proteins in human sperm cells." Journal of Cell Science 102, no. 3 (July 1, 1992): 487–94. http://dx.doi.org/10.1242/jcs.102.3.487.
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The presence and role of c-ras proteins were investigated in mature human sperm cells. The v-H-ras monoclonal antibody (mAb) against the c-ras protein, p21, reacted specifically with the acrosomal region of methanol-fixed as well as unfixed-live capacitated and non-capacitated human sperm cell in the indirect immunofluorescence technique. The v-H-ras mAb predominantly recognized c-ras protein of 21 kDa on the Western blot of lithium diiodosalicylate (LIS)-solubilized human sperm preparation. The incubation of sperm cells with v-H-ras mAb affected the sperm cell function in the human sperm penetration assay. The antibody significantly reduced the acrosome reaction and release of acrosin activity from the sperm cells. There was no effect of the mAb on percentage motility, although the mAb significantly affected various motility characteristics such as linearity, amplitude of lateral head displacement (ALH) and beat frequency, the motility parameters involved in the hyperactivation phenomenon of sperm cells leading to capacitation and acrosome reaction. These results suggest that the c-ras or c-ras-like proteins are present in mature sperm cell and may have a role in capacitation and/or acrosome reaction of human sperm cell.
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Vladić, Tomislav, and Erik Petersson. "Artificially selected human sperm morphology after swim-up processing." Canadian Journal of Zoology 90, no. 10 (October 2012): 1207–14. http://dx.doi.org/10.1139/z2012-088.
Full textAbstract:
The swim-up technique is a clinical practice used to select highly motile sperm cells from patient ejaculates to use in assisted fertilization. The aim of this study was to investigate whether the length of different sperm-cell components is related to gamete function. Thus, we explored whether swim-up technique selects for longer sperm cells than mean sperm cells from unprocessed ejaculates. Sperm midpiece, tail endpiece, and total length were measured before and after the swim-up selection by means of contrast-phase and electron microscopy. Correlations between sperm dimensions, sperm motility, and sperm concentration were also investigated. Swim-up selected cells with longer midpiece compared with the unprocessed fractions (5.8 μm (CI 5.52–6.16 μm) vs. 5.3 μm (CI 4.97–5.61 μm), p < 0.05) and shorter tail endpiece (7.8 μm (CI 7.11–8.44 μm) vs. 8.5 μm (CI 7.81–9.14 μm), p < 0.05 after meta-analysis), whereas no effect of swim-up selection was detected on the total sperm cell length. Individuals producing high sperm concentrations had longer sperm midpiece than had men producing lower sperm concentrations. It is concluded that short sperm flagellar tips with long midpieces may be used as biomarkers in infertility therapy.
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Sadler, Kirsten. "Stem cells and sperm cells – the GDNF connection." Trends in Cell Biology 10, no. 5 (May 2000): 179. http://dx.doi.org/10.1016/s0962-8924(00)01763-3.
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Burns, P. D., N. Wong, H. Arnold, N. Sirs, R. Romero, and L. Herickhoff. "61 PLANT EXTRACTS REDUCE DNA FRAGMENTATION IN FROZEN-THAWED STALLION SPERM." Reproduction, Fertility and Development 21, no. 1 (2009): 131. http://dx.doi.org/10.1071/rdv21n1ab61.
Full textAbstract:
Mares inseminated with frozen–thawed sperm have reduced pregnancy rates compared with mares inseminated with fresh sperm. Processing mammalian sperm for cryopreservation increases the concentration of free radicals and induces oxidative stress, which can result in DNA damage and may lead to lower fertility. The objective of this experiment was to examine the effects of several plant antioxidant extracts on stallion sperm post-thaw motility and DNA quality. Single ejaculates were collected from 4 stallions and the concentration of sperm cells in each ejaculate was determined spectrophotometrically. Semen was centrifuged at 300g for 10 min at room temperature and seminal plasma was removed. Sperm pellets were resuspended to a final concentration of 200 × 106 cells mL–1 in E-Z Freezin LE (ARS, Chino, CA) extender (control) or extender containing 1 of 3 plant extracts (3% v/v) from 2 different commercial sources. Extended sperm cells were loaded into 0.5-mL straws and frozen over liquid nitrogen vapor for 10 min. Straws were then plunged into liquid nitrogen and stored until further evaluation. Motility and velocity parameters were determined at 0, 30, and 60 min post-thaw using a computer-assisted sperm analyzer. DNA fragmentation was determined immediately after thawing using a single-cell gel electrophoresis (Comet) assay. Motility (total and progressive) and velocity parameters of sperm cells did not differ between controls and plant extract treatments (P > 0.05). However, total Comet length and tail length were reduced in sperm cells stored in extender containing each plant extract (P < 0.05). Tail and olive moment tended to be reduced (P < 0.10) in sperm cells stored in plant extracts. In conclusion, sperm cells stored in plant extracts had reduced post-thaw DNA damage. The addition of plant extracts to commercial freezing extenders may be a practical method for improving sperm quality.
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KRAWCZYK, ALEKSANDRA, and JADWIGA JAWORSKA-ADAMU. "Activation of sperm in the female reproductive tract in mammals." Medycyna Weterynaryjna 76, no. 09 (2020): 6445–2020. http://dx.doi.org/10.21521/mw.6445.
Full textAbstract:
The formation of a new diploidal organism is preceded by a series of mutual interactions of haploidal gametes. This process is very complicated and requires the prior activation of reproductive cells. Male gametes eventually mature in the female reproductive tract, acquiring mobility and fertilization. This process takes place in two stages. Sperms are first capacitated. This phenomenon is reversible and leads to structural, cytophysiological and biochemical changes in the sperm plasma membrane as well as to the sperm hyperactivation. Then, due to the contact with the zona pellucida of the oocyte, the irreversible acrosome reaction occurs. This process involves the fusion of the sperm plasma membrane with the outer membrane of the acrosome, the release of enzymes and exposure of the inner acrosome membrane. This enables sperm to penetrate towards the perivitelline space and oolemma. Contact with the oocyte initiates a series of interactions leading to egg activation and the fusion of gametes. Each of these stages involves many different factors that result in the recognition, attraction and adhesion of reproductive cells. Knowledge about the activation mechanisms can improve the effectiveness of supported and controlled reproduction techniques.