Journal articles: 'Culture de neurones'

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Orr, D. J., and R. A. Smith. "Neuronal maintenance and neurite extension of adult mouse neurones in non-neuronal cell-reduced cultures is dependent on substratum coating." Journal of Cell Science 91, no. 4 (December 1, 1988): 555–61. http://dx.doi.org/10.1242/jcs.91.4.555.

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Adult mouse DRG neurones have been maintained for 14 days in cultures where non-neuronal cell proliferation was inhibited by the inclusion of 5 × 10(−6) microM-cytosine arabinoside (AraC) in the medium from the onset of culture. On uncoated plastic neurone numbers significantly declined in the absence of non-neuronal cell outgrowth compared with uninhibited co-cultures. However, when neurones were maintained in the presence of AraC on certain coated surfaces this decrease in neurone numbers was not observed. Combinations of fibronectin (FN) and laminin (LAM) proved most effective for 7 and 14 days in vitro, although either was beneficial if used separately. Microexudates produced by the fibroblast line, 3T6, also significantly improved neuronal counts for 14 days in vitro. However, a microexudate derived from primary cultures of mouse hepatocytes, although advantageous for 7 days in vitro, was not effective in maintaining neurones over the 14-day culture period, reminiscent of previous observations when synthetic cationic agents were used. Electrophoretic analysis of the fibroblast exudate indicated that fibronectin was present in the substrate-attached material generated by this cell line. The reduction in non-neuronal cell growth facilitated the monitoring of neuronal structural detail by scanning electron microscopy. Examination of neurite extension, indicative of neurone differentiation, was particularly improved. FN/LAM and the fibroblast-derived exudate increased nerve fibre growth, whilst the hepatocyte exudate had little effect on neurite regeneration, and polylysine had a detrimental effect. The data demonstrate that substrata can have a significant effect on maintenance and differentiation of adult neurones in primary culture.(ABSTRACT TRUNCATED AT 250 WORDS)

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PRZYSIEZNIAK, JAN, and ANDREW N. SPENCER. "Primary Culture of Identified Neurones from a Cnidarian." Journal of Experimental Biology 142, no. 1 (March 1, 1989): 97–113. http://dx.doi.org/10.1242/jeb.142.1.97.

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Several types of neurones were dissociated from the nerve-rings of the hydrozoan jellyfish Polyorchis penicillatus, using collagenase digestion preceded, and if necessary followed, by removal of external divalent cations. The neurones were cultured for up to 2 weeks in artificial sea water, on a mesogloeal substratum. One subset of large neurones, the swimming motor neurones (SMNs; soma approx. 20×50 μm), exhibited distinct morphological features in vitro, such as large size, wide processes, clear cytoplasm and membranous inclusions around the nucleus. These neurones retained their characteristic action potential shape in culture, with spikes measuring 50±11 mV (N=18) in peak amplitude and 37 ± 11 ms in duration. SMNs could be labelled in vivo with carboxyfluorescein or Lucifer Yellow, subsequently dissociated, and identified in vitro. Two subsets of small neurones were also identifiable. One exhibited electrophysiological similarities with B system neurones, known to be presynaptic to the SMNs in vivo, showing a burstlike pattern of spikes of short duration (5.4 ± 1.4 ms; N=6) and small amplitude (25 ± 7mV). Another subset of small neurones could be labelled with antiserum against the carboxy-terminal peptide moiety, Arg-Phe-amide. Biophysical and neurotransmitter studies at the level of the single identified hydrozoan neurone will be easier in isolated cell culture. This approach will avoid problems encountered in studying the semidissected nerve-ring preparation.

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GREEN, K. A., G. B. POWELL, and A. COTTRELL. "Unitary K+ Currents in Growth Cones and Perikaryon of Identified Helix Neurones in Culture." Journal of Experimental Biology 149, no. 1 (March 1, 1990): 79–94. http://dx.doi.org/10.1242/jeb.149.1.79.

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Unitary potassium (K+) currents of several different conductances have been recorded from the growth cones of isolated Cl neurones from Helix aspersa. The isolated neurones were maintained in culture for up to 1 week. Similar unitary currents were recorded in the growth cones of other isolated Helix neurones. The activity of one type of unitary K+ current recorded from the growth cones of the Cl neurone and other neurones was very similar to that described for the S-channel of the perikarya of Aplysia sensory neurones. Another type of unitary K+ current showed fast flickering and reduced amplitude when the membrane was held at large positive potentials, which is suggestive of channel block by some agent. The conductances of the K+ channels in the growth cones of isolated Cl neurones were generally smaller than those recorded in this and in previous studies from the perikarya of Cl neurones in situ. However, unitary K+ currents recorded from the perikaryon of the Cl neurone, and from other identified neurones, in culture also had lower conductances than those recorded in situ. The mean resting potential of the isolated neurones was smaller than those from neurones in situ. This and other results suggested that reduced intracellular K+ concentration in the isolated neurones might be an important factor in deciding the conductance of the recorded channels.

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Chiquet, M., and J. G. Nicholls. "Neurite outgrowth and synapse formation by identified leech neurones in culture." Journal of Experimental Biology 132, no. 1 (September 1, 1987): 191–206. http://dx.doi.org/10.1242/jeb.132.1.191.

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After injury, neurones in the central nervous system (CNS) of the leech regenerate with a high degree of specificity. The aim of our experiments has been to study the sequential steps involved in neurite growth and synapse formation using isolated identified neurones in culture. An important requirement for sprouting of leech neurones is the substrate. Neurites grow only slowly and sparsely on polylysine or vertebrate laminin. The extracellular matrix of leech ganglion capsules contains a protease-sensitive factor which can be extracted with urea. With this material as substrate, growth proceeds rapidly in defined medium. Another neurite-promoting substrate is provided by the plant lectin concanavalin A (Con A). The activity of Con A, but not of the capsule matrix factor, is blocked by the Con A-specific hapten methyl alpha-D-mannoside. The morphology and branching pattern of the neurites in culture depend on the specific substrate and on the type of neurone. During stimulation, less Ca2+ uptake occurs into growth cones than in cell bodies. The mechanism of neurite growth seems not to depend on activity-mediated Ca2+ influx or on interactions between neuronal cell surfaces. However, even without profuse outgrowth, electrical and chemical synapses develop between neighbouring neurones. The type of synapse depends predictably on the types of neurones within the cell pair. Since the development of a synapse can be followed with time in culture, the sequential events can each be studied separately for this multi-step process.

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Newland, N. L., P. J. S. Smith, and E. A. Howes. "REGENERATING ADULT COCKROACH DORSAL UNPAIRED MEDIAN NEURONES IN VITRO RETAIN THEIR IN VIVO MEMBRANE CHARACTERISTICS." Journal of Experimental Biology 179, no. 1 (June 1, 1993): 323–29. http://dx.doi.org/10.1242/jeb.179.1.323.

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The ability of differentiated neurones to recover from disease or injury depends upon both intrinsic and extrinsic factors. Whereas most mammalian neurones have a limited capacity for regeneration, regulated, in part, by physical and chemical cues in the brain microenvironment (Bray et al. 1987; Caroni and Schwab, 1988, 1989), invertebrates, and in particular insects, exhibit a far greater capacity for repair of central neurones and circuits (Treherne et al. 1988). Studies of the cues that regulate the regenerative process are made easier by the use of individual, identified neurones, cultured under controlled conditions. Invertebrates are particularly useful in this regard; neurones from mature nervous systems of both annelids and molluscs have been grown successfully in culture and their growth can be influenced by changes in the culture conditions (Acklin and Nicholls, 1990; Dagan and Levitan, 1981; Ready and Nicholls, 1979; Syed et al. 1990). Routine and long-term culture of identified neurones from the insect central nervous system (CNS) has proved more elusive, preventing the use of neurones from these well-studied systems. Recently, however, cultures of cockroach (Howes et al. 1991), locust (Kirchoff and Bicker, 1992) and moth (Hayashi and Levine, 1992) adult neurones have been described.

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Duprat, Anne-Marie, Paulette Kan, Françoise Foulquier, and Michel Weber. "In vitro differentiation of neuronal precursor cells from amphibian late gastrulae: morphological, immunocytochemical studies, biosynthesis, accumulation and uptake of neurotransmitters." Development 86, no. 1 (April 1, 1985): 71–87. http://dx.doi.org/10.1242/dev.86.1.71.

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Neuronal differentiation has been studied in dissociated cell cultures from early neurulae of Pleurodeles waltl and Ambystoma mexicanum. Cocultures were prepared from the neural primordium and underlying chordamesoderm. NP and NF cultures were prepared from isolated neural plate and neural folds, respectively. Neuronal precursors in NP and NF cultures had distinctive aggregation properties already evident after 1–2 days in culture. After 10–15 days, mature neurones and synapses were observed by electron microscopy in the three culture types. The expression of neurofilament polypeptides and tetanus-toxin-binding sites was also present in these cultures. A small percentage of neurones contained cytochemically detectable catecholamine. Many neurones took up tritiated dopamine with a high affinity. Quantitative measurement of [3H]acetylcholine synthesis and storage from [3H]choline were negative at the early neurula stage and in 5 to 15-day-old NF cultures, and remained low in 5 to 15-day-old NP cultures. Acetylcholine production in cocultures increased linearly with time and was always much higher than in NP cultures. These results suggest that, at the early neurula stage, some neuronal precursors have acquired the capacity to express a high degree of morphological and biochemical differentiation even in the absence of further chordamesoderm influence. However, the chordamesodermal cells in the cultures increased acetylcholine synthesis.

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Fernaud-Espinosa, I., M. Nieto-Sampedro, and P. Bovolenta. "Differential effects of glycosaminoglycans on neurite outgrowth from hippocampal and thalamic neurones." Journal of Cell Science 107, no. 6 (June 1, 1994): 1437–48. http://dx.doi.org/10.1242/jcs.107.6.1437.

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Chondroitin sulphate proteoglycans are expressed in a temporally restricted pattern from embryonic day 17 to postnatal day 0 in both the thalamus and the cortical subplate, to which thalamic neurones transiently project. To study whether chondroitin sulphate proteoglycans could be specifically involved in the modulation of thalamic axon outgrowth, we compared neurite outgrowth from cultured rat embryonic hippocampal and thalamic neurones, in the presence of chondroitin sulphate type C (isolated from shark cartilage) and chondroitin sulphate type B (dermatan sulphate; isolated from bovine mucosa). When added to the culture medium, both types of glycosaminoglycan lowered the adhesion to laminin and polylysine of both hippocampal and thalamic neurones. However, only chondroitin sulphate specifically modified the pattern of thalamic but not hippocampal neurone outgrowth, promoting axon growth. The morphological changes induced by chondroitin sulphate were concentration dependent and correlated with the selective binding of chondroitin sulphate to the neuronal plasma membrane and its subsequent internalisation. Chondroitin sulphate loosely bound to the surface of hippocampal neurones, but was not internalised. These results indicate that proteoglycans, and in particular the glycosaminoglycan component of these molecules, can differentially modulate neurite outgrowth, depending on their biochemical composition and on the type of neurones they bind to; this would be a possible mechanism of controlling axon guidance in vivo.

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8

Clarke, M. J. O., and G. E. Gillies. "Comparison of peptide release from fetal rat hypothalamic neurones cultured in defined media and serum-containing media." Journal of Endocrinology 116, no. 3 (March 1988): 349–56. http://dx.doi.org/10.1677/joe.0.1160349.

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ABSTRACT Primary cultures of rat hypothalamic neurones were maintained either in a serum-supplemented medium or in a serum-free chemically defined medium for up to 6 weeks. The release of the 41 amino acid-containing peptide, corticotrophin-releasing factor (CRF-41), vasopressin (AVP) and somatostatin (SRIF) were followed using immunoassays. In response to K+ (56 mmol/l) depolarization both the quantities of peptides released and the magnitude of responses were significantly greater from cultures maintained in the fully supplemented defined medium. As a consequence, release of CRF-41 and AVP could be measured directly, without requiring the concentration step necessary for cultures grown in serum. The response to K+ depolarization increased with the age of the culture, suggesting neuronal maturation. Responses to K+ depolarization were Ca2+-dependent, and the addition of corticosterone (100 nmol/l) to the defined medium caused a significant reduction in the response of neurones secreting CRF-41 and AVP, but not those secreting SRIF, to depolarization. This suggests the retention in vitro of the responsiveness of stress-associated neuropeptides to the negative feedback effects of corticosterone. Neurones producing CRF-41 and AVP responded significantly in a dose-dependent manner to acetylcholine stimulation, whereas those producing SRIF did not. As cultures matured, the CRF-41- and AVP-producing neurones became more sensitive to acetylcholine with the maximal response at 1 nmol acetylcholine/1. In conclusion, the culture of rat hypothalamic neurones is improved in terms of peptide output when the cultures are maintained in a defined medium. Differential responses of the peptidergic neurones may be seen in the presence of corticosterone and neurotransmitters, illustrating the retention in vitro of specific receptor-mediated responses which have been observed in vivo. This model should prove useful in the further study of the physiological, pharmacological and biochemical maturation and control of peptidergic neurones. J. Endocr. (1988) 116, 349–356

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9

NICHOLLS, J. G., Y. LIU, B. W. PAYTON, and D. P. KUFFLER. "The Specificity of Synapse Formation by identified Leech Neurones in culture." Journal of Experimental Biology 153, no. 1 (October 1, 1990): 141–53. http://dx.doi.org/10.1242/jeb.153.1.141.

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The physiological and fine structural events accompanying synapse formation have been followed while identified neurones of known function make contact in tissue culture. Particular pairs of identified neurones isolated from the central nervous system (CNS) of the leech form chemical synapses; other pairs of cells form nonrectifying electrical junctions, rectifying electrical junctions, mixed chemical and electrical synapses or no synapses at all, depending upon the partners that have been paired. Moreover, certain specific regions on the cell surface (such as the soma, initial cell segment or axon tips) preferentially develop chemical or electrical synapses. Of particular interest are the large, serotonergic Retzius cells that form mixed chemical and electrical synapses in culture, as in the animal. When these cells are juxtaposed at their initial segments, it has been shown that chemical synapses can develop reliably within 6h of contact in culture. Shortly after transmission can be detected physiologically, the principal features of synaptic structure are evident. The physiological and morphological characteristics resemble those of mature synapses studied within the central nervous system. Only at later times, after the chemical synapses have been formed, do electrical connections appear. By contrast, when other specialized regions of the Retzius cells are apposed (the tips of their axons), electrical synapses appear earlier. By comparing the connections that different types of serotonergic neurones make in culture we have been able to assess the role played by the transmitter in determining specificity: the results show that the transmitter does not determine what type of synapse is made on a particular partner. For example, Retzius cells make purely chemical synapses upon the sensory P neurone in culture; other serotonergic neurones (known as DL and VL) make purely electrical connections on this same pressure sensory neurone. Together, these results demonstrate that highly specific cell-cell recognition is a necessary feature of synapse formation after neurones have grown to their appropriate destinations.

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Atterwill, Christopher K., Wendy J. Davies, and Michael A. Kyriakides. "An Investigation of Aluminium Neurotoxicity using some In Vitro Systems." Alternatives to Laboratory Animals 18, no. 1_part_1 (November 1990): 181–90. http://dx.doi.org/10.1177/026119299001800119.1.

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It has been shown that acute exposure in vitro to high concentrations of aluminium chloride does not appear to perturb neural function in terms of the electrophysiological properties of lower vertebrate leech neurones. Longer term exposure in vitro, however, both non-specifically inhibits cellular differentiation and also produces neural cytotoxicity in the rat midbrain micromass, mixed cell culture model. Furthermore, previous studies from this laboratory have demonstrated a reduction of cholinergic neuronal function in brain organotypic reaggregate cultures following long-term, but not short-term, exposure. More-immature neural cells appear to be most sensitive to the effects of aluminium. Relating these data to the tiered in vitro test system for neurotoxicants previously proposed by Atterwill (13), it is apparent that the neurotoxic effects of aluminium are detectable in a first-stage procedure using the micromass culture model, but not following acute exposure in freshly isolated, ex vivo leech neurones. Functional cholinergic toxicity was also detected in the organotypic reaggregate cultures proposed as a second level screen.

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11

Middleton, Gayle, and Alun M. Davies. "Populations of NGF-dependent neurones differ in their requirement for BAX to undergo apoptosis in the absence of NGF/TrkA signalling in vivo." Development 128, no. 23 (December 1, 2001): 4715–28. http://dx.doi.org/10.1242/dev.128.23.4715.

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Reports that apoptosis within populations of neurotrophin-dependent neurones is virtually eliminated in BAX-deficient mice and that BAX-deficient neurones survive indefinitely in culture without neurotrophins have led to the view that BAX is required for the death of neurotrophin-deprived neurones. To further examine this assertion in vivo, we have studied two populations of NGF-dependent neurones during the period of naturally occurring neuronal death in mice that lack BAX, NGF or the NGF receptor TrkA, alone and in combination. In the superior cervical ganglion (SCG), naturally occurring neuronal death and the massive loss of neurones that took place in the absence of NGF or TrkA were completely prevented by elimination of BAX. However, in the trigeminal ganglion, naturally occurring neuronal death was only partly abrogated by the elimination of BAX, and although the massive neuronal death that took place in this ganglion in the absence of NGF or TrkA was initially delayed in embryos lacking BAX, this subsequently occurred unabated. Accordingly, BAX-deficient neurones survived in defined without NGF whereas BAX-deficient trigeminal neurones died in the absence of NGF. These results indicate that whereas BAX is required for the death of SCG neurones during normal development and when these neurones are deprived of NGF/TrkA signalling in vivo, the death of trigeminal ganglion neurones occurs independently of BAX when they are deprived of NGF/TrkA signalling. We conclude that BAX is not universally required for neuronal death induced by neurotrophin deprivation, but that there are major differences for the requirement for BAX among different populations of NGF-dependent neurones.

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Howes, E. A., T. R. Cheek, and P. J. Smith. "Long-term growth in vitro of isolated, fully differentiated neurones from the central nervous system of an adult insect." Journal of Experimental Biology 156, no. 1 (March 1, 1991): 591–605. http://dx.doi.org/10.1242/jeb.156.1.591.

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A method is described for the isolation and growth in vitro of fully differentiated neurones from the thoracic ganglia of adult cockroaches. The presence of insect blood in the culture system is shown to promote growth. The morphology of the growing neurones and the plasticity of the branching processes are described and growth rates are measured. Using a fluorescent Ca2+ indicator dye, changes of intracellular calcium levels in the growing neurones in response to K+ depolarization have been measured. The results, indicating the presence of voltage-dependent Ca2+ channels on neuronal processes in vitro, show that neurones can be maintained in a functional state for several weeks by this technique. Such preparations could prove useful for studying a variety of physiological and pharmacological properties of neurones, including the mechanisms controlling growth, synapse formation and neuronal interactions with other cell types.

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13

Vidal, S., B. Raynaud, D. Clarous, and M. J. Weber. "Neurotransmitter plasticity of cultured sympathetic neurones. Are the effects of muscle-conditioned medium reversible?" Development 101, no. 3 (November 1, 1987): 617–25. http://dx.doi.org/10.1242/dev.101.3.617.

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Muscle-conditioned medium (CM) induces choline acetyltransferase (CAT) activity in primary cultures of new-born rat sympathetic neurones and depresses the development of tyrosine hydroxylase (TOH). By following these two enzymes, we have determined whether (1) the effects of CM are reversible and (2) the neurones progressively lose their sensitivity to CM with time in culture. When neurones were cultured in the presence of 50% CM (CM+ medium), TOH activity developed slowly but CAT activity developed at a high rate. When the cultures were then switched to unconditioned medium (CM- medium), CAT activity remained elevated and continued to develop at higher rate than in cultures that were never exposed to CM. On the other hand, the switch to CM- medium was accompanied by a transition from a low to a high rate of TOH development. CAT induction by CM was thus essentially irreversible, whereas the impairment of TOH development was fully reversible. Conversely, we studied the effects of altering CM- to CM+ medium at progressively later culture days. CAT remained fully inducible for at least 2 to 3 weeks. On the other hand, TOH activity, which initially developed rapidly in CM- medium, first decreased to low levels after a switch to CM+ medium and then increased again, but at a slower rate. Neuronal depolarization by elevated K+ and exposure to CM have mirror-image, and antagonistic, effects on both CAT and TOH developments (Raynaud et al. 1987a). Walicke, Campenot & Patterson (1977) showed that a previous depolarization reduced the induction of cholinergic traits by a subsequent exposure to CM. We found that (1) such a depolarization only delayed the induction of CAT by several days and did not prevent the transition to a state of low TOH expression caused by CM and (2) an exposure of the cultures to elevated K+ after exposure to CM did not cause a decline in CAT activity. These data thus suggest that a state of high TOH expression can superimpose on a previously induced state of elevated CAT expression, but that the induction of CAT caused by a delayed exposure to CM is accompanied by a transition from a high to a lower state of TOH expression. In addition, neuronal depolarization does not stabilize the noradrenergic phenotype in a permanent manner and can not reverse cholinergic expression of sympathetic neurones to a purely noradrenergic phenotype.

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Calabrese, B., and M. Pellegrino. "Remodelling of an intact neurone in the central nervous system of the leech." Journal of Experimental Biology 198, no. 9 (September 1, 1995): 1989–94. http://dx.doi.org/10.1242/jeb.198.9.1989.

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The regeneration pattern of two identified central neurones was studied in the leech Hirudo medicinalis. Anterior pagoda (AP) and mechanosensory touch-sensitive (T) neurones were stained in adult segmental ganglia, maintained in culture for 6-10 days. AP neurones, which normally project only to the contralateral nerve roots, sprouted extensively in all the available nerve paths during regeneration. Mechanosensory T cells, in the same experimental conditions, showed only a moderate growth and did not change their normal pattern of axonal projections. The observed differences in the growth pattern might account for the different electrophysiological responses to axotomy exhibited by the two types of neurone. Interruption of interganglionic connectives induced a moderate and stereotyped remodelling of the morphology of intact AP neurones, which was reminiscent of that transiently exhibited during embryonic development. This response was observed in 25% of the AP neurones we examined.

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Murray, H. E., and G. E. Gillies. "Investigation of the ontogenetic patterns of rat hypothalamic dopaminergic neurone morphology and function in vitro." Journal of Endocrinology 139, no. 3 (December 1993): 403—NP. http://dx.doi.org/10.1677/joe.0.1390403.

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ABSTRACT Using fetal rat hypothalamic cells in primary culture maintained in a serum-free defined medium we have investigated the morphological and functional development of the dopamine (DA)-containing neurones intrinsic to the hypothalamus. Immunocytochemical studies demonstrated the presence of three morphologically distinct subtypes of tyrosine hydroxylase-immunopositive neurones. On day 3 in vitro unipolar, bipolar and multipolar cell types were apparent. The latter two subtypes persisted to later days in culture and increased both in perikarya size and neurite length. All subtypes have been shown to have correlates in vivo. Biochemical studies employing [3H]DA demonstrated a time- and temperature-dependent uptake mechanism within the cultures which was significantly attenuated by the uptake inhibitors benztropine and nomifensine in a dose-dependent manner. [3H]DA was also released under both basal and 56 mmol K+/l-stimulated conditions and the magnitude of the response was reduced by exclusion of calcium from the release medium. The amount of [3H]DA accumulated and released by the cultured cells increased with the age of the culture, suggesting functional maturation of the DA-containing neurones within this preparation. The role of oestradiol-17β in regulating hypothalamic dopaminergic function was also investigated both indirectly with the use of [3H]DA and by direct measurement of endogenously synthesized DA using high-performance liquid chromatography coupled with electrochemical detection. Both uptake and release of [3H] and release of endogenous DA were significantly modulated by the concentration of steroid in the defined medium. These results demonstrate that hypothalamic dopaminergic neurones, when maintained in primary culture, undergo morphological and functional maturation which have several correlates in vivo. In addition, we have demonstrated that at least one sub-population of dopaminergic neurones within this preparation is responsive to oestradiol-17β. As DA is considered to be a vital component in the regulation of neuroendocrine activity we suggest that this model is valuable for the investigation of the functional development of the DA systems of the hypothalamus and the relationship existing between neurotransmitters, neuropeptides and neuroactive steroids. Journal of Endocrinology (1993) 139, 403–414

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Pituello, F., P. Deruntz, L. Pradayrol, and A. M. Duprat. "Peptidergic properties expressed in vitro by embryonic neuroblasts after neural induction." Development 105, no. 3 (March 1, 1989): 529–40. http://dx.doi.org/10.1242/dev.105.3.529.

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As an immediate consequence of neural induction, some neuroectodermal cells acquire the ability to develop a number of characteristic neuronal features, without requiring any subsequent embryonic cues (Duprat et al. 1987). Thus, adrenergic, cholinergic and gabaergic traits are expressed in cultures of neural fold and neural plate isolated from amphibian embryos immediately after induction and grown in a defined medium. The aim of the present study was to determine, using the same in vitro model, their abilities to develop peptidergic phenotypes. Using immunocytochemical techniques, we show that substance P-, enkephalin- (leu-enkephalin, metenkephalin), and somatostatin- like immunoreactivities are expressed in subpopulations of neurones grown in vitro, whereas VIP (vasoactive intestinal polypeptide) is not detected under the same conditions. The appearance and development of the somatostatinergic phenotype has been quantified by RIA both in cell extracts and in the culture medium. Somatostatin-like immunoreactivity (SLI) undetectable at the late gastrula stage, can be measured in cells after 4 days of culture and continues to increase over the next 10 days. In culture medium, SLI is present at a constant level from day 4 up to day 14. These data reveal that some neuronal precursor cells acquire, during neural induction, the potentiality to biosynthesize, store and release neuropeptides. Furthermore, the expression of these peptidergic phenotypes in distinct subpopulations of neurones suggests that certain neuronal precursors become committed to different metabolic pathways at the earliest steps of neurogenesis.

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Takahashi, Masanori, and Noriko Osumi. "Pax6 regulates specification of ventral neurone subtypes in the hindbrain by establishing progenitor domains." Development 129, no. 6 (March 15, 2002): 1327–38. http://dx.doi.org/10.1242/dev.129.6.1327.

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Recent studies have shown that generation of different kinds of neurones is controlled by combinatorial actions of homeodomain (HD) proteins expressed in the neuronal progenitors. Pax6 is a HD protein that has previously been shown to be involved in the differentiation of the hindbrain somatic (SM) motoneurones and V1 interneurones in the hindbrain and/or spinal cord. To investigate in greater depth the role of Pax6 in generation of the ventral neurones, we first examined the expression patterns of HD protein genes and subtype-specific neuronal markers in the hindbrain of the Pax6 homozygous mutant rat. We found that Islet2 (SM neurone marker) and En1 (V1 interneurone marker) were transiently expressed in a small number of cells, indicating that Pax6 is not directly required for specification of these neurones. We also observed that domains of all other HD protein genes (Nkx2.2, Nkx6.1, Irx3, Dbx2 and Dbx1) were shifted and their boundaries became blurred. Thus, Pax6 is required for establishment of the progenitor domains of the ventral neurones. Next, we performed Pax6 overexpression experiments by electroporating rat embryos in whole embryo culture. Pax6 overexpression in the wild type decreased expression of Nkx2.2, but ectopically increased expression of Irx3, Dbx1 and Dbx2. Moreover, electroporation of Pax6 into the Pax6 mutant hindbrain rescued the development of Islet2-positive and En1-positive neurones. To know reasons for perturbed progenitor domain formation in Pax6 mutant, we examined expression patterns of Shh signalling molecules and states of cell death and cell proliferation. Shh was similarly expressed in the floor plate of the mutant hindbrain, while the expressions of Ptc1, Gli1 and Gli2 were altered only in the progenitor domains for the motoneurones. The position and number of TUNEL-positive cells were unchanged in the Pax6 mutant. Although the proportion of cells that were BrdU-positive slightly increased in the mutant, there was no relationship with specific progenitor domains. Taken together, we conclude that Pax6 regulates specification of the ventral neurone subtypes by establishing the correct progenitor domains.

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Krieger, C., and T. A. Sears. "The development of voltege-dependent ionic conductances In murine spinal cord neurones in culture." Canadian Journal of Physiology and Pharmacology 66, no. 10 (October 1, 1988): 1328–36. http://dx.doi.org/10.1139/y88-217.

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The development of voltage-dependent ionic conductances of foetal mouse spinal cord neurones was examined using the whole-cell patch-clamp technique on neurones cultured from embryos aged 10–12 days (E10–E12) which were studied between the first day in vitro (V1) to V10. A delayed rectifier potassium conductance (IK) and a leak conductance were observed in neurones of E10.V1, E11, V1, and E12, V1 as well as in neurones cultured for longer periods. A rapidly activating and inactivating potassium conductance (IA) was seen in neurones from E11, V2 and E12, V1 and at longer times in vitro. A tetrodotoxin (TTX) sensitive sodium-dependent inward current was observed in neurones of E11 and E12 from V1 onwards. Calcium-dependent conductances were not detectable in these neurones unless the external calcium concentration was raised 10- to 20-foid and potassium conductances were blocked. Under these conditions calcium currents could be observed as early as E11, V3 and E12, V2 and at subsequent times in vitro. The pattern of development of voltage-dependent ionic conductances in murine spinal neurones is such that initially leak and potassium currents are present followed by sodium current and subsequently calcium current.

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ARECHIGA, H., M. CHIQUET, D. P. KUFFLER, and J. G. NICHOLLS. "Formation of Specific Connections in Culture by Identified Leech Neurones Containing Serotonin, Acetylcholine and Peptide Transmitters." Journal of Experimental Biology 126, no. 1 (November 1, 1986): 15–31. http://dx.doi.org/10.1242/jeb.126.1.15.

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Connections made in culture by identified leech neurones have been studied using pairs of cells that contain various transmitters. These cells were: motoneurones innervating the heart; the anterior pagoda neurones (which contain both acetylcholine and an FMRFamide-like peptide); the Retzius cells (which contain serotonin); and the pressure sensory neurones, which contain an unidentified transmitter. Heart motoneurones (HE) and anterior pagoda (AP) cells in culture reacted with antiserum against the peptide FMRFamide. The immunoreactive peptide was found in dense-core vesicles at presumptive sites of transmitter release. In culture the HE, AP and Retzius cells formed non-rectifying electrical synapses with one another within 2 days. Some pairs of cells made connections resembling those seen in normal leech ganglia; others formed novel connections seen only in culture. Electrical connections made by sensory P cells with HE and AP cells showed rectification. Confirming earlier results, P cells never established electrical connections with Retzius cells. In certain pairs of neurones, chemically mediated synaptic interactions developed. Thus, stimulation of AP cells evoked hyperpolarizing synaptic potentials that arose after a delay in Retzius cells and in P cells. Similarly, stimulation of HE cells gave rise to delayed, slow synaptic potentials in AP cells and in isolated heart muscle fibres in culture. P cells, which in culture are never presynaptic to Retzius cells, made chemical connections with AP cells. These results support the conclusion that identified leech neurones in culture make synaptic connections with certain specific target cells while ignoring others.

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Andriamampandry, C., L. Freysz, J. N. Kanfer, H. Dreyfus, and R. Massarelli. "Conversion of ethanolamine, monomethylethanolamine and dimethylethanolamine to choline-containing compounds by neurons in culture and by the rat brain." Biochemical Journal 264, no. 2 (December 1, 1989): 555–62. http://dx.doi.org/10.1042/bj2640555.

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The incubation of neurons from chick embryos in primary culture with [3H]ethanolamine revealed the conversion of this base into monomethyl, dimethyl and choline derivatives, including the corresponding free bases. Labelling with [methyl-3H]monomethylethanolamine and [methyl-3H]dimethylethanolamine supported the conclusion that in chick neuron cultures, phosphoethanolamine appears to be the preferential substrate for methylation, rather than ethanolamine or phosphatidylethanolamine. The methylation of the latter two compounds, in particular that of phosphatidylethanolamine, was seemingly stopped at the level of their monomethyl derivatives. Fetal rat neurons in primary culture incubated with [3H]ethanolamine showed similar results to those observed with chick neurones. However, phosphoethanolamine and phosphatidylethanolamine and, to a lesser extent, free ethanolamine, appeared to be possible substrates for methylation reactions. The methylation of water-soluble ethanolamine compounds de novo was further confirmed by experiments performed in vivo by intraventricular injection of [3H]ethanolamine. Phosphocholine and the monomethyl and dimethyl derivatives of ethanolamine were detected in the brain 15 min after injection.

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21

Grolleau, F., and B. Lapied. "Dorsal unpaired median neurones in the insect central nervous system: towards a better understanding of the ionic mechanisms underlying spontaneous electrical activity." Journal of Experimental Biology 203, no. 11 (June 1, 2000): 1633–48. http://dx.doi.org/10.1242/jeb.203.11.1633.

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The efferent dorsal unpaired median (DUM) neurones, which include octopaminergic neurones, are among the most intensively studied neurones in the insect central nervous system. They differ from other insect neurones in generating endogenous spontaneous overshooting action potentials. The second half of the 1980s is certain to be considered a turning point in the study of the ion channels underlying the electrical activity of DUM neurones. Recent advances made using the patch-clamp technique have stimulated an increasing interest in the understanding of the biophysical properties of both voltage-dependent and voltage-independent ion channels. Patch-clamp studies of DUM neurones in cell culture demonstrate that these neurones express a wide variety of ion channels. At least five different types of K(+) channel have been identified: inward rectifier, delayed rectifier and A-like channels as well as Ca(2+)- and Na(+)-activated K(+) channels. Moreover, besides voltage-dependent Na(+) and Ca(2+)-sensitive Cl(−) channels, DUM neurones also express four types of Ca(2+) channel distinguished on the basis of their kinetics, voltage range of activation and pharmacological profile. Finally, two distinct resting Ca(2+) and Na(+) channels have been shown to be involved in maintaining the membrane potential and in regulating the firing pattern. In this review, we have also attempted critically to evaluate these existing ion channels with regard to their specific functions in the generation of the different phases of the spontaneous electrical activity of the DUM neurone.

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22

Château, Y., G. Dorange, J. F. Clément, N. Rougier, and L. Misery. "Co-culture of neurones with Merkel cells and keratinocytes." Experimental Dermatology 13, no. 9 (June 28, 2008): 578–79. http://dx.doi.org/10.1111/j.0906-6705.2004.0212as.x.

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23

Mena, Maria Angeles, Beatriz Pardo, Carlos Luis Paino, and Justo Garcia De Yebenes. "Levodopa toxicity in foetal rat midbrain neurones in culture." NeuroReport 4, no. 4 (April 1993): 438–40. http://dx.doi.org/10.1097/00001756-199304000-00025.

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Simpson, Carol S., Heather M. Johnston, and Brian J. Morris. "Phenotypic characterisation of rat striatal neurones in primary culture." Tissue and Cell 26, no. 6 (December 1994): 929–41. http://dx.doi.org/10.1016/0040-8166(94)90042-6.

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25

Pouokam, Ervice, Matthias Rehn, and Martin Diener. "Effects of H2O2 at rat myenteric neurones in culture." European Journal of Pharmacology 615, no. 1-3 (August 2009): 40–49. http://dx.doi.org/10.1016/j.ejphar.2009.04.066.

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26

Atterwill, Christopher K. "Brain Reaggregate Cultures in Neurotoxicological Investigations: Studies with Cholinergic Neurotoxins." Alternatives to Laboratory Animals 16, no. 3 (March 1989): 221–30. http://dx.doi.org/10.1177/026119298901600304.

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The number of neurotoxicants which produce ‘lesions’ in organotypic brain reaggregate cultures in vitro, which correlate with known in vivo actions, is growing. With respect to cholinergic neurones, this includes kainic acid, organophosphorus compounds and, in our hands, ethylcholine mustard aziridinium (ECMA) and aluminium. We have demonstrated that in vitro exposure to low concentrations of ECMA (12.5μM) produces a two-stage lesion in rat whole-brain reaggregate cultures, corresponding to initial direct inhibition of choline acetyltransferase (ChAT), followed by a later loss of cholinergic neurones. Higher concentrations of ECMA (25–50μM) are more generally cytotoxic and also cause lesions in non-cholinergic cerebellar granule neurones in monolayer culture. Aluminium (0.1–0.01mM) similarly reduces ChAT activity in rat whole-brain reaggregate cultures. Both agents may be useful in providing brain cholinergic lesions in vitro analagous to those occurring in types of dementia in vivo. The use of brain reaggregates in a ‘stepwise’ procedure for testing potential neurotoxicants is also described.

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27

Sáenz, F., U. García, and U. Aréchiga. "Modulation of electrical activity by 5-hydroxytryptamine in crayfish neurosecretory cells." Journal of Experimental Biology 200, no. 23 (December 1, 1997): 3079–90. http://dx.doi.org/10.1242/jeb.200.23.3079.

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The effect of 5-hydroxytryptamine (5-HT) was tested in a population of X organ neurosecretory cells in the eyestalk of the crayfish Procambarus clarkii. Tests were conducted both in situ and on isolated neurones kept in culture. The application of 5-HT induced action potentials in silent cells. In spontaneously active neurones, 5-HT increased the firing rate and either induced firing or enhanced bursting activity. The effect of 5-HT was dose-dependent within the range 1-100 micromol l-1 in cells of the intact organ. The effect persisted for 20-30 min after 5-HT had been removed from the bathing solution. Successive applications of 5-HT onto the same neurone reduced responsiveness, suggesting that desensitization had occurred. The effects of 5-HT were blocked by prior incubation with the 5-HT antagonist methysergide. In X organ cells whose axons and branches in the neuropile had been severed, 5-HT induced a depolarisation associated with a slow inward current. In X organ neurones isolated from the eyestalk and kept in culture, 5-HT was capable of evoking bursts of action potentials and elicited a slow inward current. This effect was also blocked by methysergide (10(-4 )mol l-1). These results suggest a direct modulatory effect of 5-HT on the pattern of electrical activity in the X organ cells.

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28

Price, Jack, Brenda Williams, and Elizabeth Grove. "Cell lineage in the cerebral cortex." Development 113, Supplement_2 (April 1, 1991): 23–28. http://dx.doi.org/10.1242/dev.113.supplement_2.23.

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We have studied cell lineage in the rat cerebral cortex using retroviral mediated gene transfer. By this method, a marker gene is inserted into dividing precursor cells such that their fate can be followed. We have applied this technique to two types of experiment. First, virus was used to label precursor cells of the cerebral cortex in situ during the period of neurogenesis. Second, cortical precursor cells were grown in dissociated cell culture, and virus was used to follow their development over the culture period. These experiments showed that the majority of precursor cells generate a single cell type – neurones, astrocytes, or oligodendrocytes. Moreover, this is true both in vivo and in dissociated cell culture. The only exception is a bipotential cell, which can generate both neurones and oligodendrocytes. These data suggest that the ventricular zone – the germinal layer of the embryonic cortex – is a mosaic of precursor cells of different restricted potentials. Although precursor cells are restricted in terms of the cell types they generate, they seem not to be restricted in either the cortical laminae or cytoarchitectonic areas to which they can contribute. Both neuronal and grey matter astrocyte precursors contribute cells to multiple layers of both infra- and supragranular laminae. Moreover, in the hippocampal formation, neuronal precursors can contribute cells to more than one hippocampal field.

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29

Wang, Wengang, John H. Pizzonia, and George B. Richerson. "Chemosensitivity of rat medullary raphe neurones in primary tissue culture." Journal of Physiology 511, no. 2 (September 1998): 433–50. http://dx.doi.org/10.1111/j.1469-7793.1998.433bh.x.

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30

Díez-Guerra, F. Javier, and Jesus Avila. "MAP2 phosphorylation parallels dendrite arborization in hippocampal neurones in culture." NeuroReport 4, no. 4 (April 1993): 419–22. http://dx.doi.org/10.1097/00001756-199304000-00020.

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31

Malgrange, Brigitte, Philippe Lefebvre, Thomas R. Van de Water, Hinrich Staecker, and Gustave Moonen. "Effects of neurotrophins on early auditory neurones in cell culture." NeuroReport 7, no. 4 (March 1996): 913–17. http://dx.doi.org/10.1097/00001756-199603220-00016.

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32

Kumamoto, Eiichi, and Yuzo Murata. "GABAA-receptor channels on rat cholinergic septal neurones in culture." Neuroscience Research Supplements 19 (January 1994): S51. http://dx.doi.org/10.1016/0921-8696(94)92404-x.

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33

Lees, George, David J. Beadle, Roger P. Botham, and John S. Kelly. "Excitable properties of insect neurones in culture: A developmental study." Journal of Insect Physiology 31, no. 2 (January 1985): 135–43. http://dx.doi.org/10.1016/0022-1910(85)90018-6.

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34

Hassall, C. J. S., R. Penketh, C. Rodeck, and G. Burnstock. "The intracardiac neurones of the fetal human heart in culture." Anatomy and Embryology 182, no. 4 (October 1990): 329–37. http://dx.doi.org/10.1007/bf02433493.

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35

Silani, V., M. Braga, A. Ciammola, V. Cardin, and G. Scarlato. "Motor neurones in culture as a model to study ALS." Journal of Neurology 247, no. 13 (March 20, 2000): I28—I36. http://dx.doi.org/10.1007/s004150050554.

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36

Jackel, C., W. Krenz, and F. Nagy. "BICUCULLINE/BACLOFEN-INSENSITIVE GABA RESPONSE IN CRUSTACEAN NEURONES IN CULTURE." Journal of Experimental Biology 191, no. 1 (June 1, 1994): 167–93. http://dx.doi.org/10.1242/jeb.191.1.167.

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Neurones were dissociated from thoracic ganglia of embryonic and adult lobsters and kept in primary culture. When gamma-aminobutyric acid (GABA) was applied by pressure ejection, depolarizing or hyperpolarizing responses were produced, depending on the membrane potential. They were accompanied by an increase in membrane conductance. When they were present, action potential firing was inhibited. The pharmacological profile and ionic mechanism of GABA-evoked current were investigated under voltage-clamp with the whole-cell patch-clamp technique. The reversal potential of GABA-evoked current depended on the intracellular and extracellular Cl- concentration but not on extracellular Na+ and K+. Blockade of Ca2+ channels by Mn2+ was also without effect. The GABA-evoked current was mimicked by application of the GABAA agonists muscimol and isoguvacine with an order of potency muscimol>GABA>isoguvacine. cis-4-aminocrotonic acid (CACA), a folded and conformationally restricted GABA analogue, supposed to be diagnostic for the vertebrate GABAC receptor, also induced a bicuculline-resistant chloride current, although with a potency about 10 times lower than that of GABA. The GABA-evoked current was largely blocked by picrotoxin, but was insensitive to the GABAA antagonists bicuculline, bicuculline methiodide and SR 95531 at concentrations of up to 100 µmol l-1. Diazepam and phenobarbital did not exert modulatory effects. The GABAB antagonist phaclophen did not affect the GABA-induced current, while the GABAB agonists baclophen and 3-aminopropylphosphonic acid (3-APA) never evoked any response. Our results suggest that lobster thoracic neurones in culture express a chloride-conducting GABA-receptor channel which conforms to neither the GABAA nor the GABAB types of vertebrates but shows a pharmacology close to that of the novel GABAC receptor described in the vertebrate retina.

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37

Koizumi, Schuichi, Kayoko Fujishita, Makoto Tsuda, and Kazuhide Inoue. "Neurone-to-astrocyte communication by endogenous ATP in mixed culture of rat hippocampal neurones and astrocytes." Drug Development Research 59, no. 1 (May 2003): 88–94. http://dx.doi.org/10.1002/ddr.10206.

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38

BEADLE, DAVID J., G. HORSEMAN, Y. PICHON, M. AMAR, and T. SHIMAHARA. "Acetylcholine-Activated Ion Channels in Embryonic Cockroach Neurones Growing in Culture." Journal of Experimental Biology 142, no. 1 (March 1, 1989): 337–55. http://dx.doi.org/10.1242/jeb.142.1.337.

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Application of acetylcholine and carbamylcholine to cultured cockroach neurones held under whole-cell voltage-clamp conditions evoked an inward current that was accompanied by an increase in current noise. Fluctuation analysis of the noise revealed the existence of two Lorentzian components in acetylcholine, of corner frequencies 10 ± 0.6 Hz and 116 ± 9 Hz, and one Lorentzian component in carbamylcholine, of corner frequency 35 ± 13 Hz. Single-channel analysis of the unitary currents evoked by acetylcholine or carbamylcholine in neurones held in the cell-attached mode of the patch-clamp technique revealed the presence of two categories of channel events. The large events had mean currents of 4.77 pA with acetylcholine and 5.09 pA with carbamylcholine, and the small events 1.92 pA (acetylcholine) and 1.72pA (carbamylcholine) for a hyperpolarization of 60 mV. The reversal potentials for these currents relative to the resting potential were estimated to be - 70 mV for acetylcholine and - 68 mV for carbamylcholine, and the conductance values calculated from the I/V curves were 37 pS (large) and 19 pS (small) for acetylcholine and 52 pS (large) and 15 pS (small) for carbamylcholine. It is concluded that embryonic cockroach neurones growing in vitro possess two populations of acetylcholine-activated ion channels, and the possibility that one of these represents an embryonic receptor and the other an adult receptor is discussed.

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39

Frade, J. M. "Unscheduled re-entry into the cell cycle induced by NGF precedes cell death in nascent retinal neurones." Journal of Cell Science 113, no. 7 (April 1, 2000): 1139–48. http://dx.doi.org/10.1242/jcs.113.7.1139.

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During their early postmitotic life, a proportion of the nascent retinal ganglion cells (RGCs) are induced to die as a result of the interaction of nerve growth factor (NGF) with the neurotrophin receptor p75. To analyse the mechanisms by which NGF promotes apoptosis, an in vitro culture system consisting of dissociated E5 retinal cells was established. In this system, NGF-induced apoptosis was only observed in the presence of insulin and neurotrophin-3, conditions that favour the birth of RGCs and other neurones expressing the glycoprotein G4. The pro-apoptotic effect of NGF on the G4-positive neurones was evident after 10 hours in vitro and was preceded by a significant upregulation of cyclin B2, but not cyclin D1, and the presence of mitotic nuclei in these cells. Brain-derived neurotrophic factor prevented both the increase of cyclin B2 expression in the G4-positive neurones and the NGF-induced cell death. Finally, pharmacologically blocking cell-cycle progression using the cyclin-dependent kinase inhibitor roscovitine prevented NGF-induced cell death in a dose-dependent manner. These results strongly suggest that the apoptotic signalling initiated by NGF requires a driving stimulus manifested by the neuronal birth and is preceded by the unscheduled re-entry of postmitotic neurones into the cell cycle.

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40

Giles, D. P., and P. N. R. Usherwood. "Locust nymphal neurones in culture: A new technique for studying the physiology and pharmacology of insect central neurones." Comparative Biochemistry and Physiology Part C: Comparative Pharmacology 80, no. 1 (January 1985): 53–59. http://dx.doi.org/10.1016/0742-8413(85)90131-8.

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41

Amos, BJ, and CD Richards. "Intrinsic hydrogen ion buffering in rat CNS neurones maintained in culture." Experimental Physiology 81, no. 2 (March 1, 1996): 261–71. http://dx.doi.org/10.1113/expphysiol.1996.sp003930.

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42

Wann, K. T., P. A. Goodwin, and C. D. Richards. "High Activity K+ Channels in Rat Hippocampal Neurones Maintained in Culture." Experimental Physiology 84, no. 3 (May 1999): 501–14. http://dx.doi.org/10.1111/j.1469-445x.1999.01824.x.

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43

Ascher, P., and L. Nowak. "Quisqualate- and kainate-activated channels in mouse central neurones in culture." Journal of Physiology 399, no. 1 (May 1, 1988): 227–45. http://dx.doi.org/10.1113/jphysiol.1988.sp017077.

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44

WANN*, K. T., P. A. GOODWIN, and C. D. RICHARDS. "High activity K+ channels in rat hippocampal neurones maintained in culture." Experimental Physiology 84, no. 3 (May 1999): 501–14. http://dx.doi.org/10.1017/s0958067099018242.

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45

Jackel, Carolin, Wulf Dieter Krenz, and Frederic Nagy. "A receptor with GABAC-like pharmacology in invertebrate neurones in culture." NeuroReport 5, no. 9 (May 1994): 1097–101. http://dx.doi.org/10.1097/00001756-199405000-00019.

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46

Devaud, Jean-Marc, Brigitte Quenet, Jean Gascuel, and Claudine Masson. "A morphometric classification of pupal honeybee antennal lobe neurones in culture." NeuroReport 6, no. 1 (December 1994): 214–18. http://dx.doi.org/10.1097/00001756-199412300-00054.

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47

Malgrange, Brigitte, Jean-Michel Rigo, Paul Coucke, Shibeshi Belachew, Bernard Register, and Gustave Moonen. "β-Carbolines induce apoptotic death of cerebellar granule neurones in culture." NeuroReport 7, no. 18 (November 1996): 3041–46. http://dx.doi.org/10.1097/00001756-199611250-00049.

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48

Malgrange, Brigitte, Jean-Michel Rigo, Philippe P. Lefebvre, Paul Coucke, Frédéric Goffin, Gaël Xhauflaire, Shibeshih Belachew, Thomas R. Van De Water, and Gustave Moonen. "Diazepam-insensitive GABAA receptors on postnatal spiral ganglion neurones in culture." NeuroReport 8, no. 3 (February 1997): 591–96. http://dx.doi.org/10.1097/00001756-199702100-00003.

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49

TIAHO, Franc¸ois, Nelly CHARRIER, Janine GUEN, and Yves PICHON. "CALCIUM CURRENTS IN DEVELOPING NEURONES FROM COCKROACH BRAINS IN PRIMARY CULTURE." Biology of the Cell 86, no. 2-3 (1996): 194. http://dx.doi.org/10.1016/0248-4900(96)84816-7.

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Harrison, J. B., C. A. Leech, J. Katz, and D. B. Sattelle. "Embryonic and adult neurones of the housefly (Musca domestica) in culture." Tissue and Cell 22, no. 3 (January 1990): 337–47. http://dx.doi.org/10.1016/0040-8166(90)90008-w.

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Journal articles on the topic 'Culture de neurones'

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