Academic literature on the topic 'Axon collaterals'

1. Single axons of pontine nucleus neurons (PN axons) receiving cerebral input were stained intra-axonally with horseradish peroxidase (HRP) in the cerebellum of cats. The axonal trajectory of single PN axons was reconstructed from serial sections of the cerebellum and the brain stem. 2. Axons were penetrated in the white matter near the dentate nucleus, and, after electrophysiological identification, PN axons were injected iontophoretically with HRP. The identification criteria for the PN axons were 1) their direct responses to stimulation of the contralateral pontine nucleus (PN), 2) their synaptic activation from the contralateral cerebral cortex, and 3) the decrease in threshold for evoking direct spikes in stimulation of the PN by conditioning stimuli applied in the cerebral cortex. 3. Two hundred thirty-three axons were electrophysiologically identified as PN axons receiving the input from the cerebral cortex. Ninety-six of them were stained successfully with HRP, and reconstructions were made from 40 well-stained PN axons. All of them gave rise to mossy fibers and terminated in the granular layer of the cerebellar cortex as typical mossy fiber rosettes. Out of these, 22 gave axon collaterals to the dentate nucleus. Virtually all of the axon branches observed in the dentate nucleus were axon collaterals of mossy fibers from the PN to the cerebellar cortex. In 7 of these 22 PN axons, cell bodies were retrogradely labeled with HRP, and all of them were found in the contralateral PN. 4. The stained-stem axons arising from the PN ran medially in the pons, crossed the midline, and then ascended dorsocaudally in the branchium pontis. After passing in the white matter anterior to or lateral to the dentate nucleus, they entered into the cerebellar cortex. On their way, one to three axon collaterals were given off from parent axons to the dentate nucleus. The diameter of these collaterals was very thin (mean, 0.6 microns), compared with the large diameter of the parent axons (mean, 2.1 microns). 5. Some axon collaterals were very simple and had only one terminal branch with or without short branchlets, whereas others were more complex, and single axon collaterals ramified before forming a terminal arborization. Axon collaterals of single PN axons mainly spread mediolaterally or dorsoventrally in the frontal plane but had a very narrow rostrocaudal extension. 6. Terminal branches usually bore swellings en passant along their length and one terminal swelling at their end. The number of swellings per axon collateral ranged 23-180 (116 +/- 52, mean +/- SD).(ABSTRACT TRUNCATED AT 400 WORDS)

AbstractRetinal ganglion cells (RGCs), the output neurons of the retina, have axons that project via the optic nerve to diverse targets in the brain. Typically, RGC axons do not branch before exiting the retina and thus do not provide it with synaptic feedback. Although a small subset of RGCs with intraretinal axon collaterals has been previously observed in human, monkey, cat, and turtle, their function remains unknown. A small, more recently identified population of RGCs expresses the photopigment melanopsin. These intrinsically photosensitive retinal ganglion cells (ipRGCs) transmit an irradiance-coding signal to visual nuclei in the brain, contributing both to image-forming vision and to several nonimage-forming functions, including circadian photoentrainment and the pupillary light reflex. In this study, using melanopsin immunolabeling in monkey and a genetic method to sparsely label the melanopsin cells in mouse, we show that a subgroup of ipRGCs have axons that branch en route to the optic disc, forming intraretinal axon collaterals that terminate in the inner plexiform layer of the retina. The previously described collateral-bearing population identified by intracellular dye injection is anatomically indistinguishable from the collateral-bearing melanopsin cells identified here, suggesting they are a subset of the melanopsin-expressing RGC type and may therefore share its functional properties. Identification of an anatomically distinct subpopulation in mouse, monkey, and human suggests this pathway may be conserved in these and other species (turtle and cat) with intraretinal axon collaterals. We speculate that ipRGC axon collaterals constitute a likely synaptic pathway for feedback of an irradiance signal to modulate retinal light responses.

1. Two hundred and twelve corticospinal axons were identified by stimulation in the hindlimb representation in area 3b of the somatosensory cortex and were recorded in the left dorsolateral funiculus of the spinal cord of the cat. The mean conduction velocity was 38 m/s, range 9-113 m/s. 2. Electrical stimulation of the receptive field evoked discharge in corticospinal axons with a mean latency of 36 ms (range 9-100 ms). 3. One hundred nine of the 212 recorded axons were successfully intra-axonally labeled by iontophoretic injection of horseradish peroxidase, with the mean length of labeled axon being 4.8 mm. Seventy-three of the labeled axons issued no collaterals, and 36 issued at least one labeled collateral into the spinal gray matter along the labeled portion of the parent axon. 4. Most labeled axons issued only one labeled collateral per spinal cord segment. Fourteen collaterals from 10 units were labeled well enough to permit reconstruction of their terminal arborizations. 5. Most terminal collaterals were oriented rostrocaudally and terminated in laminae V, VI, and VII. Most collaterals terminated within large mediolateral extents of the gray matter with no apparent topographic organization. 6. No collaterals terminated in laminae I or II or within the motoneuron pools, and no apparent correlation was found between their anatomic and physiological characteristics.

Ganglion cells with intraretinal axon collaterals have been described in monkey (Usai et al., 1991), cat (Dacey, 1985), and turtle (Gardiner & Dacey, 1988) retina. Using intracellular injection of horseradish peroxidase and Neurobiotin in in vitro whole-mount preparations of human retina, we filled over 1000 ganglion cells, 19 of which had intraretinal axon collaterals and wide-field, spiny dendritic trees stratifying in the inner half of the inner plexiform layer. The axons were smooth and thin (∼2 μm) and gave off thin (<1 μm), bouton-studded terminal collaterals that extended vertically to terminate in the outer half of the inner plexiform layer. Terminal collaterals were typically 3–300 μm in length, though sometimes as long as 700 μm, and were present in clusters, or as single branched or unbranched varicose processes with round or somewhat flattened lobular terminal boutons 1–2 μm in diameter. Some cells had a single axon whereas other cells had a primary axon that gave rise to 2–4 axon branches. Axons were located either in the optic fiber layer or just beneath it in the ganglion cell layer, or near the border of the ganglion cell layer and the inner plexiform layer. This study shows that in the human retina, intraretinal axon collaterals are associated with a morphologically distinct ganglion cell type. The synaptic connections and functional role of these cells are not yet known. Since distinct ganglion cell types with intraretinal axon collaterals have also been found in monkey, cat, and turtle, this cell type may be common to all vertebrate retinas.

Here, we review our studies in rats of target recognition by developing cortical axons focusing on their innervation of the basilar pons, a major hindbrain target. The corticopontine projection develops by a ‘delayed interstitial budding’ of collaterals from layer 5 corticospinal axons, rather than by a direct ingrowth of primary axons or by bifurcation of the growth cone. Branches form de novo from the axon cylinder in the pathway overlying the basilar pons and extend directly into it. Cocultures of cortex and basilar pons in 3-dimensional collagen matrices show that a diffusible chemotropic signal released by the basilar pons directs the growth of collateral branches from layer 5 axons in a target and neuron specific manner. ‘Delayed’ co-cultures suggest that a diffusible, pontine-derived signal also initiates the selective branching of layer 5 axons. In vivo experiments support this chemotropic mechanism. First, corticospinal axons form collateral branches at novel locations directly over ectopic aggregations of basilar pontine neurons induced by X-irradiation; no branches form at positions that would normally overlie the appropriate region of basilar pons which is absent because of the X-irradiation. Thus, the basilar pons, rather than local cues in the axon pathway, appears to control the location of corticospinal axon branching. Second, in a series of experiments in which different subsets of corticospinal axons are prevented from innervating the basilar pons, remaining corticospinal axons extend collaterals in a directed manner to regions of the basilar pons deprived of cortical input, a behavior consistent with a response to a diffusible chemoattractant emanating from these regions. In conclusion, our findings suggest that a diffusible, target-derived chemotropic molecule(s) underlies target recognition in this developing system by initiating the formation and directing the growth of pontine collateral branches of primary layer 5 corticospinal axons.

To investigate intraspinal branching patterns of single corticospinal neurons (CSNs), we recorded extracellular spike activities from cell bodies of 408 CSNs in the motor cortex in anesthetized cats and mapped the distribution of effective stimulating sites for antidromic activation of their terminal branches in the spinal gray matter. To search for all spinal axon branches belonging to single CSNs in the "forelimb area" of the motor cortex, we microstimulated the gray matter from the dorsal to the ventral border at 100-micron intervals at an intensity of 150-250 microA and systematically mapped effective stimulating penetrations at 1-mm intervals rostrocaudally from C3 to the most caudal level of their axons. From the depth-threshold curves, the comparison of the antidromic latencies of spikes evoked from the gray matter and the lateral funiculus, and the calculated conduction times of the collaterals, we could ascertain that axon collaterals were stimulated in the gray matter rather than stem axons in the corticospinal tract due to current spread. Virtually all CSNs examined in the forelimb area of the motor cortex had three to seven branches at widely separated segments of the cervical and the higher thoracic cord. In addition to terminating at the brachial segments, they had one to three collaterals to the upper cervical cord (C3-C4), where the propriospinal neurons projecting to forelimb motoneurons are located. About three quarters of these CSNs had two to four collaterals in C6-T1. This finding held true for both fast and slow CSNs. About one third of the CSNs in the forelimb area of the motor cortex projected to the thoracic cord below T3. These CSNs also sent axon collaterals to the cervical spinal cord. CSNs in the "hindlimb area" of the motor cortex had three to five axon branches in the lumbosacral cord. These branches were mainly observed at L4 and the lower lumbosacral cord. None of these CSNs had axon collaterals in the cervical cord. CSNs terminating at different segments of the cervical and the thoracic cord were distributed in a wide area of the motor cortex and were intermingled. To determine the detailed trajectory of single axon branches, microstimulation was made at a matrix of points of 100 or 200 micron at the maximum intensity of 30 microA, and their axonal trajectory was reconstructed on the basis of the location of low-threshold foci and the latency of antidromic spikes.(ABSTRACT TRUNCATED AT 400 WORDS)

Axon conduction fidelity is important for signal transmission and has been studied in various axons, including the Schaffer collateral axons of the hippocampus. Previously, we reported that high-frequency stimulation (HFS) depresses Schaffer collateral excitability when assessed by whole-cell recordings from CA3 pyramidal cells but induces biphasic excitability changes (increase followed by decrease) in extracellular recordings of CA1 fiber volleys. Here, we examined responses from proximal (whole-cell or field-potential recordings from CA3 pyramidal cell somata) and distal (field-potential recordings from CA1 stratum radiatum) portions of the Schaffer collaterals during HFS and burst stimulation in hippocampal slices. Whole-cell and dual-field-potential recordings using 10–100-Hz HFS revealed frequency-dependent changes like those previously described, with higher frequencies producing more drastic changes. Dual-field-potential recordings revealed substantial differences in the response to HFS between proximal and distal regions of the Schaffer collaterals, with proximal axons depressing more strongly and only distal axons showing an initial excitability increase. Because CA3 pyramidal neurons normally fire in short bursts rather than long high-frequency trains, we repeated the dual recordings using 100–1,000-ms interval burst stimulation. Burst stimulation produced changes similar to those during HFS, with shorter intervals causing more drastic changes and substantial differences observed between proximal and distal axons. We suggest that functional differences between proximal and distal Schaffer collaterals may allow selective filtering of nonphysiological activity while maximizing successful conduction of physiological activity throughout an extensive axonal arbor.

The optic tract of the goldfish splits into two brachia just before it reaches the tectum, normal optic axons being distributed systematically between the two according to their retinal origins. The orderliness of this division, like that of the retinotectal projection itself, is conventionally attributed to a system of specific axonal guidance cues. However, the brachial distribution of regenerated axons is much less orderly; and, since there is evidence that these axons have many collateral branches in the nerve and tract, the gross order that remains after regeneration could potentially arise secondarily, in parallel with refinement of the retinotectal map, by a preferential loss of collaterals from the inappropriate brachium. The brachial paths of normal axons, and axons regenerated after optic nerve cut for periods ranging from 19 days to 5 years, were therefore studied by anterograde labelling with horseradish peroxidase from discrete retinal lesions or retrograde labelling of ganglion cells from a cut brachium. From 19 to 28 days, regenerating axons showed little or no preference for their normal brachium. During this period (which includes the first week of tectal synaptogenesis) an average of 46á3% of cells retrogradely labelled from a cut medial brachium were in dorsal retina, compared with only 1á45% in normal fish. Some preference for the normal brachium was evident at 35 days and significant order had returned by 42–70 days, when the average proportion of labelled cells in dorsal retina had fallen to 25á4% though the average number in the whole retina was unchanged. Thus a brachial refinement had occurred in parallel with refinement of the retinotectal map. These results support the idea of a selective loss of axon collaterals from the inappropriate brachium, though they do not exclude the possibility of some concurrent gain in the appropriate one. We suggest that refinement may depend on a process we term ‘sibling rivalry’: competition between different collaterals of the same axon to form a critical number of stable tectal synapses, in which the most- normally-routed branches have the best chance of succeeding and surviving. Developing normal axons might also make use of collateral formation and ‘sibling rivalry’ to generate and refine the complex interwoven patterns of the normal optic tract.

Perlmutter, S. I., Y. Iwamoto, L. F. Barke, J. F. Baker, and B. W. Peterson. Relation between axon morphology in C1 spinal cord and spatial properties of medial vestibulospinal tract neurons in the cat. J. Neurophysiol. 79: 285–303, 1998. Twenty-one secondary medial vestibulospinal tract neurons were recorded intraaxonally in the ventromedial funiculi of the C1 spinal cord in decerebrate, paralyzed cats. Antidromic stimulation in C6 and the oculomotor nucleus identified the projection pattern of each neuron. Responses to sinusoidal, whole-body rotations in many planes in three-dimensional space were characterized before injection of horseradish peroxidase or Neurobiotin. The spatial response properties of 19 neurons were described by a maximum activation direction vector (MAD), which defines the axis and direction of rotation that maximally excites the neuron. The other two neurons had spatio-temporal convergent behavior and no MAD was calculated. Collateral morphologies were reconstructed from serial frontal sections to reveal terminal fields in the C1 gray matter. Axons gave off multiple collaterals that terminated ipsilaterally to the stem axon. Collaterals of individual axons rarely overlapped longitudinally but projected to similar regions in the ventral horn when viewed in transverse sections. The number of primary collaterals in C1 was different for vestibulo-collic, vestibulo-oculo-collic, and C6-projecting neurons: on average one every 1.34, 1.72, and 4.25 mm, respectively. The heaviest arborization and most terminal boutons were seen in the ventral horn, in laminae VIII and IX. Varicosities on terminal branches in lamina IX were observed adjacent to large cell bodies—putative neck motoneurons—in counterstained tissue. Some collaterals had branches that extended dorsally to lamina VII. Neurons with different spatial properties had terminal fields in different regions of the ventral horn. Axons with type I responses and MADs near those of a semicircular canal pair had widely distributed collateral branches and numerous terminations in the dorsomedial, ventromedial, and spinal accessory nuclei and in lamina VIII. Axons with type I responses that suggested convergent canal pair input, with type II responses, and with spatio-temporal convergent behavior had smaller terminal fields. Some neurons with these more complex spatial properties projected to the dorsomedial and spinal accessory but not to the ventromedial nuclei. Others had focused projections to dorsolateral regions of the ventral horn with few branches in the motor nuclei.

There is suggestive evidence that pyramidal cell axons in neocortex may be coupled by gap junctions into an “axonal plexus” capable of generating very fast oscillations (VFOs) with frequencies exceeding 80 Hz. It is not obvious, however, how a pyramidal cell in such a network could control its output when action potentials are free to propagate from the axons of other pyramidal cells into its own axon. We address this problem by means of simulations based on three-dimensional reconstructions of pyramidal cells from rat somatosensory cortex. We show that somatic depolarization enables propagation via gap junctions into the initial segment and main axon, while somatic hyperpolarization disables it. We show further that somatic voltage cannot effectively control action potential propagation through gap junctions on minor collaterals; action potentials may therefore propagate freely from such collaterals regardless of somatic voltage. In previous work, VFOs are all but abolished during the hyperpolarization phase of slow oscillations induced by anesthesia in vivo. This finding constrains the density of gap junctions on collaterals in our model and suggests that axonal sprouting due to cortical lesions may result in abnormally high gap junction density on collaterals, leading in turn to excessive VFO activity and hence to epilepsy via kindling.

Cholinergic input to, and cholinergic mechanisms within the lower lumbar (L6) and upper sacral (S1) spinal cord of rat may influence neuronal excitability and afferent transmission (Thor et al, 2000) and may provide the environment necessary for appropriate central nervous system control of bladder and bowel function. It is unclear, however, if cholinergic terminals and receptors are present in the L6 & S1 spinal segments of rat and when this may develop. Cholinergic mechanisms have been shown to alter sensory afferent transmission, enhance motoneuron excitability, induce plateau potentials via non-linear membrane properties in motoneurons and reveal oscillations in locomotor-related interneurons. The enhanced activity of sphincter motoneurons was attributed to non-linear properties during the continence phase of distention-evoked voiding in the decerebrate cat (Paroschy & Shefchyk, 2000). Candidate neurotransmitters inducing non-linear properties in cat sphincter motoneurons are 5-HT (Paroschy & Shefchyk, 2000) and acetylcholine via motoneuron axon collaterals (Sasaki, 1994) and other spinal sources. We have established using the antibody to the vesicular acetylcholine transporter (VAChT) that cholinergic terminals are present on ventrolateral Onuf (VLO), dorsomedial Onuf (DMO) motoneurons and parasympathetic preganglionic motoneurons (PGN) in the L6 and S1 rat spinal cord segments. Muscarinic receptor (M2), nicotinic-α4 and α7 receptor subunit immunoreactivity was also present on Onuf motoneurons and in regions dorsal to the PGN. One source of the cholinergic puncta on Onuf motoneurons may be from motoneuron axon collaterals which we observed on a postnatal day 15 VLO motoneuron. Cholinergic terminals were observed on vasoactive intestinal polypeptide-immunoreactive (VIP) afferents, interneurons in the intermediolateral (IML) region and perhaps on other afferents in the lateral and medial collateral pathway of L6 and S1 spinal segments. In the ventral horn, the cholinergic puncta and receptors appear to have a mature distribution around two weeks postnatal and the cholinergic terminals appeared to have a mature distribution in the IML region by three weeks postnatal. Using whole cell patch clamp recording techniques and thick slices of the L6 and S1 rat spinal cord, we observed excitatory responses of ventral horn neurons and motoneurons to carbachol (10-50 μM), a non-specific cholinergic agonist. Ventral horn neurons (postnatal day 8- 16) exhibited prolonged firing and prolonged depolarizations (plateau potentials) beyond the duration of the applied excitatory input from cholinergic (n=6/33) and other (n= 4/37) neurotransmitter systems. In a selection of the neurons with plateau potentials, the L-type calcium current played a role in the plateau production (n=5/5) and low frequency oscillations (n=2/2) as revealed by nifedipine. Postnatally, the voiding reflex changes from a perineal-evoked reflex, to the adult bladder-bladder reflex. Cholinergic input may be responsible in part for the bursting activity of the external urethral sphincter and the activation of the bladder, which is required for complete voiding reflexes in the adult rat. Plateau potentials and enhanced excitability due to cholinergic mechanisms could render inessential a constant excitatory drive that is required in the perineal-evoked voiding reflex in the neonatal rat and may underlie changes in the voiding reflexes that occur during postnatal development.
February 2006

碩士
國立臺灣大學
生命科學系
103
Retinal structure and functional circuits have been studied for more than a century. It is well known that the information flow of retinal circuit starts form light reception by rods and cones, to horizontal cells, amacrine cells and bipolar cells, and transduces to brain by retinal ganglion cells. However, recent studies indicated that a group of melanopsin containing retinal ganglion cells, named intrinsically photosensitive retinal ganglion cells (ipRGCs), send feedback signal to specific sub-population of amacrine cells by an unknown mechanism. Recent studies showed that ipRGC contain intra-retinal axon collaterals that stratified in the inner plexiform layer, yet the morphology and functions of these collaterals remain unclear. By randomly genetic labeling of ipRGCs in mice, our study shows two morphologically distinct types of ipRGC intra-retinal axon collaterals. We also found those collaterals connect to dopaminergic amacrine cells. Our finding suggests that ipRGCs send feedback signal to amacrine cells via intra-retinal collaterals, which may modulate retinal functions.

碩士
國立臺灣大學
生命科學系
104
Ambient luminance is a vital environmental information for us to maintain our normal physiological function, but its contribution to our visual system is still poorly understood. It has been shown that our visual system primary detect contrast, while the background luminance plays little to no role for pattern forming functions. However, recent evidence has suggested that environment luminance may be involved in vision through intrinsic photosensitive retina ganglion cell (ipRGC), which is the third type of photoreceptor in mouse retina. ipRGCs express melanopsin for light sensing and transmit their signal directly to the brain for circadian photo-entrainment, pupillary light reflex and sleep regulation. Previous studies showed that ipRGCs can be divided into at least five types according to their dendritic morphology and cell body size. Furthermore, recent study showed that M1 type ipRGCs have intra retinal axons collateral innervating retrogradely to the inner plexiform layer (IPL), which could form a putative synapse with dopamine amacrine cells (DACs). Since dopamine is important for the light adaptation, we hypothesize that the M1 ipRGCs may also be involved in visual function through the connection with DACs. Using genetic mouse model and electroretinogram, our study shows that elimination of ipRGCs blocks the light adaptation of cones, while application of D1 or D4 receptor agonist can rescue the light adaptation. Together, our data indicates that ipRGCs could modulate visual function through DACs and probably be involved in higher complicated visual function.

In highly polarized cells like neurons, cytoskeleton dynamics play a crucial role in establishing neuronal connections during development and are required for adult plasticity. Actin turnover is particularly important for neurite growth, axon path finding, branching and synaptogenesis. Motoneurons establish several thousand branches that innervate neuromuscular synapses (NMJs). Axonal branching and terminal arborization are fundamental events during the establishment of synapses in motor endplates. Branching process is triggered by the assembly of actin filaments along the axon shaft giving rise to filopodia formation. The unique contribution of the three actin isoforms, α-, β- and γ-actin, in filopodia stability and dynamics during this process is not well characterized. Here, we performed high resolution in situ hybridization and qRT-PCR and showed that in primary mouse motoneurons α-, β- and γ-actin isoforms are expressed and their transcripts are translocated into axons. Using FRAP experiments, we showed that transcripts for α-, β- and γ-actin become locally translated in axonal growth cones and translation hot spots of the axonal branch points. Using live cell imaging, we showed that shRNA depletion of α-actin reduces dynamics of axonal filopodia which correlates with reduced number of collateral branches and impairs axon elongation. Depletion of β-actin correlates with reduced dynamics of growth cone filopoida, disturbs axon elongation and impairs presynaptic differentiation. Also, depletion of γ-actin impairs axonal growth and decreases axonal filopodia dynamics. These findings implicate that actin isoforms accomplish unique functions during development of motor axons. Depletions of β- and γ-actin lead to compensatory upregulation of other two isoforms. Consistent with this, total actin levels remain unaltered and F-actin polymerization capacity is preserved. After the knockdown of either α- or γ-actin, the levels of β-actin increase in the G-actin pool indicating that polymerization and stability of β-actin filaments depend on α- or γ-actin. This study provides evidence both for unique and overlapping function of actin isoforms in motoneuron growth and differentiation. In the soma of developing motoneurons, actin isoforms act redundantly and thus could compensate for each other’s loss. In the axon, α-, β- and γ-actin accomplish specific functions, i.e. β-actin regulates axon elongation and plasticity and α- and γ-actin regulate axonal branching. Furthermore, we show that both axonal transport and local translation of α-, β- and γ-actin isoforms are impaired in Smn knockout motoneurons, indicating a role for Smn protein in RNA granule assembly and local translation of these actin isoforms in primary mouse motoneurons
In stark polaren Zellen wie den Neuronen ist die Etablierung neuronaler Netzwerke ein entscheidender Faktor bei der Entwicklung des zentralen Nervensystems und spielt für die adulte Plastizität eine wesentliche Rolle. Besonders die Aktindynamik ist wichtig für das Neuritenwachstum, die axonale Wegfindung und Verzweigung, sowie die Synaptogenese. Motoneurone bilden mehrere tausend terminale Verzweigungen aus, um neuromuskuläre Endplatten (NMJ) zu innervieren. Die axonale Verzweigung ist ein fundamentales Ereignis bei Ausbildung synaptischer Verbindungen zwischen Motoneuron und innerviertem Muskel. Die Axonverzweigung geschieht durch die Polymerisierung von Aktin entlang des Axonschafts, was zur Entstehung von Filopodien und Lamellopodien führt. Allerdings ist die genaue Funktion der drei Aktin-Isoformen (α-, β- and γ-Actin), im Zusammenhang mit der Regulation der Filopodienstabilität und deren Dynamik, noch weitestgehend unbekannt. Somit konnten wir in dieser Arbeit mit Hilfe hoch sensitiver in situ Hybridisierungs- und qRT PCR Techniken zeigen, dass in primären Mausmotoneuronen alle drei Aktinisoformen (α-, β- und γ) exprimiert, und deren Transkripte entlang des axonalen Kompartiments transportiert werden. Unsere FRAP Daten weisen darauf hin, dass α-, β- und γ-Aktin sowohl im Wachstumskegel als auch an sogenannten „Translation Hot Spots“ innerhalb axonaler Verzweigungspunkte lokal synthetisiert werden. Anhand von „Live Cell Imaging“ Experimenten konnten wir dann zeigen, dass ein α-Aktin Knockdown die Dynamik axonaler Filopodien stark reduziert, und als Folge, die Anzahl von axonalen Verzweigungen und die Axonlänge verringert ist. Hingegen geht ein β-Aktin Knockdown mit reduzierter Filopodiendynamik im Wachstumskegel und betroffener Differenzierung präsynaptischer Strukturen einher. Veränderungen des axonalen Wachstum und der Filopodiendynamik sind ebenfalls bei einem γ-Aktin Knockdown zu beobachten. Diese Daten weisen darauf hin, dass die drei Aktinisoformen unterschiedliche Funktionen bei der Entwicklung von Motoraxonen haben. Darüber hinaus zeigen unsere Daten, dass die Herunterregulation einer Aktinisoform durch eine erhöhte Expression der beiden anderen Isoformen kompensiert wird. Dieser Kompensationsmechanismus erlaubt es, die gesamte Aktinmenge und somit die F-Aktin-Polymerisation in der Zelle aufrechtzuerhalten. Sehr interessant dabei ist die Beobachtung, dass nach einem α- oder γ-Actin Knockdown das G/F-Verhältnis verändert ist, so dass die Menge an β-Aktin im G-Aktin Pool steigt und im F-Aktin Pool abnimmt. Daher beruhen Polymerisation und Stabilität von β-Aktin auf den α-, und γ-Aktinisoformen. Zusammenfassend lässt sich sagen, dass alle drei Aktinisoformen übergreifende Funktionen während Wachstum und Differenzierung von Motoneuronen haben. Im Zellkörper von sich entwickelnden Motoneuronen übernehmen sie ähnliche Aufgaben und können sich somit gegenseitig kompensieren. Im Gegensatz dazu sind die Funktionen im axonalen Kompartiment wesentlich spezifischer. Hier reguliert β-Aktin axonales Wachstum und Plastizität, während α- und γ-Aktin eine entscheidende Rolle bei der Ausbildung axonaler Verzweigungen haben. Unsere Arbeit lässt nun Rückschlüsse über mögliche Funktionen des SMN Proteins beim Aufbau der sogenannten „RNA Granules“ und lokaler Proteinbiosynthese der verschiedenen Aktinisoformen in primären Mausmotoneuronen zu

The subthalamo-pallidal system constitutes the second layer of circuitry in the basal ganglia, downstream of the striatum. It consists of four nuclei. Two of them, the external segment of the globus pallidus (GPe) and subthalamic nucleus (STN), make their connections primarily within the basal ganglia. The others, the internal segment of the globus pallidus (GPi) and the substantia nigra pars reticulata (SNr), are the output nuclei of the basal ganglia. Collectively, their axons distribute collaterals to all the targets of the basal ganglia. Rare interneurons have been reported in each of them from studies of Golgi-stained preparations, but they have not so far been confirmed using more modern methods. The circuit as described here is based primarily on studies of the axonal arborizations of neurons stained individually by intracellular or juxtacellular labeling.

The mushroom body (MB) in the insect brain is composed of a large number of densely packed neurons called Kenyon cells (KCs) (Drosophila, 2200; honeybee, 170,000). In most insect species, the MB consists of two caplike dorsal structures, the calyces, which contain the dendrites of KCs, and two to four lobes formed by collaterals of branching KC axons. Although the MB receives input and provides output throughout its whole structure, the neuropil part of the calyx receives predominantly multimodal input from sensory projection neurons (PNs) of second or a higher order, and the lobes send output neurons to many other parts of the brain, including recurrent neurons to the MB calyx. Widely branching, supposedly modulatory neurons (serotonergic, octopaminergic) innervate the MB at all levels (calyx, peduncle, and lobes), including the somata of KCs in the calyx (dopamine).