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Activation of epidermal growth factor receptor mediates receptor axon sorting and extension in the developing olfactory system of the moth Manduca sexta.

During development of the adult olfactory system of the moth Manduca sexta, olfactory receptor neurons extend axons from the olfactory epithelium in the antenna into the brain. As they arrive at the brain, interactions with centrally derived glial cells cause axons to sort and fasciculate with other axons destined to innervate the same glomeruli. Here we report studies indicating that activation of the epidermal growth factor receptor (EGFR) is involved in axon ingrowth and targeting. Blocking the EGFR kinase domain pharmacologically leads to stalling of many axons in the sorting zone and nerve layer as well as abnormal axonal fasciculation in the sorting zone. We also find that neuroglian, an IgCAM known to activate the EGFR through homophilic interactions in other systems, is transiently present on olfactory receptor neuron axons and on glia during the critical stages of the sorting process. The neuroglian is resistant to extraction with Triton X-100 in the sorting zone and nerve layer, possibly indicating its stabilization by homophilic binding in these regions. Our results suggest a mechanism whereby neuroglian molecules on axons and possibly sorting zone glia bind homophilically, leading to activation of EGFRs, with subsequent effects on axon sorting, pathfinding, and extension, and glomerulus development.

Amino Acid Sequence↗

Callosal axon arbors in the limb representations of the somatosensory cortex (SI) in the agouti (Dasyprocta primnolopha).

The present report compares the morphology of callosal axon arbors projecting from and to the hind- or forelimb representations in the primary somatosensory cortex (SI) of the agouti (Dasyprocta primnolopha), a large, lisencephlic Brazilian rodent that uses forelimb coordination for feeding. Callosal axons were labeled after single pressure (n = 6) or iontophoretic injections (n = 2) of the neuronal tracer biotinylated dextran amine (BDA, 10 kD), either into the hind- (n = 4) or forelimb (n = 4) representations of SI, as identified by electrophysiological recording. Sixty-nine labeled axon fragments located across all layers of contralateral SI representations of the hindlimb (n = 35) and forelimb (n = 34) were analyzed. Quantitative morphometric features such as densities of branching points and boutons, segments length, branching angles, and terminal field areas were measured. Cluster analysis of these values revealed the existence of two types of axon terminals: Type I (46.4%), less branched and more widespread, and Type II (53.6%), more branched and compact. Both axon types were asymmetrically distributed; Type I axonal fragments being more frequent in hindlimb (71.9%) vs. forelimb (28.13%) representation, while most of Type II axonal arbors were found in the forelimb representation (67.56%). We concluded that the sets of callosal axon connecting fore- and hindlimb regions in SI are morphometrically distinct from each other. As callosal projections in somatosensory and motor cortices seem to be essential for bimanual interaction, we suggest that the morphological specialization of callosal axons in SI of the agouti may be correlated with this particular function.

Animals↗

Regeneration of retinal axons into the goldfish optic tectum.

The growth of regenerating retinal axons into the central portion of the optic tectum of adult goldfish was examined with the light and electron microscopes. Optic tracts were cut and, two days to five months later, the animals were perfused and the tecta prepared for microscopy. Regenerating axons first reached central regions of the tectum seven to ten days postoperatively. Regenerating axons appear in very large numbers and travel in fascicles in the stratum opticum (SO) and in the adjacent neuropil, the stratum fibrosum et griseum superficiale (SFGS). In the SO, the fascicles are bordered by glial cells and degenerating debris. Within the SFGS, however, the fascicles do not seem to be similarly associated with glial cells and degenerating debris. The youngest regenerating axons are very slender processes, containing microtubules but few or no neurofilaments or dense granular material. By 10 to 14 days postoperatively, neurofilaments can be seen and, in addition, large numbers of vesicles with dense cores appear. The vesicles with dense cores increase in numbers until about 28 days postoperatively and then become quite rare. That vesicles with dense cores were seen in regenerating axons in both SO and SFGS during the period of growth into the tectum but were not seen in axon terminals at any time, suggests that they may be concerned with axon elongation. During the period one month to five months postoperatively, the regenerating axons gradually increase in diameter but do not reach preoperative sizes, suggesting that the regenerative changes may still be occurring. Remyelination is delayed and proceeds slowly. Many axons remain unmyelinated for as long as five months postoperatively.

Animals↗

Developmental relationships between trigeminal ganglia and trigeminal motoneurons in chick embryos. III. Ganglion perikarya direct motor axon growth in the periphery.

The previous study in this series demonstrated that the ingrowth of the central axons of the trigeminal (V) ganglion is prerequisite to V motor axon outgrowth and somatic translocation. In the present experiment we determined whether further interactions with V ganglion cell bodies were required by V motoneurons after the V ganglion innervates the brainstem. Soon after the ganglion axons had penetrated the brainstem they were severed, and a barrier, either permeable or impermeable, was placed between the ganglion cell bodies and the metencephalon. V motor axons grew along aberrant pathways to circumvent the impermeable barriers, many rerouting to reach the V ganglion. Only those V motor nerves which contacted the V ganglion distal to the barrier reached their target musculature in the mandible. The pattern of migration of V motoneurons was normal regardless of the V motor nerve trajectory, but the cell bodies of those axons which did not reach a muscle were not fully differentiated. When permeable barriers (Millipore filters) were implanted, the nerves followed two types of trajectories. If the pore size of the filter was small (0.45 and 0.025 microns), the V motor nerves grew identically to those observed in embryos in which impermeable barriers had been implanted. If the pore size of the filter was large (8.0 and 0.08 microns), the V motor nerve grew along its normal path directly to the barrier. Small axonal bundles from these nerves frequently grew into the filter toward the distal V ganglion. These results indicate that V motor axons preferentially grow to the V ganglion perikarya after exiting from the brainstem. Contact with the V ganglion always results in V motor nerve growth to the mandible while growth of the V motor axons to aberrant target sites only occurs when the axons fail to contact the V ganglion cells distal to the barrier.

Animals↗

Numbers of axons in lateral and ventral funiculi of rat sacral spinal cord.

The present study determines the numbers of myelinated and unmyelinated axons in the ventral and lateral funiculi of rat sacral spinal cord. On average, there are 55,000 myelinated and 110,000 unmyelinated axons in the lateral funiculus and 26,000 myelinated and 9,000 unmyelinated axons in the ventral funiculus at these levels. These figures combined with data from earlier studies of the posterior funiculus and the tract of Lissauer give approximate figures of 88,500 myelinated and 131,500 unmyelinated axons for the entire white matter of one side of the rat sacral spinal cord. Thus unmyelinated axons predominate in the white matter of the rat sacral spinal cord. The majority of axons, particularly the unmyelinated axons, are located in the lateral funiculus. The axons are concentrated in the dorsolateral part of the lateral funiculus, and so the dorsal part of the lateral funiculus, often referred to as the dorsolateral funiculus, contains more than half the fibers in the white matter of the spinal cord. A small nick in the dorsal and lateral part of the lateral funiculus, which is often done for various experimental reasons, could thus remove 40% of the axons in the white matter of rat sacral spinal cord. The data reported in the present paper will serve as a basis for future studies on the white matter of the isolated spinal cord.

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The formation of the axonal pattern in the embryonic avian retina.

Both the polarity of the axonal growth and the formation of the optic fiber pattern early in retinal morphogenesis were studied in silver stained whole mounts of embryonic chick, quail, and pigeon retinae. The surface area of the retina and of the optic fiber layer increases in size exponentially, the optic fiber layer expanding faster than the retina. The optic fiber layer covers the retinal surface at E5 in quail and at E6 in chick and pigeon. In all species studied, the retinal fiber layer does not expand homogeneously with the optic nerve head as the center. Instead, the retinal fiber layer enlarges with polarities in the dorsal to ventral and nasal to temporal direction. The very first axon bearing ganglion cells appear at stage 16 in the dorsal and central portion of the retina and grow ventrally to merge at the optic disk. From stage 23 on, the optic fiber layer expands faster in the temporal than in the nasal side. Measurements on the initial polarization of young axonal processes show that the axonal growth is directed toward the optic fissure and the optic nerve head. This growth polarization is found at the onset of growth cone formation and in axons far from the nearest ganglion cells or ganglion cell axons. Therefore axon-axon interaction cannot be involved in the initial axon orientation early in retinal morphogenesis.

Animals↗

Quantitative analyses of axonal endings in the central nucleus of the inferior colliculus and distribution of 3H-labeling after injections in the dorsal cochlear nucleus.

Quantitative analyses of electron microscopic (EM) autoradiographs were used to identify the afferents from the dorsal cochlear nucleus in the central nucleus of the inferior colliculus (IC) in the cat. In order to localize the sources of radioactivity, material from axonal transport experiments was analyzed by means of a hypothetical grain procedure which takes the cross-scatter of beta particles into account. Measurements of the synaptic vesicles in axonal endings and a cluster analysis were used to identify different groups of endings. In order to determine which types of endings arise in the dorsal cochlear nucleus, axonal endings labeled after axonal transport and unlabeled endings were characterized and compared to the groups defined by the cluster analysis. Axonal endings with round synaptic vesicles were labeled with more than 2 grains/micron2 which was about 30% of the radioactivity in the central nucleus of the IC. This was six to seven times greater than if the radioactivity had been randomly distributed. Other tissue compartments usually had less radioactivity. Some myelinated and unmyelinated axons were labeled, but, as a group they had lower amounts of radioactivity than predicted by random labeling. In most cases, only low levels of activity were found in glial and postsynaptic structures. Five groups of axonal endings in the medial part of the central nucleus were identified by an analysis which clustered similar types of endings. The variance of the longest axis, the mean diameter, the variance of area, and the mean area of the synaptic vesicles were the variables most useful in distinguishing these five groups. Axonal endings with round synaptic vesicles were classified as either small, or large, or very large, while endings with pleomorphic vesicles were either large or small. Using measurements of the cross-sectional diameter of dendritic microtubules, samples of digitized axonal endings from normal and experimental cases were normalized and could be compared directly to the groups defined by the cluster analysis. Microtubules were 21.7 nm (+/- 1.6) in average diameter. After injections of 3H-leucine and/or proline in the dorsal cochlear nucleus, most of the labeled endings in the IC contained small, round vesicles (less than 47 nm in diameter) although a very small number of endings with large, round vesicles also were labeled.(ABSTRACT TRUNCATED AT 400 WORDS)

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Light microscopic observations on cat Renshaw cells after intracellular staining with horseradish peroxidase. I. The axonal systems.

Five intracellularly HRP-stained Renshaw cells were subjected to light microscopic analysis of the trajectories, branching patterns, and projections of the axonal systems. The cell bodies were located ventrally in lamina VII. In three neurons the axon originated from the cell body and in the remaining two cells from a dendrite. After a 600-870-microns distance the axons entered the ventral funiculus, where all of them continued rostrally. Two axons also gave off a caudal branch in the funiculus. The diameters of the main axons varied between 2.1 and 10.0 microns. The main axons gave off one to four first-order collaterals before entering the ventral funiculus and up to three collaterals could be seen to originate from the same node of Ranvier. In the ventral funiculus up to five first-order collaterals could be traced from the same main axon. The axon collateral trees were often very extensive and daughter branches up to the 22nd order were observed. The distance between two successive branching points varied between 4 and 410 microns. A large number of boutonlike swellings were found along (59%) or at the ends of the collateral branches. At the most, 1,278 swellings originated from a single axon collateral tree. Most of the swellings were located in lamina IX, but they also appeared ventrally and dorsolaterally in lamina VII.

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Correlation of cell body size, axon size, and signal conduction velocity for individually labelled dorsal root ganglion cells in the cat.

Measurements of cell body and peripheral and central axon sizes were made for primary sensory neurons outlined by the intracellular injection of HRP. Conduction velocities were also measured on the outlined processes. The sensory neurons were then subdivided into A and C cells on the basis of the conduction velocity of the impulses carried by the processes of these cells. Central processes of both A and C cells are smaller than the peripheral processes, but the size differential is greater for the C cells. For A cells there is a linear relation between the size of the peripheral axon and the conduction velocity of the impulses carried by these axons, but the confidence limits are wide. For C cells there is a linear relation between the size of the central process and conduction velocity of the impulses carried by the processes, but for the peripheral processes two aberrant processes resulted in no correlation between process size and conduction velocity. For A cells, the size of the central and peripheral processes and the conduction velocity of the impulses carried by the peripheral processes are linearly correlated with cell body size. By contrast no such correlations can be demonstrated for C cells. This presumably implies an important difference in that the size of the cell body is correlated with axon size and impulse conduction velocity for A cells but not for C cells. A widely accepted generalization is that large sensory cells give rise to myelinated axons and small sensory cells to unmyelinated axons. In this study, myelinated and unmyelinated are defined on the basis of impulse conduction velocity. For those cells that are clearly large (greater than 50 microns in diameter), the conduction velocity of the impulses carried by their processes is always greater than 2.5 m/s, and for those cells that are clearly small (less than 35 microns in diameter), the conduction velocity is always less than 2.5 m/s. Thus for these cells the above generalization holds. For the intermediate-sized cells (35-50 microns), however, the size of the cell body bears no predictable relation to the conduction velocity of the impulses carried by those processes, and thus to whether the axons are myelinated or unmyelinated. Thus the above generalization does not hold for this intermediate group of cells, and since there are many cells in this size range, we feel that the generalization that large cells give rise to myelinated axons and small cells to unmyelinated axons is an oversimplification.

Action Potentials↗

Axonal regeneration in spinal cord injury: a perspective and new technique.

A set of techniques is described for determining the response of mammalian spinal axons to transection. The logical selection and the advantages of these techniques are discussed. The dorsal column of guinea pig thoracic spinal cord was transected with a tungsten needle and the position of the lesion was marked by a staple-shaped wire device (Foerster: J. Comp. Neurol. 210:335-356, '82). The morphology of dorsal column axons projecting rostrally toward the lesion was examined between 1 and 50 days postlesion by anterograde staining with horseradish peroxidase, applied to a second lesion of the dorsal column two to three vertebral segments caudal to the first. Axons damaged by the original lesion were found to die back 1-2 mm from the plane of transection and at 18-20 hours were characterized by terminal club-shaped swellings attached to the proximal axon by a thin connection. At 50 days postlesion there was some evidence of limited regenerative responses in terms of growth-cone-like axon terminals, and the presence of aberrant axonal branching, but no evidence of regenerating axons approaching close to the plane of transection. These findings are in agreement with previous studies indicating little or no effective regrowth of myelinated axons in the mammalian spinal cord. These same techniques were used in a succeeding study to examine the effects of applied electric fields on the axonal response to transection.

Animals↗

Transected dorsal column axons within the guinea pig spinal cord regenerate in the presence of an applied electric field.

Using an implanted battery and electrodes, we have imposed a weak, steady electrical field across partially severed guinea pig spinal cords. We have analyzed regeneration of dorsal column axons in experimental animals and sham-treated controls at 50-60 days postinjury by anterograde filling of these axons with the intracellular marker horseradish peroxidase and by employing a marking device to identify precisely the original plane of transection (J. Comp. Neurol. 250: 157-167, '86). In response to electric field applications, axons grew into the glial scar, as far as the plane of transection in most experimental animals. In a few animals axons could be traced around the margins of the lesion (but never through it). Moreover, these fibers returned to their approximate positions within the rostral spinal cord before turning toward the brain. In sham-treated controls, ascending axons were found to terminate caudal to the glial scar, and rarely were any fibers found within the scar itself. Axons were never observed to cross into the rostral cord segment. These findings suggest that an imposed electrical field promotes growth of axons within the partially severed mammalian spinal cord, that a steady voltage gradient may be an environmental component necessary for axonal development and regeneration, and that some component(s) of the scar impede or deflect axonal growth and projection.

Animals↗

Aberrant optic axons in the retinal pigment epithelium during chick and quail visual pathway development.

Examination of a large number of retinal pigment epithelia revealed that, in a small proportion, optic axons in chick and quail eyes aberrantly entered the pigment cell layer between embryonic day (E) 7 to E14. The aberrant retinal axons originated from the main stream of retinal fibers in the optic nerve and invaded the pigment layer from various positions of the optic nerve head or fissure by growing along the basal side of the pigment epithelium. The axon bundles grew several millimeters into the epithelial sheet and arborized at the margin of the eye. As shown by electron microscopy the nerve fibers occurred as bundles of three to several hundred axons. They always were located at the basal side of the epithelium, and were enveloped by processes of epithelial cells. Very large bundles of axons, however, displaced the epithelial cells from the basal matrix. These retinal axons contacted the pigment epithelial basal lamina. The basal extracellular matrix from the retinal pigment epithelium was isolated and used as substratum for in vitro cultures of various types of neural explants. The matrix preparations consisted of a sheet of a 50 nm thick basal lamina with a central lamina densa, two laminae rarae, and a 15 micron thick stroma. Axons from avian retina explants, as well as sensory ganglia, grew on the basal lamina side of the pigment cell matrix with the same growth rate and with the same fiber density as on similarly prepared basal laminae from the neural retina. These experiments show that the matrix from the pigment epithelium of the avian eye does not have negative effects on axonal growth and indicate that a basal lamina from a normally non-innervated tissue can provide a favorable matrix for axonal growth.

Animals↗

Synaptic organization of anomalous retinal projections to the somatosensory and auditory thalamus: target-controlled morphogenesis of axon terminals and synaptic glomeruli.

These experiments examine which morphological features of axon terminals and their synaptic glomeruli are determined by afferent axons, and which by their targets. In normal, adult hamsters, electron microscopy reveals that, with respect to multiple ultrastructural features, the terminals and synaptic glomeruli of retinal afferent axons in the dorsal lateral geniculate nucleus differ from those of ascending auditory and somatosensory afferents in the medial geniculate and ventrobasal nuclei, respectively. These features include: (1) the location of specific sensory axon terminals on the somata and dendrites of their targets neurons, (2) the constitutents of the glomeruli and their synaptic relationships, (3) the number of specific sensory terminal boutons per glomerulus, (4) bouton size, (5) the number of dendritic and somatic appendages contacted by each bouton, and (6) the mitochondrial morphology of the specific sensory afferent boutons. In order to ascertain which of these features are determined by afferent axons and which by their targets, we subjected newborn Syrian hamsters to surgical procedures known to produce permanent, abnormal retinal projections to the main thalamic auditory (medial geniculate) and somatosensory (ventrobasal) nuclei. When the animals were adults, we examined the terminals and synaptic glomeruli of abnormal retino-auditory and retino-somatosensory axons that were anterogradely labeled by intraocular injection of horseradish peroxidase. With respect to all of the preceding features except mitochondrial morphology, the terminals and synaptic glomeruli of retino-medial geniculate and retino-ventrobasal axons more nearly resembled those of normal, auditory and somatosensory afferent axons, respectively, than they did those of normal, retino-lateral geniculate axons. These results demonstrate that the differentiation of all the features that we have examined, except mitochondrial morphology, is determined by factors in target neurons or their environment. This finding suggests that the differentiation of morphological features involved in contacts among neurons (including the type, number and size of interconnected neuronal elements and the loci at which they contact each other) is responsive to interactions among the connected elements, or between neural elements and their environment (e.g., glia, extracellular matrix), whereas the differentiation of structures reflecting intrinsic functions of individual neuronal elements is not responsive to such interactions.

Animals↗

Migration of Schwann cells and axons into developing chick forelimb muscles following removal of either the neural tube or the neural crest.

A study has been made of the effects of neural crest and neural tube removal at the brachial level on the migration of Schwann cells and axons into the flexor digitorum profundus (fdp) and flexor carpi ulnaris (fcu) muscles of the avian forelimb. The identification of Schwann cells was based on the assumption that antibody HNK-1 uniquely labels these cells at the growing end of limb nerves. Myotubes and nerves were identified by using antibodies to myosin and to neurofilament protein, respectively. The removal of neural crest cells at stage 13 gave a complete Schwann cell-free embryo at the brachial level. Motor axons only grew to the base of the forelimb, forming a rudimentary plexus by stage 27, and failed to penetrate the limb. Removal of the neural tube at stage 13 did not prevent sensory axons from forming a plexus at the base of the limb; these axons subsequently developed into the brachialis longus inferior (bli n) and superior (bls n) nerves. By stage 27 the bli n had branched into the interosseus nerve (in n) and the medial-ulnar nerve (m-u n) trunks. However, unlike the result in control embryos, no nerves were detected amongst the developing fdp and fcu muscles, thus indicating that sensory axons do not grow into the muscles in the absence of motor axons. In contrast, Schwann cells were observed amongst the myotubes at the level of the in n and m-u nerve trunks. The present observations show that motor axons do not enter the limb bud and innervate limb muscles in the absence of Schwann cells. Furthermore, in the absence of motor axons (neural-tube-removed embryos) sensory axons still enter the limb (behind migrating Schwann cells) but fail to innervate developing muscles even though Schwann cells are present among the developing myotubes.

Animals↗

Compartmental and topographical distributions of axons in nerves to the amphibian (Bufo marinus) glutaeus muscle.

The present work seeks to determine if axons to an amphibian muscle are segregated in nerve trunks between the spinal cord and muscle according to their primary nerve destination or their topographical projection in the muscle. The distribution of axons to different compartments and subcompartments of the amphibian (Bufo marinus) glutaeus muscle has been determined in transverse sections of spinal and limb nerves after retrogradely labelling the axons with horseradish peroxidase. Glutaeus axons were dispersed widely through spinal nerves 8 and 9 but loosely gathered together in one quadrant of the sciatic nerve after passing through the lumbar plexus. Glutaeus axons became tightly clustered to the exclusion of other axons along the length of the triceps femoris nerve after it divides from the sciatic nerve. Furthermore, axons destined for one of the two glutaeus primary nerve branches segregate from those of the other branch at the level of the triceps femoris nerve before the glutaeus nerve forms. On the other hand, motoneurones that subserve a primary branch are not segregated, but are found throughout the rostrocaudal extent of the glutaeus motoneurone pool. Injection of horseradish peroxidase under the epimysium of either the ventral or the dorsal surfaces of the glutaeus muscle labelled motoneurones preferentially in either the rostral or caudal part of the motoneurone pool, respectively. This confirms studies that have shown a topographical projection from the spinal motoneurone pool onto the glutaeus muscle. However, there was no segregation of dorsally projecting axons in the glutaeus and primary nerve branches. Thus, glutaeus axons segregate according to their muscle compartmental projections well before entering the muscle, but they show no organization in nerves with respect to their topographical projections within a compartment.

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Axon-bearing amacrine cells of the macaque monkey retina.

A new and remarkable type of amacrine cell has been identified in the primate retina. Application of the vital dye acridine orange to macaque retinas maintained in vitro produced a stable fluorescence in the somata of apparently all retinal neurons in both the inner nuclear and ganglion cell layers. Large somata (approximately 15-20 microns diam) were also consistently observed in the approximate center of the inner plexiform layer (IPL). Intracellular injections of horseradish peroxidase (HRP) made under direct microscopic control showed that the cells in the middle of the IPL constitute a single, morphologically distinct amacrine cell subpopulation. An unusual and characteristic feature of this cell type is the presence of multiple axons that arise from the dendritic tree and project beyond it to form a second, morphologically distinct arborization within the IPL; these cells have thus been referred to as axon-bearing amacrine cells. The dendritic tree of the axon-bearing amacrine cell is highly branched (approximately 40-50 terminal dendrites) and broadly stratified, spanning the central 50% of the IPL so that the soma is situated between the outermost and innermost branches. Dendritic field size increases from approximately 200 microns in diameter within 2 mm of the fovea to approximately 500 microns in the retinal periphery. HRP injections of groups of neighboring cells revealed a regular intercell spacing (approximately 200-300 microns in the retinal periphery), suggesting that dendritic territories uniformly cover the retina. One to four axons originate from the proximal dendrites as thin (less than 0.5 microns), smooth processes. The axons increase in diameter (approximately 1-2 microns) as they course beyond the dendritic field and bifurcate once or twice into secondary branches. These branches give rise to a number of thin, bouton-bearing collaterals that extend radially from the dendritic tree for 1-3 mm without much further branching. The result is a sparsely branched and widely spreading axonal tree that concentrically surrounds the smaller, more highly branched dendritic tree. The axonal tree is narrowly stratified over the central 10-20% of the IPL; it is approximately ten times the diameter of the dendritic tree, resulting in a 100 times greater coverage factor. The clear division of an amacrine cell's processes into distinct dendritic and axonal components has recently been observed in other, morphologically distinct amacrine cell types of the cat and monkey retina and therefore represents a property common to a number of functionally distinct cell types. It is hypothesized that the axon-bearing amacrine cells, like classical neurons,

Acridine Orange↗

Terminal arbors of individual "feedback" axons projecting from area V2 to V1 in the macaque monkey: a study using immunohistochemistry of anterogradely transported Phaseolus vulgaris-leucoagglutinin.

In the present study, the anterograde tracer Phaseolus vulgaris-leucoagglutinin (PHA-L) was injected into area V2 in order to demonstrate the precise morphology of individual axons from area V2 to V1. On the basis of 28 complete axon reconstructions, several characteristic features have been identified. 1) Individual axons arborize in multiple layers: 1, 2, 5, and (inconstantly) 3. A single axon may have numerous terminal clusters in layers 1 and 2, but at most one in layer 3. 2) Axons typically ascend to layer 1, turn asymmetrically in one direction, and travel for long distances in this layer (1.10-4.30 mm; dimensions uncorrected for shrinkage). A few axons (three of 28 reconstructed) were found to have a single terminal cluster (0.3-0.5 mm wide) in layers 1 and 2. 3) Collaterals in layer 5 seem to extend over shorter distances (0.60 mm or less). 4) Delicate sprays of boutons (both beads and spines) are clustered along the main trunk. Spacing is variable but usually ranges from 0.35 mm to 0.65 mm. 5) In addition to clustered boutons, there can be linear collaterals, continuously studded with boutons, parallel to the main axon in layer 1. These results indicate that axons from V2 have complex radial and tangential distributions in V1. Terminations in different layers may be directed to different sets of neurons or to different portions of the dendritic tree (for example, distal portions of pyramidal neuron apical dendrites in layers 1 and 2, but more proximal portions in layer 3). Clustered terminations over wide tangential areas may imply a divergent innervation by a single axon of multiple compartmental structures, such as ocular dominance columns or cytochrome oxidase patches.

Animals↗

Dendritic and axonal morphology of HRP-injected neurons in the inferior colliculus of the cat.

The dendritic and axonal morphology of neurons in the inferior colliculus of the cat was investigated after intracellular injection of HRP, in vivo. All injected axons gave off local collaterals, and most showed a widespread distribution and lacked a specific orientation. In contrast, the dendrites of injected neurons were distinguished by their degree of orientation and the direction of the longest axis of orientation. Dendrites showed a high, moderate, or low degree of orientation. Most highly oriented cells had their longest axis in the rostrocaudal direction with fewer in the mediolateral direction. In the central nucleus, only the rostrocaudally oriented cells correspond to the disc-shaped cells identified in Golgi preparations. Unlike most cells in our sample, the two cells that were disc-shaped had axons that were parallel to the orientation of the dendritic tree. In the dorsal cortex, rostrocaudally oriented cells also were found, but they had unoriented axons. In both the central nucleus and dorsal cortex, cells with a mediolateral axis of orientation or no specific orientation correspond to stellate cells and had axons with widespread local collaterals. These results suggest that an extensive network of local axon collaterals may contribute to neural processing within the inferior colliculus. In the central nucleus, local axons may establish connections within or across the fibrodendritic laminae. In the dorsal cortex, the local and afferent axons may form a complex reticular network. Finally, some injected cells had axons terminating locally and also entering the brachium of the inferior colliculus. This suggests that cells in the inferior colliculus may function as both interneurons and projection neurons.

Animals↗