Binocular interaction in the fetal cat regulates the size of the ganglion cell population

During fetal development of the cat's visual system there is a marked overproliferation of optic nerve axons. In utero binocular interaction contributes to the severity of fiber loss since removal of an eye during gestation attenuates axon loss in the remaining optic nerve. The purpose of the present study was to determine whether this reduced loss of optic nerve fibers is due to a failure of retraction by supernumerary axon branches or to a reduction in ganglion cell death. To resolve this issue, we compared the number of ganglion cells and optic nerve fibers in adult cats which had one eye removed at known gestational ages. Retinal ganglion cells were backfilled with horseradish peroxidase and counts were made from retinal wholemounts. The axon complement was assessed with an electron microscopic assay. In the retinas of a normal cat we estimated 151,000 and 152,000 ganglion cells. The optic nerves of two other normal cats contained approximately 158,000 and 159,000 axons. In comparison, an animal enucleated on embryonic day 42 had 180,000 ganglion cells and 178,000 optic nerve fibers, while in an animal enucleated on embryonic day 51 the corresponding estimates were 182,000 and 190,000. The close agreement between cell and fiber counts indicates that axonal bifurcation does not contribute appreciably to the axon surplus in the optic nerve of prenatally enucleated cats. These results demonstrate that prenatal binocular interaction regulates the size of the mature retinal ganglion cell population.     

Schwann cell myelin ensheathing C.N.S. axons in the nerve fibre layer of the cat retina

Summary In the retina of the cat the axons of the nerve fibre layer are unmyelinated and are provided with a C.N.S. myelin sheath only in the extraocular part of the optic nerve. The present study demonstrates that in the apparently normal cat retina close to the optic disc, some axons of the nerve fibre layer run for a short distance in the perivascular space of the retinal arteries. While coursing in the perivascular space, these C.N.S. axons become transiently myelinated by Schwann cells, which form a typical P.N.S. myelin sheath. These P.N.S. myelin sheaths terminate at a heminode in the transitional zone in which the C.N.S. axons penetrate the perivascular glial sheath in order to leave or to re-enter the nerve fibre layer. It is suggested that the Schwann cells, which elaborate the P.N.S. myelin around C.N.S. axons, are descendants of the Schwann cells of the perivascular autonomie nerves. The present study shows that Schwann cells are able to provide previously unmyelinated C.N.S. axons with a P.N.S. myelin sheath.     

The distribution of axons according to diameter in the optic nerve and optic tract of the rat

The distribution of axons according to diameter was examined in the optic nerve and optic tract of adult hooded rats. Observations were made on semithin sections, and measurements of axonal diameters were made on electron micrographs taken from various locations across thin sections through the optic nerve and tract. The distribution of axons by size differs markedly in the optic nerve and tract. Coarse (greater than 2 microns) and fine (less than or equal to 2 microns) axons are distributed throughout all regions of the optic nerve. In the optic tract, in contrast, coarse axons are especially dense dorsally, at the deep border of the tract, while they are absent ventrally, subjacent to the pial surface. No regions of the optic nerve contain densities of coarse axons as high as the deep nor as low as the superficial extremes of the optic tract. Nevertheless, even at the deep (dorsal) border of the optic tract, the coarse axons make up only a small minority (roughly 15%) of the total number of axons in that region. The axons 2 microns or smaller may be divisible into two overlapping, fine and intermediate, diameter classes, that are partially segregated within the optic tract, but not in the optic nerve: the distributions of axon diameters smaller than 2 microns are skewed to distinctly smaller diameters at the dorsal and ventral extremes of the optic tract, while in between, at mid-positions along the deep-to-superficial axis of the optic tract, the axon size distributions contain many more axons greater than 1 micron in diameter. These different axon diameter groups may arise from the morphologically distinct retinal ganglion cell types, and may underlie the components of the trimodal compound axon potential seen in the rat's primary optic pathway. Their partial segregation within the tract anticipates the partial segregation of their terminal arborizations within the laminae of the dorsal lateral geniculate nucleus. The rearrangement of axons into a partial segregation by size within the optic tract may indicate a chronology of axonal arrival during early development, proximity to the pial surface being an index of recency of arrival. As axonal outgrowth and neurogenesis appear to be directly related within the retinal ganglion cell population in mammals, the relative birthdates of the retinal ganglion cell types giving rise to the axon diameter classes in the rat may be inferred from the present results.     

Regrowth of retinal ganglion cell axons into a peripheral nerve graft in the adult hamster is enhanced by a concurrent optic nerve crush

Summary Transplantation of a segment of peripheral nerve to the retina of the adult hamster resulted in regrowth of damaged ganglion cell axons into the graft, with the fastest regenerating axons extending at 2 mm/day after an initial delay of 4.5 days (Cho and So 1987b). In this study, the effect of making 2 lesions on the same axon (the conditioning lesion effect) on the regrowth of ganglion cell axons into the peripheral nerve graft was examined. When a conditioning lesion (first lesion) was made by crushing the optic nerve 7 or 14 days before the peripheral nerve grafting (the second lesion) to the retina, the distance of regrowth achieved by the fastest regenerating axons in the graft, measured at the 7th post-grafting day, was lower than in animals with a peripheral nerve grafted to a normal eye. This indicated that in contrast to the situation in peripheral nerve axons (Forman et al. 1980) and goldfish optic axons (Edwards et al. 1981), the conditioning lesion was unable to enhance the regrowth of mammalian retinal ganglion cell axons. However, when crushing of the optic nerve was followed immediately by peripheral nerve grafting, an enhancement in axonal regrowth could be observed. The initial delay time before the axons extended into the peripheral nerve graft was reduced by 1 day while the rate of elongation of the fastest regrowing axons in the graft apparently remained unchanged. Moreover, the shortening of the initial delay could still be observed even when the sequence of performing the 2 lesions was reversed. From these data, it was concluded that the classical conditioning lesion effect was not responsible for the enhancement observed. Rather it was suggested that changes in the intra-retinal environment brought about by crushing of the optic nerve might account for it.     

Regeneration of axons in the optic nerve of the adult Browman-Wyse (BW) mutant rat

Summary We have studied the regeneration of axons in the optic nerves of the BW rat in which both oligodendrocytes and CNS myelin are absent from a variable length of the proximal (retinal) end of the nerve. In the optic nerves of some of these animals, Schwann cells are present. Axons failed to regenerate in the exclusively astrocytic environment of the unmyelinated segment of BW optic nerves but readily regrew in the presence of Schwann cells even across the junctional zone and into the myelin debris filled distal segment. In the latter animals, the essential condition for regeneration was that the lesion was sited in a region of the nerve in which Schwann cells were resident. Regenerating fibres appeared to be sequestered within Schwann cell tubes although fibres traversed the neuropil intervening between the ends of discontinuous bundles of Schwann cell tubes, in both the proximal unmyelinated and myelin debris laden distal segments of the BW optic nerve. Regenerating axons never grew beyond the distal point of termination of the tubes. These observations demonstrate that central myelin is not an absolute requirement for regenerative failure, and that important contributing factors might include inhibition of astrocytes and/or absence of trophic factors. Regeneration presumably occurs in the BW optic nerve because trophic molecules are provided by resident Schwann cells, even in the presence of central myelin, oligodendrocytes and astrocytes. All the above experimental BW animals also have Schwann cells in their retinae which myelinate retinal ganglion cell axons in the fibre layer. Control animals comprised normal Long Evans Hooded rats, BW rats in which both retina and optic nerve were normal, and BW rats with Schwann cells in the retina but with normal, i.e. CNS myelinated, optic nerves. Regeneration was not observed in any of the control groups, demonstrating that, although the presence of Schwann cells in the retina may enhance the survival of retinal ganglion cells after crush, concomitant regrowth of axons cut in the optic nerve does not take place.     

Peripheral nerve explants grafted into the vitreous body of the eye promote the regeneration of retinal ganglion cell axons severed in the optic nerve

Summary We have conducted experiments in the adult rat visual system to assess the relative importance of an absence of trophic factors versus the presence of putative growth inhibitory molecules for the failure of regeneration of CNS axons after injury. The experiments comprised three groups of animals in which all optic nerves were crushed intra-orbitally: an optic nerve crush group had a sham implant-operation on the eye; the other two groups had peripheral nerve tissue introduced into the vitreous body; in an acellular peripheral nerve group, a frozen/thawed teased sciatic nerve segment was grafted, and in a cellular peripheral nerve group, a predegenerate teased segment of sciatic nerve was implanted. The rats were left for 20 days and their optic nerves and retinae prepared for immunohistochemical examination of both the reaction to injury of axons and glia in the nerve and also the viability of Schwann cells in the grafts. Anterograde axon tracing with rhodamine-B provided unequivocal qualitative evidence of regeneration in each group, and retrograde HRP tracing gave a measure of the numbers of axons growing across the lesion by counting HRP filled retinal ganglion cells in retinal whole mounts after HRP injection into the optic nerve distal to the lesion. No fibres crossed the lesion in the optic nerve crush group and dense scar tissue was formed in the wound site. GAP-43-positive and rhodamine-B filled axons in the acellular peripheral nerve and cellular peripheral nerve groups traversed the lesion and grew distally. There were greater numbers of regenerating fibres in the cellular peripheral nerve compared to the acellular peripheral nerve group. In the former, 0.6–10% of the retinal ganglion cell population regenerated axons at least 3–4 mm into the distal segment. In both the acellular peripheral nerve and cellular peripheral nerve groups, no basal lamina was deposited in the wound. Thus, although astrocyte processes were stacked around the lesion edge, a glia limitans was not formed. These observations suggest that regenerating fibres may interfere with scarring. Viable Schwann cells were found in the vitreal grafts in the cellular peripheral nerve group only, supporting the proposition that Schwann cell derived trophic molecules secreted into the vitreous stimulated retinal ganglion cell axon growth in the severed optic nerve. The regenerative response of acellular peripheral nerve-transplanted animals was probably promoted by residual amounts of these molecules present in the transplants after freezing and thawing. In the optic nerves of all groups the astrocyte, microglia and macrophage reactions were similar. Moreover, oligodendrocytes and myelin debris were also uniformly distributed throughout all nerves. Our results suggest either that none of the above elements inhibit CNS regeneration after perineuronal neurotrophin delivery, or that the latter, in addition to mobilising and maintaining regeneration, also down regulates the expression of axonal growth cone-located receptors, which normally mediate growth arrest by engaging putative growth inhibitory molecules of the CNS neuropil.     

Regenerated synapses persist in the superior colliculus after the regrowth of retinal ganglion cell axons

Summary Synapse formation by retinal ganglion cell axons was sought in the superior colliculus of four adult rats 16–18 months after the optic nerve was transected and replaced by a peripheral nerve graft that guided regenerating RGC axons from the eye to the superior colliculus. The terminals of retinal ganglion cell axons were labelled by intravitreal injections of tritiated amino acids and studied by light and electron microscopic autoradiography. We found that (i) retinal ganglion cell axons had extended from the tips of the peripheral nerve grafts into the superior colliculus for approximately 350 ,m; (ii) within the superior colliculus, some regenerated retinal ganglion cell axons became ensheathed by CNS myelin; (iii) retinal ganglion cell terminals formed asymmetric synapses with dendrites of neurons in the superficial layers of the superior colliculus, mainly the stratum griseum superficialis.Regenerated (n=418) and normal retinal ganglion cell terminals (n=1775) in the superior colliculus were compared in terms of their size (area, perimeter, and maximum diameter), contacts per terminal, contacts per 10 m terminal perimeter, and post-synaptic structure contacted (dendritic spine, shaft, or soma). No statistically significant differences in the ultrastructural characteristics of the pre-synaptic profiles were apparent between the two groups. The post-synaptic structures contacted by axon terminals were similar in regenerated and control animals, although there were quantitative differences in the distributions of these contacts among dendritic spines and shafts.These results suggest that the regeneration of retinal ganglion cell axons in adult rats can lead to the formation of ultrastructurally normal synapses in the appropriate layers of the superior colliculus. The re-formed connections appear to persist for the life-span of these animals.A short account of this work was presented inSociety for Neurosdence Abstracts 14, 654 (1988).     

睫状神经营养因子对视神经损伤的修复作用

视神经损伤后,视网膜神经节细胞进行性损害,轴突变性坏死,再生困难。睫状神经营养因子是一种非靶源性神经营养因子,在视网膜神经节细胞的生长发育中起重要调控作用,主要表现在对损伤的视网膜神经节细胞有促进存活及轴突再生的作用。     

视神经损伤后视神经及其内毛细血管的早期超微结构的变化

目的观察视神经损伤后视神经和神经内毛细血管的超微结构变化,探讨视神经损伤的机制。方法建立家兔视神经损伤的动物模型,利用透射电子显微镜观察视神经及神经内毛细血管超微结构的变化。结果视神经损伤0.5h后,轴突部分肿胀,髓鞘疏松,微管、微丝排列出现紊乱,线粒体肿胀,毛细血管内皮细胞中的吞饮小泡和微绒毛明显减少;损伤6h后,线粒体出现髓样变和空泡样变性,血管内皮细胞肿胀,管腔变窄;损伤12h后,轴突空泡样变性,髓鞘脱失,毛细血管周围间隙增宽;损伤48h后,轴质密度增加,部分髓鞘板层完全分离,微管、微丝及线粒体发生颗粒性溶解,内皮细胞中的线粒体出现广泛变性;损伤96h时,轴索崩解呈空泡状变性,髓鞘更广泛崩解,毛细血管扩张破裂,红细胞外溢。结论视神经损伤早期轴突肿胀、空泡样变性,线粒体水肿变性,微管、微丝数量减少;视神经内毛细血管扩张,通透性增加。     

Regrowth of PNS axons through grafts of the optic nerve of the Browman-Wyse (BW) mutant rat

Summary We have examined the behaviourin vivo of regenerating PNS axons in the presence of grafts of optic nerve taken from the Browman-Wyse mutant rat. Browman-Wyse optic nerves are unusual because a 2–4 mm length of the proximal (retinal) end of the nerve lacks oligodendrocytes and CNS myelin and therefore retinal ganglion cell axons lying within the proximal segment are unmyelinated and ensheathed by processes of astrocyte cytoplasm. Schwann cells may also be present within some proximal segments. Distally, Browman-Wyse optic nerves are morphologically and immunohistochemically indistinguishable from control optic nerves.When we grafted intact Browman-Wyse optic nerves or triplets consisting of proximal, junctional and distal segments of Browman-Wyse optic nerve between the stumps of freshly transected sciatic nerves, we found that regenerating axons avoided all the grafts which did not contain Schwann cells, i.e., proximal segments which contained only astrocytes; regions of Schwann cell-bearing proximal segments which did not contain Schwann cells; junctional and distal segments (which contained astrocytes, oligodendrocytes and CNS myelin debris). However, axons did enter and grow through proximal segments which contained Schwann cells in addition to astrocytes. Schwann cells were seen within grafts even after mitomycin C pretreatment of sciatic proximal nerve stumps had delayed outgrowth of Schwann cells from the host nerves; we therefore conclude that the Schwann cells which became associated with regenerating axons within the grafts of Browman-Wyse optic nerve were derived from an endogenous population. Our findings indicate that astrocytes may be capable of supporting axonal regeneration in the presence of Schwann cells.     

Characterization of the sprouting response of axon-like processes from retinal ganglion cells after axotomy in adult hamsters: a model using intravitreal implantation of a peripheral nerve

Summary Peripheral nerves provide a favourable environment for damaged CNS axons to sprout and regenerate. It has also been demonstrated that retinal ganglion cells respond to a peripheral, nerve segment grafted to the retina by emitting axon-like processes from the somatodendritic compartment into the graft. The factors influencing the pattern of sprouting of axotomized retinal ganglion cells were explored in this study by implanting a short segment of peripheral nerve, which did not come into contact with the retina, into the vitreous body of an eye whose optic nerve was concurrently crushed. Silver staining was used to assess the morphology of the retinal ganglion cells which underwent sprouting.Some retinal ganglion cells were induced to sprout axon-like processes; these emerged primarily from dendrites and less frequently from the soma or intraretinal axon. Implantation of a nonviable graft (freezed-thawed) elicited only minimal sprouting. These results suggest that diffusible factors secreted by cells in the graft are a possible stimulus to sprouting in axotomized retinal ganglion cells.Examination of the pattern of dendritic sprouting indicates that sprouting was most intense (in terms of number of sprouts per cell) at early times post-axotomy. Moreover, a differential pattern of development of sprouts arising from individual primary dendrites of the same cell was observed; sprouts tend to arise from all primary dendrites initially but as the post-axotomy time increased, retraction of sprouts from some primary dendrites occurred. Concomitant with this retraction, however, there was an increase in the number of sprouts on those primary dendrites which were still in the active phase of sprouting. Selective stabilization of sprouts by extrinsic factors may account for this phenomenon.Changes in the area and outline (irregularity) of the somata of retinal ganglion cells with sprouts from two weeks to two months after optic nerve crush could be correlated temporally with the intensity of sprouting from the dendritic tree, suggesting that during sprouting, intrinsic mechanisms coordinate the responses of different cellular compartments.In contrast to extensive ectopic sprouting of axotomized retinal ganglion cells in the presence of an intravitreal graft, when a long peripheral nerve segment is grafted to the cut optic nerve, there is extensive axonal regeneration into the graft from retinal ganglion cells, most of which did not exhibit ectopic sprouting. Thus, a hierarchy of sprouting sites within a neuron seems to exist, with the damaged axonal tip being the most favoured site, followed by the dendrites, and then the intraretinal axon. The soma appears to be the least preferred compartment for sprout emission.     

Ultrastructure of photoreceptors and circadian pacemaker neurons in the eye of a gastropod, Bulla

Each of the paired cephalic eyes of the marine gastropod, Bulla, is about 0.5 mm in diameter and contains about 1000 large photoreceptors, small photoreceptors, numerous pigmented support cells and about 130 neurons. The photoreceptors are of three types: large (90 micron X 20-30 micron) dense ones (PRLD) with elaborate narrow microvilli and aggregates of 650 A clear vesicles in the cytoplasm; large clear ones (PRLC) with elaborate wide microvilli and aggregates of 650A clear vesicles; small slender receptors (PRS) with a tuft of microvilli and lacking vesicle aggregates. Neurons (15-25 micron) containing dense-core 1000 A vesicles are in the periphery of the retina or grouped in a collar around the neuropil below the photoreceptor layer. The 4-5 largest neurons are in the collar area. Correlation of neuron morphology with electrical activity was done with intracellular recording and Lucifer yellow injection of some of the larger neurons in the collar area whose action potentials contribute to the optic nerve impulses. Each one has an axon in the optic nerve and processes that go to the neuropil. They are the pacemaker neurons of the circadian rhythm in impulse frequency that is recorded from the optic nerve, since only their action potentials are correlated 1:1 with the optic nerve impulses. Gap junctions (with pentalaminar structure) are common between photoreceptors, glial cells, photoreceptors and glial cells, and neuronal processes in the neuropil, providing a basis for electrotonic coupling among retinal cells. There are about 2000 axons (diameter less than 3 micron) in the optic nerve, possibly one from each retinal photoreceptor and neuron plus efferent fibres from the brain. Accessory nerves, containing a few large axons, are seen in the optic nerve sheath.     

The conditioning effect of optic nerve injury upon axonal regrowth from adult rat retinal ganglion cells explanted in vitro

An in vitro assay was used to determine the effects of conditioning nerve lesions on the regeneration of adult rat retinal ganglion cell (RGC) axons from retinal explants. Following the conditioning lesion (CL) of unilateral optic nerve transection, maximal regrowth was seen from RGC explanted from ipsilateral retinae 10 days post-CL. Explants from this group initiated axonal regrowth earlier and a greater percentage regrew axons when compared with explants from normal rats. Axonal regrowth from explants of retinae contralateral to CL was also seen earlier than normal. In further experiments, the effects of both exposure of the optic nerve sheath in the orbit and the incision of the dura without injury to optic nerve axons were studied. The conditioning effect of a dural incision was found to be the same as that of optic nerve transection, whilst exposure of the optic nerve sheath had no conditioning effect on RGC axonal regrowth in vitro.     

Retrograde degeneration of myelinated axons and re-organization in the optic nerves of adult frogs (Xenopus laevis) following nerve injury or tectal ablation

Summary The optic nerve proximal to the lesion (toward the retina) was examined by light and electron microscopy in adultXenopus laevis after various types of injury to optic nerve fibres. Intraorbital resection, transection or crush of the optic nerve or ablation of the contralateral optic tectum all resulted in marked alterations in the myelinated axon population and in the overall appearance of the nerve proximal to the site of injury. Examination of the nerves from 3 days to 6 months postoperatively indicated that a progressive, retrograde degeneration of myelin and loss of large-diameter axons occurred throughout the retinal nerve stump regardless of the type of injury or distance of the injury from the retina. The retinal stump of nerves receiving resection or transection showed a nearly complete loss of myelin and large-diameter axons while the degree of degeneration was subtotal in nerves receiving crush injury or after lesions farther from the retina (i.e. tectal ablation). In addition, the entire retinal nerve stump after all types of injury was characterized by the appearance of an actively growing axon population situated circumferentially under the glia limitans. The latter fibres are believed to represent regrowing axons which are being added onto the nerve, external to the original axon population and are suspected to modify actively the glial terrain and glia limitans.     

Somatostatin-like immunoreactive material in associational ganglion cells of human retina

The retinal ganglion cell is classically viewed as the output cell of the retina, sending a single axon via the optic nerve to synapse in visual relay nuclei of the brain. However, some ganglion cells, termed associational ganglion cells, have axons which do not leave the retina and presumably serve intraretinal communication. Using high-affinity and specific monoclonal antibodies to somatostatin-14 and the avidin-biotin-peroxidase immunohistochemical procedure, somatostatin-immunoreactive associational ganglion cells are specifically stained in human retinas obtained at necropsy. These cells are more numerous in the inferior than the superior retina; they have dendrites which ramify in the inner plexiform layer; and they have sparsely branching axons, many of which can be traced over 1 cm. These axons do not enter the optic nerve. They follow remarkably straight courses at the border of the inner plexiform layer and ganglion cell layer and thereby form a gridwork of fibers covering the entire retinal area. These observations verify the existence of associational ganglion cells in the human and establish somatostatin as a neurotransmitter or neuromodulator candidate for these neurons. The morphology of these cells suggests that they are involved in long-distance interactions within the retina.     

基于对称区域生长和边缘梯度的视神经纤维的分割

在视神经横切面图像中,将每个神经纤维的内外边界进行精确分割是视神经形态分析的重要环节,提出一种基于对称区域生长和髓鞘边缘梯度的有效分割算法.该算法分两步进行,首先根据交互方式下选取的种子点,由对称区域生长算法实现轴突分割,然后在轴突轮廓模型基础上,髓鞘外轮廓在髓鞘平均边缘梯度引导下进行演化,实现自动分割.与K-均值聚类,局部阈值和水平集等其他算法的实验结果相对照显示,该算法分割获得的轴突和髓鞘轮廓与实际轮廓相吻合,其分割结果可以作为后续神经纤维形态分析的基础.     

Early distal axonopathy of the visual pathway in experimental diabetes

Diabetic retinopathy is a leading cause of acquired blindness. Visual function disorders have been observed in diabetic patients with very early retinopathy or even before the onset of retinopathy. The aim of the present work was to analyze the visual pathway in an early stage of experimental diabetes. Diabetes was induced in Wistar rats by an i.p. injection of streptozotocin. A deficit in anterograde transport from the retina to the superior colliculus was observed 6 weeks after streptozotocin injection. At this time point, morphologic studies did not reveal retinal ganglion cell loss or substantial alterations in the superior colliculus. The optic nerve was morphometrically evaluated at intraorbital (unmyelinated and myelinated) and intracranial sections. In animals that had been diabetic for 6 weeks, a large increase in astrocyte reactivity occurred in the distal (but not the intraorbital) portion, which coincided with significant axon loss. Moreover, profound myelin alterations and altered morphologic features of oligodendrocyte lineage were observed at the distal (but not the proximal) optic nerve portion. The present results suggest that axoglial alterations at the distal portion of the optic nerve could be the first structural change in the diabetic visual pathway.     

Rescuing axons from degeneration does not affect retinal ganglion cell death

After a traumatic injury to the central nervous system, the distal stumps of axons undergo Wallerian degeneration (WD), an event that comprises cytoskeleton and myelin breakdown, astrocytic gliosis, and overexpression of proteins that inhibit axonal regrowth. By contrast, injured neuronal cell bodies show features characteristic of attempts to initiate the regenerative process of elongating their axons. The main molecular event that leads to WD is an increase in the intracellular calcium concentration, which activates calpains, calcium-dependent proteases that degrade cytoskeleton proteins. The aim of our study was to investigate whether preventing axonal degeneration would impact the survival of retinal ganglion cells (RGCs) after crushing the optic nerve. We observed that male Wistar rats (weighing 200-400 g; n=18) treated with an exogenous calpain inhibitor (20 mM) administered via direct application of the inhibitor embedded within the copolymer resin Evlax immediately following optic nerve crush showed a delay in the onset of WD. This delayed onset was characterized by a decrease in the number of degenerated fibers (P<0.05) and an increase in the number of preserved fibers (P<0.05) 4 days after injury. Additionally, most preserved fibers showed a normal G-ratio. These results indicated that calpain inhibition prevented the degeneration of optic nerve fibers, rescuing axons from the process of axonal degeneration. However, analysis of retinal ganglion cell survival demonstrated no difference between the calpain inhibitor- and vehicle-treated groups, suggesting that although the calpain inhibitor prevented axonal degeneration, it had no effect on RGC survival after optic nerve damage.     

Anatomical correlations of intrinsic axon repair after partial optic nerve crush in rats.

About 15% of retinal ganglion cells survive diffuse axonal injury of the optic nerve in adult rats. Following initial blindness, discrimination of visual stimuli in behavioral tests recovers within three weeks. To investigate the mechanisms promoting this functional recovery the axonal transport and the neurofilaments were studied. Intraocularly applied MiniRuby is transported until the place of crush and accumulated in enlarged axon terminals. Three weeks after lesion the anterograde transport of MiniRuby recovers distal to the place of crush. At the same point in time the retrograde transport of surviving retinal ganglion cells is restored which was visualized by horseradish peroxidase injected into the superior colliculus. The heavy neurofilament was stained immunohistochemically and analyzed statistically up to three weeks after optic nerve crush. The stained filaments in the axon fibers of retinal ganglion cells appear wavelike and/or fragmented up to day 8, but first signs of heavy neurofilament restitution in the fibers of the optic nerve are seen at day 12 after axonal injury. Because these results cannot be explained by longlasting axon regeneration, the present results provide convincing evidence for intrinsic axon repair soon after diffuse axonal injury that correlates in time with recovery of vision.     

An immunocytochemical marker for hamster retinal ganglion cells

Summary We examined the specificity and developmental time course of the labelling of retinal ganglion cells in Syrian hamsters by a monoclonal antibody AB5. In adult hamsters, AB5 selectively labelled somata in the ganglion cell layer, dendrites in the inner plexiform layer and axons in the nerve fibre layer. When retinal ganglion cells were retrogradely labelled with Dil prior to AB5 immunocytochemistry, all of the retrogradely labelled retinal ganglion cells in the ganglion cell layer were AB5 immunoreactive, indicating that AB5 labels all classes of ganglion cell in that layer. In retinae depleted of retinal ganglion cells by neonatal optic nerve transections, AB5 did not label any somata or processes, indicating that AB5 specifically labels retinal ganglion cells. During development, AB5 labelling first appeared as a weak staining of cell bodies in the ganglion cell layer on postnatal day 12 (P12; PO=first 24 h following birth) and acquired the staining pattern seen in the adult by postnatal day 14. From the onset of AB5 immunoreactivity, AB5-labelled somata of varying sizes were present across the entire retinal surface. Although AB5 labelled retinal ganglion cell axons in the nerve fibre layer of the retina it did not label the optic nerve or retinal ganglion cell axons in the brain at any age examined. AB5 labelling was also found to be compatible with bromodeoxyuridine immunocytochemistry and, therefore, useful for determining the time of generation of hamster retinal ganglion cells.