Processes Of Ganglion Cells Research Articles

Antiserum to lucifer yellow: preparation, characterization, and use for immunocytochemical localization of dye-filled retinal neurons.

We present a new method for the preparation of antisera to Lucifer Yellow, and these antisera are here shown to be particularly suitable for immunocytochemical localization of multiple dye-injected cells in large pieces of vertebrate retina. The method involves the preparation of covalent conjugates of the VS isomer of Lucifer Yellow with keyhole limpet hemocyanin (KLH) or rabbit serum albumin (RSA), and their use as immunogens in rabbits. Both carrier protein conjugates yielded robust antibody responses. Antiserum to the KLH-LY conjugate contained precipitating antibodies against LY and KLH, although activity to the latter did not interfere with immunocytochemical staining. Rabbit antiserum to the RSA-LY conjugate contained precipitating antibody only against LY. When used for immunocytochemical staining of large retinal pieces containing many LY-filled cells, both antisera yielded well-stained, darkly filled cells similar to those seen with the Golgi technique; even very fine dendritic processes of retinal ganglion cells could be followed for long distances. LY immunocytochemistry provides a useful alternative to photooxidation for the analysis of multiple dye injected cells, especially in whole mounts. This approach may also be useful for immunocytochemical identification of cells filled with LY after tissue fixation.

Ultrastructure of human retinal cell transplants with long survival times in rats

Human fetal retinas (6–12 weeks post-conception) were obtained from elective abortions, transplanted to rat retinas and examined by electron microscopy. The oldest transplants that form the basis of this report were obtained 40 and 41 total weeks post-conception. The host rats were immunosuppressed with cyclosporin A. The transplants developed according to their intrinsic, genetically determined timetable. The development was heterogeneous with some parts showing almost normal differentiation and others, little. Both rods and cones developed with inner and outer segments and synaptic terminals. In regions corresponding to the inner plexiform layer, bipolar cell processes were seen in the typical dyad arrangement. Likewise, amacrine cell processes formed typical conventional synapses. Serial synapses were seen, engaging amacrine cell synapses as well as a few reciprocal synapses at the bipolar cell dyads. Monad-type synaptic complexes, a sign of immaturity, were common in bipolar cell processes. Similarly, incompletely differentiated synapses of both the amacrine and bipolar cell types were often observed. Ganglion cell processes could not be identified with certainty. A structure with morphological characteristics similar to the inner limiting membrane was noted to form inside the transplant. Both epi-retinal and sub-retinal transplants were obtained. Transplant cells touched host photoreceptor cells or pigment epithelium without any obvious specializations. The host pigment epithelium microvilli were absent adjacent to the graft. However, graft cells did appear in the host retina, and nerve cell processes were observed to cross the membrane separating the transplant and host.

Localization of immunoreactive cholecystokinin precursor to amacrine cells and bipolar cells of the macaque monkey retina

We used antisera that recognized precursors of the neuropeptide cholecystokinin extended at the carboxyl terminus in an immunocytochemical study of the macaque retina. A subpopulation of bipolar cells with long, obliquely oriented dendrites was labeled. Their axons terminated exclusively in the fifth stratum of the inner plexiform layer, where they contacted processes of amacrine and ganglion cells. Based on their morphology, these cells appeared to be the type that contacts short-wavelength cones selectively. Two types of amacrine cells were also labeled, and processes from both types formed dense plexuses in the second and fourth strata of the inner plexiform layer. The majority of their synaptic connections were with other amacrine cells, but they had more contacts with bipolar cell axons and retinal ganglion cell dendrites than any other peptidergic cells in the macaque retina. We studied extracts of macaque retina with gel-filtration chromatography and radioimmunoassays to confirm our immunohistochemical results. We found cholecystokinin octapeptide and other immunoreactive forms that were amidated at their carboxyl termini and were therefore likely to be biologically active. Unlike most other regions of the CNS, however, the retina had relatively low concentrations of amidated forms, and forms with extended carboxyl termini that are presumably their precursors were far more abundant. These findings suggest that the rate of cholecystokinin synthesis in the retina is quite high, as we would expect if the peptide were found in tonically active neurons.

GABA‐like immunoreactivity in the macaque monkey retina: A light and electron microscopic study

The distribution of GABA-like immunoreactivity in the macaque monkey retina was studied by using postembedding techniques on semithin and ultrathin sections. At the light microscopic level, both inner and outer plexiform layers showed strong GABA-like immunoreactivity in the central retina. All the horizontal cells, some bipolar cells, 30-40% of amacrine cells, occasional interplexiform cells, and practically all displaced amacrine cells were labeled. In the peripheral retina (beyond 5 mm eccentricity), the outer plexiform layer and the horizontal cells were not labeled, but all other cell types showed the same labeling pattern as in the central retina. Synapses of the inner plexiform layer involving a pre- or postsynaptic GABA-labeled process were studied electron microscopically. Synapses involving a GABA-labeled presynaptic amacrine cell process made up 80% of the synapses observed. These GABA-labeled amacrine processes synapsed onto amacrine, bipolar, and ganglion cell processes as well as onto amacrine and ganglion cell bodies. Synapses involving a postsynaptic GABA-labeled process made up 20% of the synapses studied. The GABA-like immunoreactive processes were postsynaptic to bipolar cells at the dyads and to amacrine cells at conventional synapses.

Synaptic analysis of amacrine cells in the turtle retina which contain tyrosine hydroxylase-like immunoreactivity

This study examined amacrine cells in the retina of the turtle Pseudemys scripta elegans, which were labelled using an antiserum directed against tyrosine hydroxylase (an enzyme participating in catecholamine synthesis). These cells were investigated using both light and electron microscopy. Labelled somata were located in the inner nuclear layer near the border of the inner plexiform layer. The dendritic arborizations of these neurons were tristratified and arborized in strata 1 and 3 and near the border between strata 4 and 5. Serial tangential sections taken through the entire inner plexiform layer of a 1 mm-2 region in mid-peripheral retina were examined. All of the synapses associated with labelled profiles were counted and classified. The majority (84%) of the synapses involving labelled processes represented output, while the remaining 16% represented synaptic input. The synaptic output of the labelled processes was as follows: 87% onto unlabelled amacrine cells, 4% onto ganglion cells, 9% onto unidentified cell processes. None of the synaptic output from labelled processes was onto bipolar cells. The synaptic input to these labelled cells was from bipolar cells (29%) and from unlabelled amacrine cells (71%). A well labelled amacrine cell was serially sectioned and examined at the ultrastructural level to analyze its synaptic connectivity. Immunoreaction product was located diffusely throughout the cytoplasm and in large vesicles. The synaptic organization of the cell was directed primarily toward output. The labelled processes were postsynaptic and presynaptic to unlabelled amacrine cell processes in strata 1 and 3 and at the border between strata 4 and 5. Synaptic input from bipolar cells was seen exclusively near the border between strata 4 and 5. Labelled processes were presynaptic to ganglion cell processes in stratum 1 and at the border between strata 4 and 5, but not in stratum 3. Quantitative studies suggested that amacrine cell inputs and outputs were evenly distributed across the dendritic arborization, while bipolar cell inputs and outputs to ganglion cells were concentrated on the distal parts of the dendritic arborization. No labelled processes were seen in the outer plexiform layer, indicating that the cells with tyrosine hydroxylase-like immunoreactivity in the turtle retina were true amacrine cells and not interplexiform cells.

Fiber counts at multiple sites along the rat ventral root after neonatal peripheral neurectomy or dorsal rhizotomy.

We hypothesized that the afferent fibers in the ventral root of the rat are the third branches of dorsal root ganglion cells; these afferent processes in the ventral root are of varying length and end bluntly along the length of the root. In the case of an injury at either the central or the peripheral processes of the dorsal root ganglion cells in the neonatal stage, these fibers sprout at the blunt endings along the length of the ventral root. We cut either the sciatic nerve or the dorsal root on one side in neonatal rats. After the rats were fully grown, the number of both myelinated and unmyelinated fibers was counted in electron photomicrographs at multiple sites along the length of the ventral root. We observed a greatly increased number of unmyelinated fibers in the ventral root after the sciatic nerve had been cut at the neonatal stage. The magnitude of increase was more at the distal than at the proximal portion of the ventral root, suggesting that added fibers originated from the distal side. Neonatal dorsal rhizotomy, however, did not produce the same result. These results are consistent with our hypothesis that peripheral nerve injury at the neonatal stage triggers sprouting of the third branches of the dorsal root ganglion cells which end bluntly along the length of the ventral root in the normal animal.

Electrophysiological evidence for an increase in the number of ventral root afferent fibers after neonatal peripheral neurectomy in the rat

Our recent study has shown that many afferent fibers in the ventral root are third branches of dorsal root ganglion cells in addition to their processes in the peripheral nerve and the dorsal root. From results of this study, we hypothesized that most of the afferent fibers in the normal ventral root are extra processes of certain dorsal root ganglion cells. To accomodate experimental findings by others, we formulated several working hypothesis in the present study as an extension of our previous hypothesis: these afferent processes in the ventral root are of varying length; they end bluntly along the length of the root; and in an event such as peripheral neurectomy in the neonatal stage, these fibers sprout at the blunt endings along the length of the ventral root. We tested the above hypotheses using electrophysiological methods. The sciatic nerve on one side in neonatal rats was cut. After the rat was fully grown, volleys of neural activity were recorded along the length of the ventral root while stimulating the dorsal root of the same segment. There was a great increase in the size of compound action potentials in the ventral root on the sciatic nerve-lesioned side. Various lines of evidence suggest that this enhancement of the evoked potentials is likely to be due to an increase in the number of afferent fibers in the ventral root in response to neonatal peripheral nerve injury. The results are consistent with our hypotheses.

Peripheral nerve regeneration induces elevated expression of GAP-43 in the brainstem trigeminal complex of adult hamsters

A polyclonal antibody was used to delineate the pattern of GAP-43 expression in central processes of trigeminal ganglion cells while their peripheral processes were undergoing regeneration. Two weeks after transection of the infraorbital nerve, levels of GAP-43 ipsilateral to the transection were greatly increased along the trajectory of infraorbital axons in the central trigeminal tract and also within the target neuropil. We conclude that elevated levels of GAP-43 in central processes of injured trigeminal ganglion cells occur in direct response to the regenerative response of the cell body and may have important implications for ‘plasticity’-related changes seen in the adult trigeminal system.

Neuropeptide Y-like immunoreactivity in neurons of the human retina

The distribution of neuropeptide Y-like immunoreactivity (NPY-LI) was investigated in wholemounts and in transverse sections of the human retina. NPY-LI was localized to the soma and axonal processes of large ganglion cells (GCs) and to the soma and dendritic arborization of amacrine cells (ACs). NPY-LI GCs were unevenly distributed across the retina, the highest density of 875 cells/mm 2 was found in the fovea centralis and the lowest density of 15 cells/mm 2 in the peripheral retina. The total number of NPY-LI GCs in the retina was estimated to be about 85,000. The soma sizes of NPY-LI GCs increased from 116 μm 2 ± 23 (s.d.) in the retinal centre to 251 μm 2 ± 57 in the retinal periphery. The soma size of NPY-LI ACs was in the range of 40 and 50 μm 2. In transverse sections NPY-LI was seen to be localized to the optic fibre layer, to the somata of GCs, to the scleral sublamina of the inner plexiform layer (AC dendrites) and to the innermost part of the inner nuclear layer (somata of ACs). The gradients of soma sizes and retinal distribution of NPY-LI GCs were taken as an indication that they correspond to the class of large to very large GCs, previously identified in the human retina by Golgi impregnation.

A peptide histidine isoleucine/peptide histidine methionine-like peptide in the rabbit retina: Colocalization with vasoactive intestinal peptide, synaptic relationships and activation of adenylate cyclase activity

Antisera against peptide histidine isoleucine and peptide histidine methionine were found to label a subpopulation of amacrine and displaced amacrine cells in the rabbit retina with processes ramifying in sublaminas 1. 3 and 5 of the inner plexiform layer. Preadsorption controls demonstrated that this immunoreactivity was specific for a peptide histidine isoleucine- or peptide histidine methionine-like (peptide histidine isoleucine peptide histidine methionine-like) peptide. and was not caused by cross-reactivity of the peptide histidine isoleucine or peptide histidine methionine antibodies with vasoactive intestinal peptide vasoactive intestinal peptide. In double-label studies, vasoactive intestinal peptide and peptide histidine isoleucine peptide histidine methionine-like immunoreactivity were colocalized in the same population of retinal neurons. Electron microscopic analysis revealed that the peptide histidine isoleucine peptide histidine methionine-labelled cells interacted with processes of bipolar cells, amacrine cells and ganglion cells. Peptide histidine methionine and peptide histidine isoleucine were slightly less potent than vasoactive intestinal peptide in stimulating adenylate cyclase activity in the rabbit retina, while the related peptides secretin, glucagon, and the C-terminal vasoactive intestinal peptide fragment, vasoactive intestinal peptide (10–28), showed little or no stimulatory activity. Stimulation of adenylate cyclase by high concentrations of vasoactive intestinal peptide and peptide histidine methionine were non-additive. These results suggest that a peptide histidine isoleucine peptide histidine methionine-like peptide may function as a neuroactive peptide in the mammalian retina, and that this peptide appears to be cosynthesized and colocalized with vasoactive intestinal peptide and to mimic the activity of vasoactive intestinal peptide through interaction with vasoactive intestinal peptide receptor-adenylate cyclase complexes.

Regeneration of adult rat retinal ganglion cell processes in monolayer culture: comparisons between cultures of adult and neonatal neurons

The aim of the present study was to develop a model culture system for the study of factors controlling regeneration of axons from injured adult mammalian central nervous system. We show, for the first time, that retinal ganglion cells (RGC) dissociated from adult rat retina, regrow processes in vitro over long distances under appropriate conditions in monolayer culture. Most importantly, adult RGC depend completely upon the presence of a preformed layer of neonatal cortical astrocytes, whereas RGC from neonatal retinae are supported by indigenous retinal glia which spread and proliferate to form a monolayer of cells upon which RGC regrow processes. Adult retinal glia fail to spread on the culture surface and proliferate in the same way and we suggest that this is a major factor in limiting the survival of adult RGC on an acellular substrate such as polylysine. Laminin does not substitute for the presence of a glial monolayer. These findings indicate that at least one type of adult CNS neuron is capable of regenerating its processes in vitro in an environment which includes a cellular component of CNS tissue.

Nicotinic antagonists enhance process outgrowth by rat retinal ganglion cells in culture.

Functional nicotinic cholinergic receptors are found on mammalian retinal ganglion cell neurons in culture. The neurotransmitter acetylcholine (ACh) can be detected in the medium of many of these retinal cultures, after release presumably from the choline acetyltransferase-positive amacrine cells. The postsynaptic effect of endogenous or applied ACh on the ganglion cells can be blocked with specific nicotinic antagonists. Here it is shown that within 24 hours of producing such a pharmacologic blockade, the retinal ganglion cells begin to sprout or regenerate neuronal processes. Thus, the growth-enhancing effect of nicotinic antagonists may be due to the removal of inhibition to growth by tonic levels of ACh present in the culture medium. Since there is a spontaneous leak of ACh in the intact retina, the effects of nicotinic cholinergic drugs on process outgrowth in culture may reflect a normal control mechanism for growth or regeneration of retinal ganglion cell processes that is exerted by ACh in vivo.

Localization of choline acetyltransferase-like immunoreactivity in the embryonic chick retina.

Putative sites of acetylcholine synthesis in the retina of the embryonic and posthatched chick were localized immunohistochemically with antisera to choline acetyltransferase; the resultant choline acetyltransferase-like immunoreactivity (ChAT-IR) was compared to demonstrated sites of acetyltransferase (AChE) activity, and changes were followed in localization during development. The results confirmed the early and rapid course of development of the chick's retinal cholinergic system described in previous biochemical and morphological studies. Immunoreactivity was first detected at embryonic day 6.5 in cells close to the retina's vitreal surface. By 8 days it was present in cells in two juxtaposed rows; by the ninth day the two rows were separated and immunoreactivity was evident in two subliminae of the inner plexiform layer. On the tenth day distribution was like that in the posthatched chicken, in type I cholinergic cells in the inner nuclear layer and in type II cells in the ganglion cell layer (Millar et al.: Neurosci. Lett. 61:311-316, '85), and similar to that of most vertebrates. Three days before hatching, a third population of weakly immunoreactive cells (type III cells) appeared within the inner nuclear layer. The onset of localizable ChAT-IR occurred in amacrine cells and in their processes, before the period of synaptogenesis. Acetylcholinesterase activity was localized at an earlier age than ChAT-IR, and at all ages was present in more cells. The results obtained support the view that "displaced" cholinergic amacrine cells begin to differentiate at the same time and in the same retinal region as type I cholinergic cells. Separation of the two groups is a consequence of the ramification of processes of amacrine and ganglion cells rather than a result of the secondary migration of cells between layers.

Motoneurone survival and neurite regeneration requirements: the role of dorsal root ganglion cells during development

The effect of dissociated dorsal root ganglion cells on the survival of motoneurones in vitro from differently aged chick embryos has been studied. Homogeneous cultures of motoneurones were prepared from 5–8-day embryos, using a cell sorter; dorsal root ganglion cells were obtained from 8-day embryos. The survival of motoneurones from 5-day and 6-day embryos was not enhanced above controls by the presence of dorsal root ganglion cells; however, the survival of motoneurones from older embryos was greatly increased, reaching a maximum of over 80% for 8-day embryonic motoneurones. In contrast, the number of motoneurones that had regenerated neurites when co-cultured with dorsal root ganglion cells for 24 h decreased with the motoneurone age at plating, from 51% at 5 days to less than 10% for 7- and 8-day motoneurones. The survival-enhancing effects were probably mediated by cell contact between the motoneurones and processes of the dorsal root ganglion cells: conditioned media from high-density cultures of dorsal root ganglion cells could not be shown to significantly enhance the survival of motoneurones above that of control levels. The possibility that the ganglion cells exert this survival enhancing effect by depolarizing the motoneurones was examined by exposing 8-day sorted motoneurones to 47 mM potassium: this did not effect the survival of the motoneurones relative to control levels. The stage dependency of the survival of motoneurones on different neurotrophic factors and the dorsal root ganglion cell is discussed.

Expression of a 48‐Kilodalton Growth‐Associated Protein in the Goldfish Retina

One of the most striking molecular correlates of optic nerve regeneration in the goldfish is the increased labeling of a 48 kilodalton (kD) acidic protein that is conveyed to the developing nerve endings from the retina by rapid axonal transport. The present study examined the biosynthesis and molecular characteristics of this protein. Retinas derived either from intact controls or from goldfish undergoing optic nerve regeneration (10-14 days postcrush) were pulse-labeled with [3H]proline or [35S]methionine, followed by subcellular fractionation and analysis of protein synthesis patterns by two-dimensional gel electrophoresis and fluorography. Synthesis of the 48-kD acidic protein (termed here GAP-48) was detected only in retinas that were undergoing axonal regeneration. Pulse-chase labeling experiments demonstrated that the protein undergoes a post-translational modification that requires 15-20 min. This processing could be selectively blocked by tunicamycin, an inhibitor of protein N-glycosylation. The protein was also found to incorporate low levels of phosphate in vitro. Thus, the differential appearance of GAP-48 in regenerating axons might be regulated either at the level of gene expression or by selective posttranslational processing in retinal ganglion cells. By the criteria of molecular weight, isoelectric point, anomalous migration properties on sodium dodecyl sulfate-polyacrylamide gels, phosphorylation, subcellular distribution, and the pattern of digestion products generated by Staphylococcus aureus V8 protease, GAP-48 appears to be equivalent to the B-50 (F-1) phosphoprotein of the mammalian brain.

Ultrastructural changes in the dorsal motor nucleus of monkey following bilateral cervical vagotomy.

The neurons of the dorsal motor nucleus (DMN) of the monkey (Macaca fascicularis) were of two main types: small (13 X 8 micron) and medium-sized (20 X 13 micron). The latter, which were the predominant form, contained a pale oval nucleus surrounded by organelle-rich cytoplasm. Between one and three long principal dendrites per section profile arose from each of the somata. Both axosomatic and axodendritic synapses were seen on these cells although the latter were more common. No structural changes were noted in the DMN 1-3 days after bilateral cervical vagotomy. Some of the dendrites of the medium-sized axotomized vagal neurons appeared darkened 5-10 days after the operation. With longer surviving intervals, i.e. 21 and 28 days after operation, darkened dendrites were more commonly seen and the cytoplasmic density of these dendrites was dramatically enhanced. Their mitochondria were pale and some of them also showed vesiculation. Both normal and degenerating axon terminals were seen to form synaptic contacts with the darkened dendrites. The degenerating axon terminals were characterized by the clumping of their round agranular vesicles. Both darkened dendrites and degenerating axon terminals were phagocytosed by hypertrophied astrocytes and activated microglial cells. Blood elements infiltrating into the DMN were a possible source for some of the neural macrophages. It was concluded from the present study that the dendrites of the vagal neurons were the first structures to degenerate in axotomy and these were subsequently removed by glial elements. Degenerating axon terminals on the darkened dendrites could represent endings of the central processes of peripheral vagal ganglion cells that had undergone transganglionic degeneration after damage to their peripheral processes.

Neurochemical stratification in the inner plexiform layer of the vertebrate retina

Most regions of metazoan nervous systems can be idealized as laminated structures in which layers of neuropil are sandwiched between layers of neuronal somas and the structure pierced by afferent or efferent fibers whose trajectories are roughly orthogonal to the layers. Within very complex systems, such as retina, mammalian cerebral cortex or invertebrate visual medulla and lobula plate, a finer lamination exists within individual layers of neuropil. The inner plexiform layer (IPL) of the vertebrate retina has long been known as a structure composed of many sublayers. As one passes along an axis normal to the plane of the IPL*, the local composition varies in terms of the kinds and numbers of synaptic complexes, axonal or dendritic processes, and glial profiles. We will consider the composition within a plane at a given level to be homogeneous, though this presumption is dependent on the scale over which one samples the composition. This review is intended to summarize current knowledge, and an extensive historical account is inappropriate. Nevertheless, the observations of Ramon y Cajal and his contemporaries are central to interpreting more recent data. Ramon y Cajal ( 1933) gives Dogiel (I 891) credit for the attribution of IPL banding to the processes of amacrine and ganglion cells but notes that many

Qualitative and quantitative observations of spiral ganglion development in the rat

The postnatal development of the spiral ganglion cells in the rat was studied from birth until the adult stage. At birth, a single population of ganglion cells is present. Some of them are surrounded by one or two layers of satellite cell processes. With maturation, the satellite cell processes increase in number around the cell body and its processes. At the end of the first postnatal week, two important events occur. The first is the appearance of myelin lamellae between the 4th and the 6th postnatal day in both ganglion cell processes, and between the 6th and the 8th day in the cell body. The second event is the appearance of a new type of cell (the Type II spiral ganglion cell) on the 6th to the 8th day postpartum. At this stage, the Type II cell is mainly characterized by densely packed neurofilamentous structures in the cytoplasm. Comparison between the myelination of the cell body and its processes reveals three main differences! 1.(1) There is a time lag of approximately 2 days between the onset of myelination in the cell body and in its processes.2.(2) The kinetics of myelination are different in the cell processes and in the cell body. The myelination of the cell body starts slowly, whereas it is very fast in the processes. Later, the kinetics of myelination decrease in the processes, and increase in the cell body.3.(3) At all stages including the adult, the fibers have a myelin sheath composed of more lamellae than the cell body. These observations are discussed with respect to development in other species.

Laminar distributions of choline acetyltransferase and acetylcholinesterase activities in the inner plexiform layer of rat retina.

Choline acetyltransferase and acetylcholinesterase activities were measured in samples taken at 7-micron increments through the inner plexiform layer of rat retina. These enzyme activities were not uniformly distributed through the depth of the inner plexiform layer. Peaks of choline acetyltransferase activity occurred at about one-third and peaks of acetylcholinesterase activity at about one-fifth of the depth into the inner plexiform layer from either side. The positions of the two peaks of choline acetyltransferase activity most likely correspond to the locations of processes from cholinergic amacrine somata in the inner nuclear layer, which spread in sublamina a, and processes from cholinergic amacrine somata "displaced" in the ganglion cell layer which spread in sublamina b of the inner plexiform layer. The peaks of acetylcholinesterase activity may in addition correspond to the processes of cholinoceptive amacrine and ganglion cells. The magnitudes of choline acetyltransferase and acetylcholinesterase activities are as high as found anywhere in rat brain, emphasizing the important role of cholinergic mechanisms in visual processing through the rat inner plexiform layer.

Peptide-like immunoreactivity in anuran optic nerve fibers.

Unilateral section, crush, or ligation of the optic nerve was performed in Rana pipiens. Following optic nerve disruption, Substance P (SP)-, leucine-enkephalin (LENK)-, cholecystokinin octapeptide (CCK8)-, and bombesin (BOM)-like immunoreactivities were analyzed in the retinae and optic nerves. Peptide-like immunoreactivity developed in the retinal stump of disrupted optic nerves within 1 hour after surgery and was retained until at least 30 days. Peptide-positive staining in the retinal stump of the optic nerves was abolished by preabsorption of each of the antibodies/antisera with the corresponding synthetic substances. No massive peptide-like immunoreactivity was observed in the cerebral stump of the ligated side, nor in the contralateral, nonoperated, optic nerve. No change in the pattern of peptide-like immunoreactivity was apparent in the retina ipsilateral or contralateral to the experimental procedure. The optic tectum contralateral to the surgical procedure displayed those changes in peptide-like immunoreactivity described previously following retinal deafferentation (Kuljis and Karten, '82a, '83a). Peptide-like immunoreactivity in the stump retinad to the surgical procedure occurred in the form of beaded and fibrillar elements often ending in an irregular expansion near the lesion site. Fluorescent double-label antibody methods demonstrated that SP-like immunoreactivity is present in different processes than those containing LENK, CCK8, or BOM. Electron microscopical immunocytochemistry revealed that peptide-like immunoreactivity is contained within unmyelinated and possibly also within myelinated axons in the stump retinad to the traumatic procedure. Radioimmunoassay studies of SP demonstrated a four- to sixfold increase in SP-like content in the retinal stump of ligated nerves, compared with both the cerebral stump and with the contralateral nonoperated optic nerves. These findings demonstrate the presence of peptide-like immunoreactivity in retinal ganglion cell processes, which is compatible with either a posttraumatic expression of previously repressed peptide-like phenotypes, or, most likely, with the existence of various classes of peptide-containing retinal ganglion cells. The latter prospect strongly suggests that peptide-specific subsets of retinal ganglion cells terminate in highly specific laminae in the optic tectum (Kuljis and Karten, '81, '82a-83a) and presumably differ in their physiological role in vision.(ABSTRACT TRUNCATED AT 400 WORDS)