Age-related functional and structural retinal modifications in the Igf1-/- null mouse.

BACKGROUND: Mutations in the gene encoding human insulin-like growth factor-I (IGF-I) cause syndromic neurosensorial deafness. To understand the precise role of IGF-I in retinal physiology, we have studied the morphology and electrophysiology of the retina of the Igf1(-/-) mice in comparison with that of the Igf1(+/-) and Igf1(+/+) animals during aging. METHODS: Serological concentrations of IGF-I, glycemia and body weight were determined in Igf1(+/+), Igf1(+/-) and Igf1(-/-) mice at different times up to 360days of age. We have analyzed hearing by recording the auditory brainstem responses (ABR), the retinal function by electroretinographic (ERG) responses and the retinal morphology by immunohistochemical labeling on retinal preparations at different ages. RESULTS: IGF-I levels are gradually reduced with aging in the mouse. Deaf Igf1(-/-) mice had an almost flat scotopic ERG response and a photopic ERG response of very small amplitude at postnatal age 360days (P360). At the same age, Igf1(+/-) mice still showed both scotopic and photopic ERG responses, but a significant decrease in the ERG wave amplitudes was observed when compared with those of Igf1(+/+) mice. Immunohistochemical analysis showed that P360 Igf1(-/-) mice suffered important structural modifications in the first synapse of the retinal pathway, that affected mainly the postsynaptic processes from horizontal and bipolar cells. A decrease in bassoon and synaptophysin staining in both rod and cone synaptic terminals suggested a reduced photoreceptor output to the inner retina. Retinal morphology of the P360 Igf1(+/-) mice showed only small alterations in the horizontal and bipolar cell processes, when compared with Igf1(+/+) mice of matched age. CONCLUSIONS: In the mouse, IGF-I deficit causes an age-related visual loss, besides a congenital deafness. The present results support the use of the Igf1(-/-) mouse as a new model for the study of human syndromic deaf-blindness.

Retinal degeneration in two lines of transgenic S334ter rats.

Aim of this study was to examine synaptic connectivity changes in the retina and the location and rate of apoptosis in transgenic S334ter line-3 and line-5 rats with photoreceptor degeneration. Heterozygous S334ter-line-3 and line-5 at P11-13, P30, P60, P90 and several control non-dystrophic rats (Long Evans and Sprague-Dawley) at P60, were studied anatomically by immunohistochemistry for various cell and synaptic markers, and by PNA and TUNEL label.- S334ter line-3 exhibited the fastest rate of degeneration with an early loss of photoreceptors, with 1-2 layers remaining at P30, and only cones left at P60. Line-5 had 4-5 layers left at P30, and very few rods left at P60-90. In both lines, horizontal cell processes (including dendrites and axon) were diminished at P11-13, showing gaps in the outer plexiform layer (OPL) at P60, and at P90, almost no terminal tips could be seen. Bipolar cells showed a retraction of their dendrites forming clusters along the OPL. Synaptic terminals of A-II amacrine cells in the IPL lost most of their parvalbumin-immunoreactivity. The apoptosis rate was different in both lines. Line-3 rats showed many photoreceptors affected at P11, occupying the innermost part of the outer nuclear layer. Line-5 showed a lower number of apoptotic cells within the same location at P13. In summary, the S334ter line-3 rat has a faster progression of degeneration than line-5. The horizontal and bipolar terminals are already affected at P11-P13 in both models. Apoptosis is related to the mutated rhodopsin transgene; the first photoreceptor cells affected are those close to the OPL.

Functional and structural modifications during retinal degeneration in the rd10 mouse.

Mouse models of retinal degeneration are useful tools to study therapeutic approaches for patients affected by hereditary retinal dystrophies. We have studied degeneration in the rd10 mice both by immunocytochemistry and TUNEL-labeling of retinal cells, and through electrophysiological recordings. The cell degeneration in the retina of rd10 mice produced appreciable morphological changes in rod and cone cells by P20. Retinal cell death is clearly observed in the central retina and it peaked at P25 when there were 800 TUNEL-positive cells per mm(2). In the central retina, only one row of photoreceptors remained in the outer nuclear layer by P40 and there was a remarkable deterioration of bipolar cell dendrites postsynaptic to photoreceptors. The axon terminals of bipolar cells also underwent atrophy and the inner retina was subject to further changes, including a reduction and disorganization of AII amacrine cell population. Glutamate sensitivity was tested in rod bipolar cells with the single cell patch-clamp technique in slice preparations, although at P60 no significant differences were observed with age-matched controls. Thus, we conclude that rod and cone degeneration in the rd10 mouse model is followed by deterioration of their postsynaptic cells and the cells in the inner retina. However, the functional preservation of receptors for photoreceptor transmission in bipolar cells may open new therapeutic possibilities.

Identification of the Photoreceptor Transcriptional Co-Repressor SAMD11 as Novel Cause of Autosomal Recessive Retinitis Pigmentosa.

Retinitis pigmentosa (RP), the most frequent form of inherited retinal dystrophy is characterized by progressive photoreceptor degeneration. Many genes have been implicated in RP development, but several others remain to be identified. Using a combination of homozygosity mapping, whole-exome and targeted next-generation sequencing, we found a novel homozygous nonsense mutation in SAMD11 in five individuals diagnosed with adult-onset RP from two unrelated consanguineous Spanish families. SAMD11 is ortholog to the mouse major retinal SAM domain (mr-s) protein that is implicated in CRX-mediated transcriptional regulation in the retina. Accordingly, protein-protein network analysis revealed a significant interaction of SAMD11 with CRX. Immunoblotting analysis confirmed strong expression of SAMD11 in human retina. Immunolocalization studies revealed SAMD11 was detected in the three nuclear layers of the human retina and interestingly differential expression between cone and rod photoreceptors was observed. Our study strongly implicates SAMD11 as novel cause of RP playing an important role in the pathogenesis of human degeneration of photoreceptors.

Morphology and distribution of neurons immunoreactive for substance P in the turtle retina.

Immunocytochemical staining procedures with the HRP-complexed antibody to substance P have been carried out on the turtle retina. Examination by light microscopy of wholemount retinas has allowed us to evaluate the morphology and distribution of the substance P immunoreactive cell types. Two amacrine cell types and two or more ganglion cell types are stained in our hands. Type A amacrines are tri-stratified wide-field amacrines. They have their major dendrites in S1 and S3 of the inner plexiform layer and they emit fine dendrites from the major dendrites that end in varicose boutons in S5 on and around cell bodies in the ganglion cell layer. Some of the dendrites in S1 radiate out in axon-like fashion for 1 mm across the retina. The type B amacrine cells are small to medium-field in dendritic extent. They have smaller cell bodies than type A and a single or, at most, two primary dendrites that pass directly to S3 before branching profusely into an intricate net-like dendritic field. The ganglion cells that are stained with substance P antibodies appear to be of several types but their exact morphologies are in doubt because only portions of their major dendrites are stained. Substance P immunoreactive axons are clearly seen to project from the cell bodies to the optic nerve head and axons are stained in the optic nerve itself. The substance P-stained ganglion cells occur in an irregular distribution that reaches a peak density in an elongated band parallel to and 1 mm below the visual streak. The type B amacrine cells reach a maximum density in the visual streak and are distributed in a highly regular mosaic decreasing in density in elliptical isodensity contours from the visual streak. In contrast the type A amacrine cells are rare or absent in the streak, being located in an irregular mosaic in peripheral retina.

Circuitry and role of substance P-immunoreactive neurons in the primate retina.

In this paper, we extend our previous light microscopic (LM) study of substance P (SP)-containing amacrine and ganglion cell types of the human retina (Cuenca et al. [1995] J. Comp. Neurol. 356:491-504) to an electron microscopic (EM) and confocal-imaging study in order to reveal synaptic circuitry and putative input and output neurons. SP-immunoreactive (-IR) amacrine cells in primate retina are typically wide-field cells with large cell bodies occurring in normal or displaced positions relative to the inner plexiform layer (IPL). Their main dendrites bear many spines and are monostratified in stratum 3 (S3) of the IPL. Axon-like processes arise from dendrites close to the cell body and run for hundreds of microns at the same level as the dendrites, thus forming a relatively dense plexus in S3 of the IPL. SP-IR axon processes also climb to S1 to surround some amacrine cell bodies, and others pass into the outer plexiform layer (OPL). Still other axons run down to the ganglion cell layer, where they encircle SP-IR ganglion cells and pass on to end in the nerve fiber layer. The SP-IR ganglion cell types have large cell bodies (20-22 microm diameter) and dendrites that costratify in S3 among the SP-IR amacrine cell processes. Double immunostaining and study by confocal microscopy reveals that SP-IR amacrine cells in the monkey colocalize gamma-aminobutyric acid (GABA). Their main plexus of dendrites in S3 of the IPL is skirted on the S2/S3 border by cone bipolar axons that stain for calbindin but intermingles primarily with glycinergic bipolar cell types of S3 and S3-S4. Strongly GABA-IR/weakly glycine-IR amacrine cell bodies, in addition to the SP-IR large-bodied ganglion cell type, are targets of encircling SP-IR axon processes. EM study of the human SP-IR amacrine cell indicates that input synapses to its dendrites are from bipolar cell axons of the S2/S3 border, S3, and the S3/S4 border of the IPL neuropil (33% of the synaptic input) and from amacrine cell processes (67% of the synaptic input). The input amacrine cells are of at least two distinct types based on cytological criteria. Synaptic output from the SP-IR amacrine cell dendrites is to bipolar cell axons as reciprocal synapses (31%), to amacrine cells (40%), and to ganglion cell profiles, primarily in S3 (29%) of the IPL. The SP-IR axons synapse upon SP-IR ganglion cell bodies and axons, upon normally placed and displaced amacrine cell bodies, and upon bipolar cell dendrites in the OPL. In addition, they appear to synapse among themselves. We shall discuss a wiring diagram and the possible role of SP-IR amacrine cells in the primate retina.

Postembedding immunocytochemistry for GABA and glycine reveals the synaptic relationships of the dopaminergic amacrine cell of the cat retina.

Postembedding electron microscope immunocytochemistry of glycine and GABA conjugated to colloidal gold has been applied to pre-embedded cat retina stained with the antibody against tyrosine hydroxylase (Toh+). Toh+ stained cells are the equivalent of A18 amacrine cells of Golgi descriptions (Kolb et al., '81). The dendrites of Toh+ cells synapse upon several different types of glycine-positive amacrine cell bodies. We suggest that these are the A8, A3/A4, and AII amacrine cell varieties by analogous immunocytochemical staining intensity, to glycine autoradiographic labeling intensity (Pourcho and Goebel, '85). The greatest number of synapses from Toh+ dendrites are directed at the least glycine-positive amacrine, which is the AII cell by all morphological criteria. A few glycine-positive profiles are also presynapatic to the Toh+ stained cell body itself. Toh+ profiles are also presynaptic to GABA-positive amacrine cell bodies. The commonest amacrine synapsed upon is very heavily labeled with GABA immunocytochemistry. We consider it to be the A17 amacrine cell, which is known to label strongly by [3H] muscimol autoradiography (Pourcho and Goebel, '83). The cell body of the Toh+ amacrine cell also receives many synapses, which appear to be GABA-positive, and Toh+ profiles running in stratum 1 of the inner plexiform layer (IPL) are both pre- and postsynaptic to GABA-positive amacrine cell profiles. In addition, the cell body and primary dendrites of the Toh+ cell receive input from a bipolar type and GABA- or glycine-negative profiles. GABA-positive profiles, belonging to the interplexiform cell (IPC), are synapsed upon by Toh+ profiles that run in the outer plexiform layer (OPL).(ABSTRACT TRUNCATED AT 250 WORDS)

Substance P-immunoreactive neurons in the human retina.

Substance P (SP) is a neuropeptide that acts as a neurotransmitter or a neuromodulator in the retina. The aim of this study was to identify the type(s) and the distribution of the SP-immunoreactive (SP-IR) cells in the human retina. We have used an antiserum to SP to immunostain neurons in postmortem human retinae. Immunostained retinae were processed with the avidin-biotin complex (ABC) to visualize the cells either whole mounted in glycerol or embedded in plastic. Some retinae were also sectioned at 20 microns in order to obtain radial views of stained cells. SP-IR amacrine cells stain intensely and appear to be of a single type in the human retina. They are large-field cells with large cell bodies (16 microns diameter) lying in normal or displaced positions on either side of the inner plexiform layer (IPL). Their sturdy, spiny, and appendage-bearing dendrites stratify in stratum 3 (S3) of the IPL, where many overlapping, fine dendrites intermingle to form a plexus of stained processes. Either cell bodies or primary dendrites emit an "axon-like" process that, typically, divides into two long, fine processes, which run in opposite directions for hundreds of micrometers in S5 and S3 before disappearing as distinct entities in the stained plexus in S3. Long, fine dendrites also pass from the dendritic plexus to run in S5 and down to the nerve fiber layer to end as large varicosities at blood vessel walls. In addition, fine processes are emitted from the dendritic plexus that runs in S1, and some pass up to the outer plexiform layer (OPL) to run therein for short distances. The SP-IR amacrine cell has many similarities to the thorny, type 2 amacrine cells described from Golgi studies. In addition to the SP-IR amacrine cells, a presumed ganglion cell type is faintly immunoreactive. Its 20-22 microns cell body gives rise to a radiate, sparsely branched, wide-spreading dendritic tree running in S3. Its dendrites and cell body become enveloped by the more intensely SP-IR processes and boutons from the SP-IR amacrine cell type. The SP-IR ganglion cell type most resembles G21 from a Golgi study.

Postnatal development of microtubules and neurofilaments in the rat optic nerve: a quantitative study.

In this paper the postnatal changes in the cytoskeleton of the rat optic nerve fibers are described and quantified. These changes are also compared with other parameters such as myelination and axonal caliber with the aim of describing a general pattern of optic nerve maturation from a morphological point of view. The results showed that during the first postnatal week microtubules outnumbered neurofilaments but between days 8 and 20 the neurofilaments rapidly increased and on day 20 were about twice as numerous as microtubules. This proportion remained almost unaltered from the end of the third week to the 44th postnatal day. The comparison with other parameters suggested that the cytoskeleton, and in particular the proportion between its components, may be a more reliable index for measuring optic nerve maturation than other variables commonly used.

Regressive and reactive changes in the connectivity patterns of rod and cone pathways of P23H transgenic rat retina.

We have used the P23H line 1 homozygous albino rat to study how progressive photoreceptor degeneration affects rod and cone relay pathways. We examined P23H retinas at different stages of degeneration by confocal microscopy of immunostained sections and electroretinogram (ERG) recordings. By 21 days of age in the P23H rat retina, there is already substantial loss of rods and reduction in rod bipolar dendrites along with reduction of metabotropic glutamate receptor 6 (mGluR6) and rod-associated bassoon staining. The cone pathway is relatively unaffected. By 150 days, when rods are absent from much of the retina, some rod bipolars remain and dendrites of rod and cone bipolar cells form synaptic complexes associated with cones and horizontal cell processes. These complexes include foci of mGluR6 and bassoon staining; they develop further by 270 days of age. Over the course of degeneration, beginning at 21 days, bipolar axon terminals atrophy and the inner retina undergoes further changes including a reduced and disorganized AII amacrine cell population and thinning of the inner plexiform layer. Electroretinogram (ERG) results at 23 days show reductions in a-wave amplitude, in rod and cone-associated b-waves (using a double flash paradigm) and in the amplitude of oscillatory potentials (OPs). By 38 days, rod scotopic a-wave responses and OPs are lost. B-wave amplitudes decline until 150 days, at which point they are purely cone-driven and remain stable up to 250 days. The results show that during the course of photoreceptor loss in the P23H rat, there are progressive degenerative changes, particularly in the rod relay pathway, and these are reflected in the changing ERG response patterns. Later reactive changes involving condensation of cone terminals and neurotransmitter receptors associated with rod and cone bipolar dendrites and with horizontal cell processes suggest that at this stage, there are likely to be complex changes in the relay of sensory information through the retina.

The localization of guanylyl cyclase-activating proteins in the mammalian retina.

PURPOSE: To explore the distribution of guanylyl cylase-activating proteins 1 and 2 (GCAP1 and GCAP2) in the mammalian retina. METHODS: Cryostat and vibratome vertical sections and wholemount retinas from mouse, rat, cat, bovine, monkey, and human eyes were prepared for immunocytochemistry and viewing by light and confocal microscopy. RESULTS: In all mammalian retinas investigated, intense GCAP1 immunoreactivity (GCAP1-IR) was seen in cone photoreceptor inner and outer segments, cell bodies, and synaptic regions. Intensity of the GCAP1-IR was strong in inner segments of rods in all species but weaker in outer segments-particularly so in primates and cats. GCAP2 immunoreactivity (GCAP2-IR) was weak in bovine, mouse, and rat cones but was intense in human and monkey cones. In all species except primates, GCAP2 staining was intense in rod inner and outer segments. In primates GCAP2-IR was intense in the rod inner segment but faint in the rod outer segment. A striking difference from the GCAP1 pattern of immunoreactivity was seen with GCAP2 antibodies as far as the inner retina was concerned. GCAP2-IR was evident in certain populations of bipolar, amacrine, and ganglion cells in all species. CONCLUSIONS: GCAP1 and GCAP2, which are involved in Ca2+-dependent stimulation and inhibition of photoreceptor guanylyl cyclase, can be detected in mammalian photoreceptor inner and outer segments, consistent with their physiological function. The occurrence of both GCAPs in the synaptic region of the photoreceptors indicates participation of these proteins in pathways other than regulation of phototransduction. The occurrence of GCAP2 in inner retinal neurons is indicative of second-messenger chemical transduction, possibly in metabotropic glutamate, gamma-aminobutyric acid (GABA) receptor, and nitric oxide-activated neural circuits.

Substance P: a neurotransmitter of amacrine and ganglion cells in the vertebrate retina.

A short history and summary of the occurrence of substance P in the vertebrate body is presented. Substance P is now generally accepted to be a neurotransmitter and can be visualized by immunocytochemistry to occur in various nerve cells in the CNS. In the retina, substance P-immunoreactivity (SP-IR) occurs in amacrine cell populations in all the species so far studied. In some vertebrates retinas SP is also apparent in one or more ganglion cell types. Anatomical investigations have revealed the morphology and connectivity of SP-IR amacrine cells: they branch in several strata of the inner plexiform layer receiving input from bipolar and amacrine cells and making synapses upon bipolar and ganglion cells. Most commonly SP-IR amacrines emit axon-like process that pass to both the outer plexiform layer and the ganglion cell and nerve fiber layers. These processes often end upon the retinal vasculature. SP-IR ganglion cells have been described in turtle, rabbit and human retinas. In turtle, intracellular dye injection has revealed the morphology of one type of SP-IR ganglion cell as being a large-field monostratified cell with a branches in the outer stratum of the inner plexiform layer. It may correspond to a "Dogiel cell" type. Intracellular investigation of SP-IR amacrine cells in turtle reveal their physiological responses to be ON-OFF in nature with some color-coding characteristics. In general SP acts as an excitatory neurotransmitter raising the spontaneous activity level of ganglion cell responses. The SP-IR ganglion cell is an OFF-center unit in the turtle retina and may be driven in the center of its receptive field by luminosity bipolar cells and in its surround by amacrine cells with color-opponent properties.

Visual experience during postnatal development determines the size of optic nerve axons.

The size of any axon is generally related to an internal programme (probably common for all neurones) that determines the number and organization of its cytoskeletal components. Here we show that axons from visually deprived rats are smaller than controls, although the number of microtubules and neurofilaments remains unchanged. Moreover the distance between both microtubules and neurofilaments and the amounts of 200 kDa, 160 kDa and 68 kDa neurofilament proteins are also diminished in the deprived axons. We suggest that cytoskeletal organization and axonal calibre are not only determined by intrinsic (genomic) factors, but that environmental stimulation is important for normal growth of nerve cells.

A compiled BASIC program for analysis of spatial point patterns: application to retinal studies.

The pattern of distribution of a population of cells is of considerable interest to biologists and neurobiologists. However, the labor involved in collecting and analyzing the data often requires a significant amount of time. This paper presents a compiled BASIC program written using the Microsoft QuickBasic compiler for Apple Macintosh to facilitate such studies. The program allows collection and analysis of data that can be introduced either with the aid of a digitizing tablet of directly imported as x,y coordinates from different sources as, for example, word processors or image analysis software. Subsequently the program provides a quick, easy and interactive way of access to statistical, mathematical and graphical techniques used in the analysis of spatial point patterns. These techniques include several measures of dispersion (quadrat count, nearest neighbor and a 2-dimensional point autocorrelogram analysis) and arrangement. Although the program has been tested on spatial organization of retinal cells, it can be used to study the distribution of other cells in the nervous system and for different projects, as for example the distribution of microtubules and neurofilaments inside the axons. This software is available from the authors.

A useful programme in BASIC for axonal morphometry with introduction of new cytoskeletal parameters.

Interest in the structure of axons and quantification of their components has been growing over the last years. However, the existing literature contains few reports of available computer programmes to facilitate such studies. This paper presents a fully comprehensive BASIC programme for the morphometric analysis of electron micrographs of cross-sectional nerve fibres. From drawings of fibre and axonal contours and dots of the microtubules and neurofilaments, the programme calculates the following parameters: area, diameter and form factor of the fibres and axons, number and density of microtubules and neurofilaments, proportion between microtubules and neurofilaments (R-proportion), myelin thickness and the diameter of the axon relative to its sheath (g-ratio). The programme also introduces three new parameters to analyse the degree of uniformity of microtubule and neurofilament distribution: distances between microtubules and between neurofilaments, equilateral index and cytoskeletal intermingling index. The programme is written in Microsoft BASIC Interpreter for Apple Macintosh (Microsoft Corporation) but can be used on other computers. Although the programme has been tested on adult rat optic nerve fibres, it can be used for different projects concerning axonal morphometry.

Choline acetyltransferase is found in terminals of horizontal cells that label with GABA, nitric oxide synthase and calcium binding proteins in the turtle retina.

In this study, we discriminated the various types of horizontal cell in the turtle retina on their content of neuroactive substances. Double label immunocytochemistry was performed on sectioned and wholemount retina using antisera to neural- and endothelial-nitric oxide synthase (nNOS, and eNOS), calretinin (CR), calbindin (CB), gamma-aminobutyric acid (GABA) and choline acetyltransferase (ChAT). H1 cells and their axon terminals label with CR, CB and GABA. Only H1 axon terminals label with eNOS. H2 cells contain CB, CR, nNOS and GABA maybe in their dendrites. H3 cells label only with nNOS. The localization of nNOS in the H2 and H3 cells is a novel finding. None of these antibodies labels H4 cells. The photoreceptor subtypes have been differentiated by different intensity of labeling with CB. The accessory member of the double cone is less intensely labeled with CB than the principal member and rods and blue cones do not appear to label at all. ChAT-IR is located in terminal boutons of H1 and H2 horizontal cells and H1 axon terminals and these boutons contact rods and all spectral types of cones. Clearly, GABA is present in H1 horizontal cells and may be used in neurotransmission between horizontal cells and possibly for feedback pathways to photoreceptors. The evidence of nNOS immunoreactivity in H2 and H3 horizontal cells, combined with available physiological evidence, suggests that NO may be involved in electrical coupling and/or modulation of synaptic input to these types of cells. Furthermore, our results raise the possibility that cholinergic synaptic transmission may occur from horizontal cell processes to photoreceptors in the outer plexiform layer of the turtle retina.

Distribution of immunoreactivity to protein kinase C in the turtle retina.

Immunocytochemical staining procedures using the HRP-complexed antibody to protein kinase C (PKC) have been carried out on the turtle retina. Wholemounts and frozen sections of retina have been studied by light microscopy to evaluate PKC immunoreactivity after stimulation of the retina with light and neurotransmitters known to be active in the vertebrate retina. The most dramatically stained sites are cone synaptic pedicles and bipolar cells under all conditions. Ganglion cells stain weakly under certain conditions. Applying the antibody to a 'control' retina under dark adapted conditions results in uniform background staining of both hyperpolarizing and depolarizing bipolar pathways, while stimulating the retina with K+ under dim light conditions results in discretely stained bipolar cells and a prominent band of staining in stratum 4 of the inner plexiform layer. Stronger stimulation of bipolar cells with their terminals contributing to strata 3 and 4 and the continuous dominant band in stratum 4 can be elicited with incubation of the retina in neurotransmitter agonists, GABA and dopamine. Incubation with dopamine, in particular, brings out the putative dopaminergic amacrine cell. The only condition in which a strong band in stratum 2 can be demonstrated is under stimulation with a flashing bar of spot of light. Thus K+ and neurotransmitter stimulation elicit PKC staining in neurons contributing to the ON or depolarizing sublamina of the IPL, while intermittent flashing light stimulus is required to elicit PKC staining in the OFF or hyperpolarizing sublamina of the IPL.

Formation and dissolution of spinules and changes in nematosome size require optic nerve integrity in black bass (Micropterus salmoides) retina.

Teleost retinas adapted to light show numerous spinules invaginated in the cone pedicles and small nematosomes in the distal horizontal cells. Darkness induces the dissolution of spinules and the presence of large and numerous nematosomes. The aim of this work is to study the influence of optic nerve integrity on spinule formation/dissolution and changes in nematosome size during light or dark adaptation of black bass (Micropterus salmoides) retinas. Eyes from fish, dark- or light-adapted, were removed and the eyecups placed in oxygenated Ringer's solution and immediately exposed to light or dark, respectively, for 1 h. The number of spinules per pedicle and the nematosome diameter were measured on electron micrographs. Isolation of eyecups in the dark, impaired both spinule formation and nematosome size reduction when they were superfused in light. In the same way, isolation of eyecups in the light, impaired both spinule dissolution and nematosome size increase when they were superfused in dark. No significant differences in spinule number and nematosome size, following dopamine superfusion, were found in comparison to retinas superfused with Ringer's solution only. Our results suggest: (1) optic nerve integrity is necessary to yield spinule formation/disruption and changes in nematosome size during light or dark adaptation. (2) dopamine does not appear to be the primary agent responsible for spinule formation.

Axon types classified by morphometric and multivariate analysis in the rat optic nerve.

Calibers of the rat optic nerve axons distribute unimodally and it is difficult to distinguish groups among them. However, these fibers arose from 3 types of ganglion cells and showed 3 conduction velocities. Performing a cluster analysis over several ultrastructural parameters we found 3 main groups of fibers. These groups are present in a very similar proportion to the ganglion cells groups described in the rat retina.

Dendrites of rod dominant ON-bipolar cells are coupled by gap junctions in carp retina.

Gap junctions are supposed to be the anatomical substrate for electrical coupling between neurons. In fish retina, bipolar cells are electrically coupled and their receptive field diameters are always larger than dendritic field size, however, gap junctions have not been described between dendrites of bipolar cells. In this paper, using immunostaining for protein kinase C, we show that every rod dominant ON-bipolar cell is connected with its neighboring dendrites by gap junctions forming a plexus in the outer plexiform layer. These dendritic processes provide the site of electrical coupling. We suggest that dendrites of bipolar cells could be involved in lateral pathways in retina.