Population Of Bipolar Cells Research Articles

A population of wide-field bipolar cells in the rabbit's retina.

We have stained an unusual population of retinal bipolar cells. When the low molecular weight tracer biocytin was injected into the vitreous body of rabbits, it subsequently accumulated in the somata and processes of a population of wide-field bipolar cells. The cells have 2-4 primary dendrites. Their dendritic arbors span a field 50 to 200 microns in diameter. The axonal arbors are sparse and often highly asymmetric. The longest dimension of the axonal arbor ranges from 100 to 300 microns. The cells are moderately evenly spaced. They make up less than 1% of the total population of bipolar cells in the rabbit retina. With the whole population stained, regularities in the spatial arrangement of nearby cells can be recognized. Their dendrites often run to a common point, where they have the appearance of making contact with each other. A similar arrangement is seen for the cells' axonal arbors, so that the whole population is spatially linked in both the outer retina and the inner. The exact nature of the points of conjunction cannot be learned from light microscopy. One possibility is that the processes run together because they contact a common target. If so, the target structures (one in the outer retina and one in the inner) must be sparse. An alternative is that the points of conjunction represent synapses or gap junctions among wide-field bipolars of this type.

Immunocytochemical analysis of bipolar cells in the macaque monkey retina

Transfer of visual information from photoreceptors to ganglion cells within the retina is mediated by specialized groups of bipolar cells. At least 10 different morphological types of bipolar cells have been distinguished in Golgi studies of primate retina. In the present study, bipolar cell populations in the macaque monkey retina were identified by their differential immunoreactivity to a spectrum of antibody markers. This enabled their spatial density and photoreceptor connections to be analysed. An antibody against the beta isozyme of protein kinase C (PKCA beta) labelled many cone bipolar cells. Invaginating (presumed ON) cone bipolar cells and rod bipolar cells were preferentially labelled with a monoclonal antibody raised against rabbit olfactory bulb. Flat (presumed OFF) bipolar cells were labelled with an antiserum against the glutamate transporter protein (GLT-1). Different populations of diffuse cone bipolar cells, which contact 5-10 cones, could be distinguished. The GLT-1 antiserum preferentially labelled the flat diffuse bipolar cell type DB2 (Boycott and Wässle, 1991, Eur. J. Neurosci. 3:1069-1088) as well as flat midget bipolar cells. Antibodies to calbindin (CaBP D-28K) labelled the flat diffuse bipolar cell type DB3 and (possibly) the invaginating diffuse bipolar cell type DB5. An antibody against the alpha isozyme of PKC labelled an invaginating diffuse bipolar cell type (DB4) as well as rod bipolar cells. Comparison of the spatial density of cone bipolar cell populations with that of photoreceptors suggests that each bipolar cell class provides a complete coverage of the cone array (each cone is contacted by at least one member of every bipolar cell class). These results support the classification scheme of Boycott and Wässle (1991) by showing that different diffuse bipolar cell classes express different patterns of immunoreactivity, and they reinforce the view that different spatial and temporal components of the signal from the photoreceptor array are processed in parallel within the primate retina.

Voltage-dependent sodium channel alpha subunit immunoreactivity is expressed by distinct cell types of the cat and monkey retina.

Polyclonal (7493 and 7317) and monoclonal (mAb3) antibodies, generated to the alpha subunit of the voltage-gated sodium channel (alpha NaCh), were employed to assess the cell types containing alpha NaCh-like immunoreactivity in the mature cat and monkey retina. Immunoblot analyses of retinal proteins in the cat revealed that the polyclonal and monoclonal antibodies we employed labeled a band in the 260-kDa region which corresponds to the molecular mass of the alpha subunit of the NaCh. In both the cat and monkey, these antibodies immunolabeled several distinct types of retinal cells. With the polyclonal antibodies immunoreactivity was observed in ganglion cells and their intraretinal axons, in horizontal cells, and unexpectedly, in cones. In addition, in both species, a limited number of heavily labeled profiles, presumed to be bipolar cells, were seen in the inner nuclear layer. In cat and monkey the monoclonal antibody labeled axons in the fiber layer, ganglion cell somata, and a continuous band of immunoreactive cell bodies (presumed bipolar cells) situated in the outer half of the inner nuclear layer. By immunolabeling isolated cells dissociated from the cat retina, it was possible to demonstrate unequivocally that a population of bipolar cells was labeled by the monoclonal and the polyclonal antibodies we employed. The differences in the labeling observed with the monoclonal antibody as compared to the polyclonal antibodies were interpreted as reflecting the presence of different alpha-subunit subtypes in the mammalian retina. Collectively, our findings suggest that alpha NaCh-like proteins are expressed by a more diverse population of retinal cells than expected on the basis of previous physiological and immunohistochemical studies.

Immunocytochemical characterization and spatial distribution of midget bipolar cells in the macaque monkey retina

Midget bipolar cells form the first distinct step in the parvocellular ( P)-pathway of the primate visual system, and are the major determinant of the receptive field properties of colour selective midget ganglion cells. This paper describes the sampling properties of the midget bipolar cell population and relates this to the processing of chromatic information in the P-pathway. Immunocytochemical markers were used to label midget bipolar cells so that their spatial density could be compared with that of cones and ganglion cells. Sections through macaque monkey retinae were immunostained with antibodies against cholecystokinin (CCK), and recoverin. In CCK-labelled sections, in addition to blue cone bipolar cells, numerous thin bipolar cell dendrites, which could be associated with individual cone pedicles are stained. CCK-immunoreactive midget bipolar cells are found throughout the retina. A different population of midget bipolar cells is revealed in recoverin-labelled sections. Based on a comparison with midget bipolar cells in Golgi-stained retinae we propose that ON-midget (invaginating) bipolars are imntunoreactive for CCK and confirm that OFF-midget (flat) bipolar cells are immunoreactive for recoverin [Milam, Dacey and Dizhoor (1993) Visual Neuroscience, 10, 1–12). The density of recoverin labelled midget bipolars matches the cone density to an eccentricity of about 10 mm; from there outwards it drops to 60% of the cone density. This suggests convergence of several cones to individual midget bipolar cells in peripheral retina. We conclude that midget bipolar cells are present throughout the entire primate retina, and could, in peripheral as well as in central retina, provide chromatically specific input to the P-pathway.

Bipolar cells of the chick retina containing alpha-bungarotoxin-sensitive nicotinic acetylcholine receptors.

Two cDNA clones for nicotinic acetylcholine receptor (nAChR) subunits sensitive to alpha-bungarotoxin (alpha-Bgt) have been isolated, the so-called alpha-Bgt binding proteins alpha 1 (or alpha 7 nAChR subunit) and alpha 2 (or alpha 8 nAChR subunit). Immunohistochemical experiments have shown that both alpha 7 and alpha 8 subunits, as well as subunits insensitive to alpha-Bgt (beta 2 and alpha 3), are present in amacrine and ganglion cells of the chick retina. However, only the alpha 8 subunit was observed in presumptive bipolar cells. The present study investigated in detail the pattern of distribution of the bipolar cells containing the alpha 8 nAChR subunit and its relation to the pattern of distribution of bipolar cells immunoreactive to protein kinase C (PKC). Presumptive alpha 8- and PKC-like immunoreactive (alpha 8-LI and PKC-LI) bipolar cells were observed sending their dendrites to the outer plexiform layers and their axons to the inner plexiform layer. Whereas alpha 8-LI bipolar cells corresponded to 40-53% of the whole population of bipolar cells, PKC-LI bipolar cells represented only 6-8% of the same population. The soma sizes of the alpha 8-LI bipolar cells were slightly smaller (mean +/- S.D.; 4.9 +/- 0.8 microns) than the soma sizes of the PKC-LI bipolar cells (5.4 +/- 0.9 microns). Double-labeling experiments indicated that probably all PKC-LI bipolar cells also contain alpha 8-LI. This indicates that two distinct groups of cholinoceptive bipolar cells exist in the chick retina, one that contains PKC-LI, and another one that does not.

Recoverin immunoreactivity in mammalian cone bipolar cells.

Human, macaque monkey, and rat retinas were immunostained with a polyclonal antibody preparation against purified recoverin, a 23-kD calcium-binding protein isolated from bovine retina that localizes to rods and cones (Dizhoor et al., 1991). In addition to immunoreactive photoreceptors, we have identified subpopulations of recoverin-positive bipolar cells in all three species. Results from immunostaining with progressive dilutions of anti-recoverin and preadsorption of the antibody with a dilution series of purified recoverin showed that photoreceptors and bipolar cells had similar affinities for the antibody and suggested that the molecule recognized by the antibody in both cell types is recoverin. Immunoreactivity for recoverin and protein kinase C, a selective marker for all rod bipolar cells, was found in separate bipolar cell populations. Recoverin immunoreactivity is therefore a characteristic of certain cone bipolar cell types. In rat retina, anti-recoverin labeled two morphologically distinct subpopulations of cone bipolar cells whose axonal arbors stratified at different depths in the inner plexiform layer (IPL). The bipolar cells labeled with anti-recoverin did not correspond to those that were reactive for calbindin, another cone bipolar cell marker. Human and monkey retinas also had two populations of cone bipolar cells that were recoverin-positive. One population showed a distinct pattern of narrow bistratification at the outer border of the IPL and a regular mosaic arrangement of its axonal arbors, suggesting that the entire population of a single cone bipolar type was labeled. Cell density, dendritic morphology, and axonal-field size and stratification indicate that anti-recoverin selectively strains the flat midget (presumed OFF-center) cone bipolar cell type observed previously in Golgi preparations. By contrast the second bipolar cell population had axonal stratification in the inner half of the IPL and showed an unusual but consistent morphology and spatial distribution. Individual cells were intensely stained but were present at an extremely low density (approximately 2-5 cells/mm2). These cells had multibranched dendritic trees characteristic of the diffuse bipolar cell class, but very small axonal fields in the size range of the midget bipolar class. Neither of the two recoverin-positive bipolar cell types in monkey was labeled with anti-calbindin or anti-cholecystokinin. An antibody preparation against bovine pineal hydroxyindole-O-methyltransferase (HIOMT) labeled photoreceptors and bipolar cells that closely resembled the recoverin-positive bipolar cells in human and rat retinas.(ABSTRACT TRUNCATED AT 400 WORDS)

Immunocytochemical staining with antibodies against protein kinase C and its isozymes in the turtle retina.

An LM immunocytochemical study has investigated the patterns of staining in turtle retina with monoclonal antibodies to the alpha, beta and gamma isozymes of protein kinase C. The protein kinase C-gamma antibody reveals cells in the ganglion cell layer, occasional amacrine cells and faint banding in strata 2 and 4 of the inner plexiform layer. The protein kinase C-beta antibody stains primarily amacrine cells that have dendrites running in strata 2, in 4 close to the 3/4 border and on the 4/5 border of the inner plexiform layer. Protein kinase C-alpha immunoreactivity is seen in a population of bipolar cells. The latter are characterized by stained axon terminals in strata 3 and 4 of the inner plexiform layer. A type of amacrine cell, different from those seen with the other antibodies, is also immunoreactive to protein kinase C-alpha. EM immunocytochemistry (using a polyclonal antibody) reveals protein kinase C immunoreactivity in photoreceptor cells, bipolar cells, amacrine cells and ganglion cells. In photoreceptors protein kinase C immunoreactivity occurs as patchy staining associated with vesicles and the plasmalemma in pedicles and telodendria. Some varieties of bipolar cell display protein kinase C reaction product throughout the entire cell. Their dendrites contact photoreceptor pedicles at wide-cleft basal junctions and ribbon and non-ribbon related narrow cleft junctions. A few lateral elements per cone or rod pedicle are always protein kinase C-immunoreactive. Amacrine and ganglion cells typically show small clumps of protein kinase C immunoreactivity around vesicles and close to the postsynaptic membranes. Synaptic boutons of some varieties of amacrine cell stain more uniformly. Protein kinase C-immunoreactive bipolar cells are most commonly presynaptic in stratum 4 of the inner plexiform layer, while protein kinase C-immunoreactive amacrine cells are both pre- and postsynaptic throughout strata 1, 2, 3 and 4. Stratum 5 appears to be almost devoid of protein kinase C-immunoreactive neural profiles.

NADPH-diaphorase positive neurons in the retina of Bufo marinus: Selective staining of bipolar and amacrine cells.

Unfixed retinal tissues of adult Bufo marinus were reacted for NADPH-diaphorase histochemistry that resulted in selective staining of populations of bipolar and amacrine cells. The number of stained bipolar cells in the retina was estimated to be 233,600 +/- 38,900 (mean +/- S.D.). These cells were non-uniformly distributed across the retina with the highest density of 4,308 cells/mm2 along the visual streak decreasing to lowest density of 1,378 cells/mm2 at the dorsal and ventral poles of the retina. The stained bipolar cells represented a morphologically uniform population. The number of labelled amacrine cells in the retina amounted to 3,116 +/- 405 including 251 cells that were displaced into the ganglion cell layer. Labelled amacrine cells were also unevenly distributed; with no stained cells in the retinal centre from where the density increased steadily to 72 cells mm2 in the retinal periphery. NADPH-diaphorase positive amacrine cells were classified on the basis of their dendritic arborization pattern into three morphological types: 1) narrow-field cells both at the periphery and the centre of the retina; 2) wide-field amacrine cells, and 3) amacrines with eccentric medium-sized dendritic field. The results of this study provide evidence for the presence of NADPH-diaphorase containing neurons in the anuran retina. Furthermore, this is the first report on the selective staining of bipolar cells for NADPH-diaphorase in the vertebrate retina.

Patterns of glutamate immunoreactivity in the goldfish retina

Postembedding silver-intensified immunogold procedures reveal high levels of glutamate immunoreactivity in "vertical" elements of the goldfish retina: (1) Red-sensitive and green-sensitive cones display strong glutamate immunoreactivity, especially in their synaptic terminals, but blue-sensitive cones are poorly immunoreactive. (2) All type Mb (on-center) and Ma (off-center) mixed rod-cone bipolar cells and all identifiable cone bipolar cells are highly glutamate immunoreactive. We find no evidence for bipolar cells that lack glutamate immunoreactivity. (3) The majority of the somas in the ganglion cell layer and certain large cells of the amacrine cell layer resembling displaced ganglion cells are strongly glutamate immunoreactive. (4) Despite their high affinity symport of acidic amino acids, the endogenous levels of glutamate in Müller's cells are among the lowest in the retina. (5) GABAergic neurons possess intermediate levels of glutamate immunoreactivity. Quantitative immunocytochemistry coupled with digital image analysis allows estimates of intracellular glutamate levels. Photoreceptors and bipolar and ganglion cells contain from 1 to 10 mM glutamate. The bipolar and ganglion cell populations maintain high intracellular glutamate concentrations, averaging about 5 mM, whereas red-sensitive and green-sensitive cones apparently maintain lower levels. Importantly, photoreceptor glutamate levels are extremely volatile, and in vitro maintenance is required to preserve cone glutamate immunoreactivity in the goldfish. GABAergic horizontal and amacrine cells contain about 0.3-0.7 mM glutamate, which matches the values predicted from the Km of glutamic acid decarboxylase. Müller's cells and non-GABAergic amacrine cells contain less than 0.1 mM glutamate. Though Müller's cells are known to possess potent glutamate symport, they clearly possess equally potent mechanisms for maintaining low intracellular glutamate concentrations.

Single cell shape and population densities of indoleamine-accumulating and displaced bipolar cells in Reeves' turtle retina

Two types of bipolar cell in the Geoclemys reevesii retina were studied quantitatively by means of specific cell labelling with an indoleamine derivative (5,6-dihydroxytryptamine, 5,6-DHT), a nucleic acid stain (4,6-diamidino-2-phenylindole, DAPI) and Lucifer yellow CH. Indoleamine-accumulating (IA) bipolar cells were selectively labelled with 5,6-DHT applied intraocularly. After the cells accumulated 5,6-DHT, the indoleamine fluorescence was photoconverted to diaminobenzidine products to allow observation of morphological details. Close examination of many cells (cell number; n = 120) showed that the IA bipolar cells consist of a single morphological type whose axon collaterals ramify sublaminae 1, 4 and 5 respectively. This terminal branching pattern corresponds to cells that hyperpolarize when their receptive field centres are illuminated (Weiler 1981). The density of IA bipolar cells was highest in the visual streak (4130 cells mm-2) and lowest at the peripheral margin (1970 cells mm-2). By applying a small amount of DAPI to the eye, nuclei located in the most proximal row of the outer nuclear layer were labelled selectively. By using selective intracellular dye injection into DAPI-labelled cells under fluorescence microscope (Tauchi & Masland 1984, 1985), these cells were found to have Landolt's clubs and single descending axons. Dye injections into more than fifty DAPI-labelled somata showed that they belonged exclusively to displaced bipolar cells. These comprised at least two subtypes that differ in the ramification pattern of their axon terminals within the inner plexiform layer: one was monostratified, whereas the other was bistratified. The displaced bipolar cell density was as high as 9400 cells mm-2 in the central retina, falling to 2000 cells mm-2 in the superior margin. In vitro Lucifer labelling revealed that the overall bipolar cell density in the central retina was as high as 39,300 cells mm-2. Both the conventionally located and displaced bipolar cells were included in this population. About 11% of the total bipolar cell population consisted of IA bipolar cells. Assuming that one half of the conventionally located bipolar cells are the centre-hyperpolarizing type, IA bipolar cells represent approximately 28% of the total. As displaced bipolar cells represent almost one quarter of the total bipolar population, the dislocation of their somata stands out morphologically, inviting investigation of possible functional correlates.

Hydroxyindole- O-methyltransferase in rat retinal bipolar cells: persistence following photoreceptor destruction

The presence ofhydroxyindole- O-methyltransferase (HIOMT) activity and localization of HIOMT immunoreactivity was examined in albino rat retinas following photoreceptor destruction. Male Sprague-Dawley rats were exposed to high intensity flourescent light for 4 consecutive days, then placed on a 14:10 h light:dark cycle for two weeks to allow for phagocytic removal of damaged cells from the retina. Histologic examination revealed almost complete destruction and removal of all photoreceptors. The damaged retinas exhibited an increase in HIOMT activity relative to controls, when expressed as activity per mg of protein. HIOMT activity in the pineal glands was not affected. When control and light damaged retinas were examined for HIOMT localization by immunocytochemistry, the control retinas displayed intense HIOMT immunoreactivity in all photoreceptors, and a somewhat lighter labeling in a population of bipolar cells, whereas the light damaged retinas (lacking photoreceptors) showed intense HIOMT immunoreactivity in bipolar cells. These results suggest that the increase in HIOMT activity following photoreceptor destruction is due to increased synthesis of this enzyme in a population of bipolar cells. Thse HIOMT-immunoreactive bipolar cells may perhaps respond in a compensatory manner to changing levels of melatonin in the retina.

HIOMT-like immunoreactivity in the vertebrate retina: A species comparison

Localization of the melatonin-synthesizing enzyme, hydroxyindole-O-methyltransferase (HIOMT)-like immunoreactivity was examined by light and electron microscopic immunocytochemistry, in the retinas of several species that have been used as animal models to study the retinal melatonin system. HIOMT-like immunoreactivity was observed in the retinal photoreceptors of rabbit, pigmented rat, guinea-pig, chick, goldfish. African clawed toad, and leopard frog. Additionally, most species displayed HIOMT immunoreactivity in a population of bipolar cells in the inner nuclear layer. At the ultrastructural level, HIOMT-like immunoreactivity was localized to the cytoplasm of rod and cone photoreceptors, and a population of cone bipolar cells. These observations are identical to earlier observations in the human retina. The similar pattern of HIOMT-like immunoreactivity among species suggests a phylogenetic conservation of the melatonin-synthesizing capability of retinal photoreceptors and some bipolar cells.

Gamma-aminobutyrate type B receptor modulation of L-type calcium channel current at bipolar cell terminals in the retina of the tiger salamander.

Bipolar-cell axon terminals receive direct synaptic input from amacrine-cell processes, suggesting a possible pathway for modulation of transmitter release. In retinal slices, bath-applied baclofen, a gamma-aminobutyrate type B (GABAB) receptor agonist, reduced a patch-clamp-recorded L-type calcium channel current in a population of bipolar cells with axon terminals that ramify along the midline of the inner plexiform layer. Lucifer yellow staining revealed that this current was found only in bipolar cells that retain axon terminals and their associated telodendria, suggesting that the current is generated at the terminal and also possibly modulated there. T-type calcium currents were found in all bipolar cells, including those without axon terminals, but were not modulated by baclofen. The baclofen-induced reduction of calcium current was enhanced by guanosine 5'-[gamma-thio]triphosphate and eliminated by guanosine 5'-[beta-thio]diphosphate added to the cytoplasm by the patch recording electrode, suggesting that the GABAB receptors act through a guanine nucleotide-binding regulatory protein (G protein). Baclofen also reduced an excitatory synaptic input to a population of amacrine cells with processes that ramify along the midline of the inner plexiform layer--cells probably postsynaptic to the bipolar terminals. This suggests that GABAB receptors modulate not only the calcium current but also transmitter release by a pathway involving G proteins and L-type calcium channels.

Enigmatic bipolar cell of rat visual cortex

Our earlier Golgi-electron microscopic study of bipolar cells in the rat visual cortex showed the axons of these neurons as forming asymmetric synapses (Peters and Kimerer; J. Neurocytol, 10:921-946, '81) in which the most common postsynaptic elements were dendritic spines. This result was unexpected, since Parnavelas (Parnavelas, Sullivan, Lieberman, and Webster: Cell Tissue Res. 183:499-517, '77) had earlier shown a bipolar cell from the same cortex to have an axon forming symmetric synapses with dendritic shafts. Here then was an enigma, strengthened by examination of neuronal components labelled by antibodies to two compounds in particular--namely, vasoactive intestinal polypeptide (VIP) and choline acetyltransferase (ChAT). Antibodies to these compounds preferentially label bipolar cells in the rat cerebral cortex, and the labelled axon terminals form symmetric synapses. Against this background the present study was performed, and it has been shown that the resolution to the enigma is that there are two different populations of bipolar cells in the rat visual cortex. Thus some Golgi-impregnated bipolar cells examined by electron microscopy after gold toning have been found to possess axons forming asymmetric synapses, and others have been found to have axons forming symmetric synapses. The axons of the bipolar cells forming asymmetric synapses most commonly synapse with dendritic spines (67%), although other terminals synapse with dendritic shafts (33%). In contrast, the bipolar cells with axons forming symmetric synapses preferentially synapse with dendritic shafts (100%). The population of bipolar cells that form symmetric synapses includes the ones that label with antibodies to vasoactive intestinal polypeptide (VIP), for the axons of VIP-labelled bipolar cells have been traced to labelled terminals forming symmetric synapses. However, examination of the population of VIP-labelled axon terminals shows that in addition to dendritic shafts, some of the labelled terminals synapse with the cell bodies of pyramidal and nonpyramidal cells. This includes bipolar cells, some of which receive large numbers of VIP-labelled axon terminals. It is also shown that some VIP-positive bipolar cells have myelinated axons. Analysis of tissue labelled with VIP antibody reveals that about 50% of the total population of bipolar cells in the rat visual cortex is VIP positive. These results are discussed in the light of information about labelling of bipolar cells with antibodies to gamma-aminobutyric acid (GABA) and to other peptides, and it is suggested that most VIP-positive bipolar cells also contain GABA.

GABA immunoreactive neurons in rat visual cortex.

An antiserum to gamma-aminobutyric acid (GABA) was used in a light and electron microscopic immunocytochemical study to determine the morphology and distribution of GABA-containing neurons in the rat visual cortex and to ascertain whether all classes of nonpyramidal neurons in this cortex are GABAergic. The visual cortex used for light microscopy was prepared in such a way that the antibody penetrated completely through tissue sections, and in these sections large numbers of GABA immunoreactive neurons were apparent. The labeled neurons could be identified as being either multipolar, bitufted, bipolar, or horizontal neurons. In layers II through VIa, GABA immunostained cells were distributed uniformly and accounted for approximately 15% of all neurons, but in layer I all neurons appeared to be immunostained. Electron microscopy of GABA immunostained visual cortex prepared to ensure good fine structural preservation confirmed the presence in layers II through VIa of numerous immunoreactive bipolar neurons, both small and large varieties, as well as multipolar and bitufted neurons. Additionally, electron microscopy reveals that astrocytes are frequently GABA immunoreactive. From a correlated light and electron microscopic evaluation of neurons in GABA immunostained visual cortex, it was possible to confirm which kinds of neurons are GABAergic and what proportion of the neuronal population they represent. Thus, from an analysis of some 950 neurons, it was found that pyramidal neurons were never immunoreactive and that except for 20% of the bipolar cell population, all examples of other types of nonpyramidal neurons encountered in this material were GABA immunoreactive.

Morphometric analysis of serotoninergic bipolar cells in the retina and its implications for retinal image processing.

The entire population of retinal serotoninergic bipolar cells in the turtle Pseudemys scripta elegans was labeled by immunocytochemical methods. This allowed a systematic analysis to be made of the morphological variabilities among a functionally homogeneous neuronal population. The analyzed morphological characteristics included: size of the Landolt club, size of the soma, lateral extension of the ramification within the outer plexiform layer, course of the axon across the inner nuclear layer, pattern of the axonal ramification within the inner plexiform layer, lateral extension of these ramifications, and density of the cells. Whereas characteristics 1-4 and 7 show a morphological variability strictly related to the location of the bipolar cell with respect to the visual streak, a fovealike structure, characteristics 5 and 6 show no such correlation. The size of the soma increases by a factor of 4 from the visual streak toward the periphery. The area covered by the ramification in the OPL increases from 330 micron 2 at the visual streak to 50,000 microns 2 at the dorsal and ventral edges of the retina. The coverage factor remains the same throughout the retina, as well as the area covered by the ramifications in the IPL, which is about 2,000 microns 2. At the visual streak and at 110 degrees ventral and 68 degrees dorsal of the visual streak, the bipolar axons cross the INL perpendicularly. In between the axons take an oblique course leading to an axon-induced shift between input and output of up to 250 microns.(ABSTRACT TRUNCATED AT 250 WORDS)

Localization of hydroxyindole-O-methyltransferase-like immunoreactivity in photoreceptors and cone bipolar cells in the human retina: a light and electron microscope study.

The localization of the melatonin-synthesizing enzyme hydroxyindole-O-methyltransferase (HIOMT) was examined by light and electron microscopic immunocytochemistry in the human retina. HIOMT-like immunoreactivity was observed in the photoreceptor layers and the inner nuclear layer (INL). The immunoreactive cells in the INL were more numerous in the central retina than in the peripheral retina and sent processes to both the outer plexiform and inner plexiform layers. The HIOMT immunoreactivity in the inner plexiform layer (IPL) appeared as punctate terminals in the proximal and distal one-thirds of that layer. At the ultrastructural level, HIOMT-like immunoreactivity was localized to the cytoplasm of rod and cone photoreceptors and to a population of cone bipolar cells. HIOMT-immunoreactive bipolar cell dendrites were observed to make both invaginating and flat synaptic contacts with cone pedicles. No immunoreactive invaginating contacts in rod spherules were observed. HIOMT immunoreactivity was observed in the bipolar cell cytoplasm in the INL, and in the bipolar synaptic terminals in the IPL. These terminals contained synaptic ribbons, which formed synaptic contacts with unlabeled cells in the IPL. HIOMT radioenzymatic assays confirmed the presence of HIOMT in the human retina. Average HIOMT activity of eight donors was determined to be 15.0 pmol/mg protein/hour +/- 7.2 S.D. The ultrastructural localization of HIOMT observed in this study, combined with reports from other laboratories, suggests that the cytoplasm of the photoreceptors and a population of cone bipolar cells may be the sites of melatonin synthesis in the human retina.

The optic tectum of gymnotiform teleosts Eigenmannia virescens and Apteronotus leptorhynchus: A golgi study

Golgi, Nissl, Biclschowsky and cholinesterase techniques have been used to analyze the optic tectum of the weakly electric teleost fish Eigenmannia virescens and Apteronotus leptorhynchus. Six layers are readily distinguished: a fairly thick stratum marginale, a narrow stratum opticum and stratum fibrosum et griseum superficiale, a well-developed stratum griseum centrale, a stratum album centrale and a compact stratum periventriculare. Fifty-six neuronal types are present. In regard to comparative aspects of tectal organization, it became apparent that although most neuronal types are similar to those reported in other teleostean fish, there are certain obvious differences such as: (i) pyramidal cell somata not confined to stratum fibrosum et griseum superficiale, but also clustered in the adjacent stratum opticum, presenting stratified or diffuse basilar dendritic arbors; and a change from vertical to oblique and almost horizontal neuronal orientation in the ventral and caudal tectum. (ii) The presence of pyramidal cells with aligned and misaligned apical and basal dendritic fields, (iii) A cell of stratum griseum centrale with an ascending axon to stratum opticum. (iv) A special projection type of fusiform cell of stratum griseum centrale, with an efferent axon of somatic origin. (v) A cell rich stratum griseum centrale, with a wider variety of multipolar and bipolar cell population than reported in other teleosts. (vi) Fourteen types of pyriform cells are present, four of which are efferent. (vii) Our observations are suggestive of regional differences in regard to the caudalmost tectum in Apteronotus: presumably this is related to the extremely sparse retinal input to this part of the tectum. A close functional correlation has been found between some multipolar and pyriform cells identified in our material with similar cells reported by Rose and Heiligenberg as multisensory cells, following recordings and horseradish peroxidase fillings of these cells. Based on the observation of patchy torus semicircularis input to stratum fibrosum et griseum superficiale, disjunct from the retinal input to this layer, it is proposed that perhaps this arrangement is the result of competition for synaptic targets during development.

Catecholamine- and indoleamine-containing neurons in the turtle retina.

We identified a population of presumed dopaminergic amacrine cells and populations of presumed serotonergic bipolar and amacrine cells in the retina of the turtle Pseudemys scripta elegans by a combination of autoradiographic, fluorescence, and immunocytochemical techniques. Antisera directed against the dopamine-synthesizing enzyme, tyrosine hydroxylase (TOH), stained perikarya located at the border of inner nuclear (INL) and inner plexiform (IPL) layers. Processes emitted by these cells arborized in sublaminae 1, 3, and 5 of the IPL. Incubation of retinas in 10(-6) M 3H-dopamine yielded a labeling pattern identical to the staining pattern achieved with TOH antisera, but when the concentration of 3H-dopamine was increased 25-fold, both amacrine and bipolar cells are labeled. Following intraocular injection of dopamine, fluorescence micrography revealed both stained amacrine and bipolar cells. The bipolar cells had Landolt's clubs, pyriform perikarya located in the distal portion of the INL, and axons that coursed horizontally in the INL, then entered the IPL, and ramified in both its superficial and deeper layers. Although no fluorescent neuronal profiles were revealed following injection of serotonin (5HT), bipolar cells identical to those described were visualized with 5HT antisera. The intensity of bipolar cell staining with 5HT antisera was improved by preinjection of the eye with exogenous 5HT. We suggest that the bipolar cell is serotonergic, but that it also can actively accumulate dopamine. The 5HT antisera also stained a population of large amacrine cells whose processes ramified in IPL sublaminae 1, 4, and 5. The same populations of presumed serotonergic bipolar and amacrine cells were labeled following incubation of the eyecup in 10(-6) M 3H-5HT.

Vasoactive intestinal polypeptide-immunoreactive neurons in rat visual cortex

An antibody to vasoactive intestinal polypeptide (VIP) was used to examine the forms of VIP-positive neurons and the synapses made by VIP-positive axon terminals. Vasoactive intestinal polypeptide-positive cells are most common in layers II and III and the majority of them are typical bipolar neurons, with two primary dendrites which emanate from the upper and lower poles of the cell body. Their somata, which have only a few symmetric and asymmetric synapses, generally have a fusiform or “tear-drop” shape and contain nuclei with a vertically oriented cleft. The dendritic trees are arranged vertically and often extend through five cortical layers. The axons are thin and extend either from the soma or from one of the primary dendrites. The axons also follow a vertical trajectory. Other VIP-positive neurons are modified bipolar cells and a few of them are multipolar cells. The synapses formed by the VIP-positive axon terminals in the neuropil are symmetric in form, and although the synaptic clefts are narrow, the junctions are usually long and continuous, rather like those described for asymmetric synapses. Most of the VIP-positive axon terminals synpase with small dendritic shafts, but a few synapse with neuronal cell bodies. Since the majority of the VIP-positive neurons are bipolar cells it is concluded that these are the source of most of the VIP-positive axon terminals. If this is so, then the VIP-positive bipolar cells form symmetric synapses. This is in contrast to the observations of Peters and Kimerer (1981. J. Neurocytol. 10, 921–946) for the bipolar cells they examined in a Golgi-electron microscopic study had axon terminals forming asymmetric synapses. It is suggested that this disparity can be reconciled if it is assumed that the bipolar cell population consists of subgroups which have different biochemical characteristics and different synaptic relationships.