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Contribution of connexin26 to electrical feedback inhibition in the turtle retina.

The first synaptic integration in the neuronal cascade of vision in vertebrates includes a feedback from horizontal cells to cones by a mechanism yet not fully understood. Recent observations in teleosts suggested an electrical feedback mechanism mediated by connexin26 (Cx26) hemichannels at the terminal dendrites of horizontal cells. By using reverse transcription-polymerase chain reaction and immunoblotting from retinal homogenate, we detected Cx26 mRNA transcripts in the turtle retina and demonstrated that they were translated into protein. Cx26 immunoreactivity was especially prominent in the outer plexiform layer. Subcellularly, immunoreactivity was located mainly between horizontal cell axon terminals and in horizontal cell dendrites forming the lateral elements at the ribbon synaptic complex of the cone pedicle. The label was confined to the horizontal cell membrane flanking the ribbon and was not found on the opposing photoreceptor membrane. No gap junctions at this location are known, so immunosignaling suggested the presence of hemichannels. Their relevance to the feedback mechanism was investigated by intracellular recordings from horizontal cells during application of the hemichannel blocker carbenoxolone. Carbenoxolone hyperpolarized the dark membrane potential by about 25 mV, decreased the amplitudes of responses to full-field light flashes, and suppressed the feedback-induced depolarizing inflexion in the response profile. These physiological findings are compatible with the involvement of hemichannels in the feedback between horizontal cells and cones and support the anatomical findings. Together, these data suggest the presence of an electrical feedback mechanism in the turtle retina, which therefore might be a common mechanism at the first visual synapse in vertebrates.

Animals↗

Pretectal and tectal afferents to the dorsal lateral geniculate nucleus of the turtle: an electron microscopic axon tracing and gamma-aminobutyric acid immunocytochemical study.

The pretectal and tectal projections to the dorsal lateral geniculate nucleus (GLd) of two species of turtle (Emys orbicularis and Testudo horsfieldi) were examined under the electron microscope by using axonal tracing techniques (horseradish peroxidase or biotinylated dextran amine) and postembedding gamma-aminobutyric acid (GABA) immunocytochemistry. After injection of tracer into the pretectum, two types of axon terminals were identified as those of pretectogeniculate pathways. Both contained pleomorphic synaptic vesicles and were more numerous in the inner part of the nucleus. They could be distinguished on the bases of size and shape of their synaptic vesicles, type of synaptic contact, and level of GABA immunoreactivity. One type had a higher density of immunolabeling and established symmetric synaptic contacts, whereas the other, less densely immunolabeled, made asymmetric synaptic contacts. In both cases, synaptic contacts were mainly with relay cells and occasionally with interneurons. We suggest that these two types of pretectogeniculate terminals originate in two separate pretectal nuclei. After injection of tracer into the optic tectum, a single population of GABA-immunonegative tracer-labeled terminals was identified as belonging to the tectogeniculate pathway. These were small, had smooth contours, contained very small round synaptic vesicles, and established asymmetric synaptic contacts with long active zones, predominantly with relay cells and less frequently with interneurons, in the inner part of the nucleus. In addition, a population of GABA-negative and occasionally GABA-positive terminals, labeled by tracer injected into either the pretectum or the tectum, was identified as retinal terminals; these were presumably labeled by the retrograde transport of tracer in collateral branches of visual fibers innervating both the GLd and the pretectum or tectum. Comparison of the present ultrastructural findings in turtles with those previously reported in mammals shows that the cytological features, synaptic morphology, and immunochemical properties of the pretectogeniculate and tectogeniculate terminals of both groups share many similarities. Nevertheless, the postsynaptic targets of these two categories of terminals display some pronounced differences between the two groups, which are discussed in terms of their possible functional significance.

Afferent Pathways↗

Synaptic patterns in the visual cortex of turtle: an electron microscopic study.

The part of turtle general cortex that receives afferent fibers from the dorsal lateral geniculate nucleus and that shows evoked potentials to light stimuli has been studied with the electron microscope. This cortex consists of an outer molecular layer, a perikaryal layer, and a subcellular layer lying on a row of ependymal cell bodies. Neurons in the perikaral lamina are characterized by long spine-bearing apical dendrites ascending through the outer molecular layer and short finer basal dendrites in the subcellular zone. Scattered neurons without apical dendrites occur in both the molecular and subcellular zones. Two types of dendritic spines can be distinguished. Some are large, have a complex irregular shape, contain a variety of membranous sacs and mitochondria, and occasionally, a single bundle of microtubules embedded in an electron-dense background [corrected] opacity. These large spines are the most common postsynaptic element in the outer third of the molecular layer, where they are located on the distal enlargement that contains only electron-dense fuzz. They are the most common post-synaptic element in the lower two-thirds of the molecular layer where they arise from the proximal portion of apical dendrites. Most synaptic contacts are found on the dendritic spines and are of the "round-asymmetrical" type. Not infrequently "flat-symmetrical" synapses are coupled to "round-asymmetrical" contacts on individual large spines. The few contacts present on spine-bearing dendritic shafts are of both types. Axo-somatic contacts are mainly of the "flat-symmetrical" variety. Thus the synaptic patterns on the principal cells of turtle visual cortex are remarkably similar to those found on pyramidal cells of mammalian neocortex. In addition, however, axon terminals, dendrites and glial (ependymal) processes were often seen to give rise to membranous pouches containing large vacuoles and invaginating into dendritic shafts or spines. Rarely, axon terminals were seen to form contacts, identical in appearance to synaptic contacts, on cell bodies in the ependymal lining. More frequently, unusual types of membrane differentiations were present at the site of apposition of the membranes of axon terminals and ependymal processes. They are interpreted as functional neuroependymal contacts.

Animals↗

A histochemical study of the innervation of cerebral blood vessels in the turtle.

Specific histochemical techniques for the demonstration of noradrenaline and of acetylcholinesterase have been used to study the distribution of adrenergic and cholinergic nerves to the cerebral blood vessels of turtle, Geoclemys reevesii. The major and medium-sized cerebral arteries were supplied with dense adrenergic nerve plexuses, the plexuses were particularly dense in the medium-sized pial arteries of very thick vascular wall, indicating the functional significance of these arteries in the cerebral circulation. The parenchymal arterioles and capillaries were also supplied with adrenergic nerves. On the other hand, the cholinergic innervation was less dense than the adrenergic one and the acetylcholinesterase activity of the nerve fibres was remarkably weak. By contrast, the parenchymal small arterioles and capillaries exhibited heavy acetylcholinesterase activity on the vascular wall and, in addition, the capillaries were supplied with the well-stained acetylcholinesterase-positive nerve fibres. These fibres and also the adrenergic fibres associating with the capillaries appear to be of central origin. It is suggested that the cholinergic mechanisms in the parenchymal small vessels also play an important role in the cerebral circulation. The basophil leucocytes observed abundantly in the blood of turtle emitted an intensive greenish yellow fluorescence after formaldehyde gas-treatment.

Acetylcholinesterase↗

Connections of the small bipolar cells with the photoreceptors in the turtle. An electron microscope study of Golgi-impregnated, gold-toned retinas.

The connections between photoreceptors and the small bipolar cells with a Landolt club in the turtle retina (Pseudemys scripta elegans) were analyzed with the electron microscope after Golgi impregnation and gold toning. The dendritic tree of several bipolars was partially or completely reconstructed from continuous series of thin sections. All bipolar cells studied established both basal and invaginating junctions with cone pedicles; in invaginating junctions, their dendrites invariably ended as the central process of triads. Dendrites of this bipolar cell type make invaginating synapses, basal junctions, or a mixture of both. In turn, each pedicle found within the dendritic field of the bipolar cell contacts it with a mixture of invaginating and basal junctions; furthermore, each pedicle synapses with more than one dendrite from the same bipolar cell. All small bipolar cells examined also synapse with rod pedicles; the geometry of the connections and the types of synapses are the same for both rods and cones. The fact that small turtle bipolars can form both basal and invaginating contacts with a single cone indicates that the polarity of the postsynaptic response can not be predicted on the basis of the structure of the photoreceptor-to-bipolar cell synapse.

Animals↗

Quantitative electron microscopic analysis of the optic nerve of the turtle, Pseudemys.

It is estimated by means of electron microscopy that the optic nerve of the turtle Pseudemys scripta elegans contains 394,900 fibers of which approximately 80% are myelinated. The total fiber count agrees well with counts obtained from electron microscopic studies on other turtle species. There are, however, differences among these species in the percentage of myelinated fibers in the optic nerve. The axon diameter distribution of the myelinated fibers (excluding myelin) has a mode at 0.87 micrometer while that of the unmyelinated fibers has a mode at 0.42 micrometer. Both distributions are unimodal and are of a similar form in all areas of the nerve sampled. The total fiber count reported here agrees well with previous reports of ganglion cell counts in Pseudemys and the characteristics of the fiber distributions are comparable to those reported for nonreptilian vertebrates.

Animals↗

Cerebellar corticonuclear projections in the red-eared turtle Pseudemys scripta elegans.

In the present study the organization of the corticonuclear projections of the cerebellum in the red-eared turtle Pseudemys scripta elegans was investigated. To find out whether a zonal pattern exists in the cerebellar cortex, a topological analysis was made of the Purkinje cell layer of a Nissl-stained series of the cerebellum. This analysis showed a distinct, longitudinally oriented zonal pattern of Purkinje cells. In each cerebellar half a medial, an intermediate, and a lateral zone could be distinguished. This pattern appeared to correspond in part to an earlier subdivision of the reptilian cerebellum suggested by Larsell ('26, '32, '67). In the experimental part of this study, the corticonuclear projections to the cerebellar nuclei and the vestibular nuclear complex were demonstrated with the horseradish peroxidase (HRP) technique. All projections of the Purkinje cells appeared to be strictly ipsilateral. After HRP slow-release gel implantations at various levels of the vestibular nuclear complex, longitudinally oriented areas of labeled Purkinje cells were found in the lateral part of the medial zone, the intermediate zone, and the lateral zone. Two strips of Purkinje cells remained unlabeled: One in the rostrolateral part of the intermediate zone, and another in the medial part of the medial zone. After HRP gel implantations in the cerebellar peduncle aimed at the medial and lateral cerebellar nuclei, respectively. Purkinje cells in the latter two strips were labeled. The present results indicate that the corticonuclear projections to the medial cerebellar nucleus arise in the medial part of the medial zone and those to the lateral cerebellar nucleus in the rostrolateral part of the intermediate zone. It can be concluded that cerebellar zones as demonstrated in the turtle Pseudemys scripta elegans, are phylogenetically very old and may represent the basic functional circuit of the cerebellum.

Animals↗

The effects of lesions of telencephalic visual structures on visual discriminative performance in turtles (Chrysemys picta picta).

Ascending thalamotelencephalic visual pathways that terminate in specific telencephalic regions have been described in all reptiles studied. Although the anatomical data suggests that such telencephalic regions may play a role in visual processing in reptiles, few behavioral data are available. In the present study, the effects of destruction of either the core nucleus (CN) of the dorsal ventricular ridge (telencephalic terminus of the tectothalamofugal pathway) or the dorsal cortex (telencephalic terminus of the retinothalamofugal pathway) on visual discriminative performance in the turtle were examined. Following extensive bilateral destruction of the CN, turtles were severely impaired in their performance of both a simultaneous pattern discrimination and a simultaneous visual intensity discrimination. The extent of the discriminative impairment was found to be specifically correlated with the amount of CN damage. In contrast to the effects of CN lesions, lesions of the dorsal cortex had no evident effect on the performance of either a simultaneous pattern discrimination or a simultaneous visual intensity discrimination. The present results suggest that, as in birds and mammals, telencephalic visual areas play an important role in visual functions in reptiles. As in at least some birds (such as pigeons), the telencephalic terminus of the tectothalamofugal visual pathway appears to play a larger, or at least more readily measurable, role in visual discrimination than does the telencephalic terminus of the retinothalamofugal pathway.

Animals↗

Some afferent and efferent connections of the vestibular nuclear complex in the red-eared turtle Pseudemys scripta elegans.

In the present study some afferent, commissural, and efferent connections of the vestibular nuclear complex in the turtle Pseudemys scripta elegans were demonstrated with the HRP tracing technique. Afferent projections to the vestibular nuclei were found to arise in the nucleus of the basal optic root, the interstitial nucleus of the fasciculus longitudinalis medialis, the medial and lateral cerebellar nuclei, the perihypoglossal nuclear complex, and the reticular formation. Distinct commissural projections appeared to arise in the dorsolateral, ventromedial, and descending vestibular nuclei. The commissural projection arising in the ventrolateral vestibular nucleus appeared to be only sparsely developed. Both ascending and descending efferent projections were demonstrated to arise from the vestibular nuclear complex. The ascending vestibulo-oculomotor projection was found to be organized in an ipsilateral pathway arising in the dorsolateral vestibular nucleus and in a contralateral pathway, arising mainly in the medial vestibular nucleus. These projections appeared to be directed to the interstitial nucleus of the fasciculus longitudinalis medialis, the oculomotor, trochlear, and abducens nuclei. Also the perihypoglossal nuclear complex appeared to be an important target of vestibular efferents. The origin and course in the brainstem of the descending vestibular projections, i.e., the lateral and medial vestibulospinal tracts, as demonstrated in previous anatomical and experimental studies in reptiles, were confirmed. However, in addition a direct projection of the vestibulospinal tracts to presumably neck motoneurons was found. THe organization of the vestibular connections observed in the turtle Pseudemys scripta elegans appeared to be basically comparable to the organization of the vestibular connections in birds and mammals.

Animals↗

The projection of three extrathalamic cell groups to the cerebral cortex of the turtle Pseudemys.

Three extrathalamic subcortical inputs to the part of the cerebral cortex that is known to receive thalamic fibers in the turtle were examined in the present study. Direct projections from the locus coeruleus, the superior medial raphe nucleus, and a wide area of the basal telencephalon that lies ventromedial to the globus pallidus were demonstrated with the horseradish peroxidase method. Fluorescence histochemistry confirmed the presence of catecholamine-containing fibers in the rostral half of dorsal cortex and also demonstrated a dense network of serotoninergic fibers. Biochemical analysis showed the concentration of both monoamines to be relatively high; the norepinephrine concentration was 709 ng/g and the serotonin concentration was 1,750 ng/g. No evidence was found to suggest the existence of either a dopamine fiber projection to cortex comparable to that of mammalian neocortex or the presence of an epinephrine pathway to turtle cortex equivalent to the epinephrine-containing fibers in the pallium of amphibians. The coexistence of the projections from the thalamus with noradrenergic projections from the locus coeruleus, serotoninergic projections from the superior medial raphe nucleus, and presumably cholinergic projections from the basal telencephalon provide at least four distinct subcortical inputs to the reptilian dorsal cortex. Neither thalamic nor similar extrathalamic inputs have been demonstrated in the dorsal pallium of amphibia. Mammalian neocortex, in contrast, has even more elaborately differentiated inputs of both types. These results support the idea that thalamic and extrathalamic inputs to cortex appear at the same time in vertebrate evolution, and that both types of inputs are required for the normal development and function of neocortex.

Animals↗

Spectral sensitivities of seven morphological types of photoreceptors in the retina of the turtle, Geoclemys reevesii.

Spectral sensitivities of photoreceptors in the turtle (Geoclemys) retina were studied by intracellular recording, and each cell was filled with Lucifer yellow (LY). Photoreceptors were classified into seven morphological types: rod, four types of single cones, and two members of a double cone. Single cones contained one of four different oil droplets: red, pale-green, orange, and clear. Double cones consisted of two apposed cones; principal members contained yellow oil droplets, while accessory members contained no oil droplet. Spectral sensitivities recorded from these seven types of photoreceptors were classified into one type of rod and three chromatic types of cones. Rods (n = 19) showed peak sensitivity at 520 nm. Single cones containing either a red (n = 51) or a pale-green (n = 9) oil droplet were red-sensitive (lambda max at 620 nm). Single cones containing an orange oil droplet (n = 14) were green-sensitive (lambda max at 540 nm). Single cones containing a clear oil droplet (n = 3) were blue-sensitive (lambda max at 460 nm). Both members of the double cone, principal (n = 22) and accessory (n = 15), were red-sensitive (lambda max at 620 nm). No diffusion of LY was detected between the apposed members of double cones. Red-sensitive cones, therefore, consisted of four different morphological types of cones, and they occupy about 70% of the photoreceptor mosaic in the turtle retina.

Animals↗

Cone synapses with Golgi-stained bipolar cells that are morphologically similar to a center-hyperpolarizing and a center-depolarizing bipolar cell type in the turtle retina.

Two types of Golgi-stained bipolar cells have been examined by light and electron microscopy to determine the ultrastructure of their synapses with photoreceptors and the spectral type of photoreceptor with which they connect in the retina of the turtle (Pseudemys scripta elegans). We have chosen bipolar cells that correspond in morphology to a center-hyperpolarizing type (H-bipolar) and a center-depolarizing type (D-bipolar) shown in Marchiafava and Weiler's paper (Proc. R. Soc. B 208:103-113, '80). The latter authors recorded intracellular responses and marked their cells with the fluorescent dye Procion yellow so that we have a clear picture of their morphology. These bipolars have been called B4 and B6, respectively, according to our recent classification scheme of Golgi-stained cells. Serial section electron microscopy of two B4 bipolars shows that they made wide-cleft, striated basal junctions with red and green cone pedicles. They connected to six or seven different cone pedicles. The three B6 bipolar cells studied made narrow-cleft, semi-invaginating basal junctions with cone pedicles. The dendrites of B6 bipolars did not become central elements at ribbon synapses although they invaginated toward the synaptic ribbon. Serial section electron microscopy indicated that B6 bipolars contacted three to five cone pedicles also of a red or green cone type. We suggest that in the case of these two varieties of bipolar cell in turtle retina, sign-conserving synapses with cones are wide-cleft basal junctions while sign-inverting synapses are narrow-cleft, semi-invaginating in morphology.

Animals↗

Morphology of neurons in the dorsal lateral geniculate complex in turtles of the genera Pseudemys and Chrysemys.

The morphology of neurons in the dorsal lateral geniculate complex of pond turtles has been studied by extracellular filling with horseradish peroxidase. The dorsal lateral geniculate complex is a rostrocaudally elongate structure that includes the nucleus ovalis and dorsal lateral geniculate nucleus of Papez. It is divided into three cytoarchitecturally distinct subnuclei: the subnucleus ovalis, the dorsal subnucleus, and the ventral subnucleus. Each subnucleus consists of a neuropile immediately internal to the optic tract and a cell plate of densely packed somata forming the medial face of the complex. The cell plate contains medial (or parvicellular) and lateral (or magnocellular) sublaminae that are separated by a cell-poor zone in subnucleus ovalis and the ventral subnucleus. The density and size of somata vary between subnuclei. Geniculate neurons fall into two distinct groups. Cell plate neurons have somata in the cell plate and vary substantially in morphology. Those in the medial sublamina have fusiform somata and dendrites that extend either mediolaterally (subnucleus ovalis) or run rostrocaudally in the cell plate (dorsal and ventral subnuclei). Those in the lateral sublamina have some dendrites that extend medially within the cell plate and others that extend into the neuropile. The latter dendrites branch and bear arbors of fingerlike, varicose branchlets in the outer half of the neuropile. By contrast, neuropile cells have fusiform somata and dendrites extending concentrically with the optic tract in the neuropile. Both groups of geniculate neurons can be retrogradely labeled by horseradish peroxidase injections in the lateral forebrain bundle. These results lead to the recognition of two principles of geniculate organization in turtles. The first is that the geniculate complex is divided into three subnuclei that vary in size and density of their neurons. The second is that the complex has a form of laminar organization different than that seen in the geniculate complex of mammals.

Animals↗

Caudal topographic nucleus isthmi and the rostral nontopographic nucleus isthmi in the turtle, Pseudemys scripta.

Isthmotectal projections in turtles were examined by making serial section reconstructions of axonal and dendritic arborizations that were anterogradely or retrogradely filled with HRP. Two prominent tectal-recipient isthmic nuclei--the caudal magnocellular nucleus isthmi (Imc) and the rostral magnocellular nucleus isthmi (Imr)--exhibited strikingly different patterns of organization. Imc cells have flattened, bipolar dendritic fields that cover a few percent of the area of the cell plate constituting the nucleus and they project topographically to the ipsilateral tectum without local axon branches. The topography was examined explicitly at the single-cell level by using cases with two injections at widely separated tectal loci. Each Imc axon terminates as a compact swarm of several thousand boutons placed mainly in the upper central gray and superficial gray layers. One Imc terminal spans less that 1% of the tectal surface. Imr cells, by contrast, have large, sparsely branched dendritic fields overlapped by local axon collaterals while distally, their axons nontopographically innervate not only the deeper layers of the ipsilateral tectum but also ipsilateral Imc. Imr receives a nontopographic tectal input that contrasts with the topographic tectal input to Imc. Previous work on nucleus isthmi emphasized the role of the contralateral isthmotectal projection (which originates from a third isthmic nucleus in turtles) in mediating binocular interactions in the tectum. The present results on the two different but overlapping ipsilateral tecto-isthmo-tectal circuits set up by Imc and Imr are discussed in the light of physiological evidence for selective attention effects and local-global interactions in the tectum.

Animals↗

Comparative distribution of dopamine D-1 and D-2 receptors in the basal ganglia of turtles, pigeons, rats, cats, and monkeys.

The distribution and density of dopamine D-1 and D-2 receptors were studied in the basal ganglia of adult turtles, pigeons, rats, cats, and monkeys. Dopamine receptors were measured in vitro by quantitative autoradiography in alternate sections processed for D-1 and D-2 receptor subtypes and compared to adjacent sections stained for acetylcholinesterase (AChE) activity. [3H]-SCH 23390 and [3H]-spiroperidol were used to label the D-1 and D-2 dopamine receptor subtypes, respectively. The anatomic distribution of both D-1 and D-2 receptors in the basal ganglia was remarkably similar across all species examined. Whereas the absolute number of D-1 and D-2 receptors in the basal ganglia varied between species, the percentage of D-1 and D-2 receptors in a region was quite similar among species. The pattern of binding to the D-1 and D-2 receptor varied among the different species. The adult turtles, pigeons, and rats demonstrated nonpatchy D-1 and D-2 receptor binding in the striatum and pallidum. The adult cat and monkey caudate nucleus and putamen demonstrated mildly heterogeneous receptor binding in a pattern that differed from that seen with AChE staining, but did occasionally demonstrate similar patterns of the D-1 and D-2 receptor subtypes. The immature cat striatum was characterized by heterogeneous D-1 receptor binding that corresponded to heterogeneous AChE rich patches, whereas D-2 receptor binding was homogeneous. Heterogeneous binding was seen in other basal ganglia structures including the nucleus accumbens, olfactory tubercle, and substantia nigra pars compacta and reticulata. Complementary D-1 and D-2 receptor binding patterns were seen in the pallidum and substantia nigra of the mammals. The results of this study indicate that both D-1 and D-2 dopamine receptors are present in the basal ganglia of five different vertebrates. A common feature of dopamine receptors in the basal ganglia is their heterogeneity in distribution and density. The heterogeneity of dopamine receptors has similarities to and differences from the distribution of presynaptic dopamine and other neurotransmitter markers of the basal ganglia.

Aging↗

Quantitative autoradiography of muscarinic and benzodiazepine receptors in the forebrain of the turtle, Pseudemys scripta.

The distribution of muscarinic and benzodiazepine receptors was investigated in the turtle forebrain by the technique of in vitro receptor autoradiography. Muscarinic binding sites were labeled with 1 nM 3H-quinuclidinyl benzilate (3H-QNB), and benzodiazepine sites were demonstrated with the aid of 1 nM 3H-flunitrazepam (3H-FLU). Autoradiograms generated on 3H-Ultrofilm apposed to tissue slices revealed regionally specific distributions of muscarinic and benzodiazepine binding sites that are comparable with those for mammalian brain. Dense benzodiazepine binding was found in the anterior olfactory nucleus, the lateral and dorsal cortices, and the dorsal ventricular ridge (DVR), a structure with no clear mammalian homologue. Muscarinic binding sites were most dense in the striatum, accumbens, DVR, lateral geniculate, and the anterior olfactory nucleus. Cortical binding sites were studied in greater detail by quantitative analysis of autoradiograms generated by using emulsion-coated coverslips. Laminar gradients of binding were observed that were specific for each radioligand; 3H-QNB sites were most dense in the inner molecular layer in all cortical regions, whereas 3H-FLU binding was generally most concentrated in the outer molecular layer and was least dense through all layers in the dorsomedial cortex. Because pyramidal cells are arranged in register in turtle cortex, the laminar patterns of receptor binding may reflect different receptor density gradients along pyramidal cell dendrites.

Animals↗

Neuropeptide Y-immunoreactive amacrine cells in the retina of the turtle Pseudemys scripta elegans.

Antiserum directed against neuropeptide Y selectively labeled certain amacrine cells in the turtle retina. The cell types, sizes, dendritic stratification, regional distribution, and degrees of immunolabeling were examined. The results indicated that three morphologically distinct cell types were labeled: types A, B, and C. Computer rotation of digitized data from camera lucida drawings was used to study dendritic stratification. The type A somata were large (11.5 micron in diameter), well-stained, and located in the third tier of the inner nuclear layer. Type A somata gave rise to well-stained processes which arborized within the inner plexiform layer in strata 1 and 3 and at the border between strata 4 and 5. Processes in stratum 1 were sparse and delicate with small boutons. Processes in stratum 3 were numerous and often coarse, with many large and small boutons. At the border between strata 4 and 5 the processes were frequently numerous but slender, with many small boutons. Occasional immunolabeled processes were found in the ganglion cell layer. The somata of the type B cells were smaller (9.0 micron in diameter) and gave rise to single labeled processes which descended into the inner plexiform layer and divided quickly into two secondary processes. These secondary processes gave rise to lightly labeled dendritic fields which arborized primarily in strata 2 and 4. The type C cells were usually observed at the periphery of the retina and had large somata (12.0 micron in diameter) with simple, but very elongated, dendritic arborizations in strata 1, 4, and 5. Observations also showed that type A and B cells were often found in close proximity to each other and suggested that dendrites of these cells made contact with each other. The labeled neurons were distributed relatively evenly throughout the retina except for the visual streak where they were sparse. This study indicates that neuropeptide Y-like immunoreactivity is found in more than one anatomically distinct class of amacrine cells in the turtle retina.

Animals↗

Organization of the inner plexiform layer of the turtle retina: an electron microscopic study.

We have performed a serial-section electron microscopic study of the inner plexiform layer (IPL) of the retina of the turtle Pseudemys scripta elegans. A qualitative and quantitative assessment of the neuropil of the IPL has been done from photomontages taken from the linear visual streak area and peripheral retina. Counts of conventional, ribbon, serial, and reciprocal synapses, of ganglion cell dendrites, and of profiles containing large, dense-cored vesicles were made in five equal-thickness strata in each montage. Averages of these different features were plotted for each stratum in the linear visual streak and compared with peripheral retina. The trend was for stratum 2 to have the highest overall absolute number of amacrine and bipolar interactions, and also of serial synapses, both in the linear visual streak and in peripheral regions. Stratum 4 tended to have the second-highest number of synapses. The total number of synapses for the entire thickness of the IPL, regardless of stratification, is higher in the streak than in the periphery. The total amacrine-to-bipolar-synapse ratio in the IPL is the highest of any vertebrate studied to date (11.0 in the streak and 14.5 in the periphery) but the number of synapses/micron 2 was found to be similar to that reported for other vertebrates. Amacrine-to-amacrine synaptic contacts greatly outnumber other types of synapses; amacrines constitute the principal input to ganglion cells, whereas bipolar output is mainly onto amacrines. The trend for higher numbers of synaptic interactions in strata 2 and 4 of the streak region of the turtle IPL can be correlated with the branching of small-field amacrine and ganglion cells described in Golgi studies (Kolb: Philos. Trans. R. Soc. Lond. B 298:355-393, '82). In peripheral retina, branching of large-field amacrines and a lower number of bipolar pathways may account for the trend for larger numbers of amacrine synapses in strata 2 and 4. Profiles having large, dense-cored vesicles tend to occur most frequently in strata 1 and 5, which correlates well with the stratification in the IPL of the processes of immunoreactive amacrine cells described in other studies.

Animals↗