Tree Shrew Retina Research Articles

Visible-Light Optical Coherence Tomography Fibergraphy of the Tree Shrew Retinal Ganglion Cell Axon Bundles.

We seek to develop techniques for high-resolution imaging of the tree shrew retina for visualizing and parameterizing retinal ganglion cell (RGC) axon bundles in vivo. We applied visible-light optical coherence tomography fibergraphy (vis-OCTF) and temporal speckle averaging (TSA) to visualize individual RGC axon bundles in the tree shrew retina. For the first time, we quantified individual RGC bundle width, height, and cross-sectional area and applied vis-OCT angiography (vis-OCTA) to visualize the retinal microvasculature in tree shrews. Throughout the retina, as the distance from the optic nerve head (ONH) increased from 0.5 mm to 2.5 mm, bundle width increased by 30%, height decreased by 67%, and cross-sectional area decreased by 36%. We also showed that axon bundles become vertically elongated as they converge toward the ONH. Ex vivo confocal microscopy of retinal flat-mounts immunostained with Tuj1 confirmed our in vivo vis-OCTF findings.

DeBruyn and Casagrande manuscripts on tree shrew retinal ganglion cells as a basis for cross-species retina research.

The purpose of this brief communication is to make publicly available three unpublished manuscripts on the organization of retinal ganglion cells in the tree shrew. The manuscripts were authored in 1986 by Dr. Edward DeBruyn, a PhD student in the laboratory of the late Dr. Vivien Casagrande at Vanderbilt University. As diurnal animals closely related to primates, tree shrews are ideally suited for comparative analyses of visual structures including the retina. We hope that providing this basic information in a citable form inspires other groups to pursue further characterization of the tree shrew retina using modern techniques.

Noninvasive imaging of the tree shrew eye: Wavefront analysis and retinal imaging with correlative histology

Tree shrews are small mammals with excellent vision and are closely related to primates. They have been used extensively as a model for studying refractive development, myopia, and central visual processing and are becoming an important model for vision research. Their cone dominant retina (∼95% cones) provides a potential avenue to create new damage/disease models of human macular pathology and to monitor progression or treatment response. To continue the development of the tree shrew as an animal model, we provide here the first measurements of higher order aberrations along with adaptive optics scanning light ophthalmoscopy (AOSLO) images of the photoreceptor mosaic in the tree shrew retina. To compare intra-animal in vivo and ex vivo cone density measurements, the AOSLO images were matched to whole-mount immunofluorescence microscopy. Analysis of the tree shrew wavefront indicated that the optics are well-matched to the sampling of the cone mosaic and is consistent with the suggestion that juvenile tree shrews are nearly emmetropic (slightly hyperopic). Compared with in vivo measurements, consistently higher cone density was measured ex vivo, likely due to tissue shrinkage during histological processing. Tree shrews also possess massive mitochondria (“megamitochondria”) in their cone inner segments, providing a natural model to assess how mitochondrial size affects in vivo retinal imagery. Intra-animal in vivo and ex vivo axial distance measurements were made in the outer retina with optical coherence tomography (OCT) and transmission electron microscopy (TEM), respectively, to determine the origin of sub-cellular cone reflectivity seen on OCT. These results demonstrate that these megamitochondria create an additional hyper-reflective outer retinal reflective band in OCT images. The ability to use noninvasive retinal imaging in tree shrews supports development of this species as a model of cone disorders.

The Second Visual System of The Tree Shrew

This review provides a historical account of the discovery of secondary visual pathways (from retina to the superior colliculus to the dorsal thalamus and extrastriate cortex), and Vivien Casagrande's pioneering studies of this system using the tree shrew as a model. Subsequent studies of visual pathways in the tree shrew are also reviewed, beginning with a description of the organization and central projections of the tree shrew retina. The organization and connectivity of second visual system components that include the retino-recipient superior colliculus, tecto-recipient pulvinar nucleus and its projections, and the tecto-recipient dorsal lateral geniculate nucleus and its projections are detailed. Potential functions of the second visual system are discussed in the context of this work and in the context of the behavioral studies that initially inspired the secondary visual system concept.

Altered gene expression in tree shrew retina and retinal pigment epithelium produced by short periods of minus-lens wear

Hyperopic refractive error is detected by retinal neurons, which generate GO signals through a direct emmetropization signaling cascade: retinal pigment epithelium (RPE) into choroid and then into sclera, thereby increasing axial elongation. To examine signaling early in this cascade, we measured gene expression in the retina and RPE after short exposure to hyperopia produced by minus-lens wear. Gene expression in each tissue was compared with gene expression in combined retina + RPE. Starting 24 days after normal eye opening, three groups of juvenile tree shrews (n = 7 each) wore a monocular −5 D lens. The untreated fellow eye served as a control. The “6h” group wore the lens for 6 h; the “24h” group wore the lens for 24 h; each group provided separate retina and RPE tissues. Group “24hC” wore the lens for 24 h and provided combined retina + RPE tissue. Quantitative PCR was used to measure the relative differences (treated eye vs. control eye) in mRNA levels for 66 candidate genes. In the retina after 6 h, mRNA levels for seven genes were significantly regulated: EGR1 and FOS (early intermediate genes) were down-regulated in the treated eyes. Genes with secreted protein products, BMP2 and CTGF, were down-regulated, whilst FGF10, IL18, and SST were up-regulated. After 24 h the pattern changed; only one of the seven genes still showed differential expression; BMP2 was still down-regulated. Two new genes with secreted protein products, IGF2 and VIP, were up-regulated. In the RPE, consistent with its role in receiving, processing, and transmitting GO signaling, differential expression was found for genes whose protein products are at the cell surface, intracellular, in the nucleus, and are secreted. After 6 h, mRNA levels for 17 genes were down-regulated in the treated eyes, whilst four genes (GJA1, IGF2R, LRP2, and IL18) were up-regulated. After 24 h the pattern was similar; mRNA levels for 14 of the same genes were still down-regulated; only LRP2 remained up-regulated. mRNA levels for six genes no longer showed differential expression, whilst nine genes, not differentially expressed at 6 h, now showed differential expression. In the combined retina + RPE after 24 h, mRNA levels for only seven genes were differentially regulated despite the differential expression of many genes in the RPE. Four genes showed the same expression in combined tissue as in retina alone, including up-regulation of VIP despite significant VIP down-regulation in RPE. Thus, hyperopia-induced GO signaling, as measured by differential gene expression, differs in the retina and the RPE. Retinal gene expression changed between 6 h and 24 h of treatment, suggesting evolution of the retinal response. Gene expression in the RPE was similar at both time points, suggesting sustained signaling. The combined retina + RPE does not accurately represent gene expression in either retina or, especially, RPE. When gene expression signatures were compared with those in choroid and sclera, GO signaling, as encoded by differential gene expression, differs in each compartment of the direct emmetropization signaling cascade.

Early development of the nervous system of the eutherian <i>Tupaia belangeri</i>

Abstract. The longstanding debate on the taxonomic status of Tupaia belangeri (Tupaiidae, Scandentia, Mammalia) has persisted in times of molecular biology and genetics. But way beyond that Tupaia belangeri has turned out to be a valuable and widely accepted animal model for studies in neurobiology, stress research, and virology, among other topics. It is thus a privilege to have the opportunity to provide an overview on selected aspects of neural development and neuroanatomy in Tupaia belangeri on the occasion of this special issue dedicated to Hans-Jürg Kuhn. Firstly, emphasis will be given to the optic system. We report rather "unconventional" findings on the morphogenesis of photoreceptor cells, and on the presence of capillary-contacting neurons in the tree shrew retina. Thereafter, network formation among directionally selective retinal neurons and optic chiasm development are discussed. We then address the main and accessory olfactory systems, the terminal nerve, the pituitary gland, and the cerebellum of Tupaia belangeri. Finally, we demonstrate how innovative 3-D reconstruction techniques helped to decipher and interpret so-far-undescribed, strictly spatiotemporally regulated waves of apoptosis and proliferation which pass through the early developing forebrain and eyes, midbrain and hindbrain, and through the panplacodal primordium which gives rise to all ectodermal placodes. Based on examples, this paper additionally wants to show how findings gained from the reported projects have influenced current neuroembryological and, at least partly, medical research.

Retinal and choroidal TGF-β in the tree shrew model of myopia: Isoform expression, activation and effects on function

A visually evoked signalling cascade, which begins in the retina, transverses the choroid, and mediates scleral remodelling, is considered to control eye growth. The ubiquitous cytokine TGF-β has been associated with alterations in ocular growth, where alterations in scleral TGF-β isoforms mediate the scleral remodelling that results in myopia. However, while the TGF-β isoforms have been implicated in the scleral change during myopia development, it is unclear whether alterations in retinal and choroidal isoforms constitute part of the retinoscleral cascade. This study characterised the retinal and choroidal TGF-β isoform profiles and TGF-β2 activation during different stages of myopia development, as induced by form deprivation, in a mammalian model of eye growth. Using quantitative real-time PCR, the mRNA for all three mammalian isoforms of TGF-β was detected in tree shrew retina and choroid. Distinct tissue-specific isoform profiles were observed for the retina (TGF-β1:TGF-β2:TGF-β3=20:2085:1) and choroid (TGF-β1:TGF-β2:TGF-β3=16:23:1), which remained constant over the development period under investigation. The active and latent pools of retinal TGF-β2 were quantified using ELISA with the majority (>94%) of total TGF-β2 found in the latent form. Unlike previous scleral data showing early and continuous decreases in TGF-β isoform expression during myopia development, the levels of the three isoforms remained within normal ranges for retinal (TGF-β1, −14 to +14%; TGF-β2, −2 to +20%; TGF-β3, −10 to +26%) and choroidal (TGF-β1, −19 to +21%; TGF-β2, −26 to +8%; TGF-β3, −11 to +28%) tissues during myopia development (induction times of 3h, 7h, 11h, 24h, and 5 days). A 40% decrease in retinal TGF-β2 activation was observed after 5 days of myopia development, however, there was no functional correlate of altered TGF-β2 activity, as assessed by the retinal ERG response. Overall, these data highlight the specific nature of TGF-β isoform expression, which reflects the differences in tissue structure and function. While TGF-β isoforms are involved in scleral regulation during myopia development in mammals, they do not have a primary role in the retinal and choroidal signals. Thus, the regulation of eye growth via the retinoscleral cascade involves more than one factor, which is likely to be tissue-specific in nature.

Temporal modulation sensitivity of tree shrew retinal ganglion cells.

Tree shrews (Tupaia belangeri) are small diurnal mammals capable of quick and agile navigation. Electroretinographic and behavioral studies have indicated that tree shrews possess very good temporal vision, but the neuronal mechanisms underlying that temporal vision are not well understood. We used single-unit extracellular recording techniques to characterize the temporal response properties of individual retinal ganglion cell axons recorded from the optic tract. A prominent characteristic of most cells was their sustained or transient nature in responding to the flashing spot. Temporal modulation sensitivity functions were obtained using a Gaussian spot that was temporally modulated at different frequencies (2-60 Hz). Sustained cells respond linearly to contrast. They showed an average peak frequency of 6.9 Hz, a high-frequency cutoff at 31.3 Hz, and low-pass filtering. Transient cells showed nonlinear response to contrast. They had a peak frequency of 19.3 Hz, a high-frequency cutoff at about 47.6 Hz, band-pass filtering, and higher overall sensitivity than sustained cells. The responses of transient cells also showed a phase advance of about 88 deg whereas the phase advance for sustained cells was about 43 deg. Comparison with behavioral temporal modulation sensitivity results suggested that transient retinal ganglion cells may underlie detection for a wide range of temporal frequencies, with sustained ganglion cells possibly mediating detection below 4 Hz. These data suggest that two well-separated temporal channels exist at the retinal ganglion cell level in the tree shrew retina, with the transient channel playing a major role in temporal vision.

Calcium channel heterogeneity among cone photoreceptors in the tree shrew retina.

Retinal photoreceptors are depolarized in darkness and release neurotransmitter tonically. They respond to light by hyperpolarization and a concomitant reduction in transmitter release. The calcium-dependent release of transmitter is coupled to graded changes in membrane potential by L-type calcium channels in the photoreceptor terminals. This paper reports the immuno-localization of an L-type channel alpha1D subunit to most, but not all, synaptic terminals of cones in the tree shrew retina. Double labelling for the alpha1D subunit and the plasma membrane Ca2+-ATPase, which has been shown to be present in all tree shrew cones, revealed a subpopulation of cone terminals that did not react with the alpha1D antibody. The nonimmunoreactive synaptic terminals represented approximately 5.8% of the total and formed a highly regular array across the retina reminiscent of the blue cones. Double-staining for the alpha1D subunit and blue cone opsin confirmed that these are the blue cones. The observed differences in calcium channel immunoreactivity between long and short wavelength cones points to previously unsuspected heterogeneity in the molecular machinery governing transmitter release from spectrally different cone types.

Localization of nitric oxide synthase in the tree shrew retina.

Nitric oxide (NO) is a novel neuronal messenger that likely influences retinal function by activating retinal guanylyl cyclase to increase levels of cGMP. In the present study, the localization of neuronal nitric oxide synthase (nNOS, Type I NOS) in the cone-dominant tree shrew retina was studied using NADPH-d histochemistry and nNOS immunocytochemistry. Both NADPH-d and nNOS-immunoreactivity (IR) labeled the inner segments of rods and the myoids of a regular subpopulation of cones, with their corresponding nuclei outlined. The labeled cone myoids were co-localized with a marker for short-wave-sensitive (SWS) cones (S-antigen) and also displayed the regular triangular packing and density (7%) characteristic of SWS cones in tree shrew and other mammalian retinas. These measures confirmed the identity of the labeled cones as SWS cones. Photoreceptor ellipsoids of all cones were strongly labeled by NADPH-d reactivity, but lacked nNOS-IR. Another novel finding in tree shrew retina was that both NADPH-d and nNOS-IR labeled Muller cells, which have not been labeled by nNOS-IR in other mammalian retinas. Consistent with findings in rod-dominant retinas, two types of amacrine cells at the vitreal edge of the inner nuclear layer and a subpopulation of displaced amacrine cells at the scleral edge of the ganglion cell layer were labeled by both NADPH-d and nNOS-IR. Processes of these labeled cells were seen to extend into the inner plexiform layer, where dense punctate label was seen, especially in the central sublamina. These results show that localization of NOS in the cone-dominant tree shrew retina shares some common properties with rod-dominant mammalian retinas, but also shows some species-specific characteristics. The new finding of nNOS localization in tree shrew SWS cones and rods, but not in other cones, raises interesting questions about the roles of NO in the earliest level of visual processing.

Starburst cholinergic amacrine cells in the tree shrew retina

In all mammalian retinae studied to date, starburst cholinergic amacrine cells are a consistently occurring cell type. Here, we show that the cone-dominated retina of the tree shrew also has starburst cells with the characteristic radially symmetric branching pattern known from other species. Dendritic field sizes increase from 150 microm in the central retina to 300 microm in the retinal periphery. The characteristic morphology is established early during postnatal development. Labelling the starburst cholinergic cells with an antibody against choline acetyltransferase (ChAT) reveals two dendritic strata in the inner plexiform layer and two corresponding soma populations in the inner nuclear layer (orthotopic) and ganglion cell layer (displaced). These features are present in the adult and in early postnatal stages. In the adult, the density of the orthotopic population as well as the displaced population peaks in the central retina at about 2,200 cells/mm2 and has a peripheral minimum of 400 cells/mm2. These properties are qualitatively similar to those of starburst cells in rod-dominated retinae. In contrast to findings in other mammals, we did not see gamma-aminobutyric acid (GABA) or glutamic acid decarboxylase 65 kDa (GAD65) immunoreactivity in tree shrew starburst cells. These cells also appear to lack synaptophysin, a ubiquitous synaptic vesicle protein detected in the starburst cells of some other mammals. However, synaptoporin, a homologous synaptic vesicle protein, appears to be present in tree shrew starburst cells.

Starburst cholinergic amacrine cells in the tree shrew retina

Starburst cholinergic amacrine cells in the tree shrew retina

Ciliogenesis in photoreceptor cells of the tree shrew retina.

Transmission electron microscopy of the retinal cones from several prenatal, young postnatal and adult tree shrews (Tupaia belangeri) reveals that the centrioles, from which the ciliary precursors of the outer segments grow out, are not transported into a pre-existing inner segment, but are positioned under the apical plasma membrane of cone precursor cells all through the inner segment formation. Ciliogenesis starts before or on embryonic day 20 and thus precedes initial formation of the inner segment by 20 days, which is half the gestation period. Thus, the maturation of the outer segment covers a considerably longer period than has been previously described. Published observations from other mammals can be interpreted as conforming with the situation in Tupaia. In other vertebrates, compared to mammals, marked heterochronies do occur. In Tupaia, the centrioles and the cilium are located close to the central longitudinal axis of the photoreceptor precursor cell from the 20-day-old embryo to the 5-day-old juvenile. In this position the microtubule apparatus originating from the centrioles should be most effective in transporting the mitochondria into the inner segment. In the 12-day-old tree shrew, when transport of the mitochondria into the inner segment has been completed, centrioles and cilium have shifted into an eccentric position and the light-collecting megamitochondria have approached the disks of the outer segment. This eccentric position is maintained in all later developmental stages. In certain of the retinal areas of the adult Tupaia, the connecting cilia of neighbouring cones are always positioned on the same side of the inner segments.

Unique distribution of somatostatin-immunoreactive cells in the retina of the tree shrew (Tupaia belangeri).

Somatostatin-like immunoreactive cells in the tree shrew retina were studied with the monoclonal antibody S8 against the neuropeptide somatostatin 14. As in some other mammals, immunoreactive somata are exclusively found in the ganglion cell layer. Immunoreactive processes form a sparse main plexus in the inner plexiform layer near the border of the inner nuclear layer; fewer additional processes are found closer to the ganglion cell layer. With retrograde labelling of retinal ganglion cells by injections of the tracer Fast Blue into the superior colliculus and lateral geniculate body and counterstaining of the retinae with S8, approximately 5% of the immunoreactive somata were double-labelled at any retinal location. The vast majority of somatostatin-like immunoreactive cells are thus displaced amacrine cells. Their somata are distributed over the entire retina. Their population density is highest in the temporal retina, with peak densities of approximately 5000 cells/mm2 near the central area and a steep density gradient. In the remaining retina densities are 200-400 cells/mm2, falling to approximately 100 cells/mm2 at the retinal margins. This is in stark contrast to the somatostatin-like immunoreactive cells in other mammalian retinae which have densities of 10-40 cells/mm2 and are confined to restricted retinal regions (inferior retina and/or retinal margin).

Müller glial cells of the tree shrew retina

The tree shrew is one of the few mammalian species whose retinae are strongly cone dominated, which is usually the case in reptilian and avian retinae. Müller cells of the tree shrew (Tupaia belangeri) retina were studied by transmission electron microscopy of tissue sections and freeze-fracture replicas, by immunolabeling of the intermediate filament protein vimentin in radial paraffin sections and in whole retinae, as well as by intracellular dye injection in slices of retinae. In addition, enzymatically isolated cells were stained by Pappenheim's panoptic staining method. The cells showed an ultrastructure that is similar to other mammalian Müller cells with two exceptions: Due to the extensive lateral fins of cone inner segments, the apical microvilli of Müller cells are arranged in peculiar palisades, and the basket-like Müller cell sheaths around neuronal somata in both nuclear layers consist of unusual multilayered membrane lamellae. Unlike Müller cells in other mammalian species studied thus far, but similar to reptilian and avian Müller cells, those of tree shrews commonly have two or more vitread processes rather than one main trunk. Müller cell densities range between some 13,000 mm-2 in the periphery and about 20,000 mm-2 in the retinal center. Neuron:(Müller)glial cell ratios were estimated to be 7.9:1 in the center and 6.2:1 in the periphery. For each Müller cell, about 1.5 (cone) photoreceptor cells, four or five interneurons of the inner nuclear layer, and about one cell of the ganglion cell layer were counted. This is a much lower number of neurons per Müller cell than in most other mammals studied.

Horizontal cells in the cone-dominated tree shrew retina: morphology, photoreceptor contacts, and topographical distribution

While most mammalian retinas are rod dominated, in the tree shrew retina 95% of the photoreceptors are cones. We studied three shrew horizontal cells to look for features associated with this unusual photoreceptor arrangement. The morphology of horizontal cells was revealed by intracellular injections of Lucifer yellow, and their photoreceptor contacts were assessed by light and electron microscopy. Horizontal cell topography was studied in material stained with a neurofilament antibody and with toluidine blue. The tree shrew has two types of horizontal cell that are basically the same as A- and B-type horizontal cells of other mammals. All the photoreceptor contacts of the larger, axonless, A-type cell and the dendritic contacts of the smaller, axon-bearing, B-type cell are with cones. Both types contact nearly all the cones in their dendritic field and both types synapse with both red and blue cones. There is no anatomical evidence for chromatic selectivity. The sparsely branched B-type horizontal cell axon probably contacts rods as in other mammals. The unusual features of the A-type cell are the profuse dendritic terminal arborizations and the large dendritic field size. These features may be related to the abundance of cones but do not justify the conclusion for a special type of horizontal cell as has previously been supposed. Both types of horizontal cell have a central-peripheral density gradient; at any location B-type cells are up to three times as numerous as A-type cells. There are detailed features of the distributions that differ from those of other mammalian horizontal cells. The density maximum of B-type cells is in inferior retina and roughly coincides with that of the cones; the A-type maximum is located more superiorly. Neither maximum is colocalized with the ganglion cell peak in the central area. The mosaic of B-type cells is much more regular than that of A-type cells.

Immunocytochemical identification of photoreceptor populations in the tree shrew retina

The presence and retinal distributions of short-wavelength-sensitive cones, long-wavelength-sensitive cones, and rods were assessed in the retina of the tree shrew Tupaia belangeri using visual pigment antibodies OS-2, COS-1 and anti-rhodopsin, respectively. Results demonstrated a clear immunocytochemical differentiation of the three photoreceptor types with each showing regional variation in density across the retina.

Retinal [ 125I]iodomelatonin binding sites in the tree shrew ( Tupaiidae)

The characteristics of melatonin-binding sites labelled by [ 125I]iodomelatonin in membrane preparations from the tree shrew retina were determined. Specific binding of [ 125I]iodomelatonin to the membrane preparations of tree shrew retina was rapid, stable, saturable, and reversible. Among the indoles tested only 6-chloromelatonin, melatonin and N-acetylserotonin had significant affinities to the [ 125I]iodomelatonin binding site. Scatchard analysis of the membrane preparations revealed a dissociation constant ( K d) of 51.0 ± 16 pM and total number of binding sites ( B max) of 1.97 ± 0.6 fmol/mg protein.

Morphology and distribution of catecholaminergic amacrine cells in the cone‐dominated tree shrew retina

The tree shrew (Tupaia belangeri) has a cone-dominated retina with a rod proportion of only 5%. This is in contrast to the usual mammalian pattern of rod-dominated retinae. Rod bipolar cells are present at relatively low densities in the tree shrew retina, suggesting that a reduced, but normal, rod pathway might be preserved. The present study investigated another common constituent of the rod pathway, the dopaminergic amacrine cells, and analysed their morphology and distribution by light and electron microscopy. Catecholaminergic (presumed dopaminergic) amacrine cells were labelled with an antibody against tyrosine hydroxylase (TH). Intense TH-immunoreactivity was found in perikarya and dendrites of a uniform amacrine cell population. TH-immunoreactive amacrine cell density varies across the retina from 10 cells/mm2 in the periphery to 40 cells/mm2 in more central regions (mean cell density about 25 cells/mm2). The relatively large cell bodies are located exclusively in the innermost part of the inner nuclear layer. The dendrites form a dense plexus at the border between the inner plexiform layer and the inner nuclear layer. The finer dendritic processes contain many varicosities and form characteristic dendritic "rings" like those seen in other mammals. TH-immunoreactive processes also run between cell bodies in the vitread inner nuclear layer; a few extend into the sclerad inner nuclear layer and occasionally reach the outer plexiform layer (possible interplexiform cells). A few TH-immunoreactive processes are seen in the middle of the inner plexiform layer. Electron microscopy of TH-immunoreactive processes revealed conventional synapses onto somata and processes of unlabelled amacrine cells.

Rod bipolar cells in the cone-dominated retina of the tree shrew Tupaia belangeri

The tree shrew has a cone-dominated retina with a rod proportion of 5%, in contrast to the common mammalian pattern of rod-dominated retinae. As a first step to elucidate the rod pathway in the tree shrew retina, we have demonstrated the presence of rod bipolar cells and studied their morphology and distribution by light and electron microscopy. Rod bipolar cells were labeled with an antiserum against the protein kinase C (PKC), a phosphorylating enzyme. Intense PKC immunoreactivity was found in perikarya, axons, and dendrites of rod bipolar cells. The cell bodies are located in the sclerad part of the inner nuclear layer, the dendrites ascend to the outer plexiform layer where they are postsynaptic to rod spherules, and an axon descends towards the inner plexiform layer (IPL). The axons branch, and terminate in the vitread third of the IPL where mammalian rod bipolar cells are known to terminate. Two amacrine cell processes are always seen as the postsynaptic elements (dyads). Dendritic and axonal arbors of rod bipolar cells are rather large, up to 100 microns in diameter. The topographical distribution of the rod bipolar cells was analyzed quantitatively in tangential sections. Their density ranges from 300 cells/mm2 in peripheral retina to 900 cells/mm2 more centrally. The distribution is rather flat with no local extremes. Consistent with the low rod proportion in tree shrew, the rod bipolar cell density is low compared to the rod-dominated cat retina for example (36,000-47,000 rod bipolar cells/mm2). Rod-to-rod bipolar cell ratios in the tree shrew retina range from smaller than 1 to about 7, and thus are also lower than in cat.