Golgi techniques have been applied to post mortem specimens of human retina. Analysis was possible on 150 human retinas processed and viewed by light microscopy as wholemounts. Camera lucida drawings and photography were used to classify the impregnated neurons into 3 types of horizontal cell, 9 types of bipolar cell, 24 basic types of amacrine cell, a single type of interplexiform cell, and 18 types of ganglion cell. We have distinguished two types of midget bipolar cell: fmB (flat) and imB (invaginating). In central retina, both types are typically single-headed, each clearly contacting a single cone. Peripherally, they may be two- or even three-headed, obviously contacting more than one cone. Two types of small-field diffuse cone bipolars occurring as flat and invaginating varieties are found across the entire retina from fovea to far periphery. The single rod bipolar type appears about 1 mm from the fovea and increases in dendritic tree diameter from there into the far periphery. The putative "ON-center" blue cone bipolar and the giant bistratified bipolar first described by Mariani are also present in human retina and we add two previously undescribed bipolar cell types: a putative giant diffuse invaginating and a candidate "OFF-center" blue cone bipolar. Taking into account the variation of cell size with eccentricity at all points on the retina, we observed three distinct varieties of horizontal cell. The HI is the well known, long-axon-bearing cell of Polyak. HII is the more recently described multibranched, wavy-axoned horizontal cell. The third variety, HIII, introduced here, has been separated from the HI type on morphological criteria of having a larger, more asymmetrical dendritic field and in contacting 30% more cones than the HI at any point on the retina. Amacrine cells proved to be most diverse in morphology. Many of the amacrine cell types that have been described in cat retina (Kolb et al., '81: Vision Res. 21; 1081-1114) were seen in this study. Where there are no equivalent cells in cat, we have adopted the descriptive terminology used by Mariani in monkey retina. Thus eight varieties of small-field amacrines (under 100 microns dendritic trees), eight varieties of medium-field cells (100-500 microns dendritic span), and eight large-field varieties (over 500 microns dendritic trees) have been classified. Often a broadly described variety of amacrine cell can be subdivided into as many as three subtypes dependent on stratification levels of their dendrites in the inner plexiform layer.(ABSTRACT TRUNCATED AT 400 WORDS)

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Morphogenesis of photoreceptor outer segment disks appears to occurby an evagination of the ciliary plasma membrane (Steinberg et al., J Comp Neurol190:501-519, '80). We tested if polymerized actin (F-actin) was necessary for the regulation of this postulated process by incubating Xenopus eyecups with 5 or 25 pM cytochalasin D for 6-28 hours. During the second hour, the incubation medium contained 3H-leucine. Both concentrations of cytochalasin resulted in: 1) dissolution of the rhodamine-phalloidin labeling pattern of photoreceptors, and 2) collapse of the calycal processes (which are normally filled with actin filaments) and disappearance of the inner segment microfilaments. In addition, the few most basal rod and cone outer segment disks appeared several times their normal diameter. These oversized disks had incorporated 3H-leucine and extended along the margin of the outer or inner segment. The nature of the overgrown disks is consistent only with a morphogenetic process involving evaginations of the ciliary plasma membrane. Deregulation by cytochalasin D was manifest by excessive growth of a few nascent disks rather than normal growth of many. Therefore, the normal network of actin filaments is apparently not necessary for continued evagination of the membrane, but it does seem to be an essential part of the mechanism that initiates the evagination of the ciliary plasma membrane andlor the mechanism that controls how far nascent disks grow.

Müller cells are subtly different in the GFAP(-/-)vim(-/-) mouse retina before detachment. The end foot region of these cells may be structurally reinforced by the presence of the intermediate filament cytoskeleton, and our data suggest a critical role for these proteins in Müller cell reaction to retinal detachment and participation in subretinal gliosis.

The vertebrate photoreceptor cell contains an elaborate cilium that includes a stack of phototransductive membrane disks. The disk membranes are continually renewed, but how new disks are formed remains poorly understood. Here we used electron microscope tomography to obtain 3D visualization of the nascent disks of rod photoreceptors in three mammalian species, to gain insight into the process of disk morphogenesis. We observed that nascent disks are invariably continuous with the ciliary plasma membrane, although, owing to partial enclosure, they can appear to be internal in 2D profiles. Tomographic analyses of the basal-most region of the outer segment show changes in shape of the ciliary plasma membrane indicating an invagination, which is likely a first step in disk formation. The invagination flattens to create the proximal surface of an evaginating lamella, as well as membrane protrusions that extend between adjacent lamellae, thereby initiating a disk rim. Immediately distal to this initiation site, lamellae of increasing diameter are evident, indicating growth outward from the cilium. In agreement with a previous model, our data indicate that mature disks are formed once lamellae reach full diameter, and the growth of a rim encloses the space between adjacent surfaces of two lamellae. This study provides 3D data of nascent and mature rod photoreceptor disk membranes at unprecedented z-axis depth and resolution, and provides a basis for addressing fundamental questions, ranging from protein sorting in the photoreceptor cilium to photoreceptor electrophysiology.photoreceptor | cilium | EM tomography | disk morphogenesis P rimary cilia detect extracellular signals via membrane receptors or channels. The most elaborate of all cilia, the cilium that forms the vertebrate photoreceptor outer segment (OS), includes a large stack of membrane disks that extends distally from the transition zone, also known as the connecting cilium (1). The OS disks contain the visual receptor, opsin, and their tight packing allows for a high concentration of opsin within a confined space, thereby limiting the trade-off between visual sensitivity and spatial resolution. Turnover of OS disk membranes occurs throughout the lifetime of an animal (2) and requires the de novo synthesis and degradation of large amounts of OS proteins; on average, 9-10 billion opsin molecules are turned over every second in each human retina (3).A key event in OS disk turnover is the formation of disk membranes from newly synthesized molecules that are trafficked as vesicles from the endoplasmic reticulum (ER)/Golgi in the inner segment to the cilium (4, 5). Disk membrane morphogenesis is essential for the survival of photoreceptor cells. Orthologs of gene mutations that disrupt disk formation in mice have been linked to various retinal degenerations in humans; for example, in the rds mouse [mutant in peripherin-2 (Prph2)], disks do not develop from the end of the connecting cilium (6, 7), and in mice whose photoreceptors express mutant human prominin-1 (...