The biosynthesis and content of gamma-aminobutyric acid in the goldifsh retina.

Goldfish retinas incubated with L-glutamate-(14)C (UL) were found to synthesize gamma-aminobutyric acid-(14)C (GABA-(14)C) The accumulation of newly synthesized GABA was enhanced by physiological stimulation of the retina with flashing light; and this increase was directly proportional to the logarithm of the light intensity. The total GABA content was also higher in light-stimulated than in dark-adapted retinas, although the glutamate content remained unchanged No differences were found in the cell-free activities of glutamate decarboxylase (EC 4 1.1 15) and GABA-glutamate transaminase (EC 2.6.1.19) extracted from light-stimulated and dark-adapted retinas. These findings, together with other physiological and morphologcal evidence, suggest that GABA plays a functional role in synaptic transmission in the goldfish retina

Biochemical studies of isolated glial (Müller) cells from the turtle retina.

A method has been developed for the preparation of large numbers of glial (Muller) cells from the turtle retina. After proteolytic dissociation of the retina, Muller cells were separated from retinal neurons by velocity sedimentation at unit gravity. Fractions containing >90 percent morphologically identifiable Muller cells were prepared by this procedure. Fractions containing only Muller cells were obtained by drawing selected cells individually into a micropipette under visual observation. Biochemical analyses of isolated Muller cells showed that (a) these cells did not synthesize and accumulate acetylcholine, gamma-aminobutyric acid, or catecholamines when incubated with appropriate radioactive precursors; (b) the specific activities of choline acetyltransferase (EC 2.3.1.6), glutamate decarboxylase (EC 4.1.1.15), and tyrosine hydroxylase (EC 1.14.16.2) in these cells were less than 2 percent of those found in the retina; (c) Muller cells, however, contained high activities of transmitter degrading enzymes-acetylcholinesterase (EC 3.1.1.7) and gamma-aminobutyrate- transamine (EC 2.6.1.19); and (d) the cells also possessed high levels of two presumably glial-specific-enzymes-glutamine synthetase (EC 6.3.1.2) and carbonic anhydrase (EC 4.2.1.1). These results, together with other findings, suggest that Muller cells are not capable of neurotransmitter syntheses but possess the enzymes necessary for two important roles in the retina: (a) the inactivation of certain transmitters after synaptic transmission by uptake and degradation, and (b) the maintenance of acid-base balance and the provision of a stable microenvironment in the retina by the removal of metabolic products such as carbon dioxide and ammonia.

An extracellular retinol-binding glycoprotein in the eyes of mutant rats with retinal dystrophy: development, localization, and biosynthesis.

Interstitial retinol-binding protein (IRBP) is a soluble glycoprotein in the interphotoreceptor matrix of bovine, human, monkey, and rat eyes. It may transport retinol between the retinal pigment epithelium and the neural retina. In light-reared Royal College of Surgeons (RCS) and RCS retinal dystrophy gene (rdy)+ rats, the amount of IRBP in the interphotoreceptor matrix increased in corresponding proportion to the amount of total rhodopsin through postnatal day 22 (P22). In the RCS-rdy+ rats, the amount increased slightly after P23. However, in the RCS rats there was a rapid fall in the quantity of IRBP as the photoreceptors degenerated between P23 and P29. No IRBP was detected by immunocytochemistry in rats at P28. The amount of rhodopsin fell more slowly. Although retinas from young RCS and RCS-rdy+ rats were able to synthesize and secrete IRBP, this ability was lost in retinas from older RCS rats (P51, P88) but not their congenic controls. The photoreceptor cells have degenerated at these ages in the RCS animals, and may therefore be the retinal cells responsible for IRBP synthesis. The putative function of IRBP in the extracellular transport of retinoids during the visual cycle is consistent with a defect in retinol transport in the RCS rat reported by others.

The ultrastructure of rat rod synaptic terminals: effects of dark-adaptation.

The ultrastructure of synaptic terminals of rat rod photoreceptors was studied in light-adapted and dark-adapted states. In the light-adapted state, horizontal cell processes embedded in the photoreceptor terminal show smooth surface contours and are ovoid in cross section. In the dark-adapted state, fingerlike protrusions of photoreceptor cytoplasm extend into the horizontal cell processes on either side of the synaptic ribbon. An electron-dense structure lies under the horizontal cell membrane that surrounds these fingerlike protrusions. This submembrane specialization is generally associated with areas of concavity of the horizontal cell membrane; it occurs in goldfish rods as well as rat rods. The protrusions and the horizontal cell specialization may be involved in membrane dynamics during synaptic activity.

Synaptic organization of neurotensin immunoreactive amacrine cells in the chicken retina.

Immunohistochemistry was utilized to investigate the light and electron microscopic localization of neurotensinlike immunoreactive (NT) amacrine cells in the chicken retina. The NT cells possess oval cell bodies (7 microns in diameter) that are located in either the second or third tier of cells from the border of the inner nuclear and inner plexiform layers. The processes of such cells extend into the inner plexiform layer where they ramify as a narrow plexus in sublamina 1 and as a broad plexus in sublaminas 3 and 4. Additionally, stained processes are observed occasionally within sublamina 5. At the ultrastructural level, NT-positive somas exhibit a rather dense and evenly distributed peroxidase reaction product throughout their cytoplasm. The nucleus of NT amacrine cells possess a round, unindented nuclear membrane. NT-immunoreactive processes in the inner plexiform layer interact synaptically only with non-NT cells. NT processes receive synaptic input mainly from the processes of amacrine cells and to a lesser degree from bipolar cells. The large majority of NT-stained varicosities form presynaptic contacts onto the processes of amacrine cells, but are also presynaptic to bipolar cell axon terminals. Moreover, each of the above synaptic relationships can be identified in each of sublaminas 1 and 3 to 4 of the inner plexiform layer. In addition, NT processes are presynaptic to processes devoid of synaptic vesicles that may originate from ganglion cells. Finally, NT processes occasionally form synaptic contacts onto somas situated in the most proximal row of the inner nuclear layer.

Localization of serotoninlike-immunoreactive amacrine cells in the larval tiger salamander retina.

Light microscopic immunocytochemistry was used to study the populations of serotoninlike-immunoreactive cells in the larval tiger salamander retina. Of 1,135 serotonin-immunostained cells observed in transverse cryosections, 87% were identified as amacrine cells, whereas 13% were tentatively designated as displaced amacrine cells. The somas of the vast majority of serotonin-amacrine cells were situated in the innermost cell row of the inner nuclear layer. Only a few serotonin-immunostained amacrine cell somas were observed in the second row of cells from the inner nuclear layer. Serotonin-immunoreactive processes generally appeared as a diffuse plexus distributed evenly throughout all levels of the inner plexiform layer. As determined in whole-mount preparations, serotonin-amacrine cells were divisible into two populations on the basis of the diameters of their somas. Large cells (45%) ranged from 16 to 19 microns in diameter with the vast majority measuring 17-18 microns. Smaller and sometimes less intensely stained cells ranged from 14 to 16 microns in diameter with the large majority measuring 15 microns. The diameters of serotonin-displaced amacrine cells ranged from 19 to 22 microns with the large majority measuring 20 microns in diameter. An examination of whole-mount retinas revealed that serotonin-immunoreactive amacrine and displaced amacrine cells were distributed throughout the center and the periphery of the retina. The density of serotonin-amacrine cells (large and small combined) was calculated to be 173 +/- 4.5 (mean +/- standard error) cells per mm2.