Choline Acetyltransferase and Cholinesterases in the Developing Xenopus Retina

To understand the developmental regulation of acetylcholine (ACh) synthesis in the Xenopus retina, the properties of choline acetyltransferase (CAT) and cholinesterase (ChE), as well as histochemical localization of ChE in the retina, were studied during development. CAT activity first became detectable in the developing eyecup at stages 35/36. This was followed by a rapid, 50-fold rise in specific activity between stages 35/36 and 44. Since this rapid rise coincided with an almost identical increase in total ACh synthesis in whole retinae found in previous studies, it is suggested that this increase was sufficient to account for the rapid increase in total ACh synthesis. Moreover, it also correlated with increased rates of synaptogenesis in both the inner and the outer plexiform layers. Total ChE was resolved into specific and nonspecific ChE by the use of tetraisopropylpyrophosphoramide. Total ChE activities first became detectable at stages 35/36. Specific ChE [acetylcholinesterase (AChE)] increased from 50% at stage 39 to 95% of total ChE activities at stage 66. Again, the most rapid increase in both total ChE and AChE activities occurred between stages 35/36 and 44. Histochemical studies showed that AChE was localized predominantly in the two plexiform layers, with the inner plexiform layer more heavily stained at all stages. Moreover, a stratified staining pattern, clearly discerned in the inner plexiform layer, also correlated with synaptogenesis during this early period of retinal development.     

Localization of Acid and Alkaline Phosphatases in Staphylococcus aureus

The localization of acid and alkaline phosphatases in Staphylococcus aureus was studied by fractionation of cells after treatment with the L-11 enzyme and by electron microscopic histochemistry. The two enzyme activities were located in distinctly different positions at the surface of the cells. Acid phosphatase appeared to be localized around the cell membrane of the bacteria, because the enzyme was recovered exclusively in the membrane fraction and because deposition of lead phosphate was detected by electron microscopic histochemistry on the inner surface of the cell membrane of intact bacteria and spheroplasts. The highest specific activity of alkaline phosphatase was also associated with the membrane fraction. However, on electron microscopic histochemistry of intact cells, the deposition of lead phosphate was only seen on the outer surface of the cell wall.     

猫视网膜年龄相关的形态学变化

取老年猫(12龄,3～3．5kg)和青年猫(1～3龄,2～2．5kg)各4只的视网膜,经4％多聚甲醛处理后,用H．E．染色以显示视网膜结构,Nissl染色显示神经节细胞,免疫组织化学ABC法染色以显示星形胶质细胞特征性标志物胶质纤维酸性蛋白(GFAP)的阳性反应细胞的分布。显微镜下观察测量视网膜厚度,计数神经节细胞、GFAP免疫反应阳性细胞数。与青年猫比较,老年猫视网膜总厚度以及外核层、外网状层、内核层和内网状层厚度均显著减小;神经节细胞层的细胞密度显著下降;GFAP免疫反应阳性细胞显著增加,GFAP阳性细胞阳性反应强,胞体明显膨胀,突起稠密粗大。推测在衰老过程中视网膜细胞有神经元丢失现象,可能是造成视觉功能衰退的重要原因之一;视网膜星形胶质细胞的功能增强可能会延缓衰老。     

Neurofibrillar long-range amacrine cells in mammalian retinae

A distinct population of wide-field, unistratified amacrine cells are shown to be selectively stained by using neurofibrillar methods in rabbit and cat retinae. Their cell bodies may be located in the inner nuclear, inner plexiform or ganglion cell layers and they branch predominantly in stratum 2 of the inner plexiform layer. Characteristically, each cell has two or more long-range distal processes which extend for 2-3 mm beyond a more symmetrical, proximal dendritic field of 0.6-0.8 mm diameter. Although the neurofibrillar long-range amacrines account for less than 1 amacrine in 500, they achieve effective coverage of the retina by both the proximal and distal dendrites.     

GABA concentration and GAD activity levels in normal and degenerated retinas from mice

Freeze-dried sections (14 m thick) were prepared from mice with normal (C57BL strain) and degenerated (C3H strain) retinas. GABA concentration and GAD activity were determined in the microsamples (1.8–20 ng dry weight) of retinal layers and sublayers, using an enzymatic amplication reaction, NADP cycling. 1) GABA was distributed over all layers of normal retina with a broad concentration peak covering both inner nuclear and plexiform layers. In contrast, GAD activity was mostly localized in the inner plexiform layer. 2) GABA concentration was similar in one-fourth of the sublayers of each inner nuclear or plexiform layer. GAD activity was highest in the innermost sublayer of the inner nuclear layer. An increasing gradient of GAD activity was present in the inward direction in the inner plexiform layer. 3) In the degenerated retina, lacking in photoreceptors, the inner nuclear and plexiform layers remained, and GABA and GAD levels in these layers were similar to those in normal retina.Special Issue dedicated to Dr. O. H. Lowry.     

On the distributive patterns of ATPase activity and its functional significance in retinae of certain birds.

The present study incorporates the details of distribution of adenosine triphosphatase amongst the various constituents of retinae of Passer, Psittacula, Streptopelia and Athene. The outer segments in all the cases are intensely positive for the enzyme. This is the part where the light strikes first and initiates the visual processes. The nuclear layers are also positive for the enzyme activity. It is interesting to note that inner plexiform layers show clear-out demarcations of various sub-synaptic layers in all the birds except Psittacula. The ganglion cells and optic nerve fibres are also positive for the enzyme.     

GABA and GAD-like immunoreactivity in the primate retina

Summary GABA immunoreactivity was studied and compared with GAD immunoreactivity in the retinae of baboon, cynomolgus monkey and man. The central and peripheral parts of the retinae were investigated separately in cynomolgus monkey and in man. The same kinds of structures were stained with both antisera. Cells with a position corresponding to amacrine cells were stained, as well as processes in the inner plexiform layer and some cells in the ganglion cell layer. The outer plexiform layer and some cells with the position and configuration of horizontal cells also appeared immunoreactive. Staining was also observed in bipolar-like cells, in man most clearly when using the GABA antiserum in sections from the central parts of the retina. It is possible that horizontal cells, as well as bipolar-like cells, may play a previously unsuspected role in GABAergic transmission in the primate retina.     

Somatostatin-like immunoreactivity in the retinae of adult and embryonic chickens

Somatostatin, a tetradecapeptide that inhibits growth hormone release, has a widespread distribution in the central and peripheral nervous systems and other cell types. In the present investigation, the chicken neural retina was studied for the presence of structures exhibiting somatostatin-like immunoreactivity by utilizing an indirect immunofluorescence technique. Controls for specificity of staining were performed on alternate sections. Several types of distinctly labeled neurons and their processes were evident in sections of adult and late embryonic retinae. Cresyl violet staining showed that these neurons, which were scattered peripherally and more numerous centrally, occupied several strata within the inner nuclear, inner plexiform, and ganglion cell layers. Labeled neurites of immunoreactive perikarya coursed within these layers as well, often approaching other immunoreactive cells and fibers. The morphology and position of the somatostatin-containing neurons indicated that these neurons were amacrine, horizontal, or ganglion associational cells. These findings indicate that somatostatin is first detectable in the retina during the late embryonic stages of the chicken.     

Identification of adipose tissue primordia in perirenal tissues of pig fetuses: utility of phosphatase histochemistry

Perirenal adipose tissue samples were obtained from fetuses removed from pregnant (crossbred) sows at 3 stages of gestation (70, 90 and 110 days). Phosphatase histochemistry, succinate dehydrogenase (SDH) histochemistry and factor VIII antigen immunocytochemistry were conducted on fresh-frozen cryostat sections. Age-associated changes in nucleosidediphosphatase (NDPase) reactions in the arteriolar system were correlated with the morphological development of the medial layer of arterioles and arteries. For instance, a strong NDPase reaction in small arterioles was associated temporally with the assumption of a normal smooth-muscle cell morphology and arrangement in the medial layer. Age-associated changes in blood vessel reactions for factor VIII antigen and alkaline phosphatase activity were not correlated with morphological development. In the youngest fetuses, alkaline phosphatase activity was evident in large and small arterioles, but in the oldest fetuses, alkaline phosphatase activity was restricted to the smallest arterioles and vessels associated with them. Arteriolar differentiation was demonstrable with either adenosine triphosphatase (ATPase) or inosine diphosphatase (IDPase) reactions. Primordial stromal cells around differentiated arterioles were reactive for ATPase but not for IDPase activities. In older fetuses, there were large areas that contained ATPase-reactive stromal cells, no adipocytes, differentiated (ATPase and IDPase) arterioles and few capillaries. Positive reactions for SDH were evident in the ATPase-reactive stromal areas that contained no adipocytes. Differentiated adipocytes were SDH- and ATPase-reactive. These data illustrate the utility of differential phosphatase histochemistry to identify adipose tissue primordia.     

Human retinal development: ultrastructure of the inner retinal layers

Studies of the developing human retina from 6.5 to 18 weeks' gestational age (16–156 mm) by light and electron microscopy are concerned with the morphogenesis of neuroblast cells, plexiform layers, and inner limiting membrane. The transient layer of Chievitz is formed posteriorly by 20 mm (7 weeks), inner plexiform by 48 mm (9 weeks), outer plexiform layer by 83 mm (12 weeks), identifiable cones by 83 mm, and rods by 120 mm (15 weeks). Mitotic activity continues posteriorly until 120 mm and was seen in inner layers of the retina until 103 mm (13 weeks). Outer neuroblastic differentiation is marked by diversification from a uniform cell population to one containing at least three cell types differing in their nuclear shape, chromatin pattern, and cytoplasmic characteristics. Differentiating ganglion cells accumulate polysomes, rough endoplasmic reticulum, Golgi complexes, microtubules, and dense bodies. Müller cell bodies in ganglion and inner nuclear layers extend processes between ganglion cells, and radial fibers, containing extensive smooth endoplasmic reticulum, to the vitreal surface. Synapses appear in the inner and outer plexiform layers by 83 mm (12 weeks), and by 120 mm (15 weeks) demonstrate a variety of conventional and ribbon forms similar to those found in the adult. Synaptogenesis therefore begins considerably before the development of photoreceptor outer segments.     

Histoenzymological studies on the distribution of certain phosphatases in the retinae of owlet (Athene brama) and house sparrow (Passer domesticus)

A comparative study on the distribution of alkaline phosphatase (AlP), acid phosphatase (AcP) and 5'-nucleotidase (5-N) amongst the different constituents of retinae of owlet and house sparrow revealed some interesting aspects of the localization of such phosphatases in both the cases. The outer segment of photoreceptors, where light strikes first, are positive for all the phosphatases. Further, areas composed of synapses, reveal activity of the three enzymes. Another interesting aspect is related to the total absence of the activity of AlP and 5-N in the ganglion cells of both the animals. Other sites of phosphatases in various layers have been also identified. The possible metabolic roles of various phosphatases at different sites have been discussed.     

LOCALIZATION OF DEHYDROGENASE AND ACID PHOSPHATASE IN THE ABSCISSION ZONE OF BEAN LEAVES

Leaf abscission in Phaseolus vulgaris L. cv. ‘Contender’ is associated with enzymatic changes during and prior to separation. Deblading resulted in a localized increase in dehydrogenase and acid phosphatase in the abscission zone. Increased enzyme activities were observed 24–48 hr after deblading. In debladed plants separation was complete in 6–8 days. At separation, dehydrogenase activity appeared to decrease and localization was specific to the protective layer, while the petiole side had no activity. In contrast, acid phosphatase activity was observed in some layers of cells on the petiole side after separation. Ethylene treatment promoted abscission and separation occurred in 24–48 hr in both debladed and intact plants. No protective layer was formed during ethylene-induced abscission. Enzymatic changes similar to those observed in debladed control plants were observed with ethylene treatment. Ethylene induced an additional abscission layer between the pulvinus and petiole, where an abscission layer normally does not form. In this ethylene-induced abscission layer, similar enzyme activities were detected.     

Immunohistochemical and quantitative analysis of 5-hydroxytryptamine in the retina of some vertebrates

5-Hydroxytryptamine immunoreactive neurons were found in retinae from chicken, pigeon, frog and goldfish. They were localized among the amacrine cells with a distribution of cell bodies and nerve fibres that varied with the species. In chicken and pigeon, bipolar-like cell bodies were also found in the middle of the inner nuclear layer, sending processes inwards to the inner plexiform layer and outwards to the horizontal cells. The signalling direction of these cells is doubtful. No 5-hydroxytryptamine immunoreactivity was found in retinae from cow, pig, cat, rabbit, guinea-pig, rat or mouse.Quantitative analyses were performed with HPLC on extracts from chicken, pigeon, frog and goldfish retinae. High concentrations were found in goldfish and frog whereas less, about 100 ng/g, was observed in chicken and pigeon.The results suggest that 5-hydroxytryptamine is the transmitter of a set of amacrine cells in cold-blooded vertebrates and perhaps also in birds. The transmitter of the indoleamine accumulating neurons of mammals remains to be further elucidated.     

Organ culture of suckling rat intestine: comparative study of various hormones on brush border enzymes

Jejunal mucosa of 6 d-old rats were cultured for 24 and 48 h in the presence of thyroxine, insulin, pentagastrin, glucagon, epidermal growth factor (EGF) or dibutyryl-A-3:5-MP cyclic with or without dexamethasone (DX). The enzymes were assayed on the purified brush borders. The various agents added alone to the basic culture medium had no effect with the exception of DX on the levels of enzyme activities. Dexamethasone alone induced sucrase, stimulated maltase, and protected other brush border enzyme activities (aminopeptidase, lactase, and alkaline phosphatase). When added to DX-supplemented medium, only the following factors modified the levels of enzymatic activities observed with DX alone. Insulin (10(-6) M) increased maltase, alkaline phosphatase, and lactase activity to a greater extent than DX at 24 h culture, the effect being maintained at 48 h on alkaline phosphatase only. At 48 h culture, both EGF (10(-8) M) and dbcAMP (10(-3) M) decreased DX-induced sucrase activity. The latter agent also depressed DX-stimulated aminopeptidase activity.     

Retinal ganglion cell topography and spatial resolution in the smelt Hypomesus japonicus (Brevoort, 1856)

The present study deals with the topography of retinal ganglion cells (GCs) and spatial resolution in the smelt Hypomesus japonicus. The eyes and retinae were examined by light microscopy and computerized tomography. DAPI labelling was used to visualize cell nuclei in the ganglion cell and inner plexiform layers. Two zones of increased GC density in the nasal and temporal retina were bridged by a horizontal streak with the GC density ranging from 5600 to 8000 cells/mm². The maximum cell density (area retinae temporalis) ranged from 9492 to 14,112 cells/mm², and the total number of GCs varied from 286 x 10³ to 326 x 10³ cells in three individuals. The theoretical anatomical spatial resolution (the anatomical estimate of the upper limit of visual acuity) was minimum in the ventral periphery (smaller fish, 1.43 cpd; larger fish, 1.37 cpd) and maximum in area retinae temporalis (smaller fish, 2.83 cpd; larger fish, 2.41 cpd). The relatively high density of GCs and presence of the horizontal streak and area retinae temporalis in the H. japonicus are consistent with its highly visual behaviour. The present findings contribute to better understanding of the factors affecting the topography of retinal ganglion cells and mechanisms of visual adaptation in fish.     

Responses of the retinal pigment epithelium of the mouse after acute loss of blood

Albino mice were bled through the hearts by cardiac puncture and 1/2, 1/3 and 1/4 mls of blood were taken out from 3 groups of animals. respectively. Half of the experimental animals were reinfused with 5% dextrose 1 h after bleeding. All were killed either 2, 5, 9, 24, 48 or 72 h after bleeding, and the phagosome numbers per 5 pigment cells counted and compared with control retinae. A severe decrease was evident after bleeding and the decrease leveled off 48 h afterwards. Reinfusion with dextrose had a positive beneficial effect.     

A comparative study on the distribution of non-specific esterase amongst the various constituents of retinas of some vertebrates

A wide coverage of the retinae of a large number of animals (Calotes, Varanus, Naja, Athene, Passer, Streptopelia, Psittacula and Funambulus) from the point of view of the histoenzymological distribution of non-specific esterase amongst the various constituents reveals mostly identical patterns. They are as follows: 1. Outer segments - positive in all cases. 2. Outer plexiform layer - equipped with enzymatic activity in all the instances. 3. Inner nuclear layer - thin cytoplasmic rim of the neurons characterized by positive activity; the nuclei of the neurons are completely negative. 4. Inner plexiform layer - this layer is endowed with the enzymatic activity. 5. Ganglion cells - negative in all cases. 6. Nerve fibres of the layer of nerve fibres, situated adjasent to ganglion cells are positive in all the animals; in case of squirrel oligodandroglial cells present in the region have demonstrated activity of a high order. On of the high-lights of the present contribution is demarcation of the inner plexiform layer into three stratified zones, equipped with enzymatic activity in Calotes, Streptopelia, Naja and Funambulus. Such stratifications are not seen in Varanus, Passer and Psittacula. The significance of the various patterns and the equipment of the enzyme in various constituents at various locals have been discussed in relation to the metabolic functions, zone-wise and interzone-wise in visual processes of various animals.     

GABA and GAD-like immunoreactivity in the primate retina

GABA immunoreactivity was studied and compared with GAD immunoreactivity in the retinae of baboon, cynomolgus monkey and man. The central and peripheral parts of the retinae were investigated separately in cynomolgus monkey and in man. The same kinds of structures were stained with both antisera. Cells with a position corresponding to amacrine cells were stained, as well as processes in the inner plexiform layer and some cells in the ganglion cell layer. The outer plexiform layer and some cells with the position and configuration of horizontal cells also appeared immunoreactive. Staining was also observed in bipolar-like cells, in man most clearly when using the GABA antiserum in sections from the central parts of the retina. It is possible that horizontal cells, as well as bipolar-like cells, may play a previously unsuspected role in GABAergic transmission in the primate retina.     

Distribution of Three Enzymes of γ-Aminobutyric Acid Metabolism in Monkey Retina

Abstract: The distributions of glutamate decarboxylase (EC 4.1.1.15), γ-aminobutyric acid transaminase (EC 2.6.1.19), and succinate semialdehyde dehydrogenase (EC 1.2.1.24) were determined in monkey retina. The decarboxylase was almost restricted to the inner plexiform layer. The transaminase was also highest in this layer, but activities were 40% as high in the adjacent third of the inner nuclear layer and in the ganglion cell and fiber layers. Succinate semialdehyde dehydrogenase was distributed very differently. Although it also showed a peak of activity in the inner plexiform layer, there was a second equal peak in the photoreceptor inner segment layer and a smaller peak in the outer plexiform layer, regions where both γ-aminobutyric acid transaminase and glutamate decarboxylase were essentially absent.     

A layered model for visual processing in avian retina

Visual processing in avian retina is interpreted by means of a layered model in which: a) outer layers provide with spatio temporal fast and retarded versions of the stimuli incident on the retina; a possibility is that horizontal cells are involved in isotropically generating the retarded version which is transversally translated; b) prominent specialization of ganglion cells is the result of local non-linear lateral interaction at the inner plexiform layer, mediated by amacrines which return, also isotropically, the translated retarded signals. Small though systematic deviations in the sites of the lateral interaction result in anisotropic but uniform receptive fields for some ganglion cells. A simple though general expressin for the model is derived which includes the various types of recorded avian ganglion retinal cells responses, which also permits a unified interpretation of visual processing in avian and cat's retinae.