Dendritic morphology and retinal distribution of tyrosine hydroxylase-like immunoreactive amacrine cells in Bufo marinus

Summary Tyrosine hydroxylase-like immunoreactive (TH-IR) amacrine cells (ACs) in the retina of metamorphosing and adult Bufo marinus were visualized, and their retinal distribution established, using immunohistochemistry on retinal wholemount and sectioned material. The somata of TH-IR ACs were located in the innermost part of the inner nuclear layer (INL). Their dendrites branched predominantly in the scieral sublamina of the inner plexiform layer (IPL), with sparse branching also in the vitreal sublamina. In the retinae of metamorphosing animals 592 ± 113 (mean ± S.D.) immunoreactive cells and in adult 5,670 ± 528 cells were found. Usually 1, 2 or 3 stem dendrites arose from the somata of TH-IR cells which branched 2 or 3 times. In the adult retinae the dendritic field sizes of immunoreactive cells were in the range of 0.059 ± 0.012 mm², which resulted in a considerable dendritic overlap across the retina. TH-IR cells were unevenly distributed over the retina, with 72 cells/mm² in the central temporal retina, 45–50 cells/mm² along the naso-temporal axis of the retina and 25 cells/mm² in the dorsal and ventral peripheral retina. The average density was 36 ± 6 cells/mm². A considerable number of TH-IR cells (range 52–133, n=4) were displaced into the ganglion cell layer (GCL) of the retina. The mean soma sizes of immunoreactive cells were significantly higher in the low density (95 ± 13 m²) than in the high cell density areas (86 ± 12 m²). There was also a slight but significant increase of the dendritic field sizes of these cells towards the low cell density areas of the retina. These observations show that the retinal distribution of TH-IR ACs parallels the non-uniform distribution of neurons of the INL demonstrated recently in Bufo marinus (Zhu et al. 1990). The class of TH-IR ACs appears to correspond to a subgroup of morphologically distinct dopaminergic ACs found in a number of other vertebrate species.On leave from Department of Anatomy, Zhanjiang Medical College, Guangdong, People's Republic of China     

Distribution of tyrosine hydroxylase and dopamine immunoreactivities in the brain of the South African clawed frog Xenopus laevis

Summary The distribution of dopamine (DA) and the biosynthetic enzyme tyrosine hydroxylase (TH) has been studied immunohistochemically in the brain of the adult South African clawed frog, Xenopus laevis. The goals of the present study are, firstly, to provide detailed information on the DA system of the brain of a species which is commonly used in laboratories as an experimental model and, secondly, to enhance our insight into primitive and derived characters of this catecholaminergic system in amphibians. Dopamine-immunoreactive cell bodies are present in the olfactory bulb, the preoptic area, the suprachiasmatic nucleus, the nucleus of the periventricular organ and its accompanying cells, the nucleus of the posterior tubercle, the posterior thalamic nucleus, the midbrain tegmentum, around the solitary tract, in the ependymal layer along the midline of the caudal rhombencephalon, and along the central canal of the spinal cord. In contrast to the DA antiserum, the TH antiserum fails to stain the liquor-contacting cells in the periventricular organ. On the contrary, the latter antiserum reveals additional immunoreactive cell bodies in the olfactory bulb, the isthmic region and the caudal brainstem. Both antisera yield an almost identical distribution of fibers. Distinct fiber plexuses are observed in the olfactory bulb, the basal forebrain, the hypothalamus and the intermediate lobe of the hypophysis. Features that Xenopus shares with other anurans are the larger number of DAi cells, which are generally smaller in size than those observed in urodeles, and the lack of DAi fibers in pallial structures. On the other hand, the paired midbrain DA cell group and the innervation of the tectum of Xenopus resemble those found in the newt rather than those in frogs. Despite the existence of these species differences, the brain of Xenopus offers an excellent model for studying general aspects of neurotransmitter interactions and the development of catecholamine systems in this class of vertebrates.     

The light microscopic localization of substance P- and somatostatin-like immunoreactive cells in the larval tiger salamander retina

Summary Light microscopic immunocytochemistry was utilized to localize the populations of substance P (SP)- and somatostatin (SOM)-like immunoreactive cells in the larval tiger salamander retina. Of 104 SP-immunostained cells observed, 82% were Type 1 amacrine cells. Another 8% of the SP-cells were classified as Type 2 amacrine cells, while 10% of the SP-cells had their cell bodies located in the ganglion cell layer and were designated as displaced amacrine cells. Each type of SP-like immunoreactive cell was observed in the central and peripheral retina. SP-immunopositive processes were observed in the inner plexiform layer as a sparse plexus in sublamina 1 and as a denser network of fibers in sublamina 5. Seventy-eight percent of the 110 somatostatin-immunopositive cells observed were designated as Type 1 amacrine cells. Another 12% of SOM-cells were classified as displaced amacrine cells, while only two SOM-immunopositive Type 2 amacrine cells were observed. Nine percent of the SOM-cells were designated as interplexiform cells, based on their giving rise to processes distributing in the outer plexiform layer as well as processes ramifying in the inner plexiform layer. Each type of SOM-immunoreactive cell was observed in the central and peripheral retina, with the exception of the Type 2 amacrine cells, whose somas were only found in the central retina. Lastly, SOM-immunopositive processes in the inner plexiform layer appeared as a fine plexus in sublamina 1 and as a somewhat denser network of fibers in sublamina 5.     

Bistratified amacrine cells in the retina of the tammar wallaby — Macropus eugenii

Summary Cajal (1911) noted that bistratified amacrine cells were common in non mammalian species and extremely rare in the mammalian retina. An examination of the marsupial retina of the tammar wallaby, stained with a modified Golgi procedure, revealed that a particular type of bistratified amacrine was frequently impregnated with the silver stain. Flat mount and transverse sections showed that the morphology of this cell did not correspond with any of the species-dependent bistratified amacrines reproduced in Cajal's drawings. Instead, the cell appeared to be almost identical to the AII or rod amacrine that has been observed in a number of mammalian retinas. The relative frequency with which the cell appears in our material, and its confirmed rod input in other species, are both consistent with the grazing habits of the tammar wallaby which is a crepuscular animal that does most of its feeding at dusk and after dark.     

Morphological characteristics of dopaminergic immunoreactive neurons in the olfactory bulb of the common marmoset monkey (Callithrix jacchus)

The present study describes the distribution of tyrosine hydroxylase (TH)-immunoreactive (IR) elements in the olfactory bulb of the common marmoset monkey (Callithrix jacchus), a primate species by immunohistochemistry. We identified six layers of the olfactory bulb of the common marmoset monkey in sections stained with cresyl violet. The majority of TH-IR cells were found in the glomerular layer. A few TH-IR cells were present in the external plexiform and granule cell layers. TH-IR fibers were identified in all layers of the olfactory bulb. The density of these nerve fibers was high in the internal plexiform and granule cell layers. The results in the olfactory bulb of the common marmoset monkey are generally similar to previous reports in some mammals. These data suggest that TH in the olfactory bulb of the common marmoset monkey may play a role in olfactory transmission via the glomeruli like in other mammals.     

The generation and changing retinal distribution of displaced amacrine cells in Bufo marinus from metamorphosis to adult.

Summary The generation and retinal distribution of displaced amacrine cells (DAs) were studied from metamorphosis to adult in the cane toad Bufo marinus. Displaced amacrine cells were identified by inducing chromatolytic changes in ganglion cells (GCs) following optic nerve section. Cells that did not chromatolyse in the ganglion cell layer (GCL) of the retina were regarded as DAs. The number of DAs increased considerably from an estimated 10000 at metamorphosis to 211000 in the adult toad, and was accompanied by a substantial decrease of average cell density. In contrast to the reported 6:1 cell density gradient of all cells of the GCL in adult toad (Nguyen and Straznicky 1989) only a shallow 1.6:1 density gradient of DAs from the visual streak to the dorsal and ventral retinal margins was detected. Consequently, the incidence of DAs increased from 15% of all cells of the GCL in the visual streak to 30% in the dorsal and ventral peripheral retina. These results indicate that the ratio of the newly generated DAs and GCs at the ciliary margin must be changing during development. More GCs are generated before and around metamorphosis then DAs, in contrast to the relative increase of the percentage of DAs generated after metamorphosis. The possible control of the numbers of DAs in the GCL is discussed.     

Morphological classification and retinal distribution of large ganglion cells in the retina ofBufo marinus

Summary The retrograde transport of horseradish peroxidase (HRP) and cobaltic-lysine complex (CLC) was used to morphologically characterize large ganglion cells (GCs) and to determine their distribution in retinal wholemounts and in sectioned material in the retina ofBufo marinus. Large GCs, amounting to about 0.5% of total GC population, were defined to be those with very large dendritic field sizes varying between 0.1 mm² to 0.6 mm² and cell soma sizes of between 100 m² to 400 m². These cells were subdivided into 3 major groups, Types I, II and III, on the basis of their dendritic field sizes, arborization patterns and the strata of dendritic branching within the inner plexiform layer (IPL). The majority of large neurons (about 90%) were classified as Type I GCs with symmetrical dendritic arbor. These cells had either bistratified branching in the scierai and vitreal sublaminae of the IPL (65% of Type I Cells) or unistratified branching in the scleral (26%) or in the vitreal (9%) sublamina. Their dendritic field sizes increased linearly from the retinal centre from 0.13 mm±0.02 mm² (mean and S.D.) to 0.58±0.11 mm² in the retinal periphery. Type II GCs (about 9% of the large GC population) were characterized by an asymmetrical dendritic arborization directed towards the ciliary margin with unistratified branching in the scierai sublamina of the IPL. The mean dendritic field sizes of these cells were 0.26±0.09 mm². Type III GCs, the least frequent (about 1%) category of large GCs had sparsely branching, elongated dendritic branching aligned approximately parallel with the nasotemporal axis of the retina. The unistratified dendritic branches of these neurons were located in the vitreal sublamina of the IPL with a mean dendritic field size of 0.42±0.11 mm². The dendritic field sizes of Types II and III GCs did not increase with retinal eccentricity. Type I GCs were distributed unevenly across the retina, the density being greatest in the visual streak, along the nasotemporal meridian of the retina. The dendritic field sizes of these cells increased towards the retinal periphery, resulting in a constant dendritic field coverage factor across the retina. Each retinal point was covered by the dendritic fields of 4–5 adjacent GCs. In contrast, Types II and III GCs had only discontinuous dendritic coverage. The identification of morphological types of large GCs with previously described functional classes of GCs in the anuran retina is discussed.On leave from the Department of Anatomy, University Medical School, Pécs, Hungary     

Tyrosine hydroxylase-like immunoreactive neurons in the olfactory bulb of the snake,Elaphe quadrivirgata,with special reference to the colocalization of tyrosine hydroxylase- and GABA-like immunoreactivities

Summary The distribution and structural features of tyrosine hydroxylase-like immunoreactive (TH-LI) neurons were studied in the olfactory bulb of a snake, Elaphe quadrivirgata, by using pre-and post-embedding immunocytochemistry at the light microscopic level. In contrast to rodent olfactory bulbs previously reported, many TH-LI neurons were seen not only in the main olfactory bulb (MOB) but also in the accessory olfactory bulb (AOB). With regard to the TH-like immunoreactivity, there appeared no appreciable differences between MOB and AOB. As in mammalian MOB, the majority of TH-LI neurons were clustered in the periglomerular region and appeared to send their dendritic branches into glomeruli, which as a whole make an intense TH-LI band in the glomerular layer (GML). In the external plexiform/mitral cell layer (EPL/ML) of MOB and AOB as well as in the outer sublamina of the internal plexiform layer (OSL) of AOB, an appreciable number of TH-LI neurons were scattered, extending dendritic processes which appeared to make a loose meshwork. TH-LI neurons in EPL/ML (including OSL) appeared to consist of at least two morphologically different types. The first had a small perikaryon and one or two smooth dendrites which usually extended to GML and were frequently confirmed to enter into glomeruli. The second had a larger perikaryon and 2–3 dendrites which branched into several varicose processes extending in EPL/ML/OSL but appeared not to enter into glomeruli. The TH-like immunoreactivity was rarely seen in the internal plexiform layer and internal granule cell layer. The colocalization of GABA-like and TH-like immunoreactivities was further studied. Almost all TH-LI neurons in both EPL/ ML/OSL and GML contained GABA-like immunoreactivity irrespectively of the type of TH-LI cells.Abbreviations in Figures AOB accessory olfactory bulb - MOB main olfactory bulb - Hem hemisphere - ON olfactory nerve layer - VN vomeronasal nerve layer - GM glomerular layer - EP/M external plexiform layer/Mitral cell layer - IP internal plexiform layer - IG internal granular layer - OS outer sublamina of the IPL of AOB - MS middle sublamina of the IPL of AOB - IS inner sublamina of the IPL of AOB     

Differential distribution of a second type of tyrosine hydroxylase immunoreactive amacrine cell in the chick retina

A second population of tyrosine hydroxylase-immunoreactive amacrine cells was demonstrated in embryonic and adult chicken retinas by immunohistochemistry techniques in whole flat-mount preparations. The populations were differentiated on a basis of different immunostaining intensities, levels of stratification in the inner plexiform layer, and topographical distributions. Cells of one type were similar to the previously described dopaminergic amacrine cells, denoted here as tyrosine hydroxylase type 1 cells. Immunoreactive neurons of the second type observed in the present work had relatively smaller somata size, and weaker immunostaining than type 1 cells, and were located preferentially in the ventral retina. These tyrosine hydroxylase type 2 cells could be visualized from embryonic day 14 to 21 days after hatching animals. The distribution of the second population was coincident with that of the targets of centrifugal fibres and with cells involved in long proprioretinal connections. We propose that the tyrosine hydroxylase type 2 amacrine cells found in the ventral retina could mediate an important pathway to the upper half of the visual field so as to aid in the detection of predators.     

Expression of aquaporin-9 immunoreactivity by catecholaminergic amacrine cells in the rat retina

Aquaporins are involved in the maintenance of ionic and osmotic balance in the central nervous system and in the eye. Whereas the expression of aquaporin-9 immunoreactivity in the brain has been described for catecholaminergic neurons and glial cells, the expression of aquaporin-9 in the neural retina is unclear. We examined the distribution of aquaporin-1 and -9 immunoreactivities in retinas of the rat. Aquaporin-9 immunoreactivity was expressed exclusively by tyrosine hydroxylase (TH) positive amacrine cells, while aquaporin-1 immunoreactivity was expressed by photoreceptor cells and by TH negative amacrine cells. The staining pattern of aquaporin-9 did not change up to 4 weeks after pressure-induced transient retinal ischemia. It is concluded that catecholaminergic, putatively dopaminergic, amacrine cells of the retina express aquaporin-9.     

Identification and characterization of SMI32-immunoreactive amacrine cells in the mouse retina

Mammalian neurons express the neural intermediate filament protein neurofilament (NF). In the retina, NFs have been detected primarily in the axons and processes of retinal ganglion and horizontal cells. We found an amacrine cell type that was immunolabeled with an antibody against SMI32, a non-phosphorylated epitope on neurofilament proteins of high molecular weight, in the mouse retina. This type of amacrine cell was non-randomly distributed, and these cells exhibited a central-peripheral density gradient. Most of these cells co-expressed GABA and ChAT, but not glycine or any other amacrine cell marker. These results suggest that some SMI32-immunoreactive amacrine cells belong to a GABAergic population, and that SMI32 can therefore be used as a marker for a subset of amacrine cells in addition to ganglion cells and horizontal cells in the mouse retina.     

The periphery effect in cat retinal ganglion cells: variation with functional class and eccentricity

Summary We have studied the responses of ganglion cells of the cat retina to visual stimulation remote from the center of their receptive field. Following previous work, this response is termed the periphery effect (PE). Cells were identified as Y-, X- or W-class from the latency of their response to optic chiasm stimulation and from their receptive field properties. The strength of the PE elicited by a rotating windmill or counterphased grating stimulus was measured for ganglion cells of all major classes. The PE was consistently stronger in Y- than in X-cells, and the strength of the effect in both X- and Y-cells increased significantly with retinal eccentricity. A PE was elicited from about 47% of W-cells studied. In some (36%) the effect was excitatory, as for X- and Y- cells; in others (11%) it was inhibitory. Despite this heterogeneity, the PE in W-cells increased significantly with eccentricity. These variations of the PE with eccentricity and cell class have implications for the circuitry of the inner plexiform layer.     

大鼠纹状体边缘区多巴胺免疫阳性反应物的分布

目的　研究纹状体边缘区多巴胺和酪氨酸免疫阳性物质的分布。方法　用免疫细胞化学ABC法检测多巴胺和酪氨酸羟化酶免疫阳性物质在大鼠纹状体边缘区的分布。结果　多巴胺和酪氨酸羟化酶免疫阳性纤维在边缘区中密集分布形成一条明显的带状 ,带的宽度和位置与边缘区一致。结论　证明了纹状体边缘区中存在密度较高的多巴胺和酪氨酸羟化酶免疫阳性纤维 ,并讨论了这些免疫阳性纤维的来源以及与学习记忆的关系。     

A double-label analysis demonstrating the non-coexistence of tyrosine hydroxylase-like and GABA-like immunoreactivities in amacrine cells of the larval tiger salamander retina.

Previous studies have localized tyrosine hydroxylase, the rate-limiting enzyme for the production of dopamine, and gamma-aminobutyric acid (GABA) to amacrine cell populations in the larval tiger salamander retina. Double-label immunocytochemistry was used to examine if tyrosine hydroxylase-like and GABA-like immunoreactivities colocalize in tiger salamander amacrine cells. A total of 2,162 tyrosine hydroxylase-like immunoreactive amacrine cells were observed in double-labelled sections. None of these cells were observed to express GABA-like immunoreactivity. Therefore, the present study demonstrates that dopamine and GABA are localized to distinct neuronal populations in the larval tiger salamander retina.     

Two modes of free migration of amacrine cell neuroblasts in the chick retina

Summary The migration of amacrine neuroblasts toward the prospective amacrine cell layer in the chick embryo retina has been studied, in Golgi-stained sections, between days 5 and 9 of embryogenesis.Two distinct populations of presumptive amacrine neuroblasts have been identified on the basis of their shape and migratory behavior. One population (smooth amacrine neuroblasts) display smooth, monopolar or bipolar contours, moving freely across the retina without major changes in the original postmitotic shape, and give processes only after reaching the primitive inner plexiform layer. The second population (multipodial amacrine neuroblasts) includes multipolar neuroblasts with abundant filiform and/or lamelliform processes sprouting in various directions; these highly plastic cells begin modifying their shapes at the time of release from the ventricular lining and continue to do so as they move toward their definitive location.Thus, the well-known heterogeneity of adult amacrine cells seems to be preluded by differences in neuroblastic migratory patterns, suggesting the existence of at least two different subsets of amacrine cell precursors.     

Two types of tyrosine hydroxylase-immunoreactive neurons in the zebrafish retina

The purpose of the present study is to identify the dopaminergic amacrine (DA) cells in the inner nuclear layer (INL) of zebrafish retina through immunocytochemistry and quantitative analysis. Two types of tyrosine hydroxylase-immunoreactive (TH-IR) cells appeared on the basis of dendritic morphology and stratification patterns in the inner plexiform layer (IPL). The first (DA1) was bistratified, with branching planes in both s1 and s5 of the IPL. The second (DA2) was diffuse, with dendritic processes branched throughout the IPL. DA1 and DA2 cells corresponded morphologically to A_on^−s1/s5 and A_diffuse⁻¹ (Connaughton et al., 2004). The average number of total TH-IR cells was 1088 ± 79 cells per retina (n = 5), and the mean density was 250 ± 27 cells/mm². Their density was highest in the mid central region of ventrotemporal retina and lowest in the periphery of dorsonasal retina. Quantitatively, 45.71% of the TH-IR cells were DA1 cells, while 54.29% were DA2 cells. No TH-IR cells expressed calbindin D28K, calretinin or parvalbumin, markers for the various INL cells present in several animals. Therefore the TH-IR cells in zebrafish are limited to very specific subpopulations of the amacrine cells.     

Localization of neurotensin-like immunoreactive amacrine cells in the larval tiger salamander retina

Summary Light microscopic immunocytochemistry was used to localize the populations of NT-like immunoreactive amacrine cells in the larval tiger salamander retina. Seventy-nine percent of NT-immunostained cells observed in transverse cryo-prepared sections were classified as Type 1 amacrine cells. Another 6% were classified as Type 2 amacrine cells, while 15% of the NT-cells had their cell bodies situated in the ganglion cell layer and were tentatively designated as displaced amacrine cells. Each type of NT-like immunoreactive cell was observed in the central and peripheral retina. NT-immunostained processes were observed to ramify in sublayers 3 and 5 of the inner plexiform layer. An examination of retinal whole mounts revealed that NT-amacrine cells were distributed throughout the center and periphery of the retina at a density of 82 ± 24 cells/ mm². The dendritic fields of NT-immunostained amacrine and displaced amacrine cells were observed to be either symmetrically or asymmetrically distributed about their somas. Symmetrical dendritic fields were generally oval-shaped and ranged in diameter from 250 to 500 m (major axis) by 150 to 250 m (minor axis). Asymmetrical dendritic fields were observed to encompass one-half or less of an imaginary circle surrounding their soma of origin and were orientated in all directions. The processes forming asymmetrical dendritic fields ranged from 75 to 260 m in length. Furthermore, partial overlap was often observed between the dendritic fields of adjacent NT-amacrine cells.     

Nogo-A在小鼠胚胎新生视网膜节细胞及其轴突的表达

目的 研究小鼠胚胎阶段Nogo-A在视网膜节细胞(RGCs)及其轴突上的表达及时程变化. 方法 取不同发育阶段的小鼠胚胎,采用免疫荧光染色,以激光扫描共焦显微镜观察Nogo-A在视觉传导通路中的表达.并采用免疫双标染色确定视网膜中表达Nogo-A蛋白的细胞类型. 结果 在视网膜发育的早期阶段(E12),Nogo-A密集表达于具有放射状形态的细胞上,Nogo-A免疫阳性产物出现在胞质、胞膜以及轴突上.Nogo-A与Tuj-1双标染色显示,此阶段的视网膜中几乎所有RGCs及其轴突都表达有Nogo-A;在稍晚的发育阶段(E13),视网膜中表达Nogo-A的RGCs数量明显减少,且仅出现在节细胞层以外的室周带和睫状体边缘区.在视网膜的神经纤维层,大部分RGCs轴突不再表达Nogo-A,仅有少量视觉纤维为Nogo-A免疫阳性;RGCs的神经发生基本完成后(E15), 视网膜中几乎检测不到Nogo-A免疫阳性的细胞,但视网膜纤维层仍有少量表达Nogo-A的节细胞轴突.与之类似,视神经盘、视茎、视交叉和视束都观察到少量Nogo-A免疫阳性的轴突.值得注意的是,视束中表达Nogo-A的纤维集中位于表浅部位,而此处恰为新近到达轴突的通过部位. 结论 Nogo-A在视网膜RGCs以及轴突上表达的时程变化和位置特点提示,新生RGCs及其轴突表达Nogo-A,成熟后RGCs内Nogo-A的表达则下调.推测新生RGCs及其轴突中表达的Nogo-A可能与减少轴突分叉等细胞的内在功能有关.