Immunocytochemical staining of cholinergic amacrine cells in rabbit retina (original) (raw)
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Neurons immunoreactive to choline acetyltransferase in the turtle retina
Vision Research, 1992
Light microscopic immunocytochemistry using anti-choline acetykransferase (ChAT) was performed to stain putative cholinergic amacrine cells in turtle retina. C&AT-immunoreactive somata lie in the inner nuclear (INL) and ganglion cell (GCL) layers. Three types of amacrine cells were found according to the location of their somata and their dendritic stratifkation pattern in the inner plexiform layer (IPL). Type I amacrines lie in the row of cells closest to the INL/IPL limits and they branch along the sl/s2 border of the IPL. Type II amacrines are displaced to the GCL and they ramify along the s3/s4 border of the IPL. Type III amacrines lie in the middle of the INL, 2-3 rows away from the IPL limits and their dendrites appear to be bi-or t&strati&d in sl and s3-4 of the IPL. The turtle ChAT-IR amacrines are thus similar to the types described in chicken retina. A regubu, non-random mosaic formed by stained type II amacrine cells was ohserved in the GCL. Their density in mid-central retina was 750 cells/mm*, tapering off to 393 cells/mm' in peripheral retina. Our study indicates that a pair of cholinergic amacrine cell types in turtle retina is arranged in mirror-image symmetry contributing to sublamina "a" and subhunina "b" of the IPL, l&e in other vertebrate retinas. Turtle retina ChAT-immunoreactive neurons Amacrine cells
Choline acetyltransferase is expressed by non-starburst amacrine cells in the ground squirrel retina
Brain Research, 2003
We have used immunostaining techniques to reveal a new type of amacrine cell that is immunoreactive for choline acetyltransferase (ChAT), the acetylcholine synthesizing enzyme, in the Ground Squirrel (Spermophilus beecheyi) retina. Cryostat sections and double immunostained wholemount preparations were examined by confocal microscopy. This new ChAT type III cell is distinct in morphology and neurotransmitter content from the well know 'starburst' amacrine cells (types I and II) that are so well represented in the ground squirrel retina [J. Comp. Neurol. 365 (1996) 173-216]. The type III cell colocalizes glycine with the acetylcholine and does not appear to be GABAergic or exhibit calcium-binding proteins like the well-known starburst type. As well, type III cells do not occur as a mirror-symmetric pair with normally placed and displaced varieties. The type III cell is probably a small field amacrine type branching broadly in upper sublamina b of the inner plexiform layer, and is most likely A6 of the Ground Squirrel retina [J. Comp. Neurol. 365 (1996) 173-216]. Type III cells are ideally placed in the architecture of the Ground Squirrel retina to influence ON directionally selective ganglion cell types. Published by Elsevier Science B.V.
Developmental Brain Research, 1991
Retinas from embryonic day 14 (El4) Sprague-Dawley rats were transplanted to the tectum of newborn (P0) recipient rats, and the distribution pattern of choline acetyltransferase immunoreactivity (ChAT-I) in developing transplants was studied and compared with those observed in the retinas of normal developing rats. In normal retinas, ChAT-I cells were first identified in restricted regions in the ganglion cell layer (GCL) at P4, but were found to cover the entire GCL by P6. A second population of ChAT-I cells was detected in the inner nuclear layer (INL) at P8, and they were observed in most parts of the INL on P10 when two immunoreactive sublaminae began to appear in the inner plexiform layer (IPL). The adult pattern of having two distinct populations of ChAT-I cells, organized in mirror symmetrical fashion in the inner retinal layers was basically established by P12. The time course of development and overall distribution pattern of ChAT-I cells in developing retinal transplants on the whole were very similar to those observed in normal retinas. The first identification of these cells and the establishment of their final distribution pattern were made at stages corresponding to P4 and P12 of normal developing retinas respectively. However, ChAT-I somata were located in the INL at a much earlier stage compared with their counterparts in the normal retina, and a transient population of immunoreactive cells with their processes extending to retinal layers other than the IPL was observed in some transplants from P6 to P10. These features were not observed in normal developing retinas. These results suggest that the development of cholinergic neurons, especially the expression of their characteristic antigen and their final distribution pattern is largely determined by programmes which arc intrinsic to the orginal retinal tissue, despite some minor deviation or variation in the developmental process which may occur under certain abnormal conditions.
Cholinergic amacrine cells of the rat retina express the δ-subunit of the GABAA-receptor
Neuroscience Letters, 1993
Antibodies directed against the ~-subunit of the GABA~,-receptor were applied to cryostat sections of rat retinae. Two narrow bands of the inner plexiform layer were strongly immunoreactive. Some cell bodies in both the amacrine-and ganglion-cell layer were weakly immunoreactive. The position of the labelled bands and the distribution of the cell bodies was strongly reminiscent of the cholinergic amacrine cells. In order to show directly that cholinergic amacrine cells express the 6-subunit of the GABAA-receptor, double immunofluorescence with an antibody against choline acetyltransferase (CHAT) and with antibodies against the ~-subunit was performed on the same cryostat sections. This showed the labelled cells to be cholinergic amacrine cells.
The Journal of neuroscience, 1996
Cholinergic regulation of the activity of rabbit retinal ganglion cells and displaced amacrine cells was investigated using optical recording of changes in intracellular free calcium ([Ca 2ϩ ] i). Labeling of neurons in the mature retina was achieved by injecting calcium green-1 dextran (CaGD) into the isolated retina. Nicotine increased ganglion cell [Ca 2ϩ ] i , affecting every loaded cell in some preparations; the pharmacology of nicotine was consistent with an action at neuronal nicotinic receptors, and specifically it was-(neuronal-)bungarotoxin-sensitive but ␣-bungarotoxin-insensitive. Muscarine also raised [Ca 2ϩ ] i , but it was less potent than nicotine, affecting only a subpopulation of ganglion cells, with an M1-like muscarinic receptor pharmacology. Neither the nicotine-nor muscarine-induced increases of ganglion cell [Ca 2ϩ ] i were blocked by the glutamate receptor antagonists 6,7-dinitroquinoxaline-2,3-dione and aminophosphonopentanoic acid. Therefore, the effects of cholinergic ago-nists on ganglion cell [Ca 2ϩ ] i were not attributable to an indirect effect mediated by glutamatergic bipolar cells. The effects of nicotine and muscarine were abolished in calcium-free solution, indicating that the responses depend on calcium influx. Displaced (Cb) cholinergic amacrine cells were also loaded with CaGD and were identified by selective labeling with the nuclear dye 4Ј,6-diamidino-2-phenyl-indole. Cb amacrine cells did not respond to either nicotine or muscarine, but responded vigorously to the glutamate receptor agonist kainic acid. There is anatomical evidence indicating that cholinergic amacrine cells make synaptic contact with each other, but the present results do not support the hypothesis that communication between these cells is cholinergic.
Intricate paths of cells and networks becoming “Cholinergic” in the embryonic chicken retina
The Journal of Comparative Neurology, 2012
Choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) are the decisive enzymatic activities regulating the availability of acetylcholine (ACh) at a given synaptic or nonsynaptic locus. The only cholinergic cells of the mature inner retina are the so-called starburst amacrine cells (SACs). A type-I SAC, found at the outer border of the inner plexiform layer (IPL), forms a synaptic subband ''a'' within the IPL, while a type-II SAC located at the inner IPL border projects into subband ''d.'' Applying immunohistochemistry for ChAT and AChE on sections of the chicken retina, we here have revealed intricate relationships of how retinal networks became dominated by AChE or by ChAT reactivities. AChE þ cells were first detectable in an embryonic day (E)4 retina, while ChAT appeared 1 day later in the very same cells; at this stage all are Brn3a þ , a marker for ganglion cells (GCs). On either side of a faint AChE þ band, indicating the future IPL, pairs of ChAT þ /AChE À / Brn3a À cells appeared between E7/8. Type-I cells had increased ChAT and lost AChE; type-II cells presented less ChAT, but some AChE on their surfaces. Direct neighbors of SACs tended to express much AChE. Along with maturation, subband ''a'' presented more ChAT but less AChE; in subband ''d'' this pattern was reversed. In conclusion, the two retinal cholinergic networks segregate out from one cell pool, become locally opposed to each other, and become dominated by either synthesis or degradation of ACh. These ''cholinergic developmental divergences'' may also have significant physiologic consequences.
2012
The inner plexiform layer (IPL) of the Vertebrate retina is a highly organized synaptic region amassed with a myriad of processes from different cell types, like ganglion cells (GCs), amacrine cells (ACs) and bipolar cells (BCs). Their dendrites stratify at different levels within this cell-free zone, e.g. so-called subbands. Further, Muller glial cells (MCs) span radially through all retinal layers. In this study, I have focused on cholinergic ACs and MCs, and their possible roles in the formation of the IPL in the embryonic chicken retina. In the first chapter I have analysed the in vivo development and differentiation of starburst amacrine cells (SACs) and their role in cholinergic stratification. Type-I and type-II SACs are mirror-symmetrically arranged ACs on both sides of the IPL, which send their processes into the latter, where they stratify as subbands ‘a’ and ‘d’. Acetylcholine is the predominant neurotransmitter of these cells, e.g. they are cholinergic. Using the choline...