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  4. Depletion of cholinergic amacrine cells does not perturb the segregation of on and off cone bipolar cell projections.

Depletion of cholinergic amacrine cells does not perturb the segregation of on and off cone bipolar cell projections.

Gunhan-Agar E, Choudary P, Landerholm TE, Chalupa LM (2000) Depletion of cholinergic amacrine cells does not perturb the segregation of on and off cone bipolar cell projections. Neuroscience 2000 Abstracts 119.3. Society for Neuroscience, New Orleans, LA.

Summary: The pathways signaling onset and offset of light are segregated in the retina with On-cone and Off-cone bipolar cells terminating on the stratified dendrites of On and Off retinal ganglion cells (RGCs). During development the axons of On and Off cone bipolar cells form two strata in the IPL in a remarkably precise manner, without any obvious refinements. Moreover, such a precise ingrowth pattern occurs even after RGCs have been depleted (J. Neurosci., 2000, 201:306-314). Here we show by immunostaining that two bands of cholinergic processes are present in the rat retina as early as P1, some 7 days before the formation of segregated bipolar inputs. Double labeling of retinal sections with the antibody to recoverin (that recognizes On and Off cone bipolar cells) and the antibody for VACHT (which labels cholinergic processes) revealed that the segregated terminals of cone bipolar cells are juxtaposed with the two bands of cholinergic fibers. These observations suggested that the cholinergic fibers could serve as a scaffold for the later ingrowing bipolar cell axons. To test this hypothesis, we devised a novel method for depleting retinal cholinergic amacrine cells with a VACHT-saporin immunotoxin. A single treatment of the developing retina with this immunotoxin was found to eliminate virtually all cholinergic cells and processes. Recoverin labeling of bipolar cells showed that the axons of these neurons still form two stratified terminal bands within the IPL. Thus, neither RGCs nor cholinergic amacrine cell processes are required for the formation of segregated ON and Off cone bipolar cell projections.

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