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. Author manuscript; available in PMC: 2012 Mar 9.
Published in final edited form as: Vis Neurosci. 2010 Jul 23;27(3-4):103–118. doi: 10.1017/S0952523810000155

Figure 8. Cellular Localization of Transmitter Associated Molecules.

Figure 8

A: ChAT staining is absent from the region surrounding the rEF terminal. Double labeling for rEF terminals and choline acetyltransferase, the synthetic enzyme for ACh, revealed no colocalization between the two. (a), ChAT staining (green) shows 2 strongly stained laminae in the IPL and regular arrays of starburst amacrine cells both above and below the IPL. The rEF terminal, labeled with antibodies against TMR (red), used to increase the signal of Fluoro-Ruby in rEF terminals, lies in a region of the INL that lacks ChAT staining. Scale bar is 20 μm. (b&c) Higher magnification images of the region surrounding the rEF shown in the merged image, (a). Scale bar is 20 μm.

B&C: Presynaptic markers for glutamate and GABA have distinct staining patterns. In both B and C the larger merged image (a) shows the entire thickness of the retina with photoreceptors at the top and the optic fiber layer at the bottom. Dotted lines indicate the INL-IPL border. The three images on the right (b-d) show higher magnification images of the region surrounding the TC so that colocalization between the two antibodies can be more closely examined. All images are compressed confocal stacks. Scale bars are 10 μm.

B: Double labeling with antibodies against rhodamine (red), identifying the rEF terminal and vGluT2 (a presynaptic marker for glutamatergic synapses, green) shows a matching pattern. In b, c, & d, individual presynaptic boutons, approximately 2μm in diameter, can often be seen in both images (arrows).

C: Double labeling with antibodies against parvalbumin (red) and GAD65/67 (green) reveals that a few small, punctate GABAergic terminals contact thin, dendritic extensions of the TC (yellow in merged images and arrows in b & c). These GABAergic terminals do not have the same staining pattern as vGluT2 in B and do not correspond to the rEF terminals.

D: TCs express high levels of NKCC. A retinal slice double labeled with antibodies against parvalbumin, used to identify the TC (a) and NKCC (b). In the merged image (c) the soma and dendritic region of the TC is seen to be strongly NKCC positive. Scale bar is 20 μm.