Serotoninergic neurons in the retina ofXenopus laevis: Selective staining, identification, development, and content

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Abstract
Uptake of 3H‐serotonin followed by autoradiography, and uptake of the serotonin analog 5,7‐dihydroxytryptamine (5,7‐DHT), with subsequent staining, were each used to define a unique set of neurons in the retina of the African clawed frog, Xenopus laevis. Both techniques demonstrated the same population of neurons, on the basis of perikaryal size, shape, and position within the retina. Two classes of amacrine cells accumulated 5,7‐DHT at the proximal (vitread) margin of the inner nuclear layer; the two classes were distinguished by the size of their perikarya. Two similar populations of cells, observed in the ganglion cell layer with lower frequency, may represent “displaced” counterparts of these two amacrine cell types. A class of bipolar cells whose perikarya were located in middle‐to‐distal regions of the inner nuclear layer also accumulated 5,7‐DHT and 3H‐serotonin. Processes of these cells contributed to a dense plexus of fine fibers that appeared evenly distributed throughout the inner plexiform layer. 3H‐Serotonin‐accumulating cells first appeared in the developing retina at stage 35/36, a time immediately after retinal stratification but before elaboration of either plexiform layer. Electron microscopic analysis permitted an identification of 3H‐serotonin‐accumulating terminals in the inner plexiform layer. Serotonin‐labeled terminals containing conventional contacts, suggestive of amacrine cells, were presynaptic to unidentified processes and postsynaptic to bipolar cells. Labeled terminals containing ribbon contacts, indicative of bipolar cells, were postsynaptic to amacrine cells. The amount of serotonin contained in isolated retinas was 15 pmol/mg protein as measured by HPLC with electrochemical detection. We attempted to stimulate the release of accumulated 3H‐serotonin from mature retinas by increasing the K+‐concentration in the bathing medium. Although preloaded glycine is readily released from 14C‐glycine‐accumulating neurons, from the same retinas there was no calcium‐dependent, K+‐stimulated release of 3H‐serotonin. This finding suggests that serotonin and glycine are processed differently by retinal neurons, the consequence of which results in differing responses to 40 mM K+.