Visually evoked behaviors mediated by the frog optic tectum require cholinergic activity, but the receptor subtypes through which acetylcholine acts are not yet identified. Using quantitative autoradiography and scintillation spectrometry, we examined the binding of [ 3 H]pirenzepine and [ 3 H]AF-DX 384 in the laminated optic tectum of the frog. In mammalian systems, these substances bind excitatory (m1 and m3 subtypes) and inhibitory (m2 and m4 subtypes) muscarinic acetylcholine receptors, respectively. Pharmacological analyses, including the use of specific muscarinic toxins, confirmed the subtype selectivity of the radioligands in the frog brain. Binding sites for [ 3 H] pirenzepine were distinct from those for [ 3 H]AF-DX 384. In the adult tectum, [ 3 H]pirenzepine demonstrated specific binding in tectal layers 5-9. [ 3 H]Pirenzepine binding was also present in tadpoles as young as stage V, but all sampled stages of tadpole tectum had significantly less binding when compared to adults. Lesioning of the optic nerve had no effect on [ 3 H]pirenzepine binding. Specific [ 3 H]AF-DX 384 binding was found in all layers of the adult tectum. All sampled tadpole stages exhibited binding sites for [ 3 H]AF-DX 384, but the densities of these sites were also significantly higher in adults than they were in developing stages. Short-term lesions of the optic nerve reduced [ 3 H]AF-DX 384 binding in all tectal layers of the deafferented lobe when compared to the afferented one. Long-term lesions decreased [ 3 H]AF-DX 384 sites in both lobes.These results indicate that multiple muscarinic acetylcholine receptor binding sites reside in the frog optic tectum at all stages of development, and their pharmacology resembles that of mammalian m1/ m3, m2 and m4 subtypes. Our data indicate that few, if any, of these receptors are likely to be located on retinal ganglion cell terminals. Furthermore, the expression of inhibitory muscarinic subtypes seems to be regulated by different mechanisms than that for excitatory subtypes.
Keywords cholinergic; visual plasticity; autoradiography; amphibiaThe optic tectum is the primary visual area of non-mammalian vertebrates and serves as a useful model for studying mechanisms of visual processing and the structural organization of topographic maps. 14, 15,18,25,84 Many studies in the brain have shown the importance of neuronal activity in maintaining the order of the visual map. 17,50,63,74 One of the factors affecting this activity is the presence of modulatory neurotransmitters within the system. *Corresponding author. Tel.: +1-859-323-9537; fax: +1-859-257-1717. E-mail address: debski@pop.uky.edu (E. A. Debski).
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Author ManuscriptNeuroscience. Author manuscript; available in PMC 2008 March 10.
Published in final edited form as:Neuroscience. 2001 ; 104(1): 161-179.
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NIH-PA Author ManuscriptAlthough neuromodulators have been shown to change visual activity in the tectum, little is known about the receptors that mediate th...