The crayfish optic lobe contains high levels of acetylcholine (ACh) and choline as measured with a chemiluminescent assay in small fragments of optic lobe tissue. The highest concentrations were found in the medulla externa and medulla interna (second and third optic neuromeres), which have ACh concentrations of 270 pmol/mg tissue. This concentration is about 16 times that measured in the photoreceptors and lamina ganglionaris (the first optic neuromere). Immunocytochemistry (based upon antisera to choline-glutaryl-BSA) revealed low levels of ACh-like reactivity in the lamina ganglionaris associated with the terminal arbors of centrifugal and/or tangential neurons. The most intense ACh- like reactivity was observed in monopolar neurons of the medulla externa and medulla interna. One monopolar neuron/medullary column (or about 2500 neurons/medullary neuropile) exhibited reactivity and an estimated cytoplasmic concentration of 8.1 mM.

The actions of acetylcholine (ACh) were examined on 4 classes of multicolumnar interneurons whose dendrites lie in close proximity to the putative cholinergic transmedullary neurons described in the companion report. ACh-elicited responses in each cell type resemble visually elicited synaptic events and persist following synaptic blockade with 20 mM CoCl2. Tangential cells exhibit a hyperpolarizing response to ACh that resembles the visual response in reversal potential and dependence on extracellular chloride. The visual response is potentiated by the anticholinesterase, neostigmine (0.1 mM). Visual and carbachol-elicited responses are blocked by nicotinic ganglionic antagonists (e.g., 10(-6) M pempidine) that are 10-100 times more potent than D-tubocurarine. Medullary amacrine cells exhibit depolarizing responses to ACh (10(-6) M) and light with similar reversal potentials. The visual response is potentiated by neostigmine. Dimming fibers respond to light and ACh with a hyperpolarization that inhibits the maintained discharge. The sustaining fiber response to ACh reflects both direct responses and indirectly elicited synaptic actions. The direct action is a hyperpolarization possibly related to the visual "off-response." It is associated with an increased conductance and a reversal potential negative to the dark potential. The off-response is abolished by curare and pempidine and potentiated by neostigmine. ACh appears to orchestrate several aspects of the dual-channel contrast detection system of the optic lobe. The actions of ACh on tangential cells, amacrine cells, and dimming fibers are all consistent with the effects of a spatially localized increment in light intensity and a corresponding local release of ACh in the retinotopic columnar array.