Abstract
We imaged the carbohydrate-selective spatial binding of 8 lectins in the ampullary organs (AOs) of electroreceptors on the rostrum of freshwater paddlefish (Polyodon spathula), by fluorescence imaging and morphometry of frozen sections. A focus was candidate sites of secretion of the glycoprotein gel filling the lumen of AOs. The rostrum of Polyodon is an electrosensory appendage anterior of the head, covered with >50,000 AOs, each homologous with the ampulla of Lorenzini electroreceptors of marine rays and sharks. A large electrosensory neuroepithelium (EN) lines the basal pole of each AO's lumen in Polyodon; support cells occupy most (97%) of an EN's apical area, along with electrosensitive receptor cells. (1) Lectins WGA or SBA labeled the AO gel. High concentrations of the N-acetyl-aminocarbohydrate ligands of these lectins were reported in canal gel of ampullae of Lorenzini, supporting homology of Polyodon AOs. In cross sections of EN, WGA or SBA labeled cytoplasmic vesicles and organelles in support cells, especially apically, apparently secretory. Abundant phalloidin+ microvilli on the apical faces of support cells yielded the brightest label by lectins WGA or SBA. In parallel views of the apical EN surface, WGA labeled only support cells. We concluded that EN support cells massively secrete gel from their apical microvilli (and surface?), containing amino carbohydrate ligands of WGA or SBA, into the AO lumen. (2) Lectins RCA120 or ConA also labeled EN support cells, each differently. RCA120-fluorescein brightly labeled extensive Golgi tubules in the apical halves of EN cells. ConA did not label microvilli, but brightly labeled small vesicles throughout support cells, apparently non-secretory. (3) We demonstrated "sockets" surrounding the basolateral exteriors of EN receptor cells, as candidate glycocalyces. (4) We explored whether additional secretions may arise from non-EN epithelial cells of the interior ampulla wall. (5) Model: Gel is secreted mainly by support cells in the large EN covering each AO's basal pole. Secreted gel is pushed toward the pore, and out. We modeled gel velocity as increasing ~11x, going distally in AOs (toward the narrowed neck and pore), due to geometrical taper of the ampulla wall. Gel renewal and accelerated expulsion may defend against invasion of the AO lumen by microbes or small parasites. (6) We surveyed lectin labeling of accessory structures, including papilla cells in AO necks, striated ectoderm epidermis, and sheaths on afferent axons or on terminal glia.
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