Immunocytochemistry of luteinizing hormone-releasing hormone in brains of bullfrogs (Rana catesbeiana) during spontaneous metamorphosis

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Abstract
The distribution of immunoreactive luteinizing hormone-releasing hormone (ir-LH-RH) in brains of bullfrogs (Rana catesbeiana) during spontaneous metamorphosis has been studied by combination of an unlabeled antibody enzyme immunocytochemical technique and an adjacent serial section approach. In prometamorphic tadpoles, immunocytochemical staining for ir-LH-RH was absent from the brain, including a structurally simple median eminence (ME) and perikarya in the anterior preoptic area (aPOA). In metamorphic tadpoles, speckled patches of immunostaining occurred over the outer layer of a modestly developed ME; coincident faint staining of a small number of medial, unpaired cell bodies was localized in the aPOA. In newly metamorphosed juvenile frogs, more diffuse and intense staining of the outer layer of the ME accompanied increased morphological differentiation of this neurohemal area; immunoreactive perikarya again were found in the aPOA, but an increased number of neurons exhibited comparatively greater (moderate) immunostaining. Changes in the quality of immunostaining and in the numbers of cells stained, therefore, were coincident with metamorphic development. Concomitant alterations of ir-LH-RH immunostaining and progressive structural development of the ME suggest a coordinated differentiation of brain neuroendocrine systems during metamorphosis of the bullfrog tadpole.