Immunocytochemical studies of vasotocin, mesotocin, and neurophysins in the Xenopus hypothalamo-neurohypophysial system.

Mesotocinergic and vasotocinergic neurons, which constitute the principal neurons in the hypothalamo-neurohypophysial system in Xenopus, were studied by immunocytochemical techniques. Antibodies that could unequivocally distinguish mesotocin, vasotocin, and their respective neurophysins were used in these studies. A monoclonal antibody directed at rat oxytocin-associated neurophysin (PS-36) detected an antigen that was colocalized with vasotocin, whereas a monoclonal antibody to rat vasopressin-associated neurophysin (PS-45) crossreacted with an antigen in mesotocinergic cells. As vasotocin is regarded as an evolutionary precursor of vasopressin, and as mesotocin is usually associated with oxytocin, we were surprised to see this apparent eptitope switch in the associated neurophysins. One interpretation of this epitope switch is that the final exons encoding for the carboxy-terminals of the mammalian neurophysins, which contain the PS-45 and PS-36 antibody epitopes, are in reversed positions in Xenopus. Approximately 4,000 mesotocinergic and vasotocinergic neurons and their fibers were topographically mapped in the Xenopus hypothalamus. The two types of neurons were intermingled and scattered throughout a large contiguous region including but not limited to the preoptic recess. Small, medium size, and large cells contained these antigens. Immunoreactive fibers were seen in the preoptic area, the neurohypophysial tract, the median eminence, and the neural lobe of the pituitary. The neurophysin-specific monoclonal antibodies have several advantages as phenotypic markers in development; i.e., high titer, low background, and affinity for the prohormone forms as well as for the fully processed neurophysin polypeptides. Their antigens are related gene products whose expression is central to the identity of the two cell types and whose expression is differentially controlled in development. This characterization of their adult distribution provides a basis for future studies of the development of peptidergic phenotype in the central nervous system of Xenopus.