Immunoreactivities to rhodopsin and rod/cone transducin antisera in the retina, pineal complex and deep brain of the bullfrog, Rana catesbeiana.

Birds and lower vertebrates are known to have extra-retinal photoreceptors in the pineal complex and deep brain. Although the photoreceptive function of the pineal complex has been investigated well, the exact location and nature of the deep brain photoreceptors are not known. In this study we tried to localize visual pigments and signal transduction proteins immunohistochemically in the brain of bullfrogs (Rana catesbeiana). The retina, and the brain with the pineal and the frontal organ were fixed with Zamboni's fixative and/or Bouin's solution. Immunoreactivities to three antisera against bovine rhodopsin (Rh-As), alpha-subunits of bovine rod (anti-pTr alpha) and cone transducin (anti-pTc alpha) were shown in the retina, pineal, frontal organ and hypothalamus. The retina and pineal were immunopositive to both Rh-As and anti-pTr alpha, whereas the frontal organ was immunopositive to only Rh-As and the hypothalamus was immunopositive to all three antisera. The cells which were immunoreactive to Rh-As, anti-pTr alpha and anti-pTc alpha were observed in the preoptic nucleus and suprachiasmatic nucleus in the hypothalamus. The shape of these immunoreactive cells in the hypothalamus was round or spindle-like with one or two immunoreactive nerve processes most of which were perpendicular to the ventricular surface. Western blot analysis of the hypothalamus, pineal and frontal organ demonstrated immunoreactive bands molecular weight of which corresponded to those of the retina (34 kDa, 38 kDa and 41 kDa). Thus, visual pigments and transducin-like proteins seem to exist in the hypothalamus as well as the pineal complex of frogs.