DIFFERENTIAL DISTRIBUTION OF SYNAPTOTAGMIN-I AND RAB-3 IN THE ANTERIOR-PITUITARY OF 4 MAMMALIAN-SPECIES

In the anterior pituitary of rat, gerbil, hamster and guinea pig, the presence and cellular distribution of the synaptic vesicle-associated proteins synaptotagmin I and rab3 were analyzed by immunoblotting and by immunocytochemical staining of serial semithin sections. Our results show that rab3 proteins are ubiquitously expressed in all endocrine cell types of both the anterior and intermediate lobe. In many cells, rab3 immunoreactivity was concentrated beneath the plasmalemma. This intracellular distribution coincided with the distribution of secretory granules, suggesting a possible association of rab3 proteins with the latter organelles. The staining patterns observed using two monoclonal rab3 antibodies with different isoform specificities are compatible with the recent suggestion that rab3B is the dominant rab3 isoform in anterior pituitary cells. However, we could demonstrate that also rab3A is present in endocrine adenohypophyseal cells, albeit at low levels. In contrast to rab3, synaptotagmin I immunoreactivity was only detected in a limited number of adenohypophyseal endocrine cells. Whereas the monoclonal synaptotagmin I antibody consistently failed to immunostain lactotrophs and endocrine cells of the intermediate lobe, other endocrine cell types displayed variable immunoreactivities towards this antibody. Although a low level of synaptotagmin I expression in the immunonegative cells cannot be excluded, the above observation may reflect a differential distribution of synaptotagmin isoforms in endocrine organs, as it has been described for the nervous system. Our study has established that endocrine cells of the anterior pituitary are endowed with proteins of the rab3 and synaptotagmin families which are generally thought to play important roles in the regulation of the trafficking and/or exocytosis of secretory organelles and, hence, probably fulfil similar functions in adenohypophyseal cells.