Calcitonin gene-related peptide (CGRP) and its effects on protein release in vitro in the developing submandibular gland of the rat

Abstract

Indirect immunohistochemical methods were used to study presence and number of CGRP-immunoreactive (CGRP-IR) nerve fibers in the submandibular gland and ganglion cells of the superior cervical, submandibular and trigeminal ganglia of the developing rat. The effect of CGRP on peroxidase and total protein release was also studied in the developing postnatal submandibular glands of 1, 5, 12 and 30-day-old, as well as adult rats by in vitro methods. The possible costimulation of CGRP with SP, NKA or carbachol on 5-day-old and adult rats was also tested. The stimulatory effects of these compounds were compared to the basic release of peroxidase and total amount of proteins from submandibular gland fragments in incubation solution. CGRP-IR nerve fibers were found in relatively high number during post-natal development, mainly around blood vessels and ducts. Some CGRP-IR nerve fibers were also detected around acini. The number of these fibers was quite low and remained constant during the post-natal development. The number of CGRP-IR trigeminal ganglion cells was higher on the 5th and 12th post-natal day than later in development and in adult animals. At the same time, superior cervical- and submandibular ganglion cells were non-reactive for CGRP, suggesting trigeminal origin of CGRP-IR nerve fibers during the development in the submandibular gland. In the secretory studies, CGRP per se stimulated both peroxidase and total protein release in the submandibular gland most effectively on 5th and 12th post-natal days, while there was no clear secretory response in the adult glands. In the 5-day-old submandibular gland CGRP in combination with SP, NKA or carbachol clearly enhanced the total protein secretory response when compared with the release caused by these substances alone. However, in the adult submandibular gland, the combinations did not enhance total protein release more than any of the substances alone. Furthermore, it can be concluded that the presence of a more dense CGRP-IR innervation during the early postnatal period in the developing submandibular gland is accompanied with an increased responsiveness of the secretory elements to CGRP.