Identification in islets of Langerhans of a new rat alpha 2-adrenergic receptor.

The islets of Langerhans are richly innervated, and an inhibitory effect on insulin secretion, mediated through alpha 2-adrenergic receptors, appears to be an important physiological neural modulator of beta-cell function. An alpha 2-receptor was cloned from isolated newborn rat islets using a polymerase chain reaction (PCR) approach. This receptor was shown by sequencing to be a new rat alpha 2-receptor very similar to the human alpha 2-C2 receptor. No other alpha 2-receptor subtype was identified in normal islets by the PCR using alpha 2-receptor primers. This was also the only alpha 2-receptor subtype present in the exocrine pancreas and liver. In contrast, in the beta-cell line, beta TC3, the alpha 2-C2 receptor was not detected, but the alpha 2-C4 and alpha 2-C10 receptor subtypes were detected. It is suggested that the alpha 2-C2 subtype may be the principal alpha 2-receptor mediating inhibitory autonomic nervous system activity in the gastrointestinal tract. A comparison of the rat islet, pancreas, and liver alpha 2-receptor sequences reported here with previously reported alpha 2-receptor sequences indicates that the rat islet alpha 2-receptor is not the rat alpha 2-C2 homologue previously denoted as RNG alpha 2, but is a new, fourth rat subtype with an even higher similarity to the human alpha 2-C2 receptor.

Protein discharge from immature secretory granules displays both regulated and constitutive characteristics.

At physiological glucose concentrations, isolated pancreatic islets release a minor portion of their newly synthesized insulin and precursors in a phase of secretion which is largely complete by 4 h of chase. Discharge during this period can be amplified by secretagogues, yet is not abolished by conditions which fully suppress regulated release from dense core secretory granules. The ability to stimulate the secretion and the biochemical profile of released proinsulin-related peptides indicate that secretion during this period originates from immature granules. The stoichiometry of release of labeled C-peptide:insulin during this phase is 1:1 at high glucose concentrations. However, at physiologic or low concentrations, C-peptide is released in molar excess of insulin as if the exocytotic vesicles carrying this secretion were budding from a post-Golgi compartment in which the lumen was composed of condensing insulin and soluble C-peptide. These findings can be explained by a model for regulated secretory protein traffic in which direct exocytosis of young granules is stimulated by higher glucose concentrations and vesicle budding from immature granules occurs at lower concentrations. Thus, insulin targeting from immature granules exhibits both regulated and constitutive-like characteristics.