The ryanodine (RY) receptors in beta-cells amplify signals by Ca2+-induced Ca2+ release (CICR). The role of CICR in insulin secretion remains unclear in spite of the fact that caffeine is known to stimulate secretion. This effect of caffeine is attributed solely to the inhibition of cAMP-phosphodiesterases (cAMP-PDEs). We demonstrate that stimulation of insulin secretion by caffeine is due to a sensitization of the RY receptors. The dose-response relationship of caffeine-induced inhibition of cAMP-PDEs was not correlated with the stimulation of insulin secretion. Sensitization of the RY receptors stimulated insulin secretion in a context-dependent manner, that is, only in the presence of a high concentration of glucose. This effect of caffeine depended on an increase in [Ca2+]i. Confocal images of beta-cells demonstrated an increase in [Ca2+]i induced by caffeine but not by forskolin. 9-Methyl-7-bromoeudistomin D (MBED), which sensitizes RY receptors, did not inhibit cAMP-PDEs, but it stimulated secretion in a glucose-dependent manner. The stimulation of secretion by caffeine and MBED involved both the first and the second phases of secretion. We conclude that the RY receptors of beta-cells mediate a distinct glucose-dependent signal for insulin secretion and may be a target for developing drugs that will stimulate insulin secretion only in a glucose-dependent manner.

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No quantitative data are available regarding the rate of occurrence of nerve cells in association with endocrine pancreas (i.e.. neuroinsular complexes type I [NICs]), or the difference in the distribution of NICs in normal and diabetic pancreas. In this report, pancreata from 20-day, 7-week, and 9-month-old lean (Umeå +/?) and obese (Umeå ob/ob) mice, as well as 10-month-old C57BL/6JBom and Umeå ob/ob mice, were analyzed with regard to the association of acetylcholinesterase (AChE)-positive and protein gene product 9.5-like (PGP-LI) immunoreactive perikarya with islets, and not in association with islets. NIC profiles were regularly observed, but were more frequent in the 20-day-old mice than in the 9-month-old +/? and ob/ob mice. The NIC profiles were often located close to a duct or blood vessel, significantly more frequently than islet profiles in general. The data did not reveal any gross abnormality in ob/ob mice as regards the frequency of NICs or the number of AChE-positive and PGP-LI perikarya. However, the 9-month-old ob/ob mice demonstrated smaller clusters of perikarya in their NIC profiles as compared to the other mice, probably reflecting the fact that the perikarya were more widely spread out in the hyperplastic islets of adult ob/ob mice. The results show that NICs are common and represent a substantial proportion of the islets in mouse pancreas, supporting the idea that they play a role in islet physiology.

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Collagenase-isolated pancreatic islets from C57BL/6J mice were cultured overnight and transplanted under the kidney capsule of non-diabetic syngeneic hosts. Cryostat sections of grafts and fresh islets were stained for acetylcholinesterase (AChE) and vasoactive intestinal polypeptide-like immunoreactivity (VIP-LI). Immediately after isolation, as well as 2-5 days after transplantation, VIP-LI- and AChE-positive nerve cell bodies were clearly seen in the periphery of the islets. Grafts 3-5 days old exhibited a transient and marked increase in VIP-LI nerve cell bodies and fibres. Seven days after transplantation VIP-LI nerve structures began to decrease in number and after 26-52 weeks they were no longer detectable. In contrast, AChE-positive nerve cell bodies and fibers, which showed a relatively constant pattern of distribution, were observed throughout the entire observation period. Restaining experiments demonstrated the coexistence of VIP-LI and AChE activity in the neurons. It is concluded that the grafts were extensively equipped with an intrinsic VIP-ergic and AChE-positive innervation. The initial, transient enhancement of VIP-LI expression probably reflects an adaptation of the neuro-insular complex to the preganglionic denervation, or to the ectopic environment, or both.