Abstract
While numerous techniques can be used to measure and analyze insulin secretion in isolated islets in culture, assessments of insulin secretion in vivo are typically indirect and only semiquantitative. The CpepSfGFP reporter mouse line allows the in vivo imaging of insulin secretion from individual islets after a glucose stimulation, in live, anesthetized mice. Imaging the whole pancreas at high resolution in live mice to track the response of each individual islet over time includes numerous technical challenges and previous reports were only limited in scope and non-quantitative. Elaborating on this previous model—through the development of an improved methodology addressing anesthesia, temperature control and motion blur—we were able to track and quantify longitudinally insulin content throughout a glucose challenge in up to two hundred individual islets simultaneously. Through this approach we demonstrate quantitatively for the first time that while isolated islets respond homogeneously to glucose in culture, their profiles differ significantly in vivo. Independent of size or location, some islets respond sharply to a glucose stimulation while others barely secrete at all. This platform therefore provides a powerful approach to study the impact of disease, diet, surgery or pharmacological treatments on insulin secretion in the intact pancreas in vivo.
Highlights
Background substractionThresholdingMasking and redirected analysis and quantification36.8 °C
We found that individual islet response to glucose is quite heterogeneous, emphasizing the impact of the inherent differences between in vitro and Scientific Reports | (2021) 11:603 |
In order to be able to track individual islets in culture over the duration of a glucose challenge, islets were immobilized in culture in a collagen gel to what has been previously used for embryonic pancreatic rudiments in c ulture[28,29,30,31,32]
Summary
36.8 °C (illustrated in Supplemental Fig. 1). We compared insulin secretion with the plastic spacers or with copper tubing and noted a ~ 1.5-fold increase in insulin secretion (Fig. 5a). Isoflurane is one of the most widely used anesthetics in mice. Induction and recovery are fast, and the risk of accidental death is low. The effects of anesthesia on insulin secretion have been previously studied[33,34,35,36,37], most anesthetic agents can blunt insulin secretion to glucose to some degree[34,35,37]. Pentobarbital has been reported to have less impact on glucose tolerance tests and insulin secretion[37].
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