The islet of Langerhans is the functional unit responsible for glucose-stimulated insulin secretion (GSIS), and thus plays a key role in blood glucose homeostasis. The importance of the islet is demonstrated by the proven ability of islet transplants to reverse Type I diabetes pathologies in human patients. Further, it has long been known that an islet yields -10 fold greater insulin response to glucose than an equivalent mass of isolated p-cells. The long-term goal of this project is to understand the multicellular mechanisms of islet function, and their role in the regulation of blood glucose under normal and pathological conditions. In many ways, the islet appears to function as a syncytium, which exhibits synchronous behavior of membrane action potentials, Ca2+ oscillations, and pulsatile insulin secretion across all p-cells in the islet. Despite the importance of the islet, we still have only a rudimentary understanding of the basic multicellular mechanisms that lead to these synchronous behaviors and to enhanced GSIS. The islet is also made up of different cell types, and very little is known about the interplay between the different cells. We hypothesize that gap junction coupling between fl cells in the islet leads to the synchronous behaviors, and this in turn, accounts for a majority of the increased insulin response from intact islets over isolated/?ce//s, although paracrine signaling between fi-cells and between the different cell types also plays a modulatory role. The validity and limits of this hypothesiswill be tested via three specific aims: 1) Determine the relative conductance strength of gap junction coupling between p-cells in terms of the KATP channel conductance;2) Determine the role of intra- and intercellular metabolism in the glucose-stimulated Ca2+ oscillations in p-cells within intact islets; and 3) Determine the metabolic, membrane potential, and Ca2+ dynamic changes of the a-cell in response to glucose, and the influences of p-cell activity on these responses. Using our unique quantitative optical imaging methods and novel microfluidic devices, the dynamics of these molecular mechanisms can be followed quantitatively in living cells within intact islets. These investigations will also utilize several available transgenic and tissue-specific knock-out mouse models with demonstrated phenotypes, as well as traditional biochemical and molecular biological approaches. Lay Summary: We propose to study the function of pancreatic islets, which contain the insulin-secreting p- cells. Two health-related motivations underlie the proposed work. First, a better understanding of pancreatic islet function is necessary to increase the success rate of islet transplants, which are a promising strategy to cure Type I diabetes. Second, understanding the cell-to-cell communication pathways may lead to new drugs that enhance insulin secretion for the treatment of Type II diabetes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK053434-12
Application #
7738490
Study Section
Cellular Aspects of Diabetes and Obesity Study Section (CADO)
Program Officer
Appel, Michael C
Project Start
1998-01-01
Project End
2011-11-30
Budget Start
2009-12-01
Budget End
2011-11-30
Support Year
12
Fiscal Year
2010
Total Cost
$305,298
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Physiology
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Elliott, Amicia D; Ustione, Alessandro; Piston, David W (2015) Somatostatin and insulin mediate glucose-inhibited glucagon secretion in the pancreatic ?-cell by lowering cAMP. Am J Physiol Endocrinol Metab 308:E130-43
Short, Kurt W; Head, W Steve; Piston, David W (2014) Connexin 36 mediates blood cell flow in mouse pancreatic islets. Am J Physiol Endocrinol Metab 306:E324-31
Benninger, Richard K P; Hutchens, Troy; Head, W Steven et al. (2014) Intrinsic islet heterogeneity and gap junction coupling determine spatiotemporal Ca²? wave dynamics. Biophys J 107:2723-33
Benninger, Richard K P; Piston, David W (2014) Cellular communication and heterogeneity in pancreatic islet insulin secretion dynamics. Trends Endocrinol Metab 25:399-406
Hang, Yan; Yamamoto, Tsunehiko; Benninger, Richard K P et al. (2014) The MafA transcription factor becomes essential to islet ?-cells soon after birth. Diabetes 63:1994-2005
Kumar, Ankur N; Short, Kurt W; Piston, David W (2013) A motion correction framework for time series sequences in microscopy images. Microsc Microanal 19:433-50
Schwetz, Tara A; Ustione, Alessandro; Piston, David W (2013) Neuropeptide Y and somatostatin inhibit insulin secretion through different mechanisms. Am J Physiol Endocrinol Metab 304:E211-21
Elliott, Amicia D; Gao, Liang; Ustione, Alessandro et al. (2012) Real-time hyperspectral fluorescence imaging of pancreatic ?-cell dynamics with the image mapping spectrometer. J Cell Sci 125:4833-40
Le Marchand, Sylvain J; Piston, David W (2012) Glucose decouples intracellular Ca2+ activity from glucagon secretion in mouse pancreatic islet alpha-cells. PLoS One 7:e47084
Head, W Steven; Orseth, Meredith L; Nunemaker, Craig S et al. (2012) Connexin-36 gap junctions regulate in vivo first- and second-phase insulin secretion dynamics and glucose tolerance in the conscious mouse. Diabetes 61:1700-7

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