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A NOVEL CONSTRUCT TO STUDY THE PULSATILITY OF INSULIN SECRETION IN SINGLE CELLS, ISLETS, AND WHOLE PANCREAS
by
Vivekanandan Shanmuganathan
A Thesis Presented to the
FACULTY OF THE GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the
Requirements for the Degree
MASTER OF SCIENCE
(BIOCHEMISTRY AND MOLECULAR BIOLOGY)
December 2008
Copyright 2008 Vivekanandan Shanmuganathan
Object Description
| Title | A novel construct to study the pulsatility of insulin secretion in single cells, islets and whole pancreas |
| Author | Shanmuganathan, Vivekanandan |
| Author email | shanmuga@usc.edu; vivekanandan85@gmail.com |
| Degree | Master of Science |
| Document type | Thesis |
| Degree program | Biochemistry & Molecular Biology |
| School | Keck School of Medicine |
| Date defended/completed | 2008-09-11 |
| Date submitted | 2008 |
| Restricted until | Unrestricted |
| Date published | 2008-12-05 |
| Advisor (committee chair) | Tokes, Zoltan A. |
| Advisor (committee member) |
Chow, Robert HP. Chan, Jonah R. Langen, Ralf |
| Abstract | Insulin is secreted in rhythmic pulses, which disappear in type 2 diabetes. Curiously, secretion periodicity differs when comparing single cells, single islets, versus whole pancreas. To test whether the different periods arise due to specific regulatory mechanisms found only at specific levels of tissue organization, it is desirable to compare secretion periods at all tissue levels with the same assay. We have designed a fluorescent vesicle reporter that is a fusion protein consisting of synaptobrevin (a vesicle SNARE) and pHluorin (a highly pH-sensitive variant of green fluorescent protein), separated by an intervening proprotein convertase 1/3 cleavage site. We demonstrate that when expressed in single AtT20 cells, it is cleaved and pHluorin is released into the supernatant. Because pHluorin is released and does not stay in the membrane after exocytosis, this reporter gives a strong signal with low background and can be used to dissect the mechanisms underlying pulsatile insulin release. |
| Keyword | exocytosis; beta cells; islets; diabetes; SynaptopHluorin; SNARE-complex |
| Language | English |
| Part of collection | University of Southern California dissertations and theses |
| Publisher (of the original version) | University of Southern California |
| Place of publication (of the original version) | Los Angeles, California |
| Publisher (of the digital version) | University of Southern California. Libraries |
| Provenance | Electronically uploaded by the author |
| Type | texts |
| Legacy record ID | usctheses-m1880 |
| Contributing entity | University of Southern California |
| Rights | Shanmuganathan, Vivekanandan |
| Repository name | Libraries, University of Southern California |
| Repository address | Los Angeles, California |
| Repository email | cisadmin@lib.usc.edu |
| Filename | etd-Shanmuganathan-2557 |
| Archival file | uscthesesreloadpub_Volume44/etd-Shanmuganathan-2557.pdf |
Description
| Title | Page 1 |
| Contributing entity | University of Southern California |
| Repository email | cisadmin@lib.usc.edu |
| Full text | A NOVEL CONSTRUCT TO STUDY THE PULSATILITY OF INSULIN SECRETION IN SINGLE CELLS, ISLETS, AND WHOLE PANCREAS by Vivekanandan Shanmuganathan A Thesis Presented to the FACULTY OF THE GRADUATE SCHOOL UNIVERSITY OF SOUTHERN CALIFORNIA In Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE (BIOCHEMISTRY AND MOLECULAR BIOLOGY) December 2008 Copyright 2008 Vivekanandan Shanmuganathan |
