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World J Gastroenterol. Dec 14, 2008; 14(46): 7112-7116
Published online Dec 14, 2008. doi: 10.3748/wjg.14.7112
Immunolocalization of nestin in pancreatic tissue of mice at different ages
Raj K Dorisetty, Sashi G Kiran, Malathi R Umrani, Sesikeran Boindala, Ramesh R Bhonde, Vijayalakshmi Venkatesan
Raj K Dorisetty, Sashi G Kiran, Sesikeran Boindala, Vijayalakshmi Venkatesan, Department of Biochemistry, National Institute of Nutrition, Hyderabad 500007, India
Malathi R Umrani, Ramesh R Bhonde, Tissue Engineering and Banking Laboratory, National Center for Cell Science, Pune University Campus, Ganeshkhind, Pune 411007, India
Author contributions: Dorisetty RK performed the research; Kiran SG captured the images and did the statistical analysis; Umrani MR contributed reagents and analytical tools; Boindala S and Bhonde RR were co-investigators and contributed towards the manuscript preparation; Venkatesan V principal investigator, designed the experiment, and drafted and finalized the manuscript.
Supported by Department of Biotechnology, Government of India Grant BT/PR 5647/MED/14/671/2004
Correspondence to: Vijayalakshmi Venkatesan, Department of Biochemistry, National Institute of Nutrition, Tarnaka, Hyderabad 500007, India. v.venkateshan@gmail.com
Telephone: +91-92-46109383 Fax: +91-40-27019074
Received: June 24, 2008
Revised: November 18, 2008
Accepted: November 25, 2008
Published online: December 14, 2008

AIM: To localize nestin positive cells (NPC) in pancreatic tissue of mice of different ages.

METHODS: Paraffin sections of 6-8 μm of fixed pancreatic samples were mounted on poly-L-lysine coated slides and used for Immunolocalization using appropriate primary antibodies (Nestin, Insulin, Glucagon), followed by addition of a fluorescently labeled secondary antibody. The antigen-antibody localization was captured using a confocal microscope (Leica SP 5 series).

RESULTS: In 3-6 d pups, the NPC were localized towards the periphery of the endocrine portion, as evident from immunolocalization of insulin and glucagon, while NPC were absent in the acinar portion. At 2 wk, NPC were localized in both the exocrine and endocrine portions. Interestingly, in 4-wk-old mice NPC were seen only in the endocrine portion, towards the periphery, and were colocalized with the glucagon positive cells. In the pancreas of 8- wk-old mice, the NPC were predominantly localized in the central region of the islet clusters, where immunostaining for insulin was at a maximum.

CONCLUSION: We report for the first time the immunolocalization of NPC in the pancreas of mice of different ages (3 d to 8 wk) with reference to insulin and glucagon positive cells. The heterogeneous localization of the NPC observed may be of functional and developmental significance and suggest(s) that mice pancreatic tissue can be a potential source of progenitor cells. NPC from the pancreas can be isolated, proliferated and programmed to differentiate into insulin secreting cells under the appropriate microenvironment.

Keywords: Nestin, Insulin, Glucagon, Immunolocalization, Mice