This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Niwa, H. Articles by Cram, D. S. Search for Related Content PubMed PubMed Citation Articles by Niwa, H. Articles by Cram, D. S.

Identification of Pancreatic ß Cell-Related Genes by Representational Difference Analysis¹

Burnet Clinical Research Unit, The Walter and Eliza Hall Institute of Medical Research, Post Office, Royal Melbourne Hospital, Parkville 3050, Australia

Address all correspondence and requests for reprints to: Dr. David Cram, Burnet Clinical Research Unit, The Walter and Eliza Hall Institute of Medical Research, Post Office, Royal Melbourne Hospital, Parkville 3050, Australia. E-mail: cram{at}wehi.edu.au

A knowledge of ß cell-specific gene expression provides a basis for identifying proteins potentially involved in ß cell function and pathology. To identify candidate ß cell-specific genes, we applied the PCR-based subtractive hybridization technique of representational difference analysis (RDA) to the mouse SV40-transformed endocrine cell lines, ßTC3 and TC1. Following three successive subtractions of TC1 complementary DNA from ßTC3 complementary DNA, difference products were cloned into pUC19 and nucleotide sequences determined. Comparison of 91 sequences against the databases identified 11 known and 8 novel genes. Known genes included previously reported ß cell-specific genes, insulin I/II and islet amyloid polypeptide, as well as other non-ß cell-specific genes such as those for insulin-like growth factor II, selenoprotein P, neuronatin, prohormone convertase, and type 1 protein kinase A regulatory subunit. By Northern blot hybridization, expression of the majority of known and novel genes was restricted to ßTC3 cells. Novel genes BA-12, -13, -14, and -18 were expressed not only in ßTC3 cells, but also in normal pancreatic islets and a limited number of other tissues. The deduced amino acid sequence of BA-14 showed significant homology with members of the cadherin superfamily indicating that BA-14 may encode a cadherin-like molecule potentially involved in ß cell adhesion events during islet ontogeny. In ßTC3 cells, none of the novel genes were regulated at the RNA level by high glucose. However, in parallel studies, transcription of BA-12 was significantly increased by both sodium butyrate and nicotinamide, suggesting that this gene may play a role in pancreatic ß cell growth and/or differentiation.

In this study, we have demonstrated that cRDA is an effective strategy for systematically mapping differences in gene expression between two related but functionally-distinct endocrine cells. Its application to experimental animal models of islet-cell regeneration may facilitate the discovery of potential factors that mediate ß cell growth and differentiation.

This article has been cited by other articles:

				J. Kohrle, F. Jakob, B. Contempre, and J. E. Dumont Selenium, the Thyroid, and the Endocrine System Endocr. Rev., December 1, 2005; 26(7): 944 - 984. [Abstract] [Full Text] [PDF]

				K. Chu and M.-J. Tsai Neuronatin, a Downstream Target of BETA2/NeuroD1 in the Pancreas, Is Involved in Glucose-Mediated Insulin Secretion Diabetes, April 1, 2005; 54(4): 1064 - 1073. [Abstract] [Full Text] [PDF]

				G. D. Sgarlato, C. L. Eastman, and H. H. Sussman Panel of Genes Transcriptionally Up-regulated in Squamous Cell Carcinoma of the Cervix Identified by Representational Difference Analysis, Confirmed by Macroarray, and Validated by Real-Time Quantitative Reverse Transcription-PCR Clin. Chem., January 1, 2005; 51(1): 27 - 34. [Abstract] [Full Text] [PDF]

				A. M Johnston, G. Naselli, H. Niwa, T. Brodnicki, L. C Harrison, and L. J. Gonez Harp (harmonin-interacting, ankyrin repeat-containing protein), a novel protein that interacts with harmonin in epithelial tissues Genes Cells, October 1, 2004; 9(10): 967 - 982. [Abstract] [Full Text] [PDF]

				T. Miura, T. Ohta, C. I. Miura, and K. Yamauchi Complementary Deoxyribonucleic Acid Cloning of Spermatogonial Stem Cell Renewal Factor Endocrinology, December 1, 2003; 144(12): 5504 - 5510. [Abstract] [Full Text] [PDF]

				J. Wang, G. Webb, Y. Cao, and D. F. Steiner Contrasting patterns of expression of transcription factors in pancreatic {alpha} and {beta} cells PNAS, October 28, 2003; 100(22): 12660 - 12665. [Abstract] [Full Text] [PDF]

				S. J. Kuerbitz, J. Pahys, A. Wilson, N. Compitello, and T. A. Gray Hypermethylation of the imprinted NNAT locus occurs frequently in pediatric acute leukemia Carcinogenesis, April 1, 2002; 23(4): 559 - 564. [Abstract] [Full Text] [PDF]

				E. M. Perera, H. Martin, T. Seeherunvong, L. Kos, I. A. Hughes, J. R. Hawkins, and G. D. Berkovitz Tescalcin, a Novel Gene Encoding a Putative EF-Hand Ca2+-Binding Protein, Col9a3, and Renin Are Expressed in the Mouse Testis during the Early Stages of Gonadal Differentiation Endocrinology, January 1, 2001; 142(1): 455 - 463. [Abstract] [Full Text] [PDF]

				E.K. Moses, K.A. Freed, J.R. Higgins, and S.P. Brennecke Alternative forms of a novel aspartyl protease gene are differentially expressed in human gestational tissues Mol. Hum. Reprod., October 1, 1999; 5(10): 983 - 989. [Abstract] [Full Text] [PDF]

				M J. Melia, N. Bofill, M. Hubank, and A. Meseguer Identification of Androgen-Regulated Genes in Mouse Kidney by Representational Difference Analysis and Random Arbitrarily Primed Polymerase Chain Reaction Endocrinology, February 1, 1998; 139(2): 688 - 695. [Abstract] [Full Text] [PDF]