Differential mRNA display analysis of two related but functionally distinct rat insulinoma (RIN) cell lines: identification of CD24 and its expression in the developing pancreas.
To identify genes that might play a role in growth and differentiation of pancreatic beta-cells, we have applied the technique of differential mRNA display to the lineage-related, but functionally distinct rat insulinoma (RIN) cell lines RIN-5AH and RIN-A12. Direct comparison of PCR-generated RIN-5AH and RIN-A12 cDNAs on DNA sequencing gels revealed 31 differentially expressed bands. By Northern blot hybridization, authentic differential expression was confirmed for three cDNAs derived from RIN-5AH cells and four cDNAs from RIN-A12 cells. Nucleotide sequences were determined for these cDNAs and database searches identified one known gene that encoded heat stable antigen CD24. Of the remaining six genes, three matched with established sequence tags from fetal tissue, and three were potentially novel. By RT-PCR analysis, five of the seven genes were expressed in normal fetal and/or adult pancreas. In a detailed survey of CD24 protein expression in the pancreas using the CD24-specific monoclonal antibody J11d, CD24 was predominantly expressed in ductal epithelial cells (E13.5-15.5), developing endocrine (alpha, beta and delta) and exocrine cells (E15.5-20.5) and mature exocrine and peripheral islet delta-cells post E20.5. The retention of CD24 expression in a large proportion of delta-cells but only in a minority of alpha- and beta-cells leads us to hypothesize that CD24 may mark a pool of precursor endocrine cells within adult islets.
Cram
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Autoimmunity and Transplantation Division, Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Parkville, VIC, Australia.
McIntosh
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Oxbrow
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Johnston
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AM
DeAizpurua
H J
HJ
D000375:Aging; D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D051927:CD24 Antigen; D005314:Embryonic and Fetal Development; D018507:Gene Expression Regulation, Developmental; D007328:Insulin / Q000235:genetics*; D007340:Insulinoma / Q000235:genetics; D007515:Islets of Langerhans / Q000196:embryology / Q000378:metabolism; D008562:Membrane Glycoproteins; D051379:Mice; D009928:Organ Specificity; D010179:Pancreas / Q000196:embryology / Q000254:growth & development / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D051927:CD24 Antigen; C497890:Cd24a protein, mouse; D007328:Insulin; D008562:Membrane Glycoproteins; D012333:RNA, Messenger