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Endocrinology Vol. 138, No. 9 3967-3975
Copyright © 1997 by The Endocrine Society

ARTICLES

Role of Prepancreatic (Pro)Insulin and the Insulin Receptor in Prevention of Embryonic Apoptosis1

Aixa V. Morales, José Serna, Cristina Alarcón, Enrique J. de la Rosa2 and Flora de Pablo2

Department of Cell and Developmental Biology, Centro de Investigaciones Biológicas, CSIC, Madrid, Spain

Address all correspondence and requests for reprints to: Dr. Flora de Pablo, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, C/Velázquez 144, E-28006 Madrid, Spain. E-mail: cibfp1f{at}fresno.csic.es

The characterization of (pro)insulin as an early embryonic growth factor requires demonstration of its expression and cellular effects in vivo. By in situ hybridization, we found widespread preproinsulin transcripts in the chick embryo throughout gastrulation and neurulation, before the beginning of preproinsulin-like growth factor I expression and pancreatic organogenesis. To analyze the prepancreatic (pro)insulin effect on apoptotic cell death, we treated embryos with antisense oligodeoxynucleotides in ovo and in vitro. The specific effect of two preproinsulin messenger RNA (mRNA) antisense oligodeoxynucleotides was confirmed by the decrease in a biosynthetically labeled protein immunoprecipitated with antiinsulin Igs. Insulin receptor mRNA antisense oligodeoxynucleotide applied in ovo increased by 2.7-fold the level of apoptosis in the 1.5-day embryo (neurulation) compared with that in its random sequence control. In a whole embryo culture, apoptosis increased by 25–35% with the addition of preproinsulin or insulin receptor mRNAs antisense oligodeoxynucleotides, respectively, whereas it decreased by 64% after 10 h in the presence of 10-8 M chicken insulin. Exogenous insulin also rescued the death induced by preproinsulin antisense oligonucleotides. These findings provide evidence for an autocrine/paracrine role of preproinsulin gene products acting through the insulin receptor in the control of cell survival/death during early embryonic development.




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