Modulation of gap and adherens junctional proteins in cultured neonatal pancreatic islets.
Fetal and neonatal pancreatic islets have lower insulin secretory responses compared with adult islets. In culture conditions and after treatment with mammosomatotropic hormones, neonatal islets undergo maturation of the secretory machinery that might involve regulation of cell-cell contacts within the islet. This study is an investigation of the effect of prolonged culturing and in vitro treatment with prolactin on the expression of the gap junction-associated connexin 43 and the adherens junction-associated beta-catenin in cultured neonatal rat islets. Pancreatic islets from neonatal Wistar rats were cultured for 24 hours or 7 days, and the treated group was exposed to 2 microg/mL prolactin daily for 7 days. Connexin 43 and beta-catenin were barely detected at the cell-cell contacts in 24-hour-cultured islets, as revealed by immunocytochemical analysis. Nevertheless, both junctional proteins were well expressed at the junctional region in islet cells cultured for 7 days and showed even greater staining in islets after long-term prolactin treatment. In accordance with the morphologic data, neonatal islets cultured for 24 hours displayed a relatively low level of connexin 43, as determined by Western blot analysis. Culturing for 7 days or combined prolactin treatment induced a significant increase in connexin 43 expression; this was 40% greater in the prolactin-treated group than in the control group. Furthermore, an enhancement of the expression of beta-catenin and translocation of this protein to the cell-cell contact site was also observed in neonatal islets cultured for 7 days compared with those cultured for 24 hours. In vitro prolactin treatment induced even greater expression of beta-catenin in islet cells. A correlation was observed between the increased expression of these junctional proteins and an increase in insulin secretion in cultured neonatal islets. In conclusion, prolonged culturing and in vitro treatment with prolactin induce the modulation of gap and adherens junctional proteins in pancreatic islets, which may be an important event in the in vitro maturation process of neonatal islet cells.
Collares-Buzato
C B
CB
Department of Histology, University of Campinas (UNICAMP), Brazil. collares@obelix.unicamp.br
Leite
A R
AR
Boschero
A C
AC
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D018031:Connexin 43 / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D011388:Prolactin / Q000494:pharmacology; D051381:Rats; D013997:Time Factors; D015534:Trans-Activators; D051176:beta Catenin
D018031:Connexin 43; C495268:Ctnnb1 protein, rat; D003598:Cytoskeletal Proteins; D007328:Insulin; D015534:Trans-Activators; D051176:beta Catenin; D011388:Prolactin