In vivo modulation of connexin 43 gene expression and junctional coupling of pancreatic B-cells.
We have explored the expression of gap junctional proteins and corresponding mRNAs by insulin-producing B-cells of native rat pancreas and of a transplantable rat insulinoma. By immunostaining cryostat sections (indirect immunofluorescence) and crude membrane preparations (Western blots) with antibodies against connexins 26, 32, and 43 and by hybridizing total islet and insulinoma RNA (Northern blot) with cRNAs for the latter two proteins, we have found that normal and tumoral B-cells express connexin 43 but do not show detectable levels of either connexin 32 or 26. By evaluating the conductance (dual patch-clamp whole-cell recording) and permeability of junctional channels (microinjection of Lucifer yellow), we have found that control B-cells show low levels of electrical and dye coupling in only a portion of the pairs studied. By studying B-cells of glibenclamide-treated rats, we have found that sustained stimulation of insulin release in vivo is associated with a two-fold increase in the level of connexin 43 gene transcripts and in the incidence of both ionic and dye coupling. These observations indicate that (1) connexin 43 is a major component of communicating channels between insulin-producing cells; (2) some but not all B-cells are electrically coupled by low conductance junctional channels; and (3) connexin 43 gene transcripts and incidence of junctional coupling are modulated in parallel during sustained stimulation of B-cell functioning in vivo.
Meda
P
P
Department of Morphology, University of Geneva, Switzerland.
Chanson
M
M
Pepper
M
M
Giordano
E
E
Bosco
D
D
Traub
O
O
Willecke
K
K
el Aoumari
A
A
Gros
D
D
Beyer
E C
EC
D000818:Animals; D002450:Cell Communication / Q000502:physiology; D002463:Cell Membrane Permeability / Q000187:drug effects; D017630:Connexins; D004553:Electric Conductivity / Q000187:drug effects; D005456:Fluorescent Dyes; D005786:Gene Expression Regulation / Q000187:drug effects; D005905:Glyburide / Q000494:pharmacology; D066298:In Vitro Techniques; D007328:Insulin / Q000096:biosynthesis; D007365:Intercellular Junctions / Q000502:physiology; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D007546:Isoquinolines; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis*; D008845:Microinjections; D015347:RNA Probes; D051381:Rats; D011919:Rats, Inbred Strains
D017630:Connexins; D005456:Fluorescent Dyes; D007328:Insulin; D007546:Isoquinolines; D008565:Membrane Proteins; D015347:RNA Probes; C017475:lucifer yellow; D005905:Glyburide