Junctional communication of pancreatic β cells contributes to the control of insulin secretion and glucose tolerance
J. Clin. Invest. Anne Charollais, et al. 106:235 doi:10.1172/JCI9398 [Go to this article.]

Figure 1
Strategy for expressing Cx32 in pancreatic β cells. (a) A transgene that contained part of the RIP, the second exon of Cx32 cDNA (Cx32; coding region limited by dots), and the coding region of human growth hormone gene (hGH) was microinjected into zygotic pronuclei, and surviving embryos were transferred into pseudopregnant foster mothers. (b) Transgenic mice were screened by Southern blot using probe 2. Compared with wild-type controls (W), which featured only the 7.6-kb endogenous gene (Cx32_e), two positive founders were identified that also expressed the 1.5-kb transgene (Cx32_t). These mice were backcrossed with C56BL/6 controls to generate the two independent lines B9 (lane 1) and D6 (lane 2). (c) The progeny were screened by PCR using primers along the hGH sequence. Heterozygous mice carrying the transgene (lanes 1, 3, and B) were distinguished from control littermates (lanes 2 and A) and wild-type controls (W) by the amplification of a 390-bp product (lane H, water; lane S, standards). (d) Heterozygous mice of the D6 line were further crossed to obtain a homozygous progeny. Mice carrying two alleles of the transgene (lanes 1 and 2) were distinguished from heterozygous littermates (lane 3) by hybridizing DNA with probe 1. Dot intensity was compared with that observed with DNA of mice from previous litters, whose homozygosity (lane C) or heterozygosity (lane B) had been biologically controlled by crossing with C57BL/6 controls.