One of the effects of neoplastic transformation by a variety of factors is a decrease in gap junctional, intercellular communication (GJIC). The investigation of junctional permeability is usually conducted through the microinjection of the fluorescent dye, Lucifer yellow, followed by observation of its migration into neighboring cells. This is a time-consuming approach, requiring expensive equipment. To overcome these problems, a novel technique was devised which takes advantage of the ability of short electric pulses to create transient "pores" on the cell membrane through which Lucifer yellow can enter, simultaneously and into large numbers of cells, with minimal disturbance to cellular metabolism. Cells were grown on a glass slide, half of which was coated with electrically conductive, optically transparent, indium-tin oxide. An electric pulse was applied in the presence of Lucifer yellow, causing its penetration into the cells growing on the conductive half of the slide, and the migration of the dye to the nonelectroporated cells growing on the nonconductive area was microscopically observed under fluorescence illumination. Using this technique, we investigated the relationship between expression of the middle tumor antigen of polyoma virus (mT) and GJIC in two representative cell systems with different responses to mT. The results show that low mT expression levels, although unable to transform rat F111 cells fully, are able to interrupt GJIC. Although parts of this mechanism might be mediated through protein kinase C (PKC), mT appears to have additional functions. PKC, however, had the opposite effect upon junctional permeability in a clone of mouse NIH-3T3 fibroblasts; intercellular communication in these cells appears to require PKC activity.

The molecular mechanisms that regulate the synthesis of the myometrial gap junction protein, connexin-43 (Cx-43), are controversial. We measured myometrial Cx-43 messenger RNA, protein and gap junction frequency, and area in myometrial samples collected from nonpregnant rats and pregnant rats at days 5, 10, 15, 17, 18, 19, 20, 21, 22, 23 (during delivery), and 1 day postpartum and correlated these data with plasma concentrations of estradiol 17 beta and progesterone. Cx-43 transcripts were low or undetectable (connexin-43:glyceraldehyde phosphate dehydrogenase ratio < 0.2) in nonpregnant rats or in rats before day 10 of pregnancy. Transcripts rose to 0.52 +/- 0.11 on day 10, increased (2.9-fold) to 1.51 +/- 0.48 on day 22, and increased a further 2.9-fold to maximal levels of 4.42 +/- 0.67 during labor. Cx-43 protein was barely detectable on day 21 [0.12 +/- 0.04 relative optical density (ROD) units], increased 2.5-fold on day 22 (0.30 +/- 0.04 ROD units), and a further 3.7-fold during delivery (1.10 +/- 0.15 ROD units), at a time when gap junctions were present in large numbers in the cell membrane. Between day 21 and delivery the increase in Cx-43 transcripts (8.2-fold) and protein (9.2-fold) were of a similar magnitude. There was a significant positive correlation between the increases in Cx-43 transcripts and the increase in the ratio of plasma estradiol to progesterone. Levels of Cx-43 transcripts, protein, and gap junctions fell rapidly postpartum. Our data demonstrate: 1) that transcripts encoding the gap junction protein, Cx-43, are at maximal levels during delivery and that this increase is temporally associated with increases in Cx-43 protein and the appearance of gap junctions; and 2) that these data, in association with changes in plasma steroid concentrations, are consistent with myometrial Cx-43 transcript levels being regulated positively by estrogen and negatively by progesterone during pregnancy.

Myometrial tissues from immature, ovariectomized mature, and pregnant rats were examined by thin-section or freeze-fracture electron microscopy for the presence of gap junctions following treatment with steroid hormones and inhibitors of prostaglandin synthesis. Gap junctions were present in the myometrium of immature rats after repeated injections of estradiol or diethylstilbestrol (500 micrograms/day). The frequency of gap junctions in immature rats after estradiol treatment was similar to that found during delivery. Concomitant administration of indomethacin or meclofenamate with estradiol (50 and 500 micrograms/day) for 3 or 4 days enhanced the effect of estradiol on gap junctions. Progesterone depressed the response of 500 micrograms/day estradiol and both 50 and 500 micrograms/day doses of estradiol when administered together with the inhibitors of prostaglandin synthesis. Gap junctions were also present in the myometria of ovariectomized mature rats after treatment with estradiol by itself or together with indomethacin, and these effects were suppressed by concomitant progesterone administration. These results indicate that the steroid hormones and prostaglandins may interact to modulate the presence of gap junctions in the myometrium.

The purpose of this study was to describe the design and evolution of a unique and successful Master of Science program in anatomical sciences at one Canadian post-secondary institution and to evaluate its long-term impact on student learning. This program prepares students to teach anatomy and design curricula in the anatomical sciences and is structured around three pillars of competency-content (disciplinary knowledge and transferable skills), pedagogy, and inquiry. Graduates of the program from the last ten years were surveyed, to better understand the knowledge, skills, and habits of mind they have adopted and implemented since completion. Interest was taken in identifying aspects of the program that students found particularly beneficial and areas that needed to be further developed. Based on the findings, this program has been a highly valuable experience for the graduates especially in helping them develop transferable skills, and grow as individuals. The hope is that other institutions that have similar programs in place or are considering developing them would benefit from this description of the program design and the sharing of the lessons learned.