The effect of ovine GH (oGH) in vivo and recombinant bovine insulin-like growth factor-I (rbIGF-I) in vitro on gill Na+,K(+)-ATPase activity was investigated in two seasonal experiments conducted during the parr-smolt transformation period of coho salmon. In 1991, when fish were held under a photoperiod of 12 h light : 12 h darkness, the stimulatory effect of oGH (1 microgram/g) on gill Na+,K(+)-ATPase in vivo decreased at the time of expected parr-smolt transformation. Gill Na+,K(+)-ATPase from control fish was insensitive to rbIGF-I in vitro from February to June, whereas GH treatment induced sensitivity to rbIGF-I (100-1000 micrograms/l) in vitro in February and March, but not later in development. In 1992, when fish were held under natural conditions, oGH (4 micrograms/g) stimulated gill Na+,K(+)-ATPase in vivo from February to July. There was, however, of pronounced developmental change in sensitivity of gill Na+,K(+)-ATPase to rbIGF-I in vitro. In February, gills from control fish were insensitive, but oGH treatment in vivo induced sensitivity to rbIGF-I in vitro (100-1000 micrograms/l). In April and May, control fish were sensitive to rbIGF-I in vitro. This sensitivity was not further potentiated by oGH treatment in vivo. In June, gills from control or oGH-treated fish were not sensitive to rbIGF-I in vitro, but in July exogenous oGH again induced gill tissue sensitivity to rbIGF-I at 1000 micrograms/l. Both studies showed that rbIGF-I stimulates gill Na(+),K(+)-ATPase directly; an ability that may depend on priming by endogenous or exogenous GH. This supports the role of IGF-I as an endocrine mediator for GH action during parr-smolt transformation.

Ovine prolactin injected subcutaneously for 2 days stimulated an increase in fluid uptake in vitro by everted gut sacs from male Sprague\p=m-\Dawley rats. Four intestinal segments from each rat were incubated, but only two of the four segments tested showed a prolactin-stimulated increase in fluid transfer in intact rats. In adrenalectomized\p=m-\nephrectomized rats all four sacs showed an increased fluid transfer after prolactin treatment. Sacs incubated for 1 h in glucose\p=m-\Ringer solution containing 0\m=.\1 mg ovine prolactin/ml did not show an increased water transfer as compared with controls incubated in medium containing albumin. Dietary changes seemed to alter the sensitivity of the gut to prolactin.

Normal endometrial luminal epithelial cells isolated from ovariectomized approximately 40-day-old BALB/cCrgl mice were purified by Percoll density gradient centrifugation and grown as primary cultures in collagen gel matrix and serum-free medium. Cells increased threefold in number during the 9-day culture period. Deletion of insulin, epidermal growth factor or bovine serum albumin resulted in decreased growth. Addition of any single factor to the unsupplemented medium had no effect. Relatively high levels of cytosolic oestrogen receptors and progestin receptors were demonstrable in the cultures. Addition of oestrogen did not enhance epithelial cell proliferation. On the contrary, all doses of oestrogen (180 fmol/l to 218 nmol/l) were inhibitory. Continuous exposure to oestradiol-17 beta (1.8 nmol/l) for 9 days in serum-free medium resulted in a decrease in cytosolic oestrogen receptors with an associated nuclear accumulation of oestrogen receptors. A corresponding increase in cytosolic progestin receptors was also observed, indicating that no qualitative modification of the oestrogen receptor system had occurred. Thus, as previously reported for vaginal epithelial cells, oestrogen, despite its stimulation of specific product synthesis (progestin receptors), did not increase proliferation of endometrial luminal epithelial cells in this culture system.

SUMMARY Mammotrophic (i.e. mammogenic and/or lactogenic) activity of mouse placentae of different stages (4–19 days of pregnancy) was examined using organ co-culture of placental explants with mammary tissue. The test mammary tissues were taken from midpregnant (11–12 days) nulliparous A/Crgl mice and cultured in a synthetic medium (Waymouth's) supplemented with insulin (5 μg/ml) and aldosterone (1 μg/ml). The responses of mammary gland to placental explants were judged histologically, and were compared with those seen after the addition of ovine prolactin (5 μg/ml). With placentae from 6- to 19-day pregnant animals, distinct mammotrophic activity was seen, with the appearance of eosinophilic secretion in the mammary alveolar lumina, whereas with 4- or 5-day-old 'placentae', no mammotrophic activity was detected. Inasmuch as growth hormone does not substitute for prolactin in mammary gland development and function in the A/Crgl mouse, it can be concluded that a prolactin-like factor is present in the mouse placenta. The influence of placentae on mammary gland was further analysed by transplantation of placental fragments to mammary fat pads. Local lobuloalveolar development was prominent in some instances in the area around the placental transplants.

Effects of prolactin on transport properties of opercular membranes from seawater-adapted tilapia, Sarotherodon mossambicus, have been examined. These membranes are high conductance (average Gt approximately 4 mS.cm-2) tissues with short-circuit currents (I) equal to net chloride secretion. Despite high Gt, nonlinear current-voltage relationships suggest that opercular membranes cannot be classified as "leaky" tissues. Variability among membranes is reflected in a linear relationship between I and Gt with a slope equal to 26 mV and the zero-current Gt intercept equal to 0.45 mS.cm-2. Prolactin injections decrease I and Gt in a dose-dependent manner. Phosphodiesterase inhibition, without effect on I in untreated fish, often partially reverses these prolactin effects. Gt-I data from prolactin-treated fish yield a slope of 18 mV and a Gt intercept of 0.10 mS.cm-2. The effects of prolactin are discussed in terms of conventional equivalent circuit analysis. Discrepancies between predictions based on this model and the actual data indicate that an alternative interpretation, based on a heterogeneous cell population, is more accurate. Analysis of this circuit suggests that the ratio of paracellular to active transport pathway conductances associated with chloride cells is constant and that differences in Gt and I are due to parallel changes in these conductances. Prolactin may effectively "remove" chloride cells from these membranes as well as inhibit (reversible by elevated cellular cAMP levels) active transport pathway conductance of remaining cells.