Effects of luteinizing hormone and prostaglandin F(2α) on gap junctional intercellular communication of ovine luteal cells throughout the estrous cycle.

Cellular interactions mediated by contact-dependent pathways may be important to maintain luteal function. The objective of the present experiment was to evaluate the role of LH and prostaglandin F(2α) (PGF) in regulation of contact-dependent, gap junctional intercellular communication (GJIC) of ovine luteal cells from several stages of luteal development. Corpora lutea (CL) obtained from superovulated ewes on days 5 (n=7), 10 (n=8), and 15 (n=9) after estrus were dispersed with collagenase and cell types were separated by elutriation. Cells were plated as a mixed population (nonelutriated), or as small or large luteal cell fractions, and incubated in serum-free media containing no hormone, LH (100 ng/mL), PGF (100 ng/mL), LH+PGF, or dibutyryl cAMP (dbcAMP; 2 mM) for 18-24 h. Media were collected for evaluation of progesterone (P4) concentrations and replaced with media containing fluorescent dye. Then the rate of GJIC was evaluated by using the fluorescence recovery after photobleaching technique and laser cytometry. The rate of GJIC was determined for selected cells: small luteal cells in contact only with small luteal (S-S) cells; large luteal cells in contact only with small luteal (L-S) cells; and large luteal cells in contact only with large luteal (L-L) cells. LH increased (p<0.01) GJIC for S-S on d 5 and 10 and for L-S cells across the estrous cycle, but did not affect GJIC for L-L cells. PGF increased (p<0.05) GJIC for L-L cells on d 10 and 15, and decreased (p<0.05) GJIC for S-S cells from d 5 and 10 of the estrous cycle. LH+PGF increased (p<0.05) GJIC for S-S cells on d 5 and 10, and for L-S and L-L cells on d 10 and 15 of the estrous cycle. In addition, PGF diminished (p<0.05) LH-stimulatory effects on GJIC for S-S cells from d 5 and 10, and for L-S cells from d 5 of the estrous cycle. Dibutyryl cAMP stimulated (p<0.05) GJIC between all evaluated cell types across the estrous cycle. LH and dbcAMP stimulated (p<0.05) P4 secretion by mixed and small luteal cell fractions, PGF alone did not affect basal P4 secretion, but LH+PGF stimulated (p<0.05) P4 production by small luteal cells across the estrous cycle. PGF diminished (p<0.05) LH-stimulatory effects on P4 production in mixed populations of luteal cells across the estrous cycle.These data demonstrate that both luteal cell types communicate with each other, and the rate of communication was affected by LH, PGF, and dbcAMP. Modulation of gap junctional contact-dependent intercellular communication may be an important mechanism by which regulatory signals are transduced during luteal growth, differentiation, and regression in sheep.