LUTEINIZING-HORMONE-RELEASING HORMONE-RECEPTOR MESSENGER-RIBONUCLEIC-ACID EXPRESSION IN THE RAT PITUITARY DURING LACTATION AND THE ESTROUS-CYCLE

To study mechanisms underlying the modulation of luteinizing hormone-releasing hormone receptor (LHRH-R) during lactation and the estrous cycle, we used a reverse transcriptase-polymerase chain reaction (RT-PCR) procedure to generate a probe for rat LHRH-R messenger RNA (mRNA). Using primers based on the mouse sequence, we amplified an approximately 300 bp fragment from rat pituitary complementary DNA. This PCR product was shown to be part of LHRH-R cDNA by direct sequencing and by comparing to the rat LHRH-R cDNA reported recently. Then, this PCR fragment was used as a probe for northern blotting analysis. The level of LHRH-R mRNA in the pituitary was significantly decreased during lactation, by approximately 80%, compared to that of ovariectomized and intact (diestrous and metestrous cycling) rats while no statistical difference in glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) mRNA level was observed between groups. During the estrous cycle, the level of LHRH-R mRNA in the pituitary was about two-fold higher on diestrous day 2 and the morning of proestrus than that on diestrous day 1 and quickly returned toward control level by noon of proestrus. In addition, we found that GAPDH mRNA levels from a so-called housekeeping gene often thought to be unchanged under different conditions, were significantly higher on proestrus while levels of 18S rRNA were not significantly changed. The large decrease in LHRH-R mRNA during lactation could account for the changes in LHRH binding previously reported.