Philip J. Brooks

Changes in preproenkephalin messenger RNA level in the rat ventromedial hypothalamus during the estrous cycle

To gain a better understanding of the relationship between the female rat reproductive system and preproenkephalin (PPE) expressing neurons under physiological conditions, we examined changes in PPE mRNA levels in the mediobasal hypothalamus during the rat estrous cycle by means of northern blotting and in situ hybridization histochemistry (ISHH). In the Northern blot studies, we found that PPE mRNA levels in the mediobasal hypothalamus were significantly increased by noon of proestrus compared to those in the morning and stayed high until diestrus day 1, and returned toward low levels on diestrous day 2. In contrast, measured as controls, glyceraldehyde-3-phosphate-dehydrogenase mRNA levels were significantly higher on proestrus regardless of time of day compared to diestrus day 2, and levels of calcineurin mRNA on proestrous and estrous were significantly lower than diestrous day 1 and day 2. ISHH studies revealed that these changes in PPE mRNA levels were specific in the ventromedial hypothalamic nucleus pars ventrolateralis (VMHVL), since we could not see any significant changes in signal in other parts including ventromedial hypothalamic nucleus pars dorsomedialis and arcuate hypothalamic nucleus. In the VMHVL, PPE mRNA levels in the afternoon of proestrous were significantly higher than those in the afternoon of diestrous day 2 whereas no significant change in PPE mRNA was observed in the caudate-putamen. The present study provides additional information relevant to possible implications of PPE gene expression in female reproductive systems, since changes in PPE mRNA levels may be associated with estrogen as well as progesterone or other hormonal concentrations during the estrous cycle.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

Prolactin receptor messenger RNA is synthesized by the epithelial cells of the choroid plexus

To identify cellular sites of prolactin receptor messenger RNA synthesis in the rat brain, we used a combined reverse transcriptase-polymerase chain reaction protocol to generate single stranded DNA probes for in situ hybridization. The results of these experiments identify the epithelial cells of the choroid plexus as a major site of prolactin receptor gene expression in the rat central nervous system.

Cell-specific expression of preproenkephalin intronic heteronuclear RNA in the rat forebrain

Using in situ hybridization with multiple probes to the rat preproenkephalin gene, we have identified a novel population of cells in the reticular thalamic nucleus and basal forebrain which express RNA derived from the preproenkephalin gene. These cells contain nuclear RNA from downstream of an alternate transcription start site in intron A of the preproenkephalin gene (Kilpatrick et al., Mol. Cell Biol., 10 (1990) 3717-3726), while in the same cells preproenkephalin exon 2 RNA is undetectable. The results suggest that in this population of cells, preproenkephalin gene transcription initiates from the intron A initiation site, and is regulated by an additional mechanism which results in the accumulation of nuclear preproenkephalin intron A-derived heteronuclear RNA. The anatomical distribution of these cells indicates that they may be involved in the control of cerebral cortical function.