Effects of prolactin on expression of the mRNAs encoding the immediate early genes zif/268 (NGF1-A), nur/77 (NGF1-B), c-fos and c-jun in the hypothalamus.

Prolactin (PRL) exerts a short-loop negative feedback effect on hypothalamic neurons which control its secretion from the anterior pituitary gland. The purpose of this study was to identify the location of hypothalamic neurons which respond to acute PRL exposure. Increasing evidence indicates that excitation of neurons often results in the rapid transcription of immediate early genes (IEGs). In the present study, quantitative in situ hybridization histochemistry (ISHH) was used to visualize the induction of mRNAs for four different IEGs: zif/268 (NGF1-A), nur/77 (NGF1-B), c-fos and c-jun. Three groups of male rats were compared: unmanipulated controls, rats injected s.c. with 2.4 mg ovine PRL (oPRL) suspended in polyvinylpyrrolidone (PVP), and PVP-injected controls. Animals were decapitated 0, 0.5, 1, 2, 3 or 4 h following injection. In all rats, the four probes labeled cells within the cortex, particularly the cingulate and piriform cortices, the hippocampus and the striatum. In the arcuate nucleus, there was a modest increase in the average number of cells/animal which expressed zif/268 mRNA following the injection of PVP and oPRL at all times studied. The average area of grains/cell representing zif/268 message also increased following the injection stimulus. The number of neurons expressing nur/77 mRNA was greater in PRL-treated rats compared with PVP-treated controls 0.5 and 1 h following injection. Nur/77-labeled neurons were co-extensive with the tuberoinfundibular dopaminergic (TIDA) neurons. The data suggest that cells located within the arcuate nucleus are involved in mediating PRL autofeedback on the brain.

Castration decreases single cell levels of mRNA encoding glutamic acid decarboxylase in the diagonal band of broca and the sexually dimorphic nucleus of the preoptic area.

Using quantitative in situ hybridization histochemistry (ISHH), we determined the effect of castration on single cell levels of glutamic acid decarboxylase (GAD) mRNA in discrete hypothalamic regions of the male rat brain associated with the control of gonadotropin secretion. A 48-base oligodeoxynucleotide probe was used to detect with equal affinity the two isoforms of GAD message, GAD65 and GAD67. GAD message also was quantitated in a number of selected areas of the brain to contrast GAD gene expression amongst several populations of GABAergic neurons. Comparison of 11 brain regions demonstrated a 9.3-fold range in the quantity of single cell GAD mRNA with levels being highest in the amygdala and the diagonal band of Broca, moderate in the piriform cortex, caudate nucleus, substantia innominata, globus pallidus, cingulate cortex and medial septal nucleus, and lowest in the lateral septal nucleus and the medial preoptic nucleus (MPN). Castration markedly reduced single cell GAD mRNA levels in the DBB and the MPN, two discrete hypothalamic structures known to contain dendritic fields, cell bodies, and axons of GnRH neurons projecting to the median eminence. A striking finding was a dense core of steroid-sensitive GABAergic neurons within the MPN comprising the sexually dimorphic nucleus of the preoptic area (SDN-POA). Similar to the MPN as a whole, the amount of GAD mRNA expressed by cells in the SDN-POA of sham operated control rats was greater than in castrated animals. GAD mRNA levels were inversely related to serum LH titers, suggesting a role for these neurons in the mechanism controlling gonadal steroid negative feedback on LH secretion. This report provides the basis for future work to determine if GAD65, GAD67 or whether both isoforms are affected by gonadal steroid input.