Cysteamine-Induced Enhancement of Growth Hormone- Releasing Factor (GRF) Immunoreactivity in Arcuate Neurons: Morphological Evidence for Putative Somatostatin/GRF Interactions within Hypothalamus*

Abstract

The episodic secretion of GH is regulated through the reciprocal release into the hypophyseal portal circulation of two hypothalamic peptides, SRIF and GRF. Recent physiological evidence suggests that, in addition to exerting their well documented opposite actions at the level of the anterior pituitary, SRIF and GRF may interact within the central nervous system to modulate GH secretion. The aim of the present study was to provide morphological evidence to support the concept that SRIF exerts an influence on the GRF-containing neuronal system within the hypothalamus. To accomplish this, we used the thiol agent cysteamine (CSH; 300 mg/kg sc) to manipulate endogenous hypothalamic SRIF and examined its effects on the immunoreactivity and mRNA content of GRF-containing arcuate (ARC) neurons, using immunohistochemistry and semi-quantitative in situ hybridization, respectively. Hypothalamic SRIF content and pulsatile GH secretion were also monitored. CSH treatment reduced hypothalamic immunoreactive SRIF concentrations (to 60% of vehicle-injected control values) and severely suppressed the spontaneous surges of GH release (mean GH peak amplitude: 24.2 +/- 2.7 vs. 168.0 +/- 27.8 ng/ml in H2O-injected controls; P less than 0.001). CSH treatment also resulted in a striking increase in the number and labeling density of GRF-immunoreactive cells detected in the ARC nucleus. Cell counts revealed an overall 126% increase, over controls, in the mean number of detectable GRF-positive ARC neurons. This increase (P less than 0.01) was apparent over all rostrocaudal levels of the ARC nucleus, but was most pronounced within the caudal tier. Administration of CSH also produced a massive qualitative increase in the intensity and extent of staining of GRF-positive terminal elements in the external zone of the median eminence. Conversely, CSH treatment resulted in a decline in GRF mRNA-associated signal in cells of the ARC nucleus, with this decline being most evident in the caudal third of the nucleus. The finding of an increase in GRF immunoreactivity in ARC neurons in association with a decrease in plasma GH, as well as with a decline in relative GRF mRNA content, suggests that CSH leads to an overall decrease in GRF release from the median eminence. These results provide morphological evidence to support the concept of a SRIF-mediated central influence on the GRF-containing ARC neuronal system. Such central regulation of GRF by SRIF may be an important mechanism in the physiological control of pulsatile GH secretion.