Expression of c-fos in bulbar nuclei involved in cardiovascular control following the electrical stimulation of sensorimotor cortex in the rat.

Previous studies have shown that electrical stimulation of the sensorimotor cortex (SMC) induces responses of the autonomic nervous system such as variations in heart rate and arterial pressure. Neuroanatomical studies have shown the existence of monosynaptic projections from the SMC to the nucleus tractus solitarius (NTS), the rostral ventrolateral medulla (RVLM) and the dorsal nucleus of the vagus nerve (DNV), which are bulbar nuclei involved in cardiovascular control. The aim of the present study was to establish whether there exists a functional connectivity between the SMC and these nuclei. Electrical stimulation applied to the SMC of 7 rats for 1 h induced the expression of c-fos-protein-like immunoreactivity in the nucleus of some neurons in NTS, RVLM and DNV. These data support the view that the SMC has functional connections with bulbar neurons involved in cardiovascular control.

Retrogradely labelled neurosecretory neurones of the rat hypothalamic arcuate nucleus express Fos protein following systemic injection of GH-releasing peptide-6.

Previously, we demonstrated that the synthetic hexapeptide GH-releasing peptide (GHRP-6) activates a subpopulation of arcuate neurones, as reflected by increased electrical activation and by the detection of Fos protein in cell nuclei. Here we set out to determine (1) what proportion of the cells activated by GHRP-6 are neurosecretory neurones and (2) whether the cells activated by GHRP-6 contain tyrosine hydroxylase (TH; a marker of dopaminergic cells in this region) or beta-endorphin. In the first study, adult male rats were injected i.v. with the retrograde tracer, Fluorogold, to detect cells which project outside the blood-brain barrier (and are therefore likely to be neurosecretory neurones). Three days later the conscious rats were injected i.v. with 50 micrograms GHRP-6 and the brains processed for the immunocytochemical detection of Fos protein. Between 68% and 82% of the arcuate neurones expressing Fos protein following GHRP-6 injection were retrogradely labelled with Fluorogold. In the second study, conscious male rats, bearing a chronically implanted jugular catheter, were killed 90 min following an i.v. injection of 50 micrograms GHRP-6 and the brains were processed for the double immunocytochemical detection of Fos protein and either TH or beta-endorphin. Less than 7% (mean +/- S.E.M. = 6.7 +/- 2.6% nuclei/section per rat) of the arcuate neurones expressing Fos protein following GHRP-6 injection were TH-containing cells. Of 143 beta-endorphin-containing arcuate cells detected only four cells were identified as containing Fos protein. Thus, the majority of arcuate neurones activated by GHRP-6 (1) project outside the blood-brain barrier (and are therefore likely to be neuro-secretory neurones) and (2) were not identified as TH- or beta-endorphin-containing cells.

Distribution of NADPHdiaphorase and calbindin-D28k neurons in the lateral septal area of the guinea pig, with special reference to the enkephalinergic hypothalamo-septal tract.

Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPHd) histochemical techniques were used to identify neurons synthesizing nitric oxide in the lateral septum of the guinea pig. Double immunocytochemical procedures were used to detect neurons immunoreactive for calbindin-D28k and enkephalinergic fibers which project to the lateral septum. The present data demonstrate that (1) the neurons containing NADPH diaphorase and the neurons immunoreactive for calbindin-D28k are observed in discrete regions of the lateral septum; (2) these populations overlap in various areas of the lateral septum including its dorsal and mediolateral parts; (3) NADPH diaphorase and calbindin-D28k are colocalized in neurons located in the overlapping areas; (4) neurons identified by the presence of calbindin-D28k, NADPH diaphorase or both substances, are surrounded by enkephalinergic fibers. These observations indicate the chemical heterogeneity of the lateral septum and suggest that the enkephalinergic hypothalamo-septal tract does not preferentially contact a subpopulation of neurons.

GH-deficient dw/dw rats and lit/lit mice show increased Fos expression in the hypothalamic arcuate nucleus following systemic injection of GH-releasing peptide-6.

In the rat, the synthetic GH secretagogue GH-releasing peptide (GHRP-6) acts centrally to activate a subpopulation of arcuate neurones as reflected by increased electrical activation and by the detection of Fos protein in cell nuclei. Since GHRP-6 also induces GH secretion via a direct action on the pituitary, we set out to determine whether the central actions of GHRP-6 are mediated by GH itself. First, we demonstrated that peripherally administered GHRP-6 induces Fos expression in the arcuate nucleus of GH-deficient animals (dw/dw rats and lit/lit mice). Secondly, in dw/dw rats, neither intracerebroventricular injection of 15 micrograms recombinant bovine GH nor 1 microgram recombinant human IGF-I resulted in an increase in the number of cells expressing Fos protein in the arcuate nucleus (or in any other hypothalamic structure studied). These results support our hypothesis that GHRP-6 has a central site and mechanism of action and provide evidence to suggest that the activation of arcuate neurones by GHRP-6 is not mediated by a central action of GH or IGF-I. Furthermore, since the lit/lit mouse pituitary does not release GH following GHRP-6 administration, our finding that the central actions of GHRP-6 remain intact in these animals suggests the possible existence of two subpopulations of putative GHRP-6 receptors.