Glucocorticoids down-regulate lipopolysaccharide-induced de novo production of neurotensin mRNA in the rat hypothalamic, paraventricular, corticotrophin-releasing hormone neurons.

OBJECTIVE: Intraperitoneal injection of the endotoxin lipopolysaccharide (LPS) produces inflammation accompanied by activation of the immune system and the secretion of cytokines. Cytokines stimulate the hypothalamo-pituitary-adrenal (HPA) axis to release the anti-inflammatory corticosterone which controls its own production by acting on the HPA axis. Upstream in the HPA axis are neuroendocrine corticotrophin-releasing hormone (CRH) neurons located in the paraventricular nucleus (PVN), whose multipeptidergic phenotype changes during inflammation: while CRH mRNA is up-regulated in these conditions, neurotensin (NT) mRNA expression is induced de novo. The negative feedback control of glucocorticoids on CRH production is well documented; however, their action on NT production in the PVN of the hypothalamus is poorly documented. The aim of this study was to determine if glucocorticoids modulate the de novo production of NT during inflammation. METHODS: Using quantitative in situ hybridization histochemistry, we examined whether the absence (adrenalectomy) or excess (corticosterone implants) of glucocorticoids modulate de novo production of NT mRNA in the PVN during inflammation induced by LPS treatment. RESULTS: A relatively low dose of LPS (50 microg/kg) that is not efficient to induce NT mRNA production in the PVN becomes efficient after adrenalectomy. Moreover, corticosterone excess reduces LPS-induced production of NT mRNA in the PVN. CONCLUSION: Glucocorticoids exert a negative control on NT mRNA production in the PVN of the hypothalamus, and this effect requires that NT mRNA production be triggered, such as during inflammation.

Seasonal variations of the beta-endorphin neuronal system in the mediobasal hypothalamus of the jerboa (Jaculus orientalis).

The distribution of neurons expressing beta-endorphin immunoreactivity was explored in the brain of adult jerboa during two distinct periods characterizing its reproductive cycle. A large presence of cell bodies displaying beta-endorphin immunoreactivity occured within different parts of the mediobasal hypothalamus along its rostrocaudal extent, from the retrochiasmatic area to the posterior arcuate nucleus. Quantitatively, the highest density of immunoreactive beta-endorphin neurons was noted at the medial level of the arcuate nucleus. Furthermore, a seasonal study showed that the number of IR-beta-endorphin neurons was highest in the anterior portion of the arcuate nucleus of jerboas sacrificed in autumn as compared to those sacrificed during spring-summer. Quantitatively, the number of beta-endorphin containing neurons in autumn was 200% in comparison to that found in spring-summer. These results suggest that beta-endorphin containing neuronal population especially localized in the anterior part of arcuate nucleus, exerts an inhibitory influence on the GnRH neurosecretory system in the jerboa, notably in autumn, probably via an increasing expression of its products. The results provide morphofunctional arguments in favour of inhibitory opioid control of GnRH neurons activity and hence the neuroendocrine events regulating reproduction in jerboa.

Paraventricular nucleus neurons producing neurotensin after lipopolysaccharide treatment project to the median eminence.

Inflammation consists in secretion of cytokines that stimulate the hypothalamo-pituitary-adrenal (HPA) axis to release the anti-inflammatory corticosterone. Upstream in this axis are corticotropin-releasing hormone (CRH) neurons in the paraventricular nucleus (PVN) whose multipeptidergic phenotype changes: both corticotropin-releasing hormone mRNAs and neurotensin mRNAs are up-regulated. Combining in situ hybridization with a retrograde neuronal marker, we demonstrated that neurotensin-containing neurons in the paraventricular nucleus project to the median eminence.

Immunocytochemical distribution of Met-enkephalin-Arg6-Gly7-Leu8 (Met-8) in the auditory system of the rat.

Methionine-enkephalin-Arg(6)-Gly(7)-Leu(8) (Met(8)) is known to act as a neurotransmitter or neuromodulator and it has been implicated in pain, cardiovascular and motor mechanisms, but its role in audition is currently unknown. In the present study we have applied an immunocytochemical technique and describe the distribution of cell bodies and fibers containing Met(8) in the auditory pathway of the rat. The main finding is that we found either Met(8)-immunoreactive fibers or cell bodies or both in virtually all nuclei of the rat auditory system except for the medial superior olive and the ventral division of the medial geniculate body in which we did not find any immunoreactivity for Met(8). This suggests that the neuropeptide Met(8) is widely distributed throughout the auditory system of the rat. Our results suggest that Met(8) could play at least two roles in hearing. It seems to be involved in the processing of the descending auditory pathway, and it may be implicated in the multisensory integration of auditory information that takes place in the non-lemniscal auditory pathway.

The endogenous cannabinoid, anandamide, activates the hypothalamo-pituitary-adrenal axis in CB1 cannabinoid receptor knockout mice.

The purpose of this study was to investigate the effects of the endogenous cannabinoid arachidonoyl-ethanolamide, anandamide (AEA), on the activity of the hypothalamo-pituitary-adrenal (HPA) axis in cannabinoid receptor (CB(1) receptor) inactivated (KO) mice. A low dose (0.01 mg/kg i.p.) of AEA significantly increased plasma corticotropin (ACTH) and corticosterone concentrations in both wild-type (+/+) and in mutant (-/-) animals. In each case, hormone levels reached their peaks at 90 min after AEA administration. In a parallel experiment, AEA administration was preceded by the injection of SR 141716A (1.0 mg/kg), a selective and potent CB(1) receptor antagonist, or of capsazepine (5.0 mg/kg), a potent vanilloid receptor of type 1 (VR1) antagonist. The latter drugs did not prevent the effects of AEA on the HPA axis. Using Fos protein immunohistochemistry, we observed that the parvocellular part of the hypothalamic paraventricular nucleus (PVN) was activated as early as 45 min after AEA injection and reached peak levels after 60 min in both +/+ and -/- mice. Furthermore, the CB(1) and VR1 receptor antagonists did not block the effects of AEA on Fos immunoreactivity. The results strongly support the view that activation of the HPA axis produced by AEA possibly occurs via a currently unknown (CB(x)) cannabinoid receptor present in PVN.

An immunocytochemical mapping of methionine-enkephalin-Arg6-Gly7-Leu8 in the cat brainstem.

The distribution of methionine-enkephalin-Arg6-Gly7-Leu8-immunoreactive cell bodies and fibres was studied in the brainstem of the cat using an indirect immunoperoxidase technique. In the mesencephalon, immunoreactive cell bodies were observed in the periaqueductal grey, the dorsal raphe nucleus, the central and pericentral nuclei of the inferior colliculus and the pericentral division of the dorsal tegmental nucleus. In the pons, immunoreactive cell bodies were observed in the dorsolateral division of the pontine nucleus; below the central division of the dorsal tegmental nucleus; above the dorsolateral division of the pontine nucleus, and close to the superior cerebellar peduncle. In the medulla oblongata, immunoreactive cell bodies were observed in the laminar spinal trigeminal nucleus and in the lateral tegmental field; the dorsal motor nucleus of the vagus; the prepositus hypoglossal nucleus; the medial nucleus of the solitary tract; the rostral division of the cuneate nucleus, and close to the parvocellular division of the alaminar spinal trigeminal nucleus. The highest (moderate) density of immunoreactive fibres was observed in the periaqueductal grey; the parvocellular and magnocellular divisions of the alaminar spinal trigeminal nucleus; the laminar spinal trigeminal nucleus; the rostral division of the cuneate nucleus; the dorsal motor nucleus of the vagus; the lateral nucleus of the solitary tract, and in the midline between the central divisions of the reticulotegmental pontine nucleus. The widespread distribution of methionine-enkephalin-Arg6-Gly7-Leu8 in the cat brainstem indicates that the peptide might be involved in several physiological functions.

A confocal microscopic study of gonadotropin-releasing hormone (GnRH) neuron inputs to dopaminergic neurons containing estrogen receptor alpha in the arcuate nucleus of GnRH-green fluorescent protein transgenic mice.

The purpose of the present study was to determine whether the septo-preoptico-tuberoinfundibular gonadotropin-releasing hormone (GnRH) pathway comes in close juxtaposition with tyrosine hydroxylase immunoreactive (TH-IR) neurons in the arcuate nucleus of female mice. Immunohistochemical staining with a TH monoclonal antibody coupled with confocal microscopy was employed on vibratome-cut brain sections of female GnRH-green fluorescent protein (GFP) transgenic mice to evaluate possible appositions between GnRH and tuberoinfundibular dopaminergic (TIDA) neurons. TH-IR neurons of the arcuate nucleus received GnRH neuronal appositions in adult female mice at proestrus and estrus stages. In contrast, no GnRH appositions were observed in adult females at diestrus. Subsequently, double immunohistochemical staining for TH and estrogen receptor-alpha (ERalpha) was performed to examine the role of estradiol on this relationship. We found that most TH-IR neurons contacted by GnRH fibers were immunoreactive for ERalpha. Our observations suggest that GnRH neurons communicate directly with TIDA neurons in the adult female. Furthermore, ERalpha activation in TIDA neurons may be involved in the formation of connections between GnRH neurons and TIDA neurons.

In vivo activation of the interleukin-6 receptor/gp130 signaling pathway in pituitary corticotropes of lipopolysaccharide-treated rats.

Adrenocorticotropic hormone (ACTH) release from anterior pituitary corticotropes is greatly increased during peripheral inflammation induced by lipopolysaccharide (LPS) administration. Interleukin-6 (IL-6) is thought to participate in LPS-induced ACTH release, but whether or not corticotropes are directly targeted by this cytokine is unclear. Therefore, we investigated the expression and activation of IL-6 signaling components in the pituitary of rats 2 and 4 h after administration of LPS (250 microg/kg). Intraperitoneal LPS treatment provoked the nuclear translocation of signal transducer and activator of transcription 3 (STAT-3) and Fos expression in the anterior pituitary lobe, as demonstrated by immunohistochemistry. By using in situ hybridization, we demonstrated that suppressor of cytokine signaling 3 (SOCS-3) and c-fos mRNAs were significantly induced by the LPS treatment in the anterior lobe of the pituitary. Dual in situ hybridization revealed that most corticotropes expressed IL-6 receptor and gp130 mRNAs, and that 2 h after LPS treatment, SOCS-3 and c-fos mRNAs were induced in corticotropes. Our results suggest that LPS-induced IL-6 could regulate the hypothalamo-pituitary-adrenal axis by directly targeting corticotropes during peripheral inflammation.

Mapping of neurokinin-like immunoreactivity in the human brainstem.

BACKGROUND: Using an indirect immunoperoxidase technique, we have studied the distribution of immunoreactive fibers and cell bodies containing neurokinin in the adult human brainstem with no prior history of neurological or psychiatric disease. RESULTS: Clusters of immunoreactive cell bodies and high densities of neurokinin-immunoreactive fibers were located in the periaqueductal gray, the dorsal motor nucleus of the vagus and in the reticular formation of the medulla, pons and mesencephalon. Moreover, immunoreactive cell bodies were found in the inferior colliculus, the raphe obscurus, the nucleus prepositus hypoglossi, and in the midline of the anterior medulla oblongata. In general, immunoreactive fibers containing neurokinin were observed throughout the whole brainstem. In addition to the nuclei mentioned above, the highest densities of such immunoreactive fibers were located in the spinal trigeminal nucleus, the lateral reticular nucleus, the nucleus of the solitary tract, the superior colliculus, the substantia nigra, the nucleus ambiguus, the gracile nucleus, the cuneate nucleus, the motor hypoglossal nucleus, the medial and superior vestibular nuclei, the nucleus prepositus hypoglossi and the interpeduncular nucleus. CONCLUSION: The widespread distribution of immunoreactive structures containing neurokinin in the human brainstem indicates that neurokinin might be involved in several physiological mechanisms, acting as a neurotransmitter and/or neuromodulator.

GABAA receptor epsilon subunit expression in identified peptidergic neurons of the rat hypothalamus.

Dual-labeling immunohistochemical or in situ hybridization studies for the recently cloned epsilon-subunit and several neuropeptides were performed in the rat hypothalamus. We revealed an extensive co-expression (>90%) with hypocretin (Hcrt), oxytocin (OT), the gonadotropin-releasing hormone (GnRH), and the melanin-concentrating hormone (MCH) peptides, whereas occasional co-expression (<10%) with cocaine-amphetamine-regulated transcript (CART) was found. Our results suggest that novel GABA(A) receptor subtypes comprising epsilon-subunit are important for metabolic and neuroendocrine functions.