Release of Atrial Natriuretic Factor from Intact and Hypophysectomized Rat Hypothalamic Expiants

Experiments examined release of atrial natriuretic factor (ANF), measured by radioimmunoassay, from acutely prepared explants of rat hypothalamus maintained in vitro by intra-arterial perfusion of artificial cerebrospinal fluid. Perfusates collected from intact preparations contained 6.1 ± 0.6 pg (mean ± SEM) of ANF per 2-min sample. Following a 3-min infusion of noradrenaline (60 μM), ANF release increased significantly (P<0.05) to 11.4±1.4 pg/sample. Media collected from hypophysectomized preparations showed the same basal ANF release (6.8 ± 0.9 pg/sample) as intact preparations, but demonstrated no significant increase after noradrenaline infusions. Levels of spontaneous ANF release were not appreciably affected by the absence of the paraventricular nuclei and/or the anteroventral third ventricle area.
Extracted material from the perfusate by reverse-phase high performance liquid chromatography revealed two main peaks of immunoreactive ANF: a small molecular weight form that coeluted with synthetic ANF (99–126) and with similar biological activity in a radioreceptor assay, and a larger molecular weight form with the same elution profile as the ANF (1–126) prohormone.
These observations indicate that the ANF released from perfused rat hypothalamic explants contains distinct contributions from the hypothalamus (sites undetermined) and the neurointermediate lobe.     

The Response to TRH, LH-RH, Metyrapone and Vasopressin in Patients with Hypothalamo-Pituitary Disorders

Abstract. The effects of slow intravenous infusion of synthetic TRH on serum TSH and of synthetic LH-RH on serum LH measured by radioimmunoassay were studied in healthy persons and in 36 cases with sellar tumours and other hypo-thalamo-pituitary disorders. The results were also compared with the responses to oral metyrapone and intravenous infusion of lysine-8-vasopressin, as tests for the hypothalamo-pituitary-adrenal regulation. 200 μ g of TRH was found to give a mean increase of serum TSH of +10.4 μ/ml in eight males and of +15.7 μ/ml in nine females. LH-RH infusion was found to give a mean increase of serum LH of -f-1.78 ng/ml in six young males. The TRH test gave a pathologically low response in one out of nine patients with an intrasellar tumour. On the other hand in patients with a craniopharyngioma, which usually causes more pronounced pituitary destruction, the response was pathologically low in five out of six cases. A normal response to TRH was obtained in seven out of nine patients with disorders mainly affecting the hypothalamus. When twelve patients with pituitary adenomas were tested post-surgery six patients gave a pathologically low TRH response. The response to LH-RH was pathologically low in five out of eight cases with a pathologically low TRH response and normal in six out of six cases with a normal TRH response. However, there was a poor correlation between the results of these two tests and the vasopressin and metyrapone administration. This difference may be explained by the direct effect of the releasing hormones on the pituitary and the need for a certain hypothalamic function to obtain a response to the vasopressin and metyrapone tests.     

The ventromedial hypothalamus mediates predator fear memory

The amygdala has been shown to be essential for the processing of acute and learned fear across animal species. However, the downstream neural circuits that mediate these fear responses differ according to the nature of the threat, with separate pathways having been identified for predator, conspecific and physically harmful threats. In particular, the dorsomedial part of the ventromedial hypothalamus (VHMdm) is critical for the expression of defensive responses to predators. Here, we tested the hypothesis that this circuit also participates in predator fear memory by transient pharmacogenetic inhibition of the VMHdm and its downstream effector, the dorsal periaqueductal grey, during predator fear learning in the mouse. Our data demonstrate that neural activity in the VMHdm is required for both the acquisition and recall of predator fear memory, whereas that of its downstream effector, the dorsal periaqueductal grey, is required only for the acute expression of fear. These findings are consistent with a role for the medial hypothalamus in encoding an internal emotional state of fear.     

The role of the brain renin–angiotensin system in hypertension: Implications for new treatment

Hypertension affects 26% of adults and is in constant progress related to increased incidence of obesity and diabetes. One-third of hypertensive patients may be successfully treated with one antihypertensive agent, one-third may require two agents and in the remaining patients will need three agents for effective blood pressure (BP) control. The development of new classes of antihypertensive agents with different mechanisms of action therefore remains an important goal. Brain renin–angiotensin system (RAS) hyperactivity has been implicated in hypertension development and maintenance in several types of experimental and genetic hypertension animal models. Among the main bioactive peptides of the brain RAS, angiotensin (Ang) II and Ang III have similar affinities for type 1 (AT1) and type 2 (AT2) Ang II receptors. Following intracerebroventricular (i.c.v.) injection, Ang II and Ang III similarly increase arginine–vasopressin (AVP) release and BP. Blocking the brain RAS may be advantageous as it simultaneously (1) decreases sympathetic tone and consequently vascular resistance, (2) decreases AVP release, reducing blood volume and vascular resistance and (3) blocks angiotensin-induced baroreflex inhibition, decreasing both vascular resistance and cardiac output. However, as Ang II is converted to Ang III in vivo, the exact nature of the active peptide is not precisely determined. We summarize here the main findings identifying AngIII as one of the major effector peptides of the brain RAS in the control of AVP release and BP. Brain AngIII exerts a tonic stimulatory effect on BP in hypertensive rats, identifying brain aminopeptidase A (APA), the enzyme generating brain Ang III, as a potentially candidate target for hypertension treatment. This has led to the development of potent orally active APA inhibitors, such as RB150 — the prototype of a new class of centrally acting antihypertensive agents.     

Dorsomedial hypothalamic nucleus neurons integrate important peripheral feeding-related signals in rats

Several studies have implicated the dorsomedial hypothalamic nucleus (DMN) in regulation of feeding behavior and body weight, but clear mechanisms by which it controls food intake are not well understood. We report the results of the present study, which showed that the DMN receives important peripheral short- and long-term feeding-related afferent signals, including gastric vagal, glycemia, and cholecystokinin (CCK) inputs, as well as from leptin, an adipostatic signal that forcefully inhibits food intake and increases metabolic rate. Among the 279 DMN neurons recorded, 173 (62.0%) responded to stimulation of gastric vagal nerves. Also, of the 123 DMN neurons responsive to gastric vagal stimulation that were tested with the administration of intravenous glucose, 75 (61.0%) were identified as being glycemia sensitive. Moreover, it is noteworthy that of the 23 DMN neurons that responded to both gastric vagal and intravenous glucose stimulation, most (19 of 23, 82.6%) were sensitive to circulating leptin, and some neurons (n = 7) were also responsive to systemic CCK, suggesting that gastric vagal, glycemic, CCK, and leptin inputs converge on single DMN neurons. Furthermore, synergistic interactions between leptin and glucose on single DMN neurons were observed (n = 6). These results demonstrate that those important peripheral feeding-related gastric vagal, glycemic, CCK and leptin signals not only reach the DMN but also interact on single DMN neurons, suggesting that the DMN may not just function as a relay station, but independently integrate the short-term and long-term feeding-associated information and actively participate in the direct regulation of feeding behavior.     

代谢综合征合并功能性下丘脑性闭经的诊断:附病例报告

目的 探讨代谢综合征合并闭经的诊断思路.方法 对一例代谢综合征合并闭经患者的临床资料及实验室检查结果进行分析,并复习相关文献,总结该病的诊断方法.结果 该代谢综合征患者存在无排卵,超声检查发现卵巢多囊样改变,但性激素检查发现可被促性腺激素释放激素(GnRH)所兴奋的低促性腺性性功能减退.结论 对于符合鹿特丹标准的闭经患者,在排除其他高雄激素病因同时,亦应注意排除其他引起排卵障碍的疾病.     

Matrix metalloproteinases MMP2 and MMP9 are upregulated by noradrenaline in the mouse neuroendocrine hypothalamus

Magnocellular neurons of the hypothalamic supraoptic nuclei (SON) are involved in the synthesis and release of two major neuropeptides: oxytocin (OT) and arginine-vassopressin (AVP). Neurochemical plasticity in this system is induced by physiological conditions such as lactation, parturition and dehydration, and may be accompanied by reversible structural plasticity affecting neurons, astrocytes and the extracellular matrix (ECM). The noradrenergic system plays a critical role in triggering this chemical plasticity associated with structural plasticity. Matrix metalloproteinases (MMPs) are good candidates for involvement in the ECM remodelling observed in structural plasticity. We investigated the possible regulation of the two gelatinases, MMP2 and MMP9, by noradrenaline (NA) in the mouse neuroendocrine hypothalamus. We looked for the presence, location and activity of MMP2 and MMP9 in the SON, using an ex vivo experimental model of mouse hypothalamic slices incubated for 4 h with 10⁻⁴  m NA. We showed that: (i) immunoreactivity for MMP2 and MMP9 was detected not only in AVP-positive and OT-positive magnocellular neurons, but also in astrocyte processes in control and NA-treated slices; (ii) the number of MMP2- and MMP9-positive cells increased after incubation with NA; (iii) MMP2 and MMP9 displayed markedly higher levels of gelatinolytic activity after NA treatment. These results suggest that both MMP2 and MMP9 are regulated by NA, and could therefore also be involved in structural plasticity within the SON.     

Morphologic evidence for intranuclear circuits in the hypothalamic arcuate nucleus

Three weeks after complete deafferentation of the medial basal hypothalamus (MBH) of adult female rats, the hypothalamic arcuate nucleus (ARCN) was examined ultrastructurally. Axodendritic and axosomatic synapses were counted in a field of 18,000 μm² in the middle part of the ARCN in each brain. Intraventricular infusion of 5-hydroxydopamine before autopsy resulted in the differentiation of two types of axon terminals. One axon terminal contained small spherical vesicles (SSVs, about 50 nm in diameter) and the other contained 5-hydroxydopamine-labeled small granular vesicles (SGVs) which were regarded as monoaminergic terminals. In the completely deafferented MBH, mean numbers of SSV and SGV synapses of the ARCN decreased to about one-half and one-fourth, respectively, compared to those of the unoperated rats. However, considerable numbers of intact SSV and SGV synapses were still encountered in the ARCN following deafferentation. There was no significant difference in the number of synapses between the large MBH island (including ARCN and ventromedial nucleus) and the small MBH island (mostly localized in ARCN). These results suggested that numerous converging nonmonoaminergic and monoaminergic fibers terminated in the ARCN and also suggested that nonmonoaminergic and monoaminergic arcuate neurons connected directly with other arcuate neurons.     

Acute stress differentially affects corticotropin-releasing hormone mRNA expression in the central amygdala of the "depressed" flinders sensitive line and the control flinders resistant line rats

Preclinical and clinical evidence suggests that neuropeptides play a role in the pathophysiology of mood disorders. In the present study, we investigated the involvement of the peptides corticotropin-releasing hormone (CRH), neuropeptide Y (NPY) and nociceptin/orphanin FQ (N/OFQ) and of their receptors in the regulation of emotional behaviours. In situ hybridization experiments were performed in order to evaluate the mRNA expression levels of these neuropeptidergic systems in limbic and limbic-related brain regions of the Flinders Sensitive Line (FSL) rats, a putative genetic animal model of depression. The FSL and their controls, the Flinders Resistant Line (FRL) rats, were subjected to one hour acute restraint and the effects of the stress exposure, including possible strain specific changes on these neuropeptidergic systems, were studied. In basal conditions, no significant differences between FSL and FRL rats in the CRH mRNA expression were found, however an upregulation of the CRH mRNA hybridization signal was detected in the central amygdala of the stressed FRL, compared to the non stressed FRL rats, but not in the FSL, suggesting a hypoactive mechanism of response to stressful stimuli in the "depressed" FSL rats. Baseline levels of NPY and N/OFQ mRNA were lower in the FSL rats compared to the FRL in the dentate gyrus of hippocampus and in the medial amygdala, respectively. However, the exposure to stress induced a significant upregulation of the N/OFQ mRNA levels in the paraventricular thalamic nucleus, while in the same nucleus the N/OFQ receptor mRNA expression was higher in the FSL rats. In conclusion, selective alterations of the NPY and N/OFQ mRNA in limbic and limbic-related regions of the FSL rats, a putative animal model of depression, provide further support for the involvement of these neuropeptides in depressive disorders. Moreover, the lack of CRH activation following stress in the "depressed" FSL rats suggests a form of allostatic load, that could alter their interpretation of environmental stimuli and influence their behavioural response to stressful situations.