R.M. Buijs

Intra- and extrahypothalamic vasopressin and oxytocin pathways in the rat.

SummaryPerfusion of rat brain followed by immersion fixation with 2.5% glutaraldehyde-1% paraformaldehyde, purification of the first antisera and application of the unlabelled antibody enzyme method were used to specifically identify vasopressin and oxytocin containing cells and fibres. The conventional sites of production of these hormones were confirmed as follows: supraoptic and paraventricular nuclei, suprachiasmatic nucleus (only vasopressin), and other cells and cell groups of the hypothalamus. Fibres from the suprachiasmatic nucleus spread out in various directions, and probably project to the nucleus praeopticus periventricularis, organum vasculosum laminae terminalis and in the direction of the supraoptic nucleus. Oxytocin and vasopressin containing pathways could be traced from the paraventricular nucleus to the lateral ventricle, the stria terminalis and the stria medullaris. Some of the oxytocin and vasopressin containing tracts appear to continue onto the septum. The possible importance of these morphological findings for the behavioural effects of vasopressin and oxytocin is discussed.

Short Sleep Duration as a Risk Factor for Hypertension Analyses of the First National Health and Nutrition Examination Survey

Depriving healthy subjects of sleep has been shown to acutely increase blood pressure and sympathetic nervous system activity. Prolonged short sleep durations could lead to hypertension through extended exposure to raised 24-hour blood pressure and heart rate, elevated sympathetic nervous system activity, and increased salt retention. Such forces could lead to structural adaptations and the entrainment of the cardiovascular system to operate at an elevated pressure equilibrium. Sleep disorders are associated with cardiovascular disease, but we are not aware of any published prospective population studies that have shown a link between short sleep duration and the incidence of hypertension in subjects without apparent sleep disorders. We assessed whether short sleep duration would increase the risk for hypertension incidence by conducting longitudinal analyses of the first National Health and Nutrition Examination Survey (n=4810) using Cox proportional hazards models and controlling for covariates. Hypertension incidence (n=647) was determined by physician diagnosis, hospital record, or cause of death over the 8- to 10-year follow-up period between 1982 and 1992. Sleep durations of ≤5 hours per night were associated with a significantly increased risk of hypertension (hazard ratio, 2.10; 95% CI, 1.58 to 2.79) in subjects between the ages of 32 and 59 years, and controlling for the potential confounding variables only partially attenuated this relationship. The increased risk continued to be significant after controlling for obesity and diabetes, which was consistent with the hypothesis that these variables would act as partial mediators. Short sleep duration could, therefore, be a significant risk factor for hypertension.

The origin of the vasopressinergic and oxytocinergic innervation of the rat brain with special reference to the lateral septum

The origin of the vasopressin-containing fibers in the rat lateral septum was studied by means of lesioning specific areas, in which vasopressin-containing cells are found, or by surgically separating the septum from the underlying structures. Following these procedures sections of the brain were stained immunocytochemically for the presence of vasopressin. In addition, retrograde labeling tracers were injected in the lateral septum. Lesioning of the paraventricular nucleus did not result in the disappearance of vasopressin fibers from the lateral septum, nor from the various other areas studied. It did, however, cause the disappearance of fibers from the nucleus of the solitary tract and the nucleus ambiguus. By contrast, after the same lesion practically the whole oxytocinergic innervation of the brain disappeared. Injection of tracers into the lateral septum revealed retrograde labeled cells, e.g. in the bed nucleus of the stria terminalis, but not in the paraventricular and supraoptic nucleus. Horizontal cuts under the lateral septum, intersecting the diagonal band of Broca, resulted in a dramatic decrease of the vasopressin fibers in the lateral septum, suggesting that the fibers enter the septum via this structure. Moreover, since the vasopressin fiber density was found to decrease drastically in the lateral septum after lesioning the bed nucleus of the stria terminalis, the vasopressin cells found in this area are probably the source of these fibers. Other areas where fibers were seen to decrease after lesions of the bed nucleus are the diagonal band of Broca, the area of the anterior amygdala, the lateral habenular nucleus, the periventricular gray, and the locus coeruleus.

Daily nighttime melatonin reduces blood pressure in male patients with essential hypertension.

Abstract—Patients with essential hypertension have disturbed autonomic cardiovascular regulation and circadian pacemaker function. Recently, the biological clock was shown to be involved in autonomic cardiovascular regulation. Our objective was to determine whether enhancement of the functioning of the biological clock by repeated nighttime melatonin intake might reduce ambulatory blood pressure in patients with essential hypertension. We conducted a randomized, double-blind, placebo-controlled, crossover trial in 16 men with untreated essential hypertension to investigate the influence of acute (single) and repeated (daily for 3 weeks) oral melatonin (2.5 mg) intake 1 hour before sleep on 24-hour ambulatory blood pressure and actigraphic estimates of sleep quality. Repeated melatonin intake reduced systolic and diastolic blood pressure during sleep by 6 and 4 mm Hg, respectively. The treatment did not affect heart rate. The day–night amplitudes of the rhythms in systolic and diastolic blood pressures were increased by 15% and 25%, respectively. A single dose of melatonin had no effect on blood pressure. Repeated (but not acute) melatonin also improved sleep. Improvements in blood pressure and sleep were statistically unrelated. In patients with essential hypertension, repeated bedtime melatonin intake significantly reduced nocturnal blood pressure. Future studies in larger patient group should be performed to define the characteristics of the patients who would benefit most from melatonin intake. The present study suggests that support of circadian pacemaker function may provide a new strategy in the treatment of essential hypertension.

Immunocytochemical localization of dopamine in the prefrontal cortex of the rat at the light and electron microscopical level

In the present study the dopaminergic innervation of the prefrontal cortex was studied by means of a recently developed anti-dopamine serum. This method can demonstrate endogenous dopamine in a specific way, and offers the opportunity to study the distribution of dopaminergic fibres in the cortex in detail in counterstained sections. Furthermore, dopaminergic nerve endings can be visualized at the electron microscopic level. Light microscopic observations demonstrated that the highest density of dopaminergic fibres in the frontal cortex is found in the prefrontal cortex and the infralimbic cortex. Within the prefrontal cortex, a good correlation is found between regional differences in distribution of dopaminergic fibres and the cytoarchitectonic parcellation of this part of the cortex. Outside the prefrontal cortex dopaminergic fibres were observed in adjacent frontal areas, the cortex surrounding the entire rhinal sulcus and the retrosplenial cortex. Electron microscopic observations demonstrated dopaminergic terminals through all cortical layers. The majority of dopaminergic terminals in the prefrontal cortex from synaptic contacts with dendritic processes. The synaptic profiles were usually symmetric and were characterized by the presence of many clear vesicles and an occasional dense-core vesicle.

Immuno-electron microscopical demonstration of vasopressin and oxytocin synapses in the limbic system of the rat

SummaryThe extensive distribution of exohypothalamic vasopressin or oxytocin containing nerve fibres is thought to be the anatomical basis for the involvement of these neuropeptides in central processes. Following light microscopic observations suggesting that these fibres terminate on other neurons, the present study was undertaken to demonstrate the existence of such endings in the limbic system, which is one of the main target areas for these peptides. For immunoelectron microscopy glutaraldehyde-paraformaldehyde perfused brains of male Wistar rats and Brattleboro rats, homozygous for diabetes insipidus, with and without postfixation in OsO4, were used. Post-embedding staining revealed false positive reaction product on all dense core vesicles, e.g., in the lateral septum. With pre-embedding staining, however, intense and specific reactions were observed for both vasopressin and oxytocin at their sites of production, as well as the neurohypophysis and in the extrahypothalamic limbic brain regions.In the lateral septum and habenular nucleus only vasopressin-containing synapses could be demonstrated, while in the medial nucleus of the amygdala synapses containing either vasopressin or oxytocin were observed. These peptide containing synapses do not seem to differ in any fundamental way from the classical transmitter-containing synapses in the brain.

Gonadal hormone actions on the morphology of the vasopressinergic innervation of the adult rat brain

Castration of adult male rats resulted in a gradual decrease in vasopressin fiber density over a period of 15 weeks to a point where hardly any fibers were found in those areas where the fibers probably are derived from the bed nucleus of the stria terminalis. The original fiber density could be restored by testosterone replacement therapy within 5 weeks. No effects of hormonal manipulations were found in the vasopressin projections of the paraventricular and the suprachiasmatic nucleus. Ovariectomy of female rats resulted in the same changes in the vasopressin fiber pathways, as did castration in males.

Vasopressin and oxytocin release in the brain; a synaptic event

In order to obtain evidence for a central release of vasopressin and oxytocin, the release of these peptides was demonstrated in various extrahypothalamic areas of the rat brain. It proved that in those areas where these peptidergic fibers terminate synaptically a vasopressin and/or oxytocin calcium-dependent release, similar to that in the neurohypophysis, could be evoked by potassium or veratridine. Such release was not found in areas in which these fibers do not exhibit synaptic specialization.

Food Intake during the Normal Activity Phase Prevents Obesity and Circadian Desynchrony in a Rat Model of Night Work

Shift work or night work is associated with hypertension, metabolic syndrome, cancer, and other diseases. The cause for these pathologies is proposed to be the dissociation between the temporal signals from the biological clock and the sleep/activity schedule of the night worker. We investigated the mechanisms promoting metabolic desynchrony in a model for night work in rats, based on daily 8-h activity schedules during the resting phase. We demonstrate that the major alterations leading to internal desynchrony induced by this working protocol, flattened glucose and locomotor rhythms and the development of abdominal obesity, were caused by food intake during the rest phase. Shifting food intake to the normal activity phase prevented body weight increase and reverted metabolic and rhythmic disturbances of the shift work animals to control ranges. These observations demonstrate that feeding habits may prevent or induce internal desynchrony and obesity.

A Suprachiasmatic Nucleus Generated Rhythm In Basal Glucose Concentrations

The daily rhythm in feeding activity in mammals, as driven by the biological clock, largely determines the daily fluctuations in basal concentrations of glucose and insulin. To investigate a possible direct impact of the suprachiasmatic nucleus (SCN) on these parameters, we subjected intact rats and SCN‐lesioned rats to a fasting regimen of 36 h, or to a scheduled feeding regimen of six identical meals equally distributed over the light:dark‐cycle. Plasma profiles of glucose and insulin in rats during the final 24 h of the 36 h of fasting, and in rats subjected to the scheduled feeding regimen were compared to profiles in rats fed ad libitum. In rats fed ad libitum, in fasted rats and in rats subjected to a scheduled feeding regimen basal glucose concentrations showed a pronounced 24‐h rhythm that was not found in rats that had been SCN‐lesioned. Basal insulin levels showed a 24‐h rhythm in 50% of the rats fed ad libitum and in 50% of the rats subjected to a scheduled feeding regimen; neither rhythms were present in SCN‐lesioned rats. However, none of the fasted rats showed a 24‐h rhythm in basal insulin concentrations. These data provide clear evidence that the SCN directly controls basal glucose concentrations independent of its influence on feeding activity. At the same time, we found no consistent evidence for a strong impact of the SCN on basal insulin concentrations.