Hypothalamic Content Research Articles

Neurotransmitter levels in the hypothalamus during postnatal development of rats.

The content of norepinephrine (NE), epinephrine (E), dopamine (DA) and serotonin (5-HT) in the rat hypothalamus was determined at four hours time interval during 24 hours period at the age of 12, 23, 27 and 75 days. It was found that the content of NE in the hypothalamus is low in 12 day-old rats, and during the postnatal period a gradual increase of NE was noted. The content of E in the hypothalamus is low in 12- to 27-day-old rats and an important increase was found in adult animals. The high values of DA and 5-HT concentration were found in the hypothalamus of 12-day-old rats and a transient decrease of 5-HT and DA at the age of 23 days was noted, further an important increase of the content of both neurotransmitters in the hypothalamus was demonstrated later in life. The presence of circadian rhythm of hypothalamic NE, E, DA and 5-HT content was found in 12-, 23-, 27-day-old and in adult rats overfed or underfed during the suckling period. However, no significant correlation was observed between the NE and E levels in the hypothalamus and the diurnal fluctuations of plasma corticosterone concentrations. In adult animals an inverse correlation between the plasma corticosterone level and hypothalamic 5-HT content was noted. In young 12- and 23-day-old rats the correlation between 5-HT in the hypothalamus and corticosterone in the plasma was positive. These results indicate that the shift of peak in plasma corticosterone levels during postnatal ontogenesis of rats observed in previous experiments is not dependent on the changes in diurnal fluctuation of hypothalamic neurotransmitter concentrations.

Effect of an acute maternal stress on the fetal hypothalamo-pituitary-adrenal system in late gestational life of the rat.

We investigated in this study the response of the fetal hypothalamo-pituitary-adrenal (HPA) system during an acute maternal stress in rats, in order to find out a possible role of developing fetal hypothalamus and to correlate its function to the androgen unbalance during the critical period of sex-specific brain differentiation. Pregnant rats of days 18-22 of gestation were subjected to an acute forced immobilization, and plasma levels of corticosterone (B) and ACTH were measured in mothers and fetuses. Hypothalamic contents of CRH and beta-endorphin (EP), pituitary content of ACTH, and plasma levels of B and ACTH were measured in mothers and fetuses under the maternal stress on day 20 of gestation. By an acute exposure to the 20 minutes' stress, plasma levels of B and ACTH elevated significantly in mothers on each day of gestation. A significant increase of fetal plasma ACTH was detected from day 18 in males, and from day 20 in females. During the maternal stress on day 20 of gestation, hypothalamic contents of CRH and EP decreased significantly in male and female fetuses, when plasma levels of B and ACTH elevated significantly. These results indicate that fetal HPA axis seems to actually respond to the maternal stress during the late gestational period. Further, a release of CRH under the stress together with an activation of EP system in the fetal hypothalamus suggests a possible mechanism regulating the androgen secretion by the fetal hypothalamus via changes of the LH levels.

Photoperiodic Condition Is Associated with Region-Specific Expression of GNRH1 mRNA in the Preoptic Area of the Male Starling (Sturnus vulgaris)1

Many seasonally breeding avian species exhibit marked changes in hypothalamic content of gonadotropin-releasing vhormone 1 (GNRH1) protein that are reflective of breeding condition. We recently cloned the GNRH1 cDNA in European starlings and demonstrated that changes in GNRH1 mRNA levels occur with a time course similar to what has been observed with GNRH1 protein. However, we did not previously resolve whether these differences were attributable to changes in the number of cells expressing the gene. Herein, we investigated photoperiod-induced changes in the number and distribution of GNRH1 mRNA-expressing cells in the preoptic area of male starlings. GNRH1 mRNA-expressing cell number was significantly greater in breeding birds than in nonbreeding birds. Starlings maintained in short nonstimulatory day length (i.e., prebreeding) showed intermediate cell numbers. Detailed analysis of the rostrocaudal and mediolateral distribution revealed that breeding birds had greater numbers of cells expressing GNRH1 mRNA in the medial intermediate, mediocaudal, and lateral intermediate preoptic area compared with prebreeding and nonbreeding birds. These data demonstrate that photoperiodic changes in reproductive state in starlings are associated with region-specific alterations in the number of cells expressing the GNRH1 gene. It remains to be determined whether these changes reflect quantitative differences in gene expression among an otherwise stable population of cells or a phenotypic switch in which cells gain or lose the ability to make GNRH1 mRNA in response to environmental cues.

Trilostane, FAD286, and the role of aldosterone in the central regulation of blood pressure: focus on “Role of central nervous system aldosterone synthase and mineralocorticoid receptors in salt-induced hypertension in Dahl salt-sensitive rats”

in a previous issue of this journal, Elise Gomez-Sanchez et al. ([4][1]) presented unequivocal evidence that intracerebroventricular (ICV) infusion of the 3β-hydroxy steroid dehydrogenase inhibitor trilostane prevented the development of hypertension in response to high-salt intake in Dahl salt-

Effects of Simulated Hypo‐ and Hyper‐Reproductive Conditions on the Characteristics of Circadian Rhythm in Hypothalamic Concentration of Serotonin and Dopamine and in Plasma Levels of Thyroxine, Triiodothyronine, and Testosterone in Japanese Quail, Coturnix coturnix japonica

In this study, hypo‐ and hyper‐reproductive conditions, as measured by concentrations of plasma testosterone in male Japanese quail held on long days LD 16:8, were experimentally simulated with injections of 5‐hydroxytryptophan (5‐HTP) and L‐dihydroxyphenylalanine, (L‐DOPA) with 8 h and 12 h phase angle differences between them in intact and melatonin‐treated birds. The effects of these treatments were assessed on the characteristics of the circadian rhythm in the hypothalamic concentration of serotonin (5‐HT), dopamine (DA), and plasma levels of thyroxine (T4), triiodothyronine (T3), and testosterone (T). These rhythms were also studied in sham‐operated (SO), pinealectomized (Px), vehicle‐ (Veh), and melatonin (Mel)‐treated birds. On the basis of the circadian mesors of the testosterone rhythms, three distinct categories could be identified: category A (i.e., normal breeding concentrations of plasma testosterone), which includes control, sham‐operated, and vehicle‐treated groups; category A+ (i.e., concentrations of plasma testosterone higher than that found in normal breeding quail), which includes 12 h, 12 h+vehicle‐treated, and Px quails; and category A− (concentrations of plasma testosterone lower than that found in normal breeding quail), which includes 8 h, melatonin‐, and 12 h+melatonin‐treated groups. It is evident that in normal and hypergonadal conditions (i.e., birds belonging to categories A and A+) the circadian rhythm in hypothalamic serotonin maintained a positive phase angle of about 16 h. In contrast, birds of category A− (i.e., in a hypogonadal condition) exhibited a negative phase angle of about 2 h. The present results clearly suggest that the internal phase relationship between the circadian rhythms in hypothalamic serotonin and dopamine might play a crucial role in strategizing and conferring a particular reproductive status to the birds. The role of circadian mechanisms involving circulating thyroid hormones in conferring reproductive status is completely ruled out, as no definite internal phase angle between these two hormonal rhythms was witnessed vis‐à‐vis different treatment groups. The testosterone peaks always occurred at the same time irrespective of breeding status of the bird, but with significant variation in its amplitude (high in hypergonadal and low in hypogonadal condition). It is suggested that administration of 5‐HTP and L‐DOPA at specific time interval and variation in pineal functions that modulate reproductive responses also alter the circadian pattern (acrophase and amplitude) of hypothalamic serotonin and dopamine, maintaining a specific phase relation between these cycles and breeding status. These findings strengthen our previous reports that a specific circadian phase relation of serotonergic and dopaminergic oscillations regulates reproduction. The present study strongly supports interdependence and specific relation of the two systems (gonadal activity and circadian pattern/phase relation of neural oscillation) in both natural and experimentally simulated conditions.

Leptin Infusion and Obesity in Mouse Cause Alterations in the Hypothalamic Melanocortin System

The objectives of this study were to identify potential alterations in gene expression of melanocortin-4 receptor (MC4-R), proopiomelanocortin (POMC), and Agouti-related protein (AgRP) in mouse hypothalamus under a chronic peripheral infusion of leptin or at early (8 weeks) and advanced (16 weeks) phases of diet-induced obesity. Control or diet-induced obesity mice (8 or 16 weeks of high-fat diet) were either treated or not treated with leptin. Metabolic features were analyzed and expression of the genes of interest was measured by quantitative reverse transcriptase-PCR (RT-qPCR) and western blot. We reported that in control mice, but not in obese mice, leptin infusion induced an increase in POMC mRNA level as well as in MC4-R mRNA level suggesting that leptin could act directly and/or through alpha-melanocyte-stimulating hormone (alpha-MSH). This hypothesis was reinforced after in vitro studies, using the mouse hypothalamic GT1-7 cell line, since both leptin and Norleucine(4), D-Phenylalanine(7)-alpha-MSH (NDP-alpha-MSH) treatments increased MC4-R expression. After 8 weeks of high-fat diet, nondiabetic obese mice became resistant to the central action of leptin and their hypothalamic content of POMC and AgRP mRNA were decreased without modification of MC4-R mRNA level. After 16 weeks of high-fat diet, mice exhibited more severe metabolic disorders with type 2 diabetes. Moreover, hypothalamic expression of MC4-R was highly increased. In conclusion, several alterations of the melanocortin system were found in obese mice that are probably consecutive to their central resistance to leptin. Moreover, when the metabolic status is highly degraded (with all characteristics of a type 2 diabetes), other regulatory mechanisms (independent of leptin) can also take place.

Melatonin reduces body weight in goldfish (Carassius auratus): effects on metabolic resources and some feeding regulators

The objective of the present study was to analyze the effects of chronic melatonin (10 microg/g body weight) on body weight and the main energetic reserves, particularly hepatic and muscle content of proteins, lipids and glycogen in goldfish. In addition, we studied plasma leptin and ghrelin, and hypothalamic content of neuropeptide Y (NPY) and monoamines after chronic melatonin treatment in order to elucidate a possible interplay between melatonin and these feeding regulators on the body weight regulation in this species. Body weight gain and specific growth rate were reduced (74% and 76%, respectively) after chronic (10 days) intraperitoneal (i.p.) treatment with melatonin. The carbohydrate and lipid metabolism was regulated by melatonin in goldfish, because this indoleamine reduced muscle glycogen stores and increased lipid mobilization. A suppressive trend, but not statistically significant, in circulating ghrelin was observed after chronic treatment with melatonin. Chronic melatonin administration significantly reduced noradrenergic metabolism and increased dihydroxiphenylacetic acid content in the hypothalamus, without significant modifications in the serotoninergic system. Thus, it could be suggested that melatonin may mediate its action on energy balance in fish, at least in part, via interactions with hypothalamic catecholaminergic system. Plasma leptin and hypothalamic NPY remained unaltered after melatonin treatment, suggesting that these feeding regulators may not be involved in the effects of melatonin on energy homeostasis in fish.

Enhanced dopamine D2 receptor function in hypothalamus and corpus striatum: their role in liver, plasma and in vitro hepatocyte ALDH regulation in ethahol treated rats

Dopamine D(2) receptors are involved in ethanol self- administration behavior and also suggested to mediate the onset and offset of ethanol drinking. In the present study, we investigated dopamine (DA) content and Dopamine D(2) (DA D(2)) receptors in the hypothalamus and corpus striatum of ethanol treated rats and aldehyde dehydrogenase (ALDH) activity in the liver and plasma of ethanol treated rats and in vitro hepatocyte cultures. Hypothalamic and corpus striatal DA content decreased significantly (P < 0.05, P < 0.001 respectively) and homovanillic acid/dopamine (HVA/DA) ratio increased significantly (P < 0.001) in ethanol treated rats when compared to control. Scatchard analysis of [(3)H] YM-09151-2 binding to DA D(2) receptors in hypothalamus showed a significant increase (P < 0.001) in B(max) without any change in K(d) in ethanol treated rats compared to control. The K(d) of DA D(2) receptors significantly decreased (P < 0.05) in the corpus striatum of ethanol treated rats when compared to control. DA D(2) receptor affinity in the hypothalamus and corpus striatum of control and ethanol treated rats fitted to a single site model with unity as Hill slope value. The in vitro studies on hepatocyte cultures showed that 10(-5) M and 10(-7) M DA can reverse the increased ALDH activity in 10% ethanol treated cells to near control level. Sulpiride, an antagonist of DA D(2), reversed the effect of dopamine on 10% ethanol induced ALDH activity in hepatocytes. Our results showed a decreased dopamine concentration with enhanced DA D(2) receptors in the hypothalamus and corpus striatum of ethanol treated rats. Also, increased ALDH was observed in the plasma and liver of ethanol treated rats and in vitro hepatocyte cultures with 10% ethanol as a compensatory mechanism for increased aldehyde production due to increased dopamine metabolism. A decrease in dopamine concentration in major brain regions is coupled with an increase in ALDH activity in liver and plasma, which contributes to the tendency for alcoholism. Since the administration of 10(-5) M and 10(-7) M DA can reverse the increased ALDH activity in ethanol treated cells to near control level, this has therapeutic application to correct ethanol addicts from addiction due to allergic reaction observed in aldehyde accumulation.

Effects of nomegestrol acetate administration on central and peripheral beta-endorphin and allopregnanolone in ovx rats

The aim of this study was to investigate the effects of nomegestrol acetate (NOMAc) on the central nervous system by analyzing the neurosteroid allopregnanolone and the opioid beta-endorphin (β-endorphin). 104 Wistar female rats were used in this study; one group of fertile and one group of ovariectomized rats were used as control. The others were ovariectomized and they underwent a 2-week oral treatment of NOMAc (0.05, 0.1, 0.2, 0.5, 1 mg/kg/day), alone or with 0.05 mg/kg/day of estradiol valerate (E2V). Allopregnanolone and β-endorphin were assessed in different brain areas and in circulation. Ovariectomy decreased allopregnanolone anywhere except in the adrenal gland and E2V reversed the effects of ovariectomy. 0.5 and 1 mg/kg/day of NOMAc increased allopregnanolone levels in hippocampus. Combined administration of 1 mg/kg/day of NOMAc plus E2V induced a further increase of allopregnanolone levels in hippocampus, hypothalamus, and anterior pituitary. NOMAc (1 mg/kg/day) decreased the adrenal content of allopregnanolone, both by itself and associated with E2V. NOMAc increased hippocampal and hypothalamic content of β-endorphin at the highest doses, and it increased positively E2V action, at 1 mg/kg/day, also in anterior pituitary and plasma. These findings reinforce the clinical data regarding the capability of NOMAc to modulate the pathways involved in mood and behaviour. In fact, due to the NOMAc action on hippocampus, hypothalamus, and anterior pituitary, our results highlight the selectivity of NOMAc on part of the limbic system and the anterior pituitary, regarding both allopregnanolone and β-endorphin.

Leptin inhibits and ghrelin augments hypothalamic noradrenaline release after stress

Metabolic conditions affect hypothalamo-pituitary-adrenal responses to stressful stimuli. Here we examined effects of food deprivation, leptin and ghrelin upon noradrenaline release in the hypothalamic paraventricular nucleus (PVN) and plasma adrenocorticotropic hormone (ACTH) concentrations after stressful stimuli. Food deprivation augmented both noradrenaline release in the PVN and the increase in plasma ACTH concentration following electrical footshocks (FSs). An intracerebroventricular injection of leptin attenuated the increases in hypothalamic noradrenaline release and plasma ACTH concentrations after FSs, while ghrelin augmented these responses. These data suggest that leptin inhibits and ghrelin facilitates neuroendocrine stress responses via noradrenaline release and indicate that a decrease in leptin and an increase in ghrelin release after food deprivation might contribute to augmentation of stress-induced ACTH release in a fasting state.

Hypothalamic pituitary adrenal axis reactivity and dehydroepiandrosterone sulfate plasma concentrations in the critically ill elderly

Hypothalamic pituitary adrenal axis reactivity and dehydroepiandrosterone sulfate plasma concentrations in the critically ill elderly

Blockade of brain histamine metabolism alters methamphetamine-induced expression pattern of stereotypy in mice via histamine H 1 receptors

The administration of methamphetamine (METH, 10 mg/kg, i.p.) to male ICR mice induced stereotyped behavior consisting of nail and/or wood chip biting (86.0%), continuous sniffing (12.0%), head bobbing (1.1%), and circling (1.0%) during the observation period of 1 h. Pretreatment of the mice with metoprine (2, 10, and 20 mg/kg, i.p.), a selective inhibitor of histamine N-methyltransferase (HMT), which metabolizes histamine in the brain, significantly increased and decreased METH-induced continuous sniffing (20.5, 51.3, and 80.3%) and nail and/or wood chip biting (77.4, 45.3, and 14.2%), respectively, in a dose-dependent manner. The hypothalamic contents of histamine and its metabolite N τ-methylhistamine were significantly increased and decreased by metoprine (10 mg/kg, i.p.), respectively. The metoprine action on METH-induced behavior was completely abolished by pyrilamine (10 and 20 mg/kg) and ketotifen (10 mg/kg), selective, centrally acting histamine H 1 receptor antagonists, but not by fexofenadine (20 mg/kg), zolantidine (10 mg/kg) and thioperamide (10 mg/kg), a peripherally acting histamine H 1 receptor antagonist and a selective, brain-penetrating antagonist for histamine H 2 and H 3 receptors, respectively. The metoprine action was mimicked by SKF 91488 (100 μg/animal, i.c.v.), another HMT inhibitor, and the action of SKF 91488 was also blocked by pyrilamine. The frequency of the expression of METH-induced total stereotypic patterns was unchanged after metoprine pretreatment. Mice pretreated with metoprine displayed no anxiety-like behavior in the elevated plus maze test. These results suggest that brain histamine, increased by agents such as metoprine and SKF 91488, binds to histamine H 1 receptors in the brain, resulting in the modulation of dopaminergic transmission associated with stereotyped behavioral patterns induced by METH.

Gender-dependent modulation of brain monoamines and anxiety-like behaviors in mice with genetic serotonin transporter and BDNF deficiencies.

1. Brain-derived neurotrophic factor (BDNF) supports serotonergic neuronal development and our recent study found that heterozygous mice lacking one BDNF gene allele interbred with male serotonin transporter (SERT) knockout mice had greater reductions in brain tissue serotonin concentrations, greater increases in anxiety-like behaviors and greater ACTH responses to stress than found in the SERT knockout mice alone. 2. We investigated here whether there might be gender differences in these consequences of combined SERT and BDNF deficiencies by extending the original studies to female mice, and also to an examination of the effects of ovariectomy and tamoxifen in these female mice, and of 21-day 17-beta estradiol implantation to male mice. 3. We found that unlike the male SERTxBDNF-deficient mice, female SERTxBDNF mice appeared protected by their gender in having significantly lesser reductions in serotonin concentrations in hypothalamus and other brain regions than males, relative to controls. Likewise, in the elevated plus maze, female SERTxBDNF-deficient mice demonstrated no increases in the anxiety-like behaviors previously found in males. 4. Furthermore, female SERTxBDNF mice did not manifest the approximately 40% reduction in the expression of TrkB receptors or the approximately 30% reductions in dopamine and its metabolites that male SERTxBDNF did. After estradiol implantation in male SERTxBDNF mice, hypothalamic serotonin was significantly increased compared to vehicle-implanted mice. These findings support the hypothesis that estrogen may enhance BDNF function via its TrkB receptor, leading to alterations in the serotonin circuits, which modulate anxiety-like behaviors. 5. This double-mutant mouse model contributes to the knowledge base that will help in understanding genexgenexgender interactions in studies of SERT and BDNF gene polymorphisms in human genetic diseases such as anxiety disorders and depression.

Intake of trans fatty acid–rich hydrogenated fat during pregnancy and lactation inhibits the hypophagic effect of central insulin in the adult offspring

Objective Using rats we examined whether maternal intake of hydrogenated fat rich in trans fatty acids affects brain fatty acid profile, hypothalamic content of insulin receptor and insulin receptor substrate-1 proteins, and the hypophagic effect of centrally administered insulin in 3-mo-old male progeny. Methods Throughout pregnancy and lactation, Wistar rats ate isocaloric/normolipidic diets with soybean oil (control) or soybean oil-derived hydrogenated fat (trans diet) as a fat source. Upon weaning, the trans offspring continued on the trans diet (trans group) or were switched to a control diet (trans-control group). Results Compared with control rats, trans rats had lower brain levels of eicosapentaenoic acid. Compared with trans rats, trans-control rats had increased levels of total polyunsaturated fatty acids and arachidonic acid and decreased levels of trans fatty acids, saturated fatty acids, and monounsaturated fatty acids. Insulin receptor and insulin receptor substrate-1 levels were significantly lower (44% and 38%, respectively) in trans than in control rats. In trans-control rats, insulin receptor was 26% lower ( P < 0.05), whereas insulin receptor substrate-1 was 50% lower, than in control rats. Insulin decreased 24-h feeding in control (27%) and trans (38%) rats but failed to do so in trans-control rats. The latter group had increased serum glucose levels. Conclusions The data suggest that the early (intrauterine/perinatal) exposure to hydrogenated fat rich in trans fatty acids programmed the hypothalamic feeding control mechanisms. As young adults, only trans-control animals showed loss of insulin-induced hypophagia, indicating that the mismatch between early and later nutritional environments was relevant. However, the trans group also showed signs of altered appetite signaling mechanisms, suggesting that the early adaptations may have deleterious consequences later in life.

Quantitative proteomics identifies a change in glial glutamate metabolism at the time of female puberty

Mammalian puberty requires activation of luteinizing hormone–releasing hormone (LHRH) neurons. In turn, these neurons are controlled by transsynaptic and glia-to-neuron communication pathways, which employ diverse cellular proteins for proper function. We have now used a high throughput relative quantitative proteomics technique to identify such proteins. We selected the method of two-dimensional liquid chromatography tandem mass spectrometry (2DLC–MS/MS) and cleavable isotope-coded affinity tags (cICAT), to both identify and quantify individual proteins within a complex protein mixture. The proteins used derived from the hypothalamus of juvenile (25-day-old) and peripubertal (first proestrus, LP) female rats, and their identity was established by analyzing their mass spectra via database searching. Five proteins involved in glutamate metabolism were detected and two of them appeared to be differentially expressed. They were selected for further analysis, because of their importance in controlling glutamate synthesis and degradation, and their preferential expression in astroglial cells. One, glutamate dehydrogenase (GDH) catalyzes glutamate synthesis; its hypothalamic content detected by 2DLC–MS/MS increases at first proestrus. The other, glutamine synthetase (GS), catalyzes the metabolism of glutamate to glutamine; its content decreases in proestrus. Western blot analysis verified these results. Because these changes suggested an increased glutamate production at puberty, we measured glutamate release from hypothalamic fragments from juvenile 29-day old rats, and from rats treated with PMSG to induce a premature proestrus surge of luteinizing hormone (LH). To determine the net output of glutamate in the absence of re-uptake we used the excitatory amino acid transporter (EAAT) inhibitor l- trans-pyrrolidine-2,4-dicarboxylic acid (PDC). PDC elicited significantly more glutamate- and LHRH-release from the proestrus hypothalamus. Thus, an increase excitatory drive to the LHRH neuronal network provided by glutamatergic inputs of glial origin, is an event contributing to the pubertal activation of LHRH secretion.

Mechanism delineating differential effect of an antiestrogen, tamoxifen, on the serum LH and FSH in adult male rats

Tamoxifen, a synthetic non-steroidal antiestrogen with residual estrogenic activity, administered to adult male rats reduces their fertility. A decrease in the circulating LH and testosterone levels with a transient rise or no change in circulating FSH levels was observed. The present study was carried out to delineate the mechanism causing the differential effect of tamoxifen on circulating gonadotropins by correlating it to changes in the hypothalamic LHRH, pituitary gonadotropins and testicular inhibin/activin. Hypothalamus, pituitary-hypothalamus complex (PHC) and intact pituitary (PI) from control and tamoxifen-treated male rats were superfused in vitro, and pulsatile release of LHRH by hypothalamus and that of LH and FSH by the PHC and PI were studied. Concomitantly, testicular immunoexpression of alpha and betaB subunits of inhibin/activin were studied by immunohistochemistry and enzyme-linked immunosorbent assays (ELISA). At 0.4 mg/kg/day dose of tamoxifen a decrease in mean hypothalamic LHRH and LH pulse frequency from PHC construct was observed. FSH pulse frequency was not affected under the same experimental conditions. At the same dose of tamoxifen, testicular expression of both alpha and betaB subunits of inhibin/activin was upregulated. The study demonstrated that reduced circulating LH levels were due to a decrease in hypothalamic LHRH concentration and in LH pulsatility following tamoxifen treatment. The lack of effect on circulating FSH under the same experimental conditions was likely due to its modulation by inhibin and activin.

Activation of brain renin-angiotensin-aldosterone system by central sodium in Wistar rats

Functional studies indicate that the sympathoexcitatory and pressor responses to an increase in cerebrospinal fluid (CSF) [Na+] by central infusion of Na+-rich artificial cerebrospinal fluid (aCSF) in Wistar rats are mediated in the brain by mineralocorticoid receptor (MR) activation, ouabain-like compounds (OLC), and AT1-receptor stimulation. In the present study, we examined whether increasing CSF [Na+] by intracerebroventricular infusion of Na+-rich aCSF activates MR and thereby increases OLC and components of the renin-angiotensin system in the brain. Male Wistar rats received via osmotic minipump an intracerebroventricular infusion of aCSF or Na+-rich aCSF, in some groups combined with intracerebroventricular infusion of spironolactone (100 ng/h), antibody Fab fragments (to bind OLC), or as control gamma-globulins. After 2 wk of infusion, resting blood pressure and heart rate were recorded, OLC and aldosterone content in the hypothalamus were assessed by a specific ELISA or radioimmunoassay, and angiotensin-converting enzyme (ACE) and AT1-receptor binding densities in various brain nuclei were measured by autoradiography using 125I-labeled 351 A and 125I-labeled ANG II. When compared with intracerebroventricular aCSF, intracerebroventricular Na+-rich aCSF increased CSF [Na+] by approximately 5 mmol/l, mean arterial pressure by approximately 20 mmHg, heart rate by approximately 65 beats/min, and hypothalamic content of OLC by 50% and of aldosterone by 33%. Intracerebroventricular spironolactone did not affect CSF [Na+] but blocked the Na+-rich aCSF-induced increases in blood pressure and heart rate and OLC content. Intracerebroventricular Na+-rich aCSF increased ACE and AT1-receptor-binding densities in several brain nuclei, and Fab fragments blocked these increases. These data indicate that in Wistar rats, a chronic increase in CSF [Na+] may increase hypothalamic aldosterone and activate CNS pathways involving MR, and OLC, leading to increases in AT1-receptor and ACE densities in brain areas involved in cardiovascular regulation and hypertension.

Photoperiodic regulation of insulin receptor mRNA and intracellular insulin signaling in the arcuate nucleus of the Siberian hamster,Phodopus sungorus

During the last 5 years it has been well established that photoperiod-induced changes in body weight in the seasonal hamster, Phodopus sungorus, are accompanied by a marked seasonal cycle in leptin sensitivity. In the present study, we investigated the possible involvement of insulin signaling in seasonal body weight regulation. We analyzed the expression pattern and relative intensity of insulin receptor (IR), phosphatidylinositol 3-kinase (PI3-kinase), and protein tyrosine phosphatase 1B (PTP1B) mRNAs by in situ hybridization in the brains of juvenile female hamsters acclimated to either long- (LD) or short-day length (SD) for 8 wk, with or without superimposed food deprivation for 48 h. Furthermore, the hypothalamic concentration and distribution of phospho-AKT, a marker of PI3-kinase activity was determined by immunoblotting and immunohistochemistry. Eight weeks of acclimation to SD led to a substantial downregulation of IR, PTP1B gene expression, and phospho-AKT concentration in this brain region, whereas PI3-kinase mRNA was unchanged. Food deprivation induced a decrease in PTP1B and a trend toward lowered IR gene expression in LD but not in SD. Additionally, a striking increase in PTP1B gene expression in the thalamus was observed after food deprivation in both photoperiods. The direction of change in neuronal insulin signaling contrasts to the central catabolic nature of this pathway described in other species. SD-induced reduction in insulin signaling may be due to decline in body fat stores mediated by enhanced central leptin sensitivity. Increased anorexigenic tone of leptin may overwrite central insulin signaling to prevent catabolic overdrive.

Involvement of brain-derived neurotrophic factor in the regulation of hypothalamic somatostatin in vivo

Brain-derived neurotrophic factor (BDNF) has been extensively studied in the central nervous system as a survival and differentiation factor and in plasticity processes. In vitro, BDNF has been shown to stimulate cellular differentiation and neurohormones synthesis and release. We demonstrated that BDNF is a potent and specific stimulatory agent of somatostatin (SRIH) synthesis in primary cultures of hypothalamic neurons. However, less information is available about its function on SRIH neurons in vivo. In the present study, we examined the effect of in vivo intracerebroventricular BDNF administration in adult non-anesthetized male rats. Two distinct experimental approaches were used: acute intracerebroventricular injection and long-term (14 days) continuous infusion (Alzet micro-pumps). We demonstrate that single intracerebroventricular BDNF injections (5 microg/rat) induce an early (60 and 180 min) decrease in the SRIH mRNA signal in the hypothalamic periventricular nucleus (PeVN) accompanied by a decrease of the hypothalamic SRIH content. 48 h after the acute injection, SRIH mRNA levels and peptide content strongly and significantly increased. After continuous intracerebroventricular BDNF administration (12 microg/day for 14 days), a significant increase in the SRIH hypothalamic content was observed. Nevertheless, the increase in peptide content was not correlated with a similar increase in the PeVN messenger level. These findings show the involvement of BDNF in the in vivo regulation of somatostatinergic neurons in adult rats, which clearly differs according to the BDNF administration mode.

Toxicokinetics of lambda-cyhalothrin in rats

The toxicokinetics of lambda-cyhalothrin after single 20 mg kg −1 oral and 3 mg kg −1 intravenous doses were studied in rats. Serial blood samples were obtained after oral and intravenous administration. Liver, brain, spinal cord, sciatic nerve, vas deferens, anococcygeus and myenteric plexus tissue samples were also collected. Plasma, liver, hypothalamus, cerebellum, medulla oblongata, frontal cortex, striatum, hippocampus, midbrain, spinal cord, vas deferens, anococcygeus, myenteric plexus and sciatic nerve concentrations of lambda-cyhalothrin were determined by HPLC. The plasma and tissue concentration–time data for lambda-cyhalothrin were found to fit a two-compartment open model. For lambda-cyhalothrin, the elimination half-life ( T 1/2β) and the mean residence time from plasma were 7.55 and 8.55 h after i.v. and 10.27 and 14.43 h after oral administration. The total plasma clearance was not influenced by dose concentration or route and reached a value of 0.060 l h −1 kg −1. After i.v. administration, the apparent volume of distribution and at steady state were 0.68 and 0.53 l kg −1, suggesting a diffusion of the pyrethroid into tissue. After oral administration, lambda-cyhalothrin was extensively but slowly absorbed ( T max, 2.69 h). The oral bioavailability was found to be 67.37%. Significant differences in the kinetic parameters between nervous tissues and plasma was observed. The maximum concentrations in hypothalamus ( C max, 24.12 μg g −1) and myenteric plexus ( C max, 25.12 μg g −1) were about 1.5 times higher than in plasma ( C max, 15.65 μg ml −1) and 1.3 times higher than in liver ( C max, 18.42 μg ml −1). Nervous tissue accumulation of lambda-cyhalothrin was also reflected by the area under the concentration curve ratios of tissue/plasma (liver). The T 1/2β for lambda-cyhalothrin was significantly greater for the nerve tissues, including neuromuscular fibres, (range 12–26 and 15–34 h, after i.v. and oral doses) than for plasma (7.55 and 10.27 h, respectively).