Glial vascular endothelial growth factor overexpression in rat brainstem under tolerable hypoxia: evidence for a central chemosensitivity.

The ventilatory response to hypoxia is mediated by peripheral inputs arising from the arterial chemoreceptors. In their absence, hypoxic adaptation can be achieved, possibly as a result of central cellular reorganization. To study this reorganization, we used chemodenervated rats to investigate the expression and localization of vascular endothelial growth factor (VEGF) in the brainstem. VEGF is a target gene of hypoxia-inducible factor (HIF) that is responsible for the morphofunctional remodeling induced by hypoxia. Intact and chemodenervated rats were subjected to normoxia or hypoxia for 6 hr (10% O(2) in N(2)). VEGF protein was quantified in micropunches of brainstem tissue. Only chemodenervated animals showed an increased VEGF expression in response to hypoxia, whereas, in normoxia, VEGF expression was not modified by chemodenervation. The same hypoxic condition was repeated for 8 days before immunocytochemical staining with anti-VEGF; antiglial fibrillary acidic protein (GFAP), a marker of astrocytes; and anti-rat endothelial cell antigen-1 (anti-RECA-1) that recognizes endothelial cells. Confocal analysis showed a cellular colocalization of GFAP and VEGF, indicating that VEGF was overexpressed predominantly in astrocytes. Increased RECA-1 immunolabeling indicated an enhanced angiogenesis in chemodenervated rats subjected to hypoxia. These results indicate that glial cells and the vascular network contribute to the brainstem remodeling. The peripheral chemodenervation reveals a central O(2) chemosensitivity involving a cascade of gene expression triggered by hypoxia, which in intact animals may act synergically with peripheral chemosensory inputs.

Developmental plasticity of the carotid chemoafferent pathway in rats that are hypoxic during the prenatal period.

The chemoreflex pathway undergoes postnatal maturation, and the perinatal environment plays a critical role in shaping respiratory control system. We investigated the role of prenatal hypoxia on the maturation of the chemoreflex neural circuits regulating ventilation in rat. Effects of hypoxia (10% O2) from the 5th to the 20th day of gestation were studied on male offspring at birth and on postnatal days 3, 7, 21 and 68. Maturation of the respiratory control system was assessed by in vivo tyrosine hydroxylase (TH) activity measurement in peripheral chemoreceptors (carotid bodies, petrosal ganglia), and in brainstem catecholaminergic cell groups (A2C2c and A1C1 areas in the medulla, A5 and A6 areas in the pons). Resting ventilation and ventilatory response to hypoxia were evaluated as functional sequelae. In peripheral structures, prenatal hypoxia reduced TH activity within the first postnatal week and enhanced it later. In contrast, in central areas, prenatal hypoxia upregulated TH activity within the first postnatal week and downregulated it later. The in vivo TH activity impairment is therefore tissue specific, with an opposite effect on the peripheral and central neural circuits. A shift of the effect of prenatal hypoxia occurred between 1 and 3 weeks, indicating a postnatal temporal effect of prenatal hypoxia. An important period in the development of the chemoafferent pathway occurred between the first and the third postnatal week. Functionally, prenatal hypoxia impaired resting ventilation and ventilatory response to hypoxia. The alterations of the catecholaminergic components of the chemoafferent pathway resulting from prenatal hypoxia might contribute to impair postnatal respiratory behaviour.

Desferrioxamine enhances hypoxic ventilatory response and induces tyrosine hydroxylase gene expression in the rat brainstem in vivo.

The iron chelator desferrioxamine (DFO) induces accumulation of the hypoxia-inducible factor (HIF-1), a transcription factor that up-regulates genes involved in adaptative responses to hypoxia. This property makes DFO a potential neuroprotector against hypoxic stress. We investigated in rats the effects of DFO on the ventilatory response to mild hypoxic tests and the expression of tyrosine hydroxylase (TH), a target gene of HIF-1. Two protocols were used, the first with repeated injections of 50 mg/kg DFO every 2 days during a 2-week period. This was aimed at define the time course of the ventilatory responses to a hypoxic test. In the second protocol, rats were given a single injection of 300 mg/kg DFO. Every day over 4 days, the hypoxic ventilatory response was recorded before the animal was sacrificed, and Western blot analysis of TH in the dorsal brainstem cardiorespiratory area was performed. DFO produced a delayed increase in the hypoxic ventilatory response, which appeared in the same time window as TH up-regulation (2-3 days after the bolus injection of DFO). This delay suggests a genic effect of the drug that improves the ventilatory response to hypoxia.

Food restriction, performance, biochemical, psychological, and endocrine changes in judo athletes.

In order to test the hypothesis that dietary restriction may have a negative influence on physiological and psychological adaptation to a judo competition, we examined the effects of weight loss induced by restricting energy and fluid intake on the physiology, psychology, and physical performance of judo athletes. Twenty male judoka were randomly assigned to one of two groups (Group A: called diet, n = 10; height 174.8 +/- 1.9 cm, body weight 75.9 +/- 3.1 kg; they were asked to lose approximately 5 % of their body weight through self-determined means during the week before the competition; Group B: called control, n = 10; height 176.4 +/- 1.1 cm, body weight 73.3 +/- 6.3 kg maintained their body weight during the week before the competition). A battery of tests was performed during a baseline period (T1), on the morning of a simulated competition (T2) and 10 min after the end of the competition (T3). The test battery included assessment for body composition, performance tests, evaluation of mood, determination of metabolic and hormonal responses. Dietary data were collected using a 7-day diet record. The nutrient analysis indicated that all the athletes followed a low carbohydrate diet whatever the period of the investigation. For the Group A, the food restriction (- 4 MJ per day) resulted in significant decreases of the body weight and altered the mood by increasing Fatigue, Tension and decreasing Vigour. Dietary restriction had also a significant influence on metabolic and endocrine parameters and was associated with poor performance. After the competition, significant decreases of the levels in testosterone, T/C ratio, alkali reserve, and free fatty acid were observed in both groups, whereas the plasma concentrations in insulin, ammonia, urea, and uric acid were increased. In conclusion, our results suggest that the combination of energy restriction and intense exercise training, which causes weight reduction before a competition, adversely affects the physiology and psychology of judo athletes and impairs physical performance before the competition. Our data are the first to demonstrate that a competition including five 5-min bouts induced the same changes of physiological and psychological variables and performance whatever the dietary intake (dietary restriction or not) during the seven days before the competition.

Skeletal muscle HSP72 and norepinephrine response to static magnetic field in rat.

The present work was undertaken in order to investigate the noradrenergic system and skeletal muscle heat shock protein 72 (HSP72) response to static magnetic field (MF) in male rats. At thermoneutrality (25 degrees C), the exposition of rats 1 hour/day for 5 consecutive days to MF of 128 mT (m tesla) induced an increase in norepinephrine content in gastrocnemius muscle (+25%, p < 0.05) but had no effect at 67 mT (+1%, p > 0.05), indicating a stimulatory effect of sub-acute MF exposure on the noradrenergic system activity. Moreover, exposed rats to MF displayed a non-significant increase of HSP72 levels in gastrocnemius muscles (+29%, p > 0.05). The results indicate that noradrenergic systems in rat's gastrocnemius muscles are affected by MF exposure. Interestingly, sub-acute exposure insufficiency increased HSP72 levels in gastrocnemius muscles.

Ret deficiency in mice impairs the development of A5 and A6 neurons and the functional maturation of the respiratory rhythm.

Although a normal respiratory rhythm is vital at birth, little is known about the genetic factors controlling the prenatal maturation of the respiratory network in mammals. In Phox2a mutant mice, which do not express A6 neurons, we previously hypothesized that the release of endogenous norepinephrine by A6 neurons is required for a normal respiratory rhythm to occur at birth. Here we investigated the role of the Ret gene, which encodes a transmembrane tyrosine kinase receptor, in the maturation of norepinephrine and respiratory systems. As Ret-null mutants (Ret-/-) did not survive after birth, our experiments were performed in wild-type (wt) and Ret-/- fetuses exteriorized from pregnant heterozygous mice at gestational day 18. First, in wt fetuses, quantitative in situ hybridization revealed high levels of Ret transcripts in the pontine A5 and A6 areas. Second, in Ret-/- fetuses, high-pressure liquid chromatography showed significantly reduced norepinephrine contents in the pons but not the medulla. Third, tyrosine hydroxylase immunocytochemistry revealed a significantly reduced number of pontine A5 and A6 neurons but not medullary norepinephrine neurons in Ret-/- fetuses. Finally, electrophysiological and pharmacological experiments performed on brainstem 'en bloc' preparations demonstrated impaired resting respiratory activity and abnormal responses to central hypoxia and norepinephrine application in Ret-/- fetuses. To conclude, our results show that Ret gene contributes to the prenatal maturation of A6 and A5 neurons and respiratory system. They support the hypothesis that the normal maturation of the respiratory network requires afferent activity corresponding to the A6 excitatory and A5 inhibitory input balance.

Age-related sensitivity to lung oxidative stress during ozone exposure.

As immature and aged rats could be more sensitive to ozone (O(3))-linked lung oxidative stress we have attempted to shed more light on age-related susceptibility to O(3) with focusing our interest on lung mitochondrial respiration, reactive oxygen species (ROS) production and lung pro/antioxidant status. For this purpose, we exposed to fresh air or O(3) (500 ppb 12 h per day, for 7 days) 3 week- (immature), 6 month- (adult) and 20 month-old rats (aged). We determined, in lung, H(2)O(2) release by mitochondria, activities of major antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT)], heat shock protein (HSP(72)) content and 8-oxodG and dG-HNE nDNA contents, as DNA oxidative damage markers. In adult rats we did not observe alteration of pro/antioxidant status. In contrast to adults, immature rats exposed to O(3) higher nDNA 8-oxodG content and HSP(72) and without antioxidant enzymes modification. Aged rats displayed mild uncoupled lung mitochondria, increased SOD and GPx activities, and higher 8-oxodG content after O(3) exposure. Thus, in contrast to adults, immature and aged rats displayed lung oxidative stress after O(3) exposure. Higher sensitivity of immature to O(3) was partly related to ventilatory parameters and to the absence of antioxidant enzyme response. In aged rats, the increase in cytosolic SOD and GPx activities during O(3) exposure was not sufficient to prevent the impairment in mitochondrial function and accumulation in lung 8- oxodG. Finally, we showed that mitochondria seem not to be a major source of ROS under O(3) exposure.

Role of catecholamines in glucagon-induced thermogenesis.

The present work was undertaken in order to investigate whether the observed thermogenesis following glucagon injection requires the participation of catecholamines. Our experiments aim at studying the effects of intraperitoneal injection of glucagon on metabolic rates, plasma catecholamine and fuel metabolites in guanethidine-treated ducklings reared at thermoneutrality (25 degrees C). The chronic guanethidine treatment induced a marked decrease in catecholamines levels in peripheral tissues (heart, muscle and intestine) but not in adrenals. At thermoneutrality, intraperitoneal injection of glucagon had lower thermogenic effects in guanethidine-treated compared to control ducklings. Glucagon injection elicited a concomitant increase of plasma norepinephrine, metabolic rate and energy metabolites in control ducklings, whereas in guanethidine-treated ducklings, the plasma catecholamines and metabolic rate did not undergo any consistent change. The thermogenic action of glucagon in birds involves at least the mobilization of lipids and catecholaminergic system stimulation.

Effects of aging on the cardiac remodeling induced by chronic high-altitude hypoxia in rat.

Effects of chronic high-altitude hypoxia on the remodeling of right ventricle were examined in three age groups of rats: 2, 6, and 18 mo. The extent of right ventricular (RV) hypertrophy (RVH) showed an age-associated diminution. RV cell size and pericellular fibrosis showed a significant increase in the 2- and 6-mo-old exposed rats but not in the 18-mo-old exposed rats compared with control. A hyperplasic response was underscored in the three exposed age groups but appeared less pronounced in the 18-mo-old rats. A significant decrease in the transient outward potassium current (Ito) density was observed in RV cell only in the 2-mo-old exposed group compared with the control group. In the control group, there was a clear tendency for Ito density to decrease as a function of age. The sustained outward current density was modified neither by the hypoxia condition nor by the age. Neither the cytochrome c oxidase activity nor the heat shock protein 72 content in the RV was altered after hypoxic exposure regardless of age. The norepinephrine content in the RV was significantly decreased in each age group exposed to hypoxia when compared with their age-matched control group. Our findings indicate that the remodeling (at morphological and electrophysiological levels) induced by chronic hypoxia in the RV can be decreased by the natural aging process.

Sub-chronic exposure to toluene at 40 ppm alters the monoamine biosynthesis rate in discrete brain areas.

Effects of long-term exposure to a sub-toxic concentration of toluene (40 ppm, 104 h per week, 16 weeks) have been studied on monoamine biosynthesis rate in rat. The activities of the rate limiting enzymes in catecholamine and 5-hydroxytryptamine biosynthesis tyrosine and tryptophan hydroxylase, respectively, were estimated in vivo by measuring the accumulation of l-dihydroxyphenylalanine and 5-hydroxytryptophan after pharmacological blockade of l-aromatic acid decarboxylases by NSD-1015 (100 mg kg(-1) ip). The sub-chronic exposure to toluene led to a significant and gender dependent alteration in both catecholamine and 5-hydroxytryptamine biosynthesis rate in brainstem catecholaminergic cell groups and hypothalamus. In females exposed to toluene, changes in tryptophan hydroxylation were found in rostral subset of A2C2 (+41%) and in A1C1 (+41%) while a decrease in A5 (-44%) and an increase in A2C2 (+28%) were found in tyrosine hydroxylation. In males, an increase in tryptophane hydroxylation was observed in rostral subset of A2C2 (+35%) while a decrease was observed in ventro-median hypothalamus (-17%). These results suggest that toluene exposure to a dose generally recognized as sub-toxic (40 ppm, no observed adverse effect level) leads to adverse effects on monoaminergic systems. Therefore, the neurotoxicity of toluene should be carefully re-evaluated taking into account not only the exposure level but also the duration.

Phox2a gene, A6 neurons, and noradrenaline are essential for development of normal respiratory rhythm in mice.

Although respiration is vital to the survival of all mammals from the moment of birth, little is known about the genetic factors controlling the prenatal maturation of this physiological process. Here we investigated the role of the Phox2a gene that encodes for a homeodomain protein involved in the generation of noradrenergic A6 neurons in the maturation of the respiratory network. First, comparisons of the respiratory activity of fetuses delivered surgically from heterozygous Phox2a pregnant mice on gestational day 18 showed that the mutants had impaired in vivo ventilation, in vitro respiratory-like activity, and in vitro respiratory responses to central hypoxia and noradrenaline. Second, pharmacological studies on wild-type neonates showed that endogenous noradrenaline released from pontine A6 neurons potentiates rhythmic respiratory activity via alpha1 medullary adrenoceptors. Third, transynaptic tracing experiments in which rabies virus was injected into the diaphragm confirmed that A6 neurons were connected to the neonatal respiratory network. Fourth, blocking the alpha1 adrenoceptors in wild-type dams during late gestation with daily injections of the alpha1 adrenoceptor antagonist prazosin induced in vivo and in vitro neonatal respiratory deficits similar to those observed in Phox2a mutants. These results suggest that noradrenaline, A6 neurons, and the Phox2a gene, which is crucial for the generation of A6 neurons, are essential for development of normal respiratory rhythm in neonatal mice. Metabolic noradrenaline disorders occurring during gestation therefore may induce neonatal respiratory deficits, in agreement with the catecholamine anomalies reported in victims of sudden infant death syndrome.

Electrical conduction through nerve and DNA.

The aim of the present study was to analyse electric resistivity at different ambient temperatures between 300 to 20K in the frog sciatic nerve and salmon sperm DNA. When the electrical contacts were leaned just into the sciatic nerve, an increase of the sciatic nerve resistivity was observed for 240 K < T < 300 K and a rise of electrical conductivity was apparent below 240 K. This dependence is generally associated with a semiconductor behaviour. Once the sciatic nerve temperature was driven below 250K, the resistivity abruptly decreased and then at temperatures lower than 234 K, it remained constant and close to one tenth of its ambient temperature value. By contrast, when the electrical contacts were leaned into Salmon sperm DNA, the resistivity remained constant between 300K to 20K, showing a high electrical stability at low temperature. Thus, we report the existence of a new form of electric conductivity in the sciatic nerve at low ambient temperature, which in turn has many electric similarities with inorganic or organic superconductors, whereas temperature failed to alter DNA electrical properties until 20K.

Sciatic nerve monoaminergic system response to cold acclimatization in Muscovy duckling.

The effect of cold acclimatization on the monoamines in duckling sciatic nerve was studied. In thermoneutral (TN) ducklings, the high dopamine (DA) to norepinephrine (NE) ratios in the right (R, 0.14) or in the left (L, 0.20) sciatic nerves at 6 weeks of age suggests the presence of non-precursor specific dopaminergic pools. DA, DOPAC (3, 4-dihydroxyphenylacetic acid) levels and the ratio DA to NE were similar in cold-acclimated (CA) and TN ducklings sciatic nerves. Moreover, serotonin and its metabolite 5-hydroxyindoleacetic acid (5-HIAA) remained unchanged in the sciatic nerve, indicating that this system is not affected with cold exposure. The contents of NE were higher (R = +44.46%; L = +43.73%) in sciatic nerve of CA than in TN ducklings. The results indicate that noradrenergic systems in the ducklings sciatic nerve are markedly affected by cold exposure. Interestingly, chronic exposure to cold failed to alter the dopaminergic and the serotonergic system in duckling sciatic nerve.

Resistance to obesity in Lou/C rats prevents ageing-associated metabolic alterations.

AIMS/HYPOTHESIS: Ageing is associated with metabolic alterations characterised by changes in energy expenditure, obesity, leptin and insulin resistance. The Lou/C rat, an inbred strain of Wistar origin, is presented as both an obesity-resistant rat and a model of healthy ageing. METHODS: To characterise the mechanisms underlying obesity resistance in Lou/C rat, we measured food intake and energy expenditure by indirect calorimetry at 1, 6, 12, 18, and 24 months of age. Moreover, plasma insulin and leptin concentrations were assessed by radioimmunoassay in Lou/C and Wistar rats throughout their life span. RESULTS: Compared to Wistar rats, Lou/C rats presented a higher food intake only at 24 months of age and they had a higher energy expenditure at 6 and 12 months of age (+21% and +14%, respectively). Plasma insulin concentration increased markedly in 18- and 24-month-old Wistar rats, but remained stable during ageing in Lou/C rats. From the age of 6 months, the plasma leptin concentrations in Wistar rats were higher than in Lou/C rats of the same age (four-, seven-, five- and threefold higher at 6, 12, 18, 24 months of age, respectively). CONCLUSION/INTERPRETATION: Compared to Wistar rats, Lou/C rats did not develop insulin resistance as confirmed by a higher glucose infusion rate during the hyperinsulinaemic-euglycaemic clamp. These data provide evidence that insulin resistance is associated with the excess of adipose tissue in Wistar rats. Not only Lou/C rats present a higher median life span than Wistar rats (+20%), but they also show a healthy ageing process considering fat accretion and insulin resistance.

Neurochemical development of the brainstem catecholaminergic cell groups in rat.

The postnatal development of tyrosine hydroxylase activity has been studied in the brainstem catecholaminergic cell groups (A1C1, A2C2, A5, A6, A7), involved in cardiorespiratory control. In rat, at birth and at postnatal days P3, P7, P14, P21 ant P68, we used a microdissection technique followed by in vivo measurement of the tyrosine hydroxylase (TH) activity, the rate-limiting enzyme in catecholamine synthesis. There is two successive marked increases in TH activity: at P3 in every catecholaminergic cell groups (A1C1, +225%; A2C2, +300%; A5, +190%; A6, +205% compared to birth) and during the third postnatal week with a peak of TH activity at P14 (A6, +90% above the P7 level) or at P21 (A1C1, +715%; caudal A2C2, +585%; rostral A2C2, +15%; A5, +445%; A7, +180% compared to P7). The data suggest the existence of two temporal windows during the neurochemical development of the catecholaminergic cell groups, which correspond to two metabolic transitions. The first one could be related to the intra-, extrauterine transition and the second one, to a deep energetic phase of maturation in the rat brain, closely related to the maturation of cardiorespiratory processes.

Psychobiologic responses to 4 days of increased training and recovery in cyclists.

The psychobiologic status of cyclists after 4 days of training and the kinetics of recovery were assessed by measuring the sympatho-adrenal level, the central noradrenergic activity and the cortisol/testosterone status by non-invasive methods. For this purpose, urinary excretion of methoxyamines (metanephrine [MN], normetanephrine [NMN]), which are metabolites of circulating catecholamines, 3-methoxy-4-hydroxyphenyl glycol sulfate (MHPG-S), a metabolite of brain norepinephrine, and salivary output of cortisol and testosterone were measured in twelve national cyclists (aged 19.5 +/- 4.5 years), just before (T 1 ) and at the end of the training (T 2 ), and during the three following recovery days (R 1, R 2, R 3 ). Urinary and salivary samples were also collected during a period of relative rest, in order to get reference values (T 0 ). At T 0, T 1 and T 2, mood states, as measured by the Profile of Mood States, and rating of perceived muscle soreness were assessed. The overall mood and muscle soreness levels were not affected by the training. The load increased by 187 % as an average between the first and the fourth day of training. A significant increase in NMN levels and a decrease in T:F ratio were observed at T 2, while MHPG-S excretion remained unchanged. Persistent high urinary output of NMN and MN were observed during the post-training recovery period for 24 h (R 1 ) and 48 h (R 2 ), respectively. After 72 h of recovery (R 3 ), MN levels had returned to baseline while NMN output was lower than the control level. T:F values returned to their control levels within 48 h of recovery. The strenuous training seems to induce an alteration in peripheral neuro-endocrine parameters without modifications of central factors. The hormonal status remained altered for at least 1 day of post-training recovery and seemed to be achieved within 3 days.

Activity patterns of cardiac vagal motoneurons in rat nucleus ambiguus.

Extracellular recordings were made in the right nucleus ambiguus of urethane-anesthetized rats from 33 neurons that were activated at constant latency from the craniovagal cardiac branch. Their calculated conduction velocities were in the B-fiber range (1.6-13.8 m/s, median 4.2), and most (22/33) were silent. Active units were confirmed as cardiac vagal motoneurons (CVM) by the collision test for antidromic activation and by the presence of cardiac rhythmicity in their resting discharge (9/9). Brief arterial pressure rises of 20-50 mmHg increased the activity in five of five CVM by 0.1 +/- 0.02 spikes. s(-1). mmHg(-1) from a resting 3.8 +/- 1.2 spikes/s; they also recruited activity in two of four previously silent cardiac branch-projecting neurons. CVM firing was modulated by the central respiratory cycle, showing peak activity during inspiration (8/8). Rat CVM thus show firing properties similar to those in other species, but their respiratory pattern is distinct. These findings are discussed in relation to mechanisms of respiratory sinus arrhythmia.

Comparison of passive heat or exercise-induced dehydration on renal water and electrolyte excretion: the hormonal involvement.

The effects of hydromineral hormones and catecholamines on renal water and electrolyte excretion were examined during and after dehydration induced by either passive heat or exercise. Eight healthy young Caucasian subjects participated in three separate trials, each including three consecutive phases. Phases 1 and 3 involved a 90-min period at rest in a thermoneutral environment, while phase 2 involved a 120-min period designed to provide: (1) euhydration (control trial), (2) passive heat-induced dehydration of 2.8% body mass, or (3) exercise-induced dehydration of 2.8% body mass. During the two dehydration procedures, the decreases in urine flow and sodium excretion were more marked during exercise (P < 0.05). An increase in plasma catecholamines occurred only during exercise, together with a reduction in creatinine clearance and more marked increases in plasma renin and aldosterone than during passive heat exposure (P < 0.05). Although plasma vasopressin was elevated during the two dehydration procedures, urine osmolality did not change and, moreover, free water clearance increased during exercise (P < 0.05). Plasma levels of atrial natriuretic peptide increased markedly only during exercise compared to the other trials (P < 0.05). After the dehydration procedures, urine flow decreased again and urine osmolality increased markedly (P < 0.05), while plasma vasopressin remained elevated. These results suggest that sympathoadrenal activation during exercise plays a major role in the more marked reduction in diuresis and natriuresis than during passive heat exposure. Despite high plasma vasopressin concentrations during the two dehydrating events, the observed antidiuresis was not due to an increased renal concentrating ability, and the vasopressin was more effective after the dehydration procedures.

Responsiveness of plasma catecholamines to intracerebroventricular injection of glucagon in Muscovy ducklings.

Recent investigations have demonstrated a modulatory action of glucagon on shivering via the central nervous system in ducklings. The aim of this study was to investigate the effects of intracerebroventricular injection (i.c.v.) of glucagon on metabolic rates (MR) and plasma catecholamines in control ducklings (TN) and in ducklings exhibiting nonshivering thermogenesis after chronic glucagon treatment: (GT). At thermoneutrality (25 degrees C, Ta), i.c.v. injection of glucagon had no thermogenic effects in TN and GT ducklings. At cold (+4 degrees C, Ta), i.c.v. glucagon injection elicited a concomitant decrease of plasma norepinephrine (NE) and MR in TN ducklings, whereas in GT ducklings, the plasma catecholamines and the MR remained unchanged. These results indicate that glucagon treatment rendered the catecholaminergic system of GT ducklings insensitive to cold or i.c.v. glucagon injection.

Selective cardiorespiratory and catecholaminergic areas express the hypoxia-inducible factor-1alpha (HIF-1alpha) under in vivo hypoxia in rat brainstem.

Under severe oxygen deprivation, all cells are able to express the transcription factor HIF-1, which activates a wide range of genes. Under tolerable hypoxia, chemosensory inputs are integrated in brainstem areas, which control cardiorespiratory responses. However, the molecular mechanisms of this functional acclimatization are unknown. We investigated when and where the inducible HIF-1alpha subunit is expressed in the rat brainstem in vivo, under physiological hypoxia. The regional localization of HIF-1alpha mRNA and protein was determined by in situ hybridization and immunocytochemistry in adult male rats exposed to moderate hypoxia (10% O2) for 1-6 h. HIF-1alpha protein was found in cell types identified by immunocytochemistry as catecholaminergic neurons. Hypoxia induced HIF-1alpha mRNA and protein in only some parts of the brainstem located dorsomedially and ventrolaterally, which are those involved in the cardiorespiratory control. No labelling was detected under normoxia. The protein was detected in glia and neurons after 1 and 6 h of hypoxia, respectively. A subset of A2C2 and A1C1 catecholaminergic neurons colocalized tyrosine hydroxylase and HIF-1alpha proteins under hypoxia, but no HIF-1alpha was detected in more rostral catecholaminergic areas. In contrast to cardiorespiratory areas, HIF-1alpha protein was already present under normoxia in glial cells of brainstem tracts but was not overexpressed under hypoxia, although HIF-1alpha mRNA was up-regulated. In conclusion, there appear to be two regulatory mechanisms for HIF-1alpha expression in the brainstem: hypoxic induction of HIF-1alpha protein in cardiorespiratory-related areas and constitutive protein expression unaffected by hypoxia in brainstem tracts.