Immunocytochemical localization of the insulin-responsive glucose transporter 4 (Glut4) in the rat central nervous system.

We have previously reported that the insulin-responsive glucose transporter GLUT4 is strongly expressed by discrete areas of the rat brain (Leloup et al. [1996] Molec. Brain Res. 38:45-53). In the present study, a sensitive immunocytochemical technique has been used to analyze extensively the anatomical and ultrastructural localizations of GLUT4 in the rat central nervous system in order to gain insight into the physiological role of this transporter. We confirm that GLUT4 is expressed by numerous neurons of the brain and spinal cord, whereas glial cells are more scarcely labeled. In both light and electron microscopy, we observe that the immunoreactivity for GLUT4 is localized mainly in the somatodendritic portion of neurons, where some cisterns of rough endoplasmic reticulum, ribosomal rosettes, certain Golgi saccules, and some intracytoplasmic vesicles are labeled. In contrast, axons and nerve terminals are only occasionally immunostained in certain brain regions such as the neocortex and the ventricular surfaces for example. The GLUT4-immunoreactive structures appear concentrated and most prominently immunostained in motor areas, such as the sensorimotor cortex, most basal ganglia and related nuclei, the cerebellum and deep cerebellar nuclei, a number of reticular fields, motor nuclei of cranial nerves, and motor neurons of the ventral horn of the spinal cord. The labeled regions, which also include some sensory nuclei, are often those in which Vissing et al. ([1996] J. Cerebral Blood Flow Metab. 16:729-736) have shown that exercise stimulates local cerebral glucose utilization, so that GLUT4 might be involved in this effect. On the other hand, the fact that the anatomical localizations of GLUT4 reported here generally agree with the distribution of insulin- or insulin-receptor- related receptors is important since it indicates that the translocation of GLUT4 might also be regulated by insulin in the central nervous system.

Light and electron microscopic immunocytochemical visualization of 5-HT1B receptors in the rat brain.

Specific antipeptide antibodies were used for the immunohistochemical visualization of 5-HT1B receptors in the rat brain. A dense, specific 5-HT1B receptor-like immunoreactivity was found in the globus pallidus, the dorsal subiculum and the substantia nigra. At the light microscope level, immunostaining was diffuse within the neuropil but absent from cell bodies. Observations at the electron microscope level in the substantia nigra showed immunoperoxidase staining in fine unmyelinated axons and nerve terminals.

[Effect of halothane on ventilation and arterial blood gases in rats with and without diaphragmatic paralysis]. / Effets de l'halothane sur la ventilation et les gaz du sang arteriel chez le rat avec ou sans paralysie diaphragmatique.

Some patients with diaphragmatic paralysis or dysfunction maintain ventilation by use of other muscles. Anaesthesia, in modifying the performance of these muscles, presents a potential risk to such patients. To evaluate this risk, the effects of halothane on ventilation and arterial blood gases were studied on a model of bilateral diaphragmatic paralysis, the phrenectomized rat. The study was performed on 43 rats. Success of phrenectomy was confirmed at laparotomy, which did not result in blood gas changes. Laparotomy was performed in 23 rats and a carotid artery was catheterized. In 11 control rats, phrenic nerves were exposed but not sectioned, and in 12 other rats, the phrenic nerves were sectioned. Ventilation was measured by plethysmography in awake rats before and after surgery and in the same rats anaesthetized with halothane 1.1%. In the 23 rats, a decrease in weight and core temperature was observed after operation and this was more marked in phrenectomized than in control rats. In the 11 control rats, ventilation increased postoperatively without change in blood gases. In these rats, halothane caused a decrease in minute ventilation and PaO2 and an increase in PaCO2. Phrenectomy in awake rats led to an increase in minute ventilation, hypoxaemia and hypercapnia. In these rats, halothane led to death in three and a decrease in minute ventilation, with hypercapnia and hypoxaemia in the nine other rats. Blood gas changes were greater than in anaesthetized controls. In the intact rat, halothane leads to blood gas changes comparable to those observed in other species and humans. The present study confirms the effects of halothane on respiratory muscles other than the diaphragm and demonstrates the severe respiratory risk of anaesthesia in patients whose ventilation is maintained by these muscles.

[The effect of halothane on blood gases and arterial acid-base equilibrium in intact rats and in chemo-denervated rats]. / Effets de l'halothane sur les gaz du sang et l'équilibre acido-basique artériels chez le rat intact et chez le rat chémodénervé.

Halothane decreases the ventilatory response to hypoxia and the activity of peripheral arterial chemoreceptors, resulting in "chemical chemodenervation." In order to evaluate the role of this halothane-induced "chemical denervation" in acid-base and arterial blood gas changes, these values were measured in intact and chemodenervated rats, awake and under anaesthesia. Since the depth of anaesthesia could be modified by the anatomical chemodenervation, the ED50 of inspired halothane was determined in six rats before and after anatomical chemodenervation. To prevent haemodynamic changes due to halothane and/or anatomical chemodenervation from interfering with the results, systemic arterial blood pressure and heart rate were measured in six intact rats, awake and then anaesthetized, and in the same rats after chemodenervation, awake and then anaesthetized. In nine intact rats and in 19 chemodenervated rats, arterial pH, arterial bicarbonate concentration, and arterial blood gases (PaO2 and PaCO2) were measured before and after administration of halothane. Anatomical chemodenervation modified neither the inspired ED50 (1.1%), nor the mean arterial blood pressure or heart rate. The haemodynamic effects of halothane were comparable in intact and in chemodenervated rats. Changes in arterial blood gases and acid-base balance due to halothane in intact rats and due to chemodenervation in awake rats were not different, but there was a decrease in PaO2 and pHa, and an increase in PaCO2. In chemodenervated rats, halothane caused a further decrease in PaO2 and a further increase in PaCO2. The fact that halothane and anatomical chemodenervation have similar effects on arterial blood gases and acid-base balance favours a "chemical chemodenervating" action of halothane. However, the additional effects of halothane in the anatomically chemodenervated animal show that the action of halothane on blood gases and acid-base balance is the result of multiple sites of impact on the respiratory system.

[Respiratory effects of moderate hypothermia (36 degrees C-28 degrees C) in dogs under halothane anesthesia]. / Effets ventilatoires d'une hypothermie modérée (36 degrees C-28 degrees C) chez le chien anesthésie à l'halothane.

Changes in systemic haemodynamic variables (mean arterial pressure, MAP; heart rate, HR; cardiac output, Qc), in oxygen consumption, VO2, and in ventilation (minute ventilation, V; respiratory frequency, f; tidal volume, VT; and arterial blood gases) with particular attention to respiratory times (duration of inspiration, TI; duration of expiration, TE; duration of the breathing cycle, TTOT), to respiratory timing (TI/TTOT) and respiratory drive (VT/TI) were studied during moderate progressive hypothermia (36 degrees C to 28 degrees C) during stable halothane anaesthesia (MAC = 1.5) in six dogs. MAP, HR and Qc decreased; V and f decreased, the decrease in f being correlated with that in temperature (r = 0.66; P < 0.01). Tidal volume did not change. The PaO2 and pHa decreased while PaCO2 increased slightly. The decrease in ventilation was related to changes in respiratory times (TI and TE) which increased (TE more than TI) and in respiratory drive (VT/TI which decreased due to the increase in TI). The relation between VT/TI and TI/TTOT changes was not constant during cooling. Changes in respiratory times and drive could be due to the effect of cold on medullar respiratory control.

[The measurement of pH and gases in the blood using microanalysis: the effect of storage at 4 degrees C for an hour. A study in the rat]. / Mesure du pH et des gaz du sang sur des micro-echantillons: effets de la conservation à 4 degrees C pendant une heure. Etude chez le rat.

The effects of one hour storage at 4 degrees C on micro blood gas samples (150 microliters) were studied for a wide range of values (pH: 7.11-7.58; PCO2: 26-97 mmHg; PO2: 31-503 mmHg) in 20 rats with indwelling carotid artery catheters. Blood gas values were modified by varying the composition of inspired gases: normoxia, hypocapnic hypoxia, hyperoxia, hypercapnia (in this case eight animals were anaesthetized with halothane 1.1%). One hundred and eight double micro-samples were taken. For each double sample, one was analysed immediately (H0) and compared with the second sample after one hr storage at 4 degrees C (H1). The Bland and Altman method was used for the statistical analysis of results. After one hr storage at 4 degrees C, the PCO2 was slightly higher than at H0 (mean difference +/- SD: +1.08 +/- 1.7 mmHg) and arterial pH was slightly lower (mean difference +/- SD: -0.016 upH +/- 0.011 upH). These results show that for these two variables, in the range studied, one hour storage at 4 degrees C had little effect. In contrast, for arterial PO2 the mean difference between all measurements between H1 and H0 was -17 +/- 25 mmHg. If results lower than 200 mmHg (56 double samples) are considered separately, the mean difference between values at H1 and H0 was only -0.98 +/- 5.3 mmHg. For PaO2 greater than 200 mmHg (52 double samples), the mean difference was -34 +/- 26.3 mmHg; this may be due to low reproducibility of measurements of elevated PO2 levels and to the effects of cellular metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)

Enkephalin-containing nerve cell bodies in the substantia nigra of the rat: demonstration by immunohistochemistry and in situ hybridization.

The use of a new and very sensitive immunohistochemical method, combined with intracerebral injections of colchicine, has allowed us to show that a number of nerve cell bodies immunoreactive for Met-enkephalin are present in several mesencephalic nuclei of the rat, including the different subdivisions of the substantia nigra (SN). The existence of numerous neuronal somata of this kind in the medial part of the SN pars compacta and in the lateral half of the pars reticulata is rather new. The latter has been ascertained by demonstrating a perikaryal immunoreactivity for synenkephalin in the same regions of the SN. In addition, by in situ hybridization, we have shown that neuronal cell bodies expressing the preproenkephalin A (PPA) gene are also present in the same regions of the SN. However, the fact that a strong radioautographic reaction was found only in rats which received an intranigral injection of 6-hydroxydopamine indicates that these neurons are probably not dopaminergic and that an induction of the PPA gene occurs in these animals.

[Protein composition of the hemolymph of Biomphalaria glabrata (Mollusca, Gastropoda). Influence of the method of sampling and relation to the protein composition of different organs]. / Composition protéique de l'hémolymphe de Biomphalaria glabrata (Mollusque Gastéropode). Influence de la méthode de prélèvement et relation avec la composition protéique des différents organes.

The blood composition of B. glabrata is analyzed by electrophoresis. Its proteic composition differs in relation with the blood sampling method. Most of the supplementary fractions also have been found in the gut, some proteins possibly come from the hepatopancreas, whereas others could have their origin in the albumen gland. The sampling method must be selected according to these observations.