RESUMEN
The distribution of vasopressin receptors in the brain of the jerboa (Jaculus orientalis) was studied using tritiated arginin vasopressin ([3H]vasopressin). beta-Particles emitted from tritiated ligand bound to brain sections were detected by a newly developed beta-radio imager to generate a light spot which was read by a charge coupled device camera. The number and coordinate of the center of gravity of the light spot were recorded. After summation in pixels of the counts collected during 2-20 h, an image was produced representing the distribution of [3H]vasopressin bound to brain sections. Specific vasopressin binding was detected in various brain regions such as the cerebral cortex, islands of Calleja, pallidum, amygdala and the hippocampus as well as in the pituitary gland. The intensity of the binding was quantified directly from the images obtained and expressed in decays/min/surface unit. The linearity of this method of detection allowed a relevant measurement of non-specific binding, therefore its subtraction from images representing the total binding. Three-dimensional reconstructions of labeled structures were also performed. The presence of numerous vasopressin receptors in the jerboa hippocampus suggests a major role for this neuropeptide in this part of the brain.
Asunto(s)
Autorradiografía/métodos , Encéfalo/metabolismo , Procesamiento de Imagen Asistido por Computador , Vasopresinas/metabolismo , Animales , Sitios de Unión , Hipocampo/metabolismo , Masculino , Roedores , Distribución Tisular , TritioRESUMEN
The distribution of vasopressin receptors was studied in the brain of a photoperiodic animal, the Siberian hamster. Attention was focused on [3H]vasopressin binding sites located in the hypothalamic ventromedial nucleus, medial tuberal nucleus, and ventral premammillary nucleus in males or females kept in long or short photoperiod conditions. Displacement experiments with structural analogs suggested that vasopressin receptors in the hamster hypothalamus are of the vasopressor (V1) type. Quantitative data obtained with a gaseous detector of beta-particles indicated that in the ventromedial nucleus and in the ventral premammillary nucleus of animals in long photoperiod, the number of beta-particles emitted per unit area was significantly greater in males than in females. In the ventromedial hypothalamic nucleus, in both males and females, the number of beta-particles emitted was significantly lower in short than in long photoperiod conditions. In the ventral premammillary nucleus, shortening of the photoperiod had a significant effect in reducing the amount of [3H]vasopressin bound in females, but not in males. These data suggest that, in the hamster, the control of the expression of vasopressin receptors differs among various hypothalamic nuclei and may depend on the sex and/or on the level of circulating gonadal steroids.
Asunto(s)
Hipotálamo/fisiología , Luz , Receptores de Angiotensina/metabolismo , Vasopresinas/metabolismo , Animales , Autorradiografía , Cricetinae , Oscuridad , Femenino , Hipotálamo/metabolismo , Radioisótopos de Yodo , Masculino , Receptores de Vasopresinas , Caracteres Sexuales , TritioRESUMEN
Quantitative analysis of tritium polymer standards and of brain sections labeled with tritiated vasopressin was carried out by using a gaseous detector of beta particles designed for this purpose. The gaseous detector showed major advantages compared with film autoradiography: the linearity and the large dynamic range of intensity measurements as well as the short time needed for data acquisition.
Asunto(s)
Encéfalo/citología , Receptores de Angiotensina/análisis , Receptores de Vasopresinas , Animales , Arginina Vasopresina/metabolismo , Autorradiografía/métodos , Técnica de Dilución de Radioisótopos , Ratas , TritioRESUMEN
The multiplication in gases of ionization electrons, by the effect of the electric fields between parallel electrodes, leads to the emission of light from the molecules excited in the avalanche process. The optical imaging of this light, with intensifiers, on charge-coupled devices permits the localization, in the gaseous volume, of the entrance points of the beta-particles emitted by radioactive compounds placed close to or at the cathode electrode. Thin slices of anatomical samples labeled with 3H show detailed structures 30 microns in size. Gels carrying 32P or 35S are imaged with accuracies of the order of 0.5 mm (full width at half maximum). In comparison with photographic emulsion, the gain in time for data taking is close to a factor of 100, with the advantage of linearity and wider dynamic range in the intensity measurement and a greatly improved signal-to-noise ratio.