J. M. Riesco

Coexistence of NADPH-diaphorase with tyrosine hydroxylase in hypothalamic magnocellular neurons of the rat

The presence and distribution of NADPH-diaphorase (ND) neurons as well as tyrosine hydroxylase-immunoreactive (TH) neurons in the hypothalamus are well established. Previous studies have shown the coexistence of ND with neuroactive substances such as calbindin, somatostatin, vasopressin and oxytocin in neurons of this region of the brain. As the tópographical patterns of distribution of ND and TH coincide in many cases, the aim of this study was to determine the possible coexistence of both substances in the main hypothalamic magnocellular nuclei of the albino rat. Histochemical-immunocytochemical double labelling was employed on the same sections as well as a morphometric study. NADPH-diaphorase and tyrosine hydroxylase neurons were observed in all the nuclei under study (supraoptic, paraventricular and accessory nuclei), although most neurons showing the coexistence of both substances were mainly located in the supraoptic nucleus, isolated neurons with double labelling being found in the magnocellular parts of the paraventricular nucleus and in some of the accessory nuclei. Although both substances have previously been shown to be modified in hypothalamic neurons after osmotic stimuli, the range of functions of ND in the CNS is only beginning to be understood. Further studies are needed to elucidate the functional role that ND/TH neurons play in the nervous system.

Morphometric changes of specific located vasopressin-reacting parvicellular neurons in the paraventricular nucleus of the rat after adrenalectomy.

The morphological-morphometric consequences of bilateral adrenalectomy on vasopressin-reacting neurons of the paraventricular nucleus of the rat hypothalamus were analyzed. Bilateral adrenalectomy led to a dramatic increase in the cellular area as well as the number of immunoreactive cells (when compared to those obtained in normal colchicine-treated animals) in the neurons located in the anterior, medial and periventricular parvicellular subdivisions of the paraventricular nucleus. By contrast, no changes were observed in either the dorsal or lateral parvicellular subdivisions or in any of the magnocellular subdivisions of the paraventricular nucleus.

Morphological and functional study of the GH-immunoreactive adenohypophyseal cells in ovariectomized rats

SummaryIn view of the existence of a different secretion pattern of growth hormone (GH) between male and female rats, the aim of the present study was to analyse the role played by ovarian steroid hormones in the modulation of such secretion. To do so, postpuberal female rats were ovariectomized and killed at 30 days after the operation. The basal serum levels of growth hormone, together with cell area, cytoplasmic area and nuclear area of the hypophyseal somatotropic cells of normal and ovariectomized rats were compared. The results obtained show that ovariectomy induces a significant decrease (p<0.05) in the basal serum levels of GH, accompanied by an increase in cellular and cytoplasmic areas, with no significant differences in nuclear area. Overiectomy was also accompanied by an increase in reaction intensity and the number of GH-immunoreactive cells (p<0.01). These findings point to the shift towards a masculine secretory and morphological pattern following ovariectomy and supports the hypothesis that ovarian steroids intervene in the establishment of a different pattern in females compared to males.

Estrogenic modulation of met-enkephalin-induced prolactin secretion in rats.

SummaryIn order to elucidate the possible role of estrogens in the sex-dependent modulation of met-enkephalin-induced prolactin secretion, and the cellular response at adenohypophyseal level, analyses of serum prolactin levels together with a morphometric study of prolactin-immunoreactive cells were carried out following intraventricular administration of the opioid to estrogenprimed male rats. Administration of the opioid led to an increase in the serum levels of the hormone and, morphometrically, a significant increase in the cellular, cytoplasmic and nuclear areas was observed. When naloxone was administered previously, the above, mentioned changes did not appear, and the values found were similar to those observed in the animals treated only with estrogens. The results suggest that the met-enkephalin-induced prolactin secretion is modulated by the gonadal steroid environment.

Morphofunctional study of mammotropic cells following intraventricular administration of met-enkephalin

SummaryAn ultrastructural and morphometric study was carried out on the adenohypophyseal mammotropic cells of rats treated intraventricularly with an acute dose (150 μg) of Met-enkephalin. In the female rats, clear features of cellular hyperactivity appeared after opioid administration. The changes affected the Golgi complex, the rough endoplasmic reticulum, the mature and immature secretory granules and the images of exocytosis. Such changes did not appear when naloxone was administered before the opioid, and naloxone induced an increase in the numerical density of lysosomal dense bodies with lipoid inclusions. In the male animals, administration of an identical dose of Metenkephalin caused only a few significant changes, similar to those observed in the controls. It is concluded that Metenkephalin administered intraventricularly causes evident modifications in the mammotropic cells of female rats whereas such changes in the male animals are not significant.

Glial fibrillary acidic protein-like immunoreactive ependymal elements in the third ventricle of the rat : a study at different stages of development

Using the peroxidase-antiperoxidase method a study was made of the cells immunoreactive to glial fibrillary acidic protein (GFAP) anti-serum in the ependyma of the third ventricle of the rat at different stages of growth. Most of the ependymal cells of the third ventricle were seen to be unreactive to this protein; however, it was sometimes possible to observe some GFAP-immunoreactive ependymocytes and occasionally other immunoreactive cellular types, such as tanycytes and supraependymal cells. Despite this, the most frequent localization of the elements immunoreactive to the protein adopted the shape of an immunoreactive subependymal band situated parallel to the ventricular wall. As the weights of the animals increased an increase in the elements immunoreactive to this protein could be observed in all the zones considered, there being no differences between the male and female animals.

Morphometrical variations of prolactin cells in response to prolonged and systemic administration of Met-enkephalin in female rats

SummaryA stimulatory effect on prolactin secretion had been describe after acute and systemic administration of met-enkephalin, but the effects of this opioid after chronic administration has not been reported, and the response of mammotroph cells is not clear. As a complement to previous studies, a morphometric analysis (light and electron microscopy) was carried out on prolactin cells from female rats treated chronically with met-enkephalin. Clear features of cellular hyperactivity appeared after chronic and systemic administration of the opioid, and these persisted for two weeks. The changes consisted in increases of cellular, cytoplasmic and nuclear areas, volume and surface densities of the Golgi complex and rough endoplasmic reticulum, as well as the numbers of exocytotic figures. These morphological alterations were paralleled by an increase in serum prolactin levels as detected by RIA. It is concluded that the increase in the synthesis and secretory activity of prolactin cells following chronic and systemic administration of met-enkephalin is very similar to those observed after acute and intraventricular administration.

Morphometric analysis of GH-immunoreactive cells after GH release induced by treatment with estrogen in adult male rats

In order to elucidate the response of somatotropic cells to the influence of gonadal steroids on the regulation of growth hormone secretory patterns, a morphometric analysis was carried out on the GH-immunoreactive cells of adult male rats treated chronically with intramuscular injections of estradiol valerate. The morphometric and morphological results obtained were correlated to the serum levels of GH at the moment of sacrifice. Treatment with a daily dose of 125 micrograms of estradiol vaterate over 15 d was seen to lead to an increase (p less than 0.01) in the serum GH values accompanied by a decrease in the intensity of reaction of the GH-cells and, morphometrically, a reduction in their size (p less than 0.01) due to a decrease in the cytoplasmic area (p less than 0.01), but without significant changes in the nuclear area. Our results suggest that in male rats estrogens enhance the release of the intracellular GH pool but that they do not affect hormone synthesis to a great extent.

Sexually Dependent Changes in Rat Somatotropic Cells following Bilateral Adrenalectomy

In order to elucidate the existence of gender-related variations in both growth hormone (GH) release and the activity of somatotropic cells following bilateral adrenalectomy, a morphometric analysis was performed on GH-immunoreactive cells from adult male and female rats after bilateral adrenalectomy, correlating the findings with the serum levels of the hormone. The results obtained were compared to those found in untreated animals. Bilateral adrenalectomy was seen to induce a decrease in serum GH levels (p less than 0.01) in male rats; this was accompanied by a significant decrease in cellular area (p less than 0.01), cytoplasmic area (p less than 0.05) and nuclear area (p less than 0.01) and by a decrease in the cytoplasmic immunoreaction intensity of GH cells. By contrast, the above-mentioned changes did not appear in the female rats. These results suggest that the action of glucocorticoids on the synthesis and release of GH depends on the sex of the animal.

Sex-specific effects of MET-enkephalin treatment on vasopressin immunoreactivity in the rat supraoptic nucleus

The supraoptic nucleus of male and female rats treated with met-enkephalin or naloxone and met-enkephalin was examined with light microscopical immunocytochemistry for Arginine-vasopressin. Both genders exhibited the same distribution of immunostained magnocellular neurons. Met-enkephalin treatment caused an increase in number of immunostained vasopressin neurons. This effect was more pronounced in females than in males. Naloxone treatment diminished immunoreactive cytoplasmic vasopressin in males more effectively than in females. In enkephalin-treated animals numerous vasopressin immunoreactive varicosities appeared within the supraoptic nucleus, but were mostly absent in naloxone-treated animals and in controls. Our results indicate that met-enkephalin treatment either stimulates vasopressin synthesis or inhibits secretion. It is likely that steroid hormones mediate the action of enkephalin on vasopressin secretion in a specific manner.