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Biomedical subjects

W E Stumpf

Publications and source records attributed to W E Stumpf.

At least 109 records · Page 6Linked to original sources
PubMed1987

Ontogeny of gonadotropin releasing hormone and gonadotropin immunoreactivity in brain and pituitary of normal and estrogen-treated guppies, Poecilia reticulata Peters.

Gonadotropin releasing hormone (GnRH) and gonadotropic hormone (GTH) were identified by immunohistochemistry in the brains and pituitaries of neonate, juvenile and adult guppies. GTH was present in some cells of the pars intermedia (pi) and proximal pars distalis (ppd) of all animals. GnRH was found in the perikarya of the nucleus olfactoretinalis. In the pituitaries of juvenile 30-day-old guppies, GnRH-immunoreactive cells existed in a "juvenile pattern", whereas in adult animals GnRH was recognized in only a few cells. GnRH-immunoreactive fibers were seen in the pituitaries of animals that were 30 days or older. In adult guppies, the ventral and lateral ppd (the gonadotropic region) contained a dense network of GnRH-immunoreactive fibers. Pituitary cells staining for either GnRH or GTH were located in different places. After immunohistochemical double staining of adult pituitaries, none of the GnRH-immunoreactive cells were LH-immunoreactive, although both cell types were often found in close proximity. After 20 days or more of ethinylestradiol treatment, less immunoreactive GnRH was detected in the pituitary cells of juvenile guppies, and fewer animals exhibited the "juvenile pattern" of GnRH-immunoreactive pituitary cells, when compared with untreated controls. The results indicate that GnRH-immunoreactive pituitary cells in the guppy are distinct from gonadotropes and that these cells are involved in regulatory processes along the juvenile brain-pituitary-gonad axis.

Aging↗
PubMed1987

Vitamin D sites of action in the pituitary studied by combined autoradiography-immunohistochemistry.

Adult male and female mice under normal diet were injected with 3H 1,25(OH)2 vitamin D3 and sacrificed 3.5 h afterwards. Autoradiograms were prepared according to our thaw-mount technique and stained with antibodies to pituitary hormones. Thyrotropes showed strong and extensive nuclear concentration of radioactivity: about 90% of the immunostained thyrotropes were labeled. Lactotropes, somatotropes and gonadotropes showed no or only weak nuclear radioactivity: a subpopulation of 5%-10% of each of these immunostained cell types displayed nuclear labeling that was weak when compared to thyrotropes. Neural lobe pituicytes also showed weak to intermediate nuclear labeling. The results indicate a presence of nuclear receptors for 1,25(OH)2 vitamin D3 in pituitary cell types and suggest direct but differential genomic effects of 1,25(OH)2 vitamin D3 on pituitary hormone secretion. Evidence further suggests the existence of a vitamin D regulated brain-pituitary-thyroid axis.

Animals↗
PubMed1987

Nuclear receptors for 1,25(OH)2 vitamin D3 in thymus reticular cells studied by autoradiography.

After injection of 3H 1,25(OH)2 vitamin D3 to rats fed a vitamin D-deficient diet, nuclear concentration and retention of radioactivity exists in reticular cells of the thymus medulla and cortex, as well as outer cells of developing Hassal's corpuscles. Lymphocytes do not show nuclear concentration of radioactivity. Nuclear concentration in reticular cells is prevented by prior injection of excess 1,25(OH)2 vitamin D3. The results indicate that reticular-endothelial cells contain nuclear receptors for 1,25(OH)2 vitamin D3 and suggest that effects of 1,25(OH)2 vitamin D3 on immune response and lymphocyte differentiation are indirect and mediated through genomic modulation of reticular cell functions such as messenger secretion.

Animals↗
PubMed1987

1,25 (OH)2 vitamin D3 sites of action in the brain. An autoradiographic study.

After injection of 3H 1,25 (OH)2 vitamin D3 to adult rats and mice, under normal or vitamin D deficient diet, the hormone was found to be accumulated in nuclei of neurons in certain brain regions. Nuclear concentration was prevented or diminished, when excess unlabeled 1,25 (OH)2 vitamin D3 was injected before 3H 1,25 (OH)2 vitamin D3, while excess 25 (OH) vitamin D3 did not prevent nuclear labeling. Highest nuclear concentration of 3H 1,25 (OH)2 vitamin D3 is observed in certain neurons in the nucleus interstitialis striae terminalis, involving its septo-preoptic pars dorsolateralis and its anterior hypothalamic-thalamic portion, and in the nucleus centralis of the amygdala, all constituting a system of target neurons linked by a component of the stria terminalis. Nuclear concentration of 3H 1,25 (OH)2 vitamin D3 is also found in neurons in the periventricular nucleus of the preoptic-hypothalamic region, including its extensions, the parvocellular paraventricular and arcuate nucleus, in the ventromedial nucleus, supramammillary nucleus, reticular nucleus of the thalamus, ventral hippocampus, caudate nucleus, pallium, in the midbrain-pontine central gray, dorsal raphe nucleus, parabrachial nuclei, cranial motor nuclei, substantia gelatinosa of the sensory nucleus of the trigeminus, Golgi type II cells of the cerebellum, and others. The extensive distribution of target neurons suggests that 1,25 (OH)2 vitamin D3 regulates the production of several aminergic and peptidergic messengers, and influences the activity of certain endocrine-autonomic, sensory and motor systems.

Animals↗
PubMed1987

Autoradiographic studies with 3H 1,25 dihydroxyvitamin D3 in thyroid and associated tissues of the neck region.

Rats and mice fed a vitamin D-deficient or vitamin D-complete diet were injected with 3H 1,25 (OH)2 vitamin D3. Autoradiograms prepared from cross sections through the neck region revealed nuclear concentration of radioactivity strongest in parathyroid chief cells, occasionally in thyroid follicular epithelial and interfollicular cells, in the epithelium of tubular remnants of the ultimobranchial body, in epithelium of the esophagus, in chondrocytes of tracheal cartilage, and in myoepithelial cells of tracheal glands. In the thyroid, most of the follicle epithelial cells did not show nuclear concentration of radioactivity which occurred only occasionally and predominantly in follicles located in marginal positions. Thyroglobulin in lumina of thyroid follicles contained varying amounts of radioactivity that correspond to the diameter of the follicles, with relatively high amounts in large follicles and little or no radioactivity in small follicles. Competition with excess of unlabeled 1,25 (OH)2 vitamin D3 abolished nuclear radioactivity, but not the radioactivity in the colloid, while 25 (OH) vitamin D3 did not affect either. When a combination of autoradiography and immunohistochemistry was applied, follicular and parafollicular C-cells positive for calcitonin antibodies, did not show nuclear concentration of radioactivity. Tubular remnants of ultimobranchial bodies, however, showed distinct nuclear labeling, but did not stain, or only weakly stain, with antibodies to calcitonin. When 3H 25 (OH) vitamin D3 was injected, no nuclear concentration of radioactivity was noted in any of the tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

High resolution autoradiography at the regional topographic level with [14C]2-deoxyglucose and [3H]2-deoxyglucose.

After injection of 2-deoxyglucose (2-DG) labeled with tritium or carbon-14, autoradiograms were produced by thaw-mounting 4 micron frozen sections of rat brains on nuclear emulsion-coated slides. The results show that the distribution of radioactivity among different brain regions was similar and that the resolution at the regional topographical level was virtually identical for both compounds. The resolution obtained with the thaw-mounting of thin frozen sections onto nuclear emulsion was considerably greater than the resolution demonstrated in published results in the literature, when carbon-14 or tritium-labeled 2-deoxyglucose were used with 20 micron frozen sections and X-ray film or tritium-sensitive film. The results indicate that section thickness, detection medium and intimacy of contact between section and photographic emulsion influence resolution. At the regional level, the detection medium apparently influences resolution to a greater extent than the energy differences of the beta particles emitted from 14C or 3H. Although diffusion of radiolabeled 2-deoxyglucose and metabolites during the thaw-mounting process precludes single cell resolution of these autoradiograms, the improvement of visualizing regional topographic detail demonstrates that the described technique is a valuable approach with which to study regional 2-DG uptake.

Animals↗
PubMed1987

1,25-Dihydroxyvitamin D3 target cells in immature pancreatic islets.

Target cells of 1,25-dihydroxyvitamin D3 were identified by autoradiography in islets from rats of different ages. Nuclei of pancreatic islet cells selectively concentrated 1,25-[3H]dihydroxyvitamin D3 but not 25-[3H]hydroxyvitamin D3 or 24,25-[3H]dihydroxyvitamin D3. Developmental studies of pancreatic islets indicated that target cells, as revealed by significant nuclear concentration of 1,25-[3H]dihydroxyvitamin D3, are present in islet cells of fetal rats. The percentage of islet cells that concentrated 1,25-[3H]dihydroxyvitamin D3 increased from 10 to 15% in the fetus to 60% at 1 day of age. Immunocytochemical staining indicated that insulin-containing cells but not glucagon or somatostatin cells concentrated 1,25-[3H]dihydroxyvitamin D3. Peak uptake of 1,25-[3H]dihydroxyvitamin D3 was calculated to be 400 pmol/mg DNA, with no significant difference in nuclear accumulation between islet cells from neonatal and adult rats or between islets in vivo and isolated islets in vitro. The results of these studies indicate that 1,25-[3H]dihydroxyvitamin D3 target cells are present in islets before pancreatic beta-cells are morphologically or functionally mature; islet beta-cells concentrate 1,25-dihydroxyvitamin D3, but not 25-hydroxyvitamin D3 or 24,25-dihydroxyvitamin D3. We conclude that only the 1,25-dihydroxyvitamin D3 metabolite of vitamin D is accumulated by nuclei of developing and mature beta-cells and suggest that 1,25-dihydroxyvitamin D3 plays a role in the maturation of islet beta-cells.

Animals↗
PubMed1987

Utility of [16 alpha-125I] iodoestradiol for autoradiography for the study of cellular and regional distribution of receptors.

We demonstrate the utility of [16 alpha-125I]iodoestradiol for thaw-mount autoradiography with 2 micron and 4 micron thick sections of rat and mouse uterus, pituitary, and brain after in vivo administration. Under the conditions of the experiments, short-term autoradiography with exposure times between 3 and 14 days provides optimal cellular resolution, whereas long-term autoradiography with 1-2 months of exposure may be used to obtain topographic-regional surveys of distribution.

Animals↗
PubMed1986

Sites of aromatization of [3H]testosterone in forebrain of male, female and androgen receptor-deficient Tfm mice: an autoradiographic study.

The distribution of radioactivity after injection of [3H]testosterone was studied in the forebrain of adult mice by thaw-mount autoradiography. Nuclear labeling was high in neurons in the dorsal part of the medial nucleus of the amygdala and in the dorsocaudal part of the bed nucleus of the stria terminalis. Low nuclear uptake occurred in the medial preoptic nucleus, in mediobasal hypothalamic nuclei and in the ventromedial amygdala. Nuclear concentration of radioactivity was not influenced by competition with dihydrotestosterone and was present in androgen receptor deficient Tfm mice. It was totally abolished by competition with estradiol. This indicates that in the brain [3H]testosterone is converted to estrogenic metabolites which bind to estrogen receptors. Nuclear labeling after [3H]testosterone was restricted to a few of the brain nuclei, known to contain estrogen receptors indicating that aromatization occurs only in select regions. The results suggest that testosterone acts on the brain via estrogen receptors.

Amygdala↗
PubMed1986

Estradiol target sites immunoreactive for beta-endorphin in the arcuate nucleus of rat and mouse hypothalamus.

Combined autoradiography and immunocytochemistry was performed to examine estradiol-concentrating neurons of the arcuate nucleus in the hypothalamus of ovariectomized rats and mice for beta-endorphin-like immunoreactivity. A high number of [3H]estradiol-labeled cells occurred in the arcuate nucleus of both species. Neurons immunoreactive for beta-endorphin could be visualized throughout the arcuate nucleus as well as ventrolaterally outside of this nucleus. Colocalization of [3H] estradiol nuclear labeling and beta-endorphin-like cytoplasmic immunoreactivity could be found scattered throughout the arcuate nucleus, most frequently in the ventromedial part, although most of the [3H]estradiol target neurons did not react with the antibodies. The results suggest that a subpopulation of beta-endorphin neurons is directly addressed by estradiol and that the arcuate nucleus contains different groups of estradiol receptive neurosecretory cells.

Animals↗
PubMed1986

The adrenal: a new target organ of the calciotropic hormone 1,25-dihydroxyvitamin D3.

Target cells for 1,25-dihydroxyvitamin D3 were demonstrated in the adrenal medulla by frozen-section autoradiography. The appearance of these target cells was age-dependent in neonatal mice. Immunocytochemical staining for phenylethanolamine-N-methyltransferase revealed that both epinephrine and non-epinephrine cells concentrate 1,25-dihydroxyvitamin D3 in their nuclei. In contrast, immunocytochemical staining for "vitamin D-dependent calcium-binding protein" (D-CaBP) demonstrated that D-CaBP immunoreactivity is localized in only a small percentage of adrenomedullary cells, in mice and rats. Comparison of PNMT and D-CaBP immunoreactivities in sequential sections showed that epinephrine-producing cells do not contain D-CaBP. These results indicate that adrenal medullary cells have receptors for 1,25-dihydroxyvitamin D3 and that 1,25-dihydroxyvitamin D3 may directly affect certain functions of these endocrine cells.

Adrenal Medulla↗
PubMed1986

Immunocytochemical evidence for CDD/ANP-like peptides in strands of myoendocrine cells associated with the ventricular conduction system of the rat heart.

Immunohistochemical investigations with different antisera against cardiodilatin 99-126 or alpha atrial natriuretic polypeptide revealed the presence of cardiac hormones not only in the atria of rats but also in strands of myoendocrine cells located in subendocardial regions of the ventricular septum. The localization of CDD-IR (cardiodilatin immunoreactivity) in the ventricle is associated with the location of the conduction system in the rat. The significance of the morphological relationship between cardiodilatin and the conduction system of the rat heart is discussed.

Aging↗
PubMed1986

Autoradiographic studies with 3H dihydrotestosterone in the brain of sex reversed mice, heterozygous for androgen insensitive testicular feminization (Tfm). A comparison with normal female mice and sex reversed male mice.

Specific binding sites for 3H dihydrotestosterone are demonstrated by autoradiography in brain nuclei of sex reversed mice heterozygous for testicular feminization (Tfm) which are phenotypically intersexes with testes and accessory sex glands that consist of a mosaic of androgen insensitive Tfm cells which lack specific dihydrotestosterone binding and androgen sensitive normal cells. The nuclear group evaluated include: nucleus (n.) septi lateralis, n. interstitialis striae terminalis, n. medialis amygdalae, the hypothalamic n. arcuatus, n. ventromedialis lateralis, n. pre-mammillaris ventralis, n. preopticus medialis, and nuclei of the cranial nerves VII, X, and XII. In the sex reversed males and the female, used as controls, the frequency of neurons with specific DHT binding show a distinct male-female difference in the caudal part of the arcuate nucleus. In the sex reversed Tfm heterozygotes, in all brain nuclei studied, the frequency of labeled neurons is reduced. The extent of reduction of androgen binding in the different brain nuclei varies among as well as within individual sex reversed Tfm heterozygotes, suggesting variations of the ratio of normal to Tfm neurons in sex reversed Tfm heterozygotes. The differentially reduced androgen binding of different brain systems corresponds to a differentially reduced androgen dependent behaviour reported in the literature.

Amygdala↗
PubMed1986

Coexistence of atrial natriuretic factor (ANF) and oxytocin in neurons of the rat hypothalamus.

Sections of rat hypothalamus were examined immunocytochemically for coexistence of atrial natriuretic factor (cardiodilatin) and oxytocin. These biosynthetically unrelated neuropeptides were demonstrated using the peroxidase-antiperoxidase method followed by immunofluorescence. Neurons immunoreactive for atrial natriuretic factor were found in the supraoptic, periventricular and paraventricular nuclei. A sub-population of these neurons contained oxytocin-like immunoreactivity. They constitute only a small portion of the total number of oxytocinergic neurons.

Animals↗
PubMed1986

Gonadotropin-releasing hormone immunoreactive neurons with access to fenestrated capillaries in mouse brain.

Gonadotropin-releasing hormone (GnRH) producing neurons which have access to fenestrated capillaries were identified through a combination of indirect immunofluorescence for GnRH with a fluorescein-taged second antibody and histochemical demonstration of the localization of blood borne and retrogradely transported horseradish peroxidase. In the mouse, GnRH positive neurons were present in the septum, which includes neurons originating from the nervus terminalis, the nucleus medialis and triangularis septi, and the nucleus of the diagonal band. Also, GnRH immunoreactive neurons could be seen in the lateral anterior hypothalamus, the nucleus preopticus medianus, the rostral nucleus periventricularis hypothalami and, to a lesser extent, the nucleus preopticus medialis. Single GnRH positive neurons were found in the nucleus supraopticus, the bed nucleus of the stria terminalis and the cingulate cortex. GnRH neurons which showed uptake of horseradish peroxidase were located in all of these regions and intermingled with unlabeled GnRH neurons. No preferential topographical concentration of GnRH neurons with access to the fenestrated vasculature was apparent. In animals in which GnRH secretion was stimulated by castration for 2 weeks, 65% of all GnRH neuronal perikarya contained horseradish peroxidase. This was reduced to 35% after a 2-week treatment of ovariectomized animals with 10 micrograms/day estradiol while the total number of immunoreactive GnRH cells remained unchanged. No differences in the number of GnRH-horseradish peroxidase positive cells was seen when the dose of horseradish peroxidase of the survival time were increased. While the presence of certain GnRH neurons with dual actions via collaterals cannot be excluded, the results suggest that there are two populations of GnRH neurons, one with access to fenestrated capillaries which is probably related to neurosecretory endocrine regulation of anterior pituitary gonadotropin secretion, and one without access to fenestrated capillaries which is probably related to intracerebral neurotransmission only.

Animals↗
PubMed1986

High resolution autoradiographic determination of the topographic distribution of radioactivity in the hippocampal formation after injection of [1-14C]glucose or 2-deoxy[14C]glucose.

Using high resolution autoradiography, the accumulation of radioactivity after intravenous injection of [1-14C]glucose was measured in the corpus callosum, hippocampus, dorsal hippocampal commissure, somatosensory cortex, inferior colliculus and pontine periaqueductal grey. Autoradiograms were prepared by thaw-mounting 4 micron frozen sections on nuclear emulsion-coated slides, and were evaluated quantitatively with a computer-assisted video system for automated counting of silver grains. In all brain regions examined, silver grain densities were greater in rats killed 30 min after injection of [1-14C]glucose compared to rats killed 10 min after injection. After intravenous injection of [1-14C]glucose or 2-deoxy[14C]glucose, the relative uptake and retention of radioactivity in different hippocampal subregions was compared. Striking differences were found in the hippocampus between 2-deoxy[14C]glucose and [1-14C]glucose autoradiograms. After injection of 2-deoxy[14C]glucose, there were large variations in the uptake and retention of radioactivity among different pyramidal cell fields. The CA 3 pyramidal cell field retained considerably more radioactivity than other pyramidal cell fields after injection of 2-deoxy[14C]glucose, while after injection of [1-14C]glucose, the retention of radioactivity was similar in all pyramidal cell fields. After [1-14C]glucose injection, the dentate gyrus contained relatively high levels of radioactivity and more 14C accumulated in the granular layer, compared to the molecular layer. In contrast, after 2-deoxy[14C]glucose injection, there was uniformly less radioactivity throughout the dentate gyrus when compared to rats injected with [1-14C]glucose and there was no preferential accumulation of 2-deoxy[14C]glucose in the granular layer compared to the molecular layer.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗