A 66-kDa protein of bovine hypophyseal Pars tuberalis induces luteinizing hormone release from rat Pars distalis

In this study, evidence for a factor secreted by bovine hypophyseal pars tuberalis that stimulates luteinizing hormone (LH) release from rat pars distalis cells is shown. The secretion products of bovine pars tuberalis cells into the culture medium were assayed on dispersed rat pars distalis cells in 30 min incubations and superfusion experiments. The culture medium from pars tuberalis total cell populations, added at a dose of 6 microg per tube, induced the greater LH release from pars distalis cells, without effect on follicle stimulating hormone (FSH) release. After pars tuberalis cells separation on a discontinuos Percoll gradient, only the culture medium of cells from 50 and 60% strength Percoll were able to release LH from rat pars distalis cells. Therefore, cell fractions from 50 and 60% strenght Percoll were cultured together. To elicit maximal LH release (6 times the basal output), with the addition of 2 microg of pars tuberalis protein was required, suggesting that these cells produce the factor or factors which affect pars distalis gonadotrope cells. After applying the pars tuberalis culture medium on 12% SDS-PAGE, the band with biological activity was that of 66-kDal. Fifty ng protein of its eluate released almost 9 times the basal output of LH from pars distalis cells. Results suggest a modulating effect of a protein from the bovine pars tuberalis on rat cultured gonadotrope cells from the pars distalis.

Localization of protein-protein interactions in live cells using confocal and spectral imaging FRET microscopy.

Microscopy has become an essential tool for cellular protein investigations. The development of new fluorescent markers such as green fluorescent proteins generated substantial opportunities to monitor protein-protein interactions qualitatively and quantitatively using advanced fluorescence microscope techniques including wide-field, confocal, multiphoton, spectral imaging, lifetime, and correlation spectroscopy. The specific aims of the investigation of protein dynamics in live specimens dictate the selection of the microscope methodology. In this article confocal and spectral imaging methods to monitor the dimerization of alpha enhancer binding protein (C/EBPalpha) in the pituitary GHFT1-5 living cell nucleus have been described. Also outline are issues involved in protein imaging using light microscopy techniques and the advantages of lifetime imaging of protein-protein interactions.

Pituitary cell type-specific electrical activity, calcium signaling and secretion

All secretory anterior pituitary cells exhibit spontaneous and extracellular calcium-dependent electrical activity, but differ with respect to the patterns of firing and associated calcium signaling and hormone secretion. Thus, somatotrophs and lactotrophs fire plateau-bursting action potentials spontaneously and without coupling to calcium release from intracellular stores, which generate calcium signals of sufficient amplitude to keep steady hormone release. In these cells, both spontaneous electrical activity and basal hormone secretion can be further amplified by activation of Gq/11 and Gs-coupled receptors and inhibited by Gi/o/z-coupled receptors. In contrast, gonadotrophs fire single, high-amplitude spikes with limited ability to promote calcium influx and exocytosis, whereas activated Gq/11-coupled receptors in these cells transform single-action potential spiking into the plateau-bursting type of electrical activity and trigger periodic high-amplitude calcium signals and exocytosis of prestored secretory vesicles. Here, we review biochemical and biophysical aspects of spontaneous and receptor-controlled electrical activity, calcium signaling, and hormone secretion in pituitary cells.

Prolactin-releasing activity of GHRP-5 (Momany peptide) on lactotrophs in vivo and in vitro

In the present study the in vivo and in vitro effects of GHRP-5 on the PRL-releasing activity in correlation with the morphological changes of lactotroph cells and their transcriptional activity were evaluated. The in vivo treatment (12 micrograms/100 g BW/day for 3 days) of male rats with GHRP-5 does not induce any significant changes in serum PRL levels. In contrast, the addition of GHRP-5 to pituitary cell cultures increased significantly the release of PRL. This effect is enhanced in cell cultures of enriched lactotrophs, increasing significantly the secretion of PRL, the concentrations of which were 50 higher than that of untreated control cells. The administration of GHRP-5 provokes several changes in the fine structure of lactotrophs, compatible with an increased secretory activity. After the GHRP-5 treatment the different lactotroph subtypes persist but the subtype I displaying secretory granules of larger size (500-900 nm) and a significant development of the Golgi apparatus and RER were more frequently observed. These results can be correlated with a significant augmentation in PRL mRNA after the GHRP-5 treatment. In spite of that no variations in serum PRL levels were observed in vivo, following GHRP-5 treatment, the lactotroph population experienced evident fine structure modifications, concordant with an upsurge of PRL synthesis. These observations confirmed a direct action of GHRP-5 on receptors expressed by lactotrophs. The differential actions of GHRP-5 on in vivo and in vitro designs confirm a different effectiveness of this secretagogue to induce PRL secretion.

Effects of oncostatin M on hormone release of rat pituitary cells in primary culture

It has become increasingly clear that cytokines play an important role in modulating neuroendocrine regulation, especially in the secretion of corticotropin (ACTH) in the pituitary. Oncostatin M (OSM), a cytokine of IL-6 family has been reported to increase ACTH secretion and pro-opiomelanocortin (POMC) transcription in murine corticotroph pituitary tumor cells (AtT20 cells). The present study was undertaken to determine the effects of OSM on hormonal release in primary culture of rat pituitary cells. Growth hormone or prolactin release was not affected by OSM. OSM (1 nM) stimulated ACTH release (35.1% increase versus control, p>0.001) in dispersed pituitary cells of rat to a lesser extent than in AtT20 cells. Corticotropin releasing hormone (CRH) (10 nM) also induced a 2.3-fold increase of ACTH secretion (p>0.001), but co-treatment of OSM and CRH did not exhibit any synergistic effect on ACTH secretion. We conclude OSM has a stimulatory effect on ACTH secretion in normal rat pituitary cell cultures, and OSM acts mainly on corticotroph, supporting the potential role of OSM to modulate immune-endocrine regulation in the pituitary.

Effects of oncostatin M on hormone release of rat pituitary cells in primary culture

It has become increasingly clear that cytokines play an important role in modulating neuroendocrine regulation, especially in the secretion of corticotropin (ACTH) in the pituitary. Oncostatin M (OSM), a cytokine of IL-6 family has been reported to increase ACTH secretion and pro-opiomelanocortin (POMC) transcription in murine corticotroph pituitary tumor cells (AtT20 cells). The present study was undertaken to determine the effects of OSM on hormonal release in primary culture of rat pituitary cells. Growth hormone or prolactin release was not affected by OSM. OSM (1 nM) stimulated ACTH release (35.1% increase versus control, p>0.001) in dispersed pituitary cells of rat to a lesser extent than in AtT20 cells. Corticotropin releasing hormone (CRH) (10 nM) also induced a 2.3-fold increase of ACTH secretion (p>0.001), but co-treatment of OSM and CRH did not exhibit any synergistic effect on ACTH secretion. We conclude OSM has a stimulatory effect on ACTH secretion in normal rat pituitary cell cultures, and OSM acts mainly on corticotroph, supporting the potential role of OSM to modulate immune-endocrine regulation in the pituitary.

Role of temperature in regulation of ovarian cycle in bull frog Rana tigerina.

Effect of temperature on the ovarian cycle was studied in R. tigerina by exposing them to (1) constant low (22 degrees C) temperature during preparatory (active vitellogenic growth) phase (March-May) when the mean ambient temperature ranged from 26 degrees-28 degrees C and (2) to constant high (30 degrees +/- 1 degrees C) temperature during postbreeding regression phase (August-November) when the mean ambient temperature ranged from 22 degrees-26 degrees C. The ovaries of initial controls (biopsy samples taken prior to the commencement of the experiment) in March contained only first growth phase (FGP) oocytes with a maximum size range of 361-480 microns in diameter. In the frogs exposed to constant low temperature for 2 months, only 7% of FGP oocytes were recruited to second growth phase (SGP) with a mean largest diameter of 631 microns compared to 31% large SGP oocytes with a mean diameter of 1114 microns in the frogs collected from natural fields. The number of atretic follicles (AF) was lower and fat body weights were significantly higher in low temperature exposed frogs. The exposure of the frogs to constant high temperature during postbreeding months caused an increase in the mean diameter and number of large FGP oocytes, numerical increase in AF and decrease in fat body weights over corresponding controls maintained at room temperature. The pituitary gonadotrophs of these frogs showed stimulatory changes such as increase in cell size and appearance of secretory granules in the cytoplasm. The results suggest that in R. tigerina high temperature stimulates oocyte growth while low temperature retards it.(ABSTRACT TRUNCATED AT 250 WORDS)

Aspecto morfológico da glândula pituitária do gambá sul-americano Didelphis azarae / Morphologic aspect of the pituitary gland of the south-american opossum Didelphis azarae

Na hipófise do gambá sul-americano Didelphis azarae, o processo infundibular (neuro-hipófise) está envolvido pela adeno-hipófise e separa-se do lúmen hipofisário por uma camada de células pavimentosas que é contínua cocm a adeno-hipófise. A pars tuberalis forma um colar de células em otrno da haste infundibular e é rica em vasos sanguíneos. Na adeno-hipófise agrupamentos de células parenquimatosas estäo envolvidos por uma delicada malha de fibras conjuntivas associada a capilares

Polichromic staining of five distinct cellular types in human and rat pituitaries

Se describe un método de tinción pentacrómico que permite identificar los diferentes tipos celulares en tejidos adenohipofisiario humano y de rata y en células dispersas de rata. Esta técnica mostró un grado suficiente de selectividad que hizo posible distinguir 5 tipos celulares: lactotropos (rojos), somatotropos (amarillos), tirotropos (azul intenso), corticotropos (violeta) y gonadotropos (verde). De las células dispersas de rata un 90% del total analizado estuvo representado por los 2 grupos de células acidófilas, mientras que el 10% restante correspondió a: tirotropos 1.7%, corticotropos 1.1% y gonadotropos casi 7.0%. En la estirpe acidófila las proporciones de los dos tipos celulares obtenidas mediante esta tinción (PRL 63.7%, CH 26.7%) fueron comparables a los datos informados en la literatura utilizando métodos inmunohistoquímicos. Cada uno de los cinco grupos de células estudiados fue a su vez subdividido en tres categorías dependiendo de su tamaño. El bajo costo y la selectividad del método permiten proponerlo como de gran utilidad para aquellas instituciones que carecen de instalaciones para realizar estudios inmunocitoquímicos o de microscopía electrónica