• The Korean Journal of Physiology and Pharmacology
• /
• v.7 no.2
• /
• pp.79-84
• /
• 2003

We have previously reported that expression of the somatostatin receptor subtypes, sst1-5, is differentially regulated by growth hormone (GH)-releasing hormone (GHRH) and forskolin (FSK), in vitro. GHRH binds to membrane receptors selectively located on pituitary somatotropes, activates adenylyl cyclase (AC) and increases sst1 and sst2 and decreases sst5 mRNA levels, without significantly altering the expression of sst3 and sst4. In contrast FSK directly activates AC in all pituitary cell types and increases sst1 and sst2 mRNA levels and decreases sst3, sst4 and sst5 expression. Two explanations could account for these differential effects: 1) GHRH inhibits sst3 and sst4 expression in somatotropes, but this inhibitory effect is masked by expression of these receptors in unresponsive pituitary cell types, and 2) FSK inhibits sst3 and sst4 expression levels in pituitary cell types other than somatotropes. To differentiate between these two possibilities, somatotropes were sequentially labeled with monkey anti-rat GH antiserum, biotinylated goat anti-human IgG, and streptavidin-PE and subsequently purified by fluorescent-activated cell sorting (FACS). The resultant cell population consisted of 95% somatotropes, as determined by GH immunohistochemistry using a primary GH antiserum different from that used for FACS sorting. Purified somatotropes were cultured for 3 days and treated for 4 h with vehicle, GHRH (10 nM) or FSK ($10{\mu}M$). Total RNA was isolated by column extraction and specific receptor mRNA levels were determined by semi-quantitative multiplex RT-PCR. Under basal conditions, the relative expression levels of the various somatostatin receptor subtypes were sst2＞sst5＞sst3=sst1＞ sst4. GHRH treatment increased sst1 and sst2 mRNA levels and decreased sst3, sst4 and sst5 mRNA levels in purified somatotropes, comparable to the effects of FSK on purified somatotropes and mixed pituitary cell cultures. Taken together, these results demonstrate that GHRH acutely modulates the expression of all somatostatin receptor subtypes within GH-producing cells and its actions are likely mediated by activation of AC.

• Korean Journal of Veterinary Research
• /
• v.38 no.3
• /
• pp.488-496
• /
• 1998

Somatotropes, mammotropes and somatomammotropes of the Korean native goat hypophysis were studied by double immunoelectron microscopy using antisera to growth hormone(GH) and prolactin(PRL), and protein A-gold particles of different sizes. Mammotropes were round or oval in shape, and contained round and electron dense secretory granules. The size of secretory granules was variable from 460nm to 680nm in diameter. Somatotropes were elliptical or triangular in shape and the oval nucei were located eccentrically at the periphery of the cell. Secretory granules of the cell were oval in shape and clearly distinguished from round granules of mammotropes. The size of granules was 320~680nm in diameter, smaller than that of mammotropes. Somatomammotropes contained round or oval secretory granules. The granules had intermediate size between somatotropes and mammotropes. Some of granules contained both GH and PRL, while the others contained only one of them.

• Korean Journal of Veterinary Research
• /
• v.38 no.3
• /
• pp.497-507
• /
• 1998

The localizations and morphological characteristics of immunoreactive cells for prolactin(PRL) and growth hormone(GH) antisera were studied with double immunohistochemistry in the adenohypophysis of the Korean native goat. PRL immunoreactive cells(mammotropes) and GH immunoreactive cells(somatotropes) were present in the pars distalis and pars tuberalis, but not in the pars intermedia. A few cells were stained with both PRL and GH antisera (somatomammotropes). The possessional percentages of mammotropes, somatotropes and somatomammotropes were 37.0%, 32.6%, 1.0% in females and 35.6%, 32.6%, 1.1% in males, respectively. Mammotropes were oval or round in shape, and $10{\sim}20{\mu}m{\times}17{\sim}25{\mu}m$ in size. These cells were distributed throughout the pars distalis, but were more abundant on the dorsal part adjacent to the hypophyseal cavity and along the lateral and ventral peripheral regions. Somatotropes were elliptical, triangular or polygonal in shape, and $8{\sim}17{\mu}m{\times}17{\sim}18{\mu}m$ in size. The distribution pattern of somatotropes was similar to that of mammotropes. Some somatomammotropes were intercalated between clusters of mammotropes while the others were dispersed independently among the secretory cells.

• Proceedings of the Zoological Society Korea Conference
• /
• 1998.10b
• /
• pp.230.1-230
• /
• 1998

No Abstract, See Full Text

• Development and Reproduction
• /
• v.2 no.2
• /
• pp.189-196
• /
• 1998

Several lines of evidence indicate that some neuropeptides classically associated hypothalamus have been found in pituitary gland, suggesting the existence of local regulation of pituitary function. Among the hypothalamic releasing hormones, genes for TRH and GnRH are expressed in the rat anterior pituitary gland. The present study was carried out to investigate the expression of the GHRH gene in rat anterior pituitary and the pituitary-derived cell lines. The presence of GHRH transcripts in pituitary tissue was shown by 3'rapid amplification of cDNA end (3'-RACE) analysis. In reverse transcription-polymerase chain reaction (RT-PCR) study, GHRH cDNA fragments were amplified from two pituitary-derived cell lines, $\alpha$T3 cells originated from mouse gonadotrope and GH3 cells from rat somatolactotrope. Immunoreactive GHRH was detected in large and medium-sized pituitary cells by immunocytochemistry. Significant amounts of GHRH-like molecules were found in the GH3 cell extracts. In RNase protection assay, the level of pituitary GHRH mRNA was augmented by ovariectomy. These results demonstrate that GHRH gene is expressed in the rat gonadotropes and somatotropes, and suggest that the pituitary GHRH could be participated in the paracrine and/or autocrine regulation of cell proliferation, as well as promoting growth hormone secretion.