Prolactin gene expression in human ovarian follicular cells.
ABSTRACT To investigate prolactin gene expression in human ovarian follicular cells.
RNA was isolated from follicular cells obtained at the time of transvaginal oocyte retrieval from patients after controlled ovarian hyperstimulation. The RNA was subjected to reverse transcription and polymerase chain reaction (RT-PCR) using prolactin-specific primers. The RT-PCR products were analyzed by gel electrophoresis, followed by Southern blot analysis using prolactin-specific probes.
Department of gynecology and obstetrics research laboratory.
Women undergoing controlled ovarian hyperstimulation followed by transvaginal oocyte retrieval.
Controlled ovarian hyperstimulation followed by transvaginal oocyte retrieval. Expression of prolactin mRNA by follicular cells.
The presence of prolactin-specific cDNA amplified by RT-PCR from RNA isolated from human follicular cells was confirmed by Southern blot analysis.
Human ovarian follicular cells are an extrapituitary site of prolactin gene expression.
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ABSTRACT: The role of prolactin in the regulation of ovarian folliculogenesis and corpus luteal function and in particular its relationship to atresia in these structures is as yet unclear. We established a model of apoptosis in which to examine the actions of prolactin. Granulosa cells collected from IVF-flush were cultured at 0.1-0.3 x 10(6) cells/well in growth media for 48 h, placed into serum-free media for 24 h prior to dosing for 24 h. Dose responses to C2-ceramide and prolactin were performed. Cells were then treated with an apoptotic dose of C2-ceramide alone, prolactin (100 ng/ml) alone or a combination of the two. Cell death was assessed by Trypan Blue cell counting and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; Thiazolyl Blue] assay and apoptosis confirmed by morphological assessment and flow cytometry. C2-ceramide (0-40 micro mol/l) induced a dose-dependent increase in cell death (63.8% increase at 40 micro mol/l) and, morphologically, cells exhibited classical features of apoptosis. Prolactin alone had no effect on metabolic activity or total cell number. On co- incubation, prolactin alone had no effect on cell death, whereas C2-ceramide induced an approximately 62.6% increase in apoptosis, which was inhibited in the presence of prolactin. Prolactin may contribute significantly to early corpus luteum formation and survival by acting as a potent antiapoptotic factor for human granulosa cells.Human Reproduction 01/2004; 18(12):2672-7. · 4.67 Impact Factor
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ABSTRACT: It is known that hyperprolactinaemia can cause galactorrhoea and irregular cycles or even amenorrhoea. High serum prolactin (PRL) can disturb follicular maturation and corpus luteum function. Treatment of hyperprolactinaemia in patients with resulting bleeding anomalies is established, but the question is how to manage normal cyclic hyperprolactinaemic women? Studies have shown that in a subgroup of asymptomatic patients the serum contains mainly high molecular weight form (big big PRL), which has a low bioactivity, called macroprolactinaemia. It is evident that macroprolactin does not affect the control of pituitary PRL secretion via the short loop feedback mechanism or the secretion of gonadotrophins as does monomeric PRL. Identification of macroprolactinaemia is therefore clinically important to prevent unnecessary examinations and inappropriate treatment. Prolactinoma can be associated with macroprolactinaemia. Performance of pituitary imaging in asymptomatic patients with hyperprolactinaemia may therefore be justified, but further studies are needed to evaluate the relation of costs and benefit. An unsolved problem is the differentiation between inactive and PRL-secreting tumours. Caution should be exercised concerning medical treatment in unstimulated patients and also in patients during ovarian stimulation alone or in combination with intrauterine insemination or in-vitro fertilization. The potential clinical significance of hyperprolactinaemia/macroprolactinaemia in asymptomatic women must be further evaluated.Reproductive biomedicine online 12/2005; 11(5):581-8. · 2.68 Impact Factor
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ABSTRACT: Apart from the well established role of prolactin (PRL) in the control of mammary development and lactation, this hormone appears to possess a variety of physiological functions and evidence exists about its expression in many extra-pituitary sites. This experimental work was undertaken to gain knowledge about PRL and its receptor presence in the porcine antral follicle. In particular, we investigated the expression and local production of PRL in follicular fluid, theca and granulosa cells cultured in standard conditions and in hypoxia. Then, we also investigated its modulatory effect on several parameters mainly involved in granulosa cell function, namely redox status and steroidogenesis. In order to verify an involvement of PRL in the control of ovarian angiogenesis, a process strictly linked to follicle growth and development, we have verified possible PRL effects on granulosa cell production of Vascular Endothelial Growth Factor (VEGF) and nitric oxide as well as its modulatory role on the angiogenic activity of endothelial cells. Our data demonstrate that in the swine PRL is expressed in both components of the antral follicle, theca and granulosa layer, and it is produced by granulosa cells. Moreover, the hormone represents a relevant modulatory factor on key processes underlying follicular growth and development, such as steroidogenesis and angiogenesis.Regulatory Peptides 01/2014; · 2.06 Impact Factor