Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P30536 (PBS)
 9,886 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Searching for the best procedure for simultaneous estimation of the anterior pituitary hormones, extraction efficiencies of various media, additives such as urea and triton X-100, and physical treatments such as freezing-thawing (F-T) and sonication, were examined by measuring prolactin (PRL), growth hormone (GH), lutropin (LH), follitropin (FSH), and thyrotropin (TSH) in the extracts. Ethanolic media (60% EtOH) gave high yields of PRL at neutral to alkaline pH, but poor extraction of GH accompanied by a marked loss of its immunoreactivity during storage. Ethanolic media also gave a poor yield of LH even at high pH. Aqueous media like PBS at various pH, 0.1 M acetic acid and distilled water were considerably effective in the extraction of GH, LH, FSH and TSH if they were coupled with F-T and sonication. However, high yields of PRL could not be obtained with these aqueous media even with F-T and sonication. Hartree's 40% EtOH-6% ammonium acetate, pH 5.1, solubilized considerable amounts of glycoprotein hormones, but yielded almost no GH and only a small amount of PRL. The addition of triton X-100 to PBS (pH 7) at 0.1% resulted in the maximum extraction of glycoprotein hormones with homogenization and F-T, but further sonication was necessary for GH and PRL. When the anterior pituitaries were homogenized and frozen-thawed in PBS (pH 7) containing 1 M urea, yields of PRL, GH, LH, FSH, and TSH were maximum, and sonication did not cause any additional extraction, indicating that this procedure, i.e. homogenization and F-T in 1 M urea-PBS, would be the best for the simultaneous estimation of these anterior pituitary hormones.
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PMID:Choice of extraction procedure for estimation of anterior pituitary hormone content. 343 4

Anterior pituitaries from primiparous lactating C3H/He mice cultured in the medium containing 0.1% dimethyl sulfoxide (DMSO) for 48 hours and pituitaries from lactating mice given subcutaneous injections of 0.05 ml DMSO twice daily for two days and once on the morning of the third day were used in the studies of the in vitro and the in vivo effects of DMSO, respectively. Phosphate buffer saline was used in the control. Synthesis and release of growth hormone and prolactin were estimated by the incorporation of [14C]leucine into each hormone during three hours' incubation of the pituitaries pre-exposed to DMSO or PBS. The values in the medium represented released hormone and sum of the values in the medium and the pituitary represented the synthesized hormone. DMSO stimulated synthesis of GH and synthesis and release of prolactin in vitro. Meanwhile, in the vivo study, synthesis of GH and prolactin were lower in the DMSO-injected mice than in the control. The results suggest that the effects of DMSO on the pituitary secretion of GH and prolactin are adverse in vitro and in vivo. In vivo exposure of pituitary to DMSO resulted in the suppression of lactation.
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PMID:The in vitro and in vivo effects of dimethyl sulfoxide on the pituitary secretion of growth hormone and prolactin in mice. 619 45

Male Sprague-Dawley rats were castrated and given daily sc. injections of estradiol (E2, 10 micrograms/day), testosterone propionate (TP, 1.0 mg/day), dihydrotestosterone (DHT, 1.0 mg/day) or sesame oil (SO, 0.2 ml/day). A group of sham castrate males received daily sc. injections of SO (0.2 ml/day). On day 8 of steroid treatment animals were decapitated and anterior pituitaries were removed and hemisected. Each half was homogenized in PBS buffer (0.01 M Na2HPO4-NaH2PO4; 0.14 M NaCl; 0.1% bovine serum albumin) at either pH 7.6 or 10.6. Homogenates were chromatographed on Sephadex G-100 columns and eluted fractions were assayed for prolactin (PRL) by RIA. Four immunoreactive forms of PRL, designated as "void volume," "big big," "big" and "little," were eluted from the pituitary homogenates of each experimental group. Homogenates obtained at pH 7.6 contained a greater percentage of PRL in the "void volume" and less activity in the "big" and "little" forms than pH 10.6 homogenates in all experimental groups. Pituitaries from SO- and TP-treated castrate animals contained significantly greater percentages of activity in the "void volume" at pH 7.6 compared to the other groups. At pH 10.6, the pituitary homogenates from the E2-treated group eluted a significantly greater percentage of "big" PRL and a smaller percentage of "little" PRL compared to all other groups. These findings suggest that androgenic and estrogenic steroids may play a role in the pituitary PRL molecular size profile of the male rat.
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PMID:The effect of steroids on the size heterogeneity of pituitary prolactin in castrate male rats. 669 10

For characterization and determination of recombinant bovine GH (rbGH) eight monoclonal antibodies (MAb) were produced against rbGH from Monsanto. The various MAb showed different affinities to rbGH, pituitary bovine GH (pbGH), and pituitary ovine GH (poGH). With epitope analysis several MAb were shown to recognize different epitopes of rbGH. The MAb MUC-rbGH-3A11 and MUC-rbGH-1E5 were used to develop a Sandwich ELISA. By checking the specificity of the assay no cross reactivity was found with pituitary porcine GH, pituitary human GH, bovine or ovine prolactin and little cross reactivity with poGH could be found. The Sandwich ELISA detected various rbGH (Monsanto, Elanco, Cyanamid) with different N-terminal amino acids and discriminated between rbGH and pituitary bovine GH by an affinity factor of 2.0. The detection level was 2 ng rbGH per ml PBS buffer. The recovery was about 86% in bovine serum. It might therefore be possible to detect rbGH-treated cows using a Sandwich ELISA, but this would need a field study.
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PMID:Identification of antigenic differences of recombinant and pituitary bovine growth hormone using monoclonal antibodies. 751 99

Studies were undertaken to identify intracellular mediators of prolactin inhibition of glucocorticoid-induced apoptosis in Nb2 lymphoma cells. A short-term assay was implemented that quantitates fragmented DNA released from the genome by reaction with diphenylamine. Induction and inhibition of internucleosomal DNA cleavage (indicative of apoptosis) was verified by agarose gel electrophoresis of extracted cellular DNA. Synchronized Nb2 cells (G0/G1) exhibited increased DNA fragmentation after 4-hr incubation with dexamethasone (DEX) (25-100 nM) which was inhibited by ovine prolactin (oPRL) (0.1-1 ng/ml), the glucocorticoid receptor antagonist, RU486 (500 nM), and the nuclease inhibitor, aurintricarboxylic acid (100 microM). Signals previously implicated in prolactin induction of mitogenesis in Nb2 cells were investigated for their role in prolactin inhibition of apoptosis including: protein kinase C activation, arachidonic acid metabolism, polyamine production, tyrosine phosphorylation, and extracellular calcium. Protein kinase C agonists, phorbol-12-myristate-13-acetate, and 1,2-dioctanoyl-sn-glycerol, +/- the calcium ionophore, A23187 (200 nM), did not mimic oPRL inhibition of DEX-induced DNA fragmentation. Protein kinase C inhibitors, gossypol and quercetin, did not block prolactin action. Arachidonic acid did not mimic prolactin protection against DEX-induced DNA fragmentation. Inhibitors of arachidonic acid metabolism, 5,8,11,14-eicosatetraynoic acid, nordihydroguaiaretic acid, and indomethacin did not block prolactin action. The polyamine, spermine, inhibited DEX-induced DNA fragmentation at 1.5 to 2.5 mM. However, inhibition of polyamine synthesis with alpha-difluoromethyl ornithine or methylglyoxal bis(guanylhydrazone) did not inhibit prolactin action. Prolactin action was not blocked by inhibitors of tyrosine kinase activation, genistein and tyrphostin-47. On the other hand, pervanadate, a potent tyrosine phosphatase inhibitor, consistently inhibited DEX-induced DNA fragmentation. Prolactin action and DEX-induced apoptosis both occurred in calcium-free PBS. In summary, protein kinase C activation and eicosanoid production do not appear to mediate this prolactin action. Although spermine could block DNA fragmentation, blockade of the polyamine cascade did not inhibit prolactin action, suggesting that polyamines do not mediate this prolactin effect. While inhibitors of tyrosine kinase activation did not block prolactin action, tyrosine phosphatase inhibition in the presence of basal tyrosine kinase activity mimicked prolactin action, suggesting tyrosine phosphorylation participation in the anti-apoptotic effect. Extra-cellular calcium was not required for prolactin or DEX action.
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PMID:Investigation of intracellular signals mediating the anti-apoptotic action of prolactin in Nb2 lymphoma cells. 777 88

Male rats were treated for 21 days with drugs known to affect prolactin secretion, in order to assess the effects of these drugs on mitochondrial benzodiazepine receptors (MBRs). Sulpiride, a selective dopamine D2 receptor antagonist and hyperprolactinemic agent, decreased MBR density in the adrenal gland (49%; P < 0.005), whereas metoclopramide, another dopamine antagonist with a preference for dopamine D2 receptors, increased adrenal gland MBR density (31%; P < 0.05). Bromocriptine, a specific dopamine agonist, increased MBR density in this organ (87%; P < 0.001). None of the three agents influenced kidney or testicular MBRs. These data indicate that the mechanism of organ-specific alterations in MBRs seems to be prolactin independent.
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PMID:Adrenal mitochondrial benzodiazepine receptors are sensitive to agents active at the dopamine receptor. 838 54

The purpose of this study was to examine the role of vasoactive intestinal peptide (VIP) in prolactin (PRL) release in young turkey poults. Poults obtained from hens immunized with keyhole limpet hemocyanin (KLH)-chicken VIP (cVIP-KLH) conjugate or with KLH alone were used in this study. Plasma VIP antibody was tested by means of monoiodinated cVIP. Plasma samples were prediluted (1:100) in 0.05 EDTA PBS and used as a source of primary antibody in a cVIP-binding assay. Antibody levels averaged 50.8 +/- 10.3% at hatch and then declined with age to a level of 5.0 +/- 2.1% after 3 wk. Plasma PRL concentration was lower in cVIP-KLH-immunized poults (p < 0.05) than in KLH-immunized birds. Exogenous VIP administration at 7 days of age (7.8 micrograms/kg) increased plasma PRL level (p < 0.05) to a peak value of 359 +/- 32 ng/ml in the KLH-immunized birds. A smaller increase (p < 0.05) was obtained when the KLH-immunized poults received injections at 2 and 3 wk of age. The PRL response to cVIP administration was not observed in 1- and 2-wk-old poults maternally immunized with cVIP-KLH. Similarly, electrical stimulation of the ventromedial nucleus of the hypothalamus, at 1 and 8 days of age, induced a significant increase in the plasma PRL level of maternally KLH-immunized birds but not in maternally cVIP-KLH-immunized birds. These findings suggest that cVIP is a PRL-releasing factor in young turkey poults, similar to the finding in adult turkey hens.
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PMID:Effect of maternal passive immunization against vasoactive intestinal peptide on prolactin secretion in turkey poults. 883 90

Prostaglandins primarily of uterine origin play an important role in parturition. Hysterectomy of nongravid pigs early in the luteal phase maintains luteal function until about Day 150, whereas the duration of normal pregnancy is about 114 days. A precisely timed peak release of relaxin and coincident decrease in progesterone secretion in unmated hysterectomized gilts are similar to hormonal changes that occur a few hours before parturition. It is hypothesized that prostaglandin F2alpha (PGF2alpha) in hysterectomized pigs mimics abrupt changes in ovarian and pituitary hormone secretion seen before normal parturition and in early lactation. Unmated Yorkshire gilts were hysterectomized on Days 6-8 of a normal estrous cycle, and at 1200 h on Day 113, they were given an i.m. injection of 30 mg PGF2alpha-trihydroxymethylaminomethane (THAM) salt or PBS. None of these gilts expressed behavioral estrus immediately after PGF2alpha or vehicle treatment. On Day 113, PGF2alpha increased peak relaxin (60 ng/ml) compared with that of controls (34 ng/ml; p < 0.01), whereas progesterone decreased abruptly (4 vs. 16 ng/ml in PGF2alpha and PBS; p < 0.01). Prolactin remained at < 5 ng/ml from Day 98 to 120 in controls but peaked at 33 ng/ml immediately after PGF2alpha treatment on Day 113, and then decreased to levels similar to those of controls on Day 120. Sequential bleeding revealed an acute growth hormone release (4.5 ng/ml) immediately after PGF2alpha injection and return to basal levels (< 0.6 ng/ml) on Days 114-120. PGF2alpha induced abrupt shifts in progesterone, relaxin, prolactin, and growth hormone secretion in hysterectomized gilts that mimicked hormone changes seen in late pregnancy, parturition, and early lactation. These findings provide new insight into the role of PGF2alpha in abruptly changing hormone secretions by aging corpora lutea and the pituitary gland even in the absence of conceptuses or the uterus in the pig.
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PMID:Prostaglandin F2alpha-induced luteolysis of aging corpora lutea in hysterectomized pigs. 954 36

Serotonin is known to stimulate prolactin secretion by decreasing tyrosine hydroxylase (TH) activity in the tuberoinfundibular dopaminergic (TIDA) neurons. However, the effects of aging on the responsiveness of TIDA neurons to serotonin are not known. An effective way to increase serotonergic activity is to administer 5-hydroxytryptophan (5-HTP), a serotonin precursor. The present study was done to investigate the effects of 5-HTP on TIDA neuronal activity in aging animals. Middle-aged (10-12 mo), old (18-20 mo), and very old (22-24 mo) female Sprague-Dawley rats were bilaterally ovariectomized. Ten days later, they were injected iv with 50 mg/kg body wt of 5-HTP or the vehicle for 5-HTP (PBS-HCI). Twenty minutes later, m-hydroxybenzylhydrazine (NSD), a DOPA decarboxylase inhibitor, was administered. Ten minutes later, the animals were killed, and tyrosine hydroxylase (TH) activity was determined by measuring L-DOPA accumulation in the stalk median eminence by HPLC-EC. In all three groups, administration of 5-HTP increased serum prolactin levels significantly. In control middle-aged rats, TH activity (L-DOPA pg/ microg protein) was 33.0+/-5.6. Treatment with 5-HTP decreased TH activity by 60%. Similarly, 5-HTP treatment decreased TH activity by 52 and 56% in 18- to 20- and 22- to 24-mo-old rats, respectively, compared to the control rats. The magnitudes of the 5-HTP-induced decreases in TH activities in middle-aged, old, and very old rats were not different from each other. These results indicate that TIDA neuronal responsiveness to serotonin does not change with age and that 5-HTP is capable of stimulating PRL release even in very old rats.
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PMID:Responsiveness of tuberoinfundibular dopaminergic neurons to 5-hydroxytryptophan: effects of aging. 979 28

Antibody against porcine relaxin (antipRLX540; 1:950,000) was produced in sheep and used to determine the effect on relaxin and progesterone secretion, and on parturition in late pregnant pigs. In group 1, Yorkshire gilts with normal estrous cycles were bred on the second observed estrus and fitted with an indwelling jugular cannula and an intraperitoneal cannula on day 100 of pregnancy. Gilts were infused at 6-h intervals with antipRLX540 (n = 10) or PBS (n = 10) beginning on day 103 until parturition. From days 103 to 120, daily blood samples (10 ml) were collected for RIA of relaxin, progesterone, and prolactin. In group 2, bred gilts were randomly assigned to antipRLX540 (n = 11), relaxin (n = 5), and PBS (n = 8) treatment on days 111, 113, and 115. Blood was collected twice daily from day 108 to 120, and every 20 min on days 111, 113, and 115 beginning 60 min before treatment and continuing 180 min. Parturition in gilts given antipRLX540 occurred on day 112.7 compared with day 114.0 in relaxin-treated gilts and day 114.3 in PBS controls (P < 0.05). Duration of delivery from first to last piglet was greatly delayed in antipRLX540 gilts (240 min) compared with PBS controls ([117 min] P < 0.005). Average number of stillborns was greater in antipRLX540- than in PBS-treated controls (2.4 vs. 1.0; P < 0.05). Relaxin concentration in peripheral plasma was lower in antipRLX540-treated gilts from day 105 to 110, but on day 113 the antipRLX540-treated group had a greater relaxin peak release compared with PBS-treated animals (P < 0.05). Plasma progesterone concentrations were similar in antipRLX540- and PBS-treated gilts throughout the period of the study. In group 2, by day 113, progesterone decreased in antipRLX540-treated gilts compared with relaxin- and PBS-treated gilts. Prolactin levels were similar in both antipRLX540- and PBS-treated gilts; however, from 1 to 3 days postpartum the antipRLX540 group had higher prolactin concentration (P < 0.05). The results indicate that antipRLX540 decreased circulating plasma concentrations of unbound or free relaxin during the last 10 days of pregnancy in Yorkshire gilts. AntipRLX540 markedly increased both the duration of delivery of piglets and the average number of stillbirths in this litter-bearing species compared with PBS-treated controls. This study provides strong evidence that increasing circulating concentrations of relaxin during late pregnancy is crucial for unimpaired parturition in the pig.
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PMID:Antiporcine relaxin (antipRLX540) treatment decreases relaxin plasma concentration and disrupts delivery in late pregnant pigs. 982 4


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