Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.4.21.4 (trypsin)
42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

An inhibitor for luteinizing hormone (LH) receptor site binding has been partially purified from aqueous extracts of ovaries from pseudo-pregnant or pregnant rats. The LH receptor binding inhibitor (LH-RBI) was heat-resistant, partially inactivated by trypsin or pronase digestion, and had a molecular weight of approximately 3800 daltons. The LH-RBI was not found in the ovary of mature non-pregnant rats or immature rats, nor was it found in testis or liver extracts. The LH-RBI strongly inhibited the in vitro binding of 125I-labeled ovine LH to ovarian LH receptors but did not inhibit 125I-labeled ovine prolactin binding to ovarian PRL receptors.
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PMID:Characterization of an inhibitor for luteinizing hormone receptor site binding. 17 92

There is strong evidence that the hypophyseal neurointermediate lobe (NIL) mediates 17 beta-estradiol (E2)-induced PRL secretion in rats. Our laboratory has previously demonstrated that E2 stimulates NIL cells to release an activity that acutely increases the relative abundance of PRL-releasing cells in anterior pituitary (AP) cell cultures. We later found that secretory products of the NIL melanotropes, specifically the acetylated forms of alpha MSH and beta-endorphin (beta END), can account for this activity. Given that blood from the NIL initially perfuses the region of the AP proximal to the NIL, we tested the hypothesis that this specific area was preferentially responsive to the lactotrope recruitment activities. AP glands from ovariectomized rats were dissected into an inner zone, proximal to the NIL, and the remaining outer zone of the gland, then dispersed with trypsin. The resulting cells were cultured for 16 h, either alone or in coculture with NIL cells, and subsequently treated with medium alone (control) or with alpha MSH, beta END, or E2 (all at 1 x 10(-7) M) for 3 h and then subjected to reverse hemolytic plaque assays for PRL release. Under control conditions, the proportion of PRL-secreting cells was significantly greater in cultures from the outer zone of the AP than in those from the corresponding inner zone of the gland. Treatment of AP cells from the inner zone with alpha MSH, beta END, or the E2-induced NIL activity significantly increased the percentage of PRL secretors by about 8% of all AP cells. In contrast, the fraction of PRL-secreting cells in cultures from the outer zone was not affected by these treatments. We conclude that the recruitment of PRL-secreting cells in response to products of the NIL occurs only in that region of the AP proximal to the NIL.
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PMID:Neurointermediate lobe peptides recruit prolactin-secreting cells exclusively within the central region of the adenohypophysis. 133 43

We have previously established a somatomammotroph cell line (rPC0) derived from normal rat pituitary that secretes GH and PRL. In this study we report that conditioned medium from rPC0 cells (CM-rPC0) exhibited a stimulatory effect on the growth of rat mammary epithelial cells in culture. Fractionation of CM-rPC0 revealed that the growth-promoting activity of CM-rPC0 was associated with a fraction eluting from a diethylaminoethyl-Sephacel column at 0.5 M NaCl. The growth-promoting activity of this fraction was abolished by trypsin and was resistant to dithiothreitol treatment. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the predominant components of the 0.5-M NaCl fraction were proteins migrating at the 14-18K region of the gel. The 0.5-M NaCl fraction did not contain immunodetectable GH or PRL. Further fractionation of CM-rPC0 on a heparin-agarose column showed that the growth-promoting activity eluted at 0.9 M NaCl. The two predominant proteins in this fraction had apparent mol wt of 14.5K and 18.2K. Based on the structural and biological properties, several known hormones and growth factors were excluded from consideration as potential candidates responsible for the growth-promoting effect of the 14-18K proteins. It is postulated that this group of 14-18K proteins contains a new factor(s) that affects the growth of mammary epithelial cells.
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PMID:Mitogenic effect of a factor from rat somatomammotrophs on mammary epithelial cells. 137 23

The progression from nonpregnancy through late lactation is associated with an increase in the proportion of anterior pituitary cells that secrete PRL and a comparable decrease in the percentage of cells that release GH. These fluctuations result from variations in both the number of cells that release GH or PRL alone and mammosomatotropes, cells that release both hormones concurrently. However, it has not been determined whether this reciprocal shift in PRL and GH secretors during the onset of lactation is readily reversible. In the present study, anterior pituitaries from adult virgin, late lactating, or postweaning female rats (4, 6, or 8 days) were dispersed with trypsin and subsequently assayed for PRL and GH release using reverse hemolytic plaque assays. We found that separating lactating females from their pups for only 4 days induced a reciprocal shift in the proportions of GH and PRL cells back to levels found in virgin females. Simultaneous plaque assays were then performed to determine whether this post-weaning shift in the percentages of GH and PRL secretors was due to changes in the abundance of cells that secrete each hormone alone or in the proportion of mammosomatotropes. The overall changes in GH and PRL cell proportions consisted of variations only in the fraction of cells that secreted either GH or PRL alone; no differences were observed in the percentage of mammosomatotropes or in the overall abundance of acidophils. Our results demonstrate that the reciprocal shifts in the proportions of PRL- and GH-secreting cells associated with lactation are rapidly reversible. Moreover, these results are consistent with our hypothesis that PRL and GH secretors are functionally interconvertible and further suggest that this process is bidirectional.
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PMID:Evidence for bidirectional interconversion of mammotropes and somatotropes: rapid reversion of acidophilic cell types to pregestational proportions after weaning. 187 67

There is extensive evidence that the posterior pituitary (PP) participates in the regulation of PRL secretion. We recently reported that a putative PRL-releasing factor is localized, and possibly produced, in the intermediate lobe of the PP. The aim of the present investigation was to determine whether cultured PP cells affect anterior pituitary (AP) function in terms of cell content and cumulative release of PRL. Anterior and posterior pituitaries from adult male rats were dispersed with trypsin and cultured either alone or together for 4 and 8 days in serum-free medium. The concentrations of PRL, GH, and LH in cell extracts and culture media were measured by RIA. Coculturing of AP and PP cells at different plating densities resulted in a 2-fold rise in PRL cell content after 4 days. The cumulative release of PRL in the cocultures was significantly increased only after 8 days. LH and GH were affected slightly, or not at all. Medium conditioned by PP cells mimicked the effects of coculture on the cumulative release of PRL, but not on the cell content. Short-term incubation with TRH induced a much larger release of PRL from AP + PP cocultures than from AP cells cultured alone. In conclusion, these data suggest that 1) PP cells stimulate the production and release of PRL in a hormone-specific manner, and 2) coculturing of AP + PP cells augments the responsiveness of lactotrophs to secretagogues such as TRH. We propose that at least two factors, one of which might be PRL-releasing factor, are involved in these effects.
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PMID:Coculture of anterior and posterior pituitary cells: selective stimulation of lactotrophs. 190 68

Expression of tyrosine hydroxylase (TH) in cultured cells of the ventral hypothalamus-midbrain of fetal rats has been investigated. TH mRNA and TH were quantified by an S1 nuclease protection assay and an immunoblot assay, respectively. Dihydroxyphenylalanine (DOPA) and dopamine secretion were evaluated using their rates of accumulation in the culture medium. The rate of accumulation of DOPA was 2-3 times that of dopamine. Inhibitors of TH activity caused a dose-dependent reduction in DOPA secretion. During an 11-week culture of dissociated cells, TH mRNA increased from 1.6 to 2.8 attomole/well between the first and fourth week of culture, remained steady to the ninth week, and then declined. TH increased from 12 to 105 fmol/well between the first and seventh week and then declined. DOPA secretion increased until the sixth week and then remained steady to the tenth week. An extract of rat pituitaries stimulated DOPA secretion by the cultures in a dose-dependent manner. This activity, attributed to a cytotropic factor (CTF), was inactivated by heating for 10 min in a boiling water bath, but was unaffected by trypsin digestion. Incubation with CTF for 24, 48, 72, and 96 h resulted in a day by day increase in the secretion of DOPA. After 96 h of culture with CTF, the amount per well of TH mRNA, but not TH, was significantly (P less than 0.01) greater than the control value. Pituitary CTF is probably not PRL, since rat PRL did not appreciably affect DOPA secretion or the amount of TH mRNA or TH in the cells. Withdrawal of CTF from CTF-stimulated cells resulted in a marked reduction in DOPA secretion as well as a decrease in TH mRNA. These results support the hypothesis that the pituitary gland contains a cytotropic factor that stimulates TH expression in fetal brain cells of the hypothalamus-midbrain.
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PMID:Expression of tyrosine hydroxylase in cultured brain cells: stimulation with an extractable pituitary cytotropic factor. 197 Feb 92

This laboratory has previously demonstrated that PRL-secreting cells are virtually nonexistent on day 3 but appear in appreciable numbers on day 4 of neonatal life. The purpose of the present study was to determine whether this explosive appearance of PRL cells is due to maternal influences specific to the first 4 days of lactation. Litters of 1-day-old rat pups were placed with foster mothers that had been lactating for either 1 or 4 days. Four days later (at 5 days of age), the anterior pituitaries from these pups were removed, dispersed into individual cells with trypsin, and subjected to reverse hemolytic plaque assays for PRL and GH release. We found that the proportion of PRL-releasing cells in pituitaries from pups placed with day 1 mothers (4.2 +/- 0.6%; mean +/- SE; n = 4 experiments) was similar to that in 5-day-old pups that had not been fostered (3.4 +/- 0.6%; n = 3 experiments). In contrast, placing 1-day-old pups with day 4 mothers significantly reduced (P less than 0.01) the fraction that released PRL to 0.6 +/- 0.2% of all pituitary cells (n = 4 experiments). This effect appeared to be specific to PRL cells, since the percentage of all pituitary cells that secreted GH was not different between litters fostered onto day 1 or day 4 dams (40.0 +/- 3.8% and 41.1 +/- 3.9%, respectively; n = 3 experiments). Furthermore, the suppression of PRL cell expression did not result from a gross nutritional deficit, since the rates of body wt gain during the foster period were not different between the two groups (12.4 +/- 0.5 and 12.9 +/- 0.8 g/litter.day, respectively; n = 4 experiments). In a second design, newborn pups were placed with females that had been lactating for 0, 2, 4, or 7 days and analyzed for PRL- and GH-releasing cells at 5 days of age. In two trials of this experiment, the fractions of pituitary cells that secreted PRL were 5.2, 6.3, 5.9, and 1.2%, respectively for trial 1 and 3.3, 4.1, 0.7, and 0.6%, respectively for trial 2. As in the first experiment, the proportion of GH-releasing cells and the rate of growth were not different among the four groups. Taken together, these results indicate that normal differentiation of PRL cells in neonatal rats is triggered by a maternal signal present during the first few days of lactation, and that the magnitude of this signal declines with the progression of lactation.
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PMID:Normal differentiation of prolactin cells in neonatal rats requires a maternal signal specific to early lactation. 198 61

Our laboratory has provided substantial evidence for the presence of PRL-releasing factor (PRF) in the posterior pituitary. The objectives of this study were 1) to determine the distribution of PRF activity between the neural and intermediate lobes, and 2) to assess the PRF activity of cultured posterior pituitary cells. Posterior pituitaries from adult male rats were dispersed with trypsin and cultured for 1-7 days. Cultured cells or intact posterior pituitaries were extracted with acid and lyophilized. PRF activity was determined by the ability of reconstituted extracts to increase PRL release from cultured anterior pituitary cells. Upon dissection of the posterior pituitary, PRF activity was primarily present in the intermediate lobe. There was minimal contamination between the two lobes, as indicated by the localization of 90% of the total oxytocin in the neural lobe and 95% of alpha MSH in the intermediate lobe. Extracts from intact posterior pituitaries and posterior pituitary cells cultured for 4 days stimulated PRL secretion in a similar dose-dependent manner. Cultured liver and cerebral cortex cells had very low PRF activity. Both oxytocin and dopamine, two neuronal markers, were reduced to less than 5% of their original values within 1 week of cell culture. There was also a significant reduction in the cell content of alpha MSH. On the other hand, PRF activity was relatively stable during culture. Incubation of posterior pituitary cells for 4 days with either cycloheximide or PRL caused a 55-60% reduction of the PRF activity of the cells. We conclude the following. 1) PRF is localized, almost exclusively, in the intermediate lobe of the pituitary. 2) PRF activity is present within nonneuronal cells, either melanotrophs or a small subpopulation of nonopioid-producing cells. 3) PRF is tissue specific, and its presence in cultured posterior pituitary cells depends at least in part on de novo synthesis. 4) The synthesis and/or release of PRF may be subjected to short loop negative feedback regulation by PRL.
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PMID:Prolactin-releasing factor: cellular origin in the intermediate lobe of the pituitary. 211 57

Serum concentrations of PRL and GH increase and decrease, respectively, during the progression from nonpregnancy through lactation. However, it is unknown whether the secretory capacities and/or relative abundance of cells that release PRL or GH are altered during these physiological states. In the present study anterior pituitaries from adult virgin, gestating, and early or late lactating female rats were dispersed with trypsin and subsequently assayed for PRL and GH release using reverse hemolytic plaque assays. We found that the relative abundance of PRL-secreting cells was greater and that of GH cells lower in pituitaries from lactating females than in those from virgins. Moreover, the relative amounts of both PRL and GH released per cell were diminished in gestating and lactating females. For PRL, this decrease could be accounted for by an increase in the number of cells that released small quantities of hormone. We then performed simultaneous plaque assays to determine whether the shifts in the relative proportions of PRL and GH secretors were due to changes in the percentages of cells that secrete each hormone alone or in the fraction that releases both PRL and GH concurrently. Variations in both single and dual hormone-secreting cells appear to contribute to the overall fluctuations in the relative abundance of PRL and GH cells during these physiological transitions. We conclude that the additional PRL secretors present during lactation may arise from cells that previously released only GH, and that this functional interconversion of GH and PRL secretors might involve an intermediate cell type, the mammosomatotrope.
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PMID:Is the mammosomatotrope a transitional cell for the functional interconversion of growth hormone- and prolactin-secreting cells? Suggestive evidence from virgin, gestating, and lactating rats. 212 41

It is well established that the suckling stimulus sensitizes or primes the anterior pituitary to PRL-releasing stimuli. It is also recognized that PRL-secreting cells from a given animal are not all alike but instead exhibit a considerable degree of functional heterogeneity. The goal of the present study was to determine whether the suckling-induced priming phenomenon is manifest at the cellular level by shifts in the relative abundance of various mammotrope subpopulations. This was accomplished by using reverse hemolytic plaque assays to evaluate the secretory characteristics of individual PRL secretors derived from lactating rats either before or after the transient application of a suckling stimulus. Groups of day 10 lactating rats separated from their litters for 4 h were either killed immediately or were reunited briefly (10 min) with their pups before death. Adenohypophyseal cells obtained after trypsin dispersion were then subjected to plaque assays for PRL. Mammotropes derived from suckled rats were, on average, considerably more responsive to the stimulatory actions of TRH and angiotensin II and less susceptible to inhibition by dopamine. Mammotropes from nonsuckled rats exhibited a bimodal frequency distribution in which plaques from the second mode were roughly 6-8 times larger (released considerably more PRL) than those from the first. Superimposition of suckling (or in vitro treatment with dopamine) caused the second mode to disappear. Suckling also enhanced greatly the fraction of PRL cells that shifted from the first to the second mode (i.e. released more hormone) after treatment with TRH or angiotensin II. Taken together, our results demonstrate that the suckling-induced sensitization of pituitary tissue to PRL-releasing stimuli is manifest at the cellular level as proportional shifts toward those cells most responsive to stimulatory secretagogues and away from those most susceptible to inhibition by dopamine.
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PMID:Suckling increases the proportions of mammotropes responsive to various prolactin-releasing stimuli. 222 1


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