Biomedical subjects

A Barnea

Publications and source records attributed to A Barnea.

At least 73 records · Page 4Linked to original sources

PubMed1986

Copper stimulation of LHRH release from median eminence explants. III. A process dependent on extracellular sodium.

Copper, complexed to histidine (CuHis), stimulates LHRH release from explants of the median eminence area (MEA). To gain further understanding of the mechanism of copper action, in this study, we assessed the Na+ and energy requirements for CuHis stimulation of LHRH release. MEA explants, obtained from adult male rats, were incubated at 37 degrees C for 15 min with 100 microM CuHis and then for 45 min in CuHis-free medium (Krebs-Ringer-phosphate buffer, pH 7.4). LHRH released into the medium was evaluated by RIA. When the incubation buffer contained 143 mM Na+, CuHis stimulated the release of LHRH from a basal level of 17.2 +/- 1.26 (mean +/- SEM, n = 7) to 74.5 +/- 6.2 pg/60 min per MEA. When [Na+] was reduced to 16 mM Na+ (by substituting with Li+), CuHis-stimulated LHRH release was inhibited by 80% (p less than 0.001); indicating a requirement for Na+. In addition, we found that CuHis-stimulated LHRH release was a saturable function of Na+ concentration; saturation achieved with about 100 mM Na+. To assess the requirement for Na+ transport, we evaluated the effect of 1 mM ouabain, 10 microM tetrodotoxin (TTX), or 100 microM amiloride on CuHis stimulation of LHRH release. Ouabain inhibited CuHis stimulation of LHRH release by 80%, whereas TTX and amiloride were ineffective. In addition, we observed that CuHis did not stimulate LHRH release when incubation was carried out at 4 degrees C or at 37 degrees C in the presence of 5 mM KCN.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1986

Evidence that a short-lived effect of copper leads to amplification of prostaglandin E2 stimulation of the release of gonadotropin-releasing hormone from median eminence explants.

We have previously shown that copper (Cu) amplifies prostaglandin E2 (PGE2) stimulation of LHRH release from explants of the median eminence area (MEA) incubated under in vitro conditions. In this study, we have carried out an extensive comparative study of the kinetics of the process of PGE2 stimulation and C alpha-amplified PGE2 (Cu/PGE2) stimulation of LHRH release. MEA explains obtained from adult male rats were incubated under in vitro conditions, and LHRH released into the medium was assayed by RIA. Kinetic parameters were established by varying the time of exposure of the MEA to Cu or PGE2 and the dose of Cu or PGE2. Cu action requires a minimal 5-min period of exposure of the MEA, is rapidly manifested (less than 5 min after transfer of Cu-free medium), and is reversible (half-life, 10-15 min). Cu amplification of PGE2 action is a saturable function of the concentration of both Cu and PGE2; half-saturation and saturation were achieved with 120 and 200 microM copper, respectively, and with 0.6 and 10 microM PGE2, respectively. A 5-min exposure of Cu-treated MEA to PGE2 has the following time course of PGE2 action: the maximal rate of LHRH release is attained within 5 min; release is maintained at the maximal rate for another 5 min and then declines. Importantly, this decline is not prevented by a longer (15-min) exposure of the MEA to PGE2, indicating desensitization to PGE2 action. When MEA treated with Cu (5 min) and PGE2 (15 min) is allowed to recover for 45 min in buffer, the tissue regains its responsiveness to PGE2. All of the kinetic parameters of the process induced by PGE2 alone are similar to those described above for Cu/PGE2, except that the magnitude of the maximal rate of PGE2-stimulated release is 4- to 6-fold lower and that release is maintained at this rate for about 45 min. We also examined the PG structure specificity for stimulation of LHRH release and found that PGD2 by itself or after Cu pretreatment is much less effective than PGE2 in stimulating LHRH release. In summary, Cu amplification of PGE2 stimulation of LHRH release from LHRH neuronal terminals (i.e. MEA explants) involves rapid activation of a short-lived component.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1986

Extracellular calcium is required for copper-amplified prostaglandin E2 stimulation of the release of gonadotropin-releasing hormone from median eminence explants.

Prostaglandin E2 (PGE2) is known to stimulate the release of LHRH from hypothalamic tissue incubated under in vitro conditions. We have previously shown that a short preincubation of explants of the median eminence area with copper(II) [Cu(II)] complexed to histidine (CuHis) markedly amplifies PGE2 stimulation of LHRH release. In this study, we wished to ascertain if Cu-amplified PGE2 stimulation of LHRH release is dependent on influx of extracellular calcium (Ca) and, if so, whether it is the copper and/or PGE2 action that is Ca dependent. Median eminence area explants were incubated for 5 min with 200 microM CuHis, then for 15 min with 10 microM PGE2, and finally for 45 min in buffer (Cu/PGE2). Controls were incubated with CuHis or PGE2. In the presence of Ca (1.8 mM), Cu/PGE2 stimulation of LHRH release reached a peak value within 15 min of exposure to PGE2 (accelerated phase), after which the rate of release declined, with a half-life of about 20 min (decelerated phase). In the absence of Ca, Cu/PGE2 stimulation of LHRH release was inhibited by 70% during the accelerated phase, and it was completely inhibited during the decelerated phase. When incubation was carried out in the presence of Ca and 100 microM verapamil, Cu/PGE2 stimulation of LHRH release was similar to that seen in the absence of extracellular Ca. When Ca was omitted from the incubation medium during the exposure to CuHis but was included during and after the exposure to PGE2, the response to PGE2 was fully restored. In addition, we evaluated the extracellular Ca requirement for stimulation of LHRH release by PGE2 alone (without pretreatment with CuHis) and found that release was inhibited by 65% in the absence of Ca. These results indicate that the process of PGE2 stimulation of LHRH release is partially dependent on extracellular Ca, that the process of Cu-amplified PGE2 stimulation of release is highly dependent on influx of extracellular Ca, and that PGE2 action, but not Cu action, is dependent on Ca influx. Since cAMP has been implicated in Ca2+-dependent secretion in other cells, we propose that Cu amplifies the functional state of a postreceptor component involved in the Ca-cAMP secretory pathway stimulated by PGE2.

Animals↗

PubMed1985

Castration of male rats reduces the capacity of granules isolated from the median eminence to secrete luteinizing hormone-releasing hormone in response to copper.

The effect of castration of male rats on the secretory function of median eminence area (MEA) granules containing luteinizing hormone-releasing hormone (LHRH) was examined, using copper as a test substance. LHRH granules, isolated from MEA of sham or castrated rats 1, 2, or 12 weeks postoperatively, were incubated with various concentrations of copper complexed to histidine (CuHis) and the kinetic constants of LHRH release estimated. CuHis-stimulated release of LHRH was found to be a saturable function of the concentration of CuHis. Castration did not alter the apparent Km for CuHis-stimulated release of LHRH; the Km being 3 and 16.9 microM copper for granules obtained from 1-2 weeks and 12 weeks sham-operated rats, respectively, and 3.7-5.6 and 10.2 microM copper for the granules of castrated rats, respectively. Moreover, castration did not alter the fractional amount of LHRH released in response to CuHis. In contrast, castration markedly reduced the actual amount of LHRH released in response to CuHis; the Vmax being 1,933 and 2,942 pg LHRH released/6 min/MEA for 1-2 weeks and 12 weeks sham-operated rats, respectively, and 782 and 757 pg for the castrated rats, respectively. Similarly, the LHRH content of the MEA of castrated rats was lower than that of the shams. These results are suggestive that castration reduces the capacity of the MEA granules to release LHRH in response to a secretion stimulus such as copper and that this is due not to alterations in the biochemical parameters underlying the release process itself but to a reduced level of MEA LHRH available for release.

Androgens↗

PubMed1985

Effects of castration and maturational age of male rats on the process of copper-stimulated release of luteinizing hormone releasing hormone from median eminence explants: evidence that androgens increase the affinity of the copper-interactive sites for copper.

We have previously shown that chelated copper stimulates the release of luteinizing hormone releasing hormone (LHRH) from explants of the median eminence area (MEA) incubated under in vitro conditions and that this stimulation involves a ligand-specific interaction. In this study, we addressed the question: do testicular steroids regulate the secretory response of LHRH neurons to copper? MEA, obtained from immature, mature, immature castrated and sham-operated rats, were incubated in the presence of various concentrations of copper for 15 min and then in the absence of copper for an additional period of 30 min. We noted that after a lag period of 5 min of incubation, the rate of LHRH release increased in a linear fashion for a period of 15 min. In addition, the rate of LHRH release as well as the rate at which LHRH release was accelerated were saturable functions of the concentration of copper. When incubation was carried out in the presence of a nonsaturating concentration of copper (50 microM), the fractional amount (percent of the total MEA content) of LHRH released from the MEA of castrated rats was significantly (p less than 0.001) lower than that from sham-operated rats; stimulated release being 0.9% and 1.4%, respectively. Similarly, copper-stimulated release from the MEA of immature rats was lower than that from the MEA of mature rats. However, when incubation was carried out in the presence of saturating concentrations of copper (100 or 200 microM), the percentage of stimulated release from the MEA of castrated rats was similar to that of sham-operated rats and significantly lower than that of mature and sham-operated rats, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

PubMed1985

A role for extracellular copper in modulating prostaglandin E2 (PGE2) action: facilitation of PGE2 stimulation of the release of gonadotropin-releasing hormone (LHRH) from median eminence explants.

Copper is present in high concentrations in axonal terminals of hypothalamic neurons (1). We have previously postulated that copper is released from axonal terminals of hypothalamic neurons (2). In this study, we addressed the question: Does extracellular copper facilitate PGE2 action, specifically, the stimulation of LHRH release? The median eminence area (MEA) of male rats was incubated under in vitro conditions, and LHRH release into the medium was quantified by RIA. When MEA were incubated for 5 min with 100 microM copper and then for 15 min with 50 microM PGE2 (Cu/PGE2), Cu/PGE2-stimulated release of LHRH was significantly (P less than 0.001) greater (2 times) than the sum of copper- and PGE2-stimulated release. When MEA were first incubated for 15 min with PGE2 and then for 5 min with copper (PGE2/Cu), PGE2/Cu-stimulated release equalled the sum of PGE2- and copper-stimulated release of LHRH. Thus, copper facilitates PGE2 stimulation of LHRH release from the MEA, and the characteristics of this facilitation are consistent with an enhancement of PGE2 binding to its receptor. These results support the proposition that extracellular copper can serve as a modulator of PGE2 action.

Animals↗

PubMed1984

A ligand-specific action of chelated copper on hypothalamic neurons: stimulation of the release of luteinizing hormone-releasing hormone from median eminence explants.

We have previously shown that chelated copper stimulates the release of luteinizing hormone-releasing hormone (LHRH) from isolated hypothalamic granules. In this study, we wished to ascertain if chelated copper acts on hypothalamic neurons to stimulate LHRH release and, if so, what is the ligand specificity of this interaction. An in vitro system of explants of the median eminence area (MEA) was established and characterized. MEA explants were exposed for 15 min to 50 microM copper, and then they were incubated for 75 min in copper-free medium. Copper led to a transient increase in the rate of LHRH release; the maximal rate was attained 15 min after transfer of the MEA to copper-free medium. In addition, we found that copper complexed to histidine (Cu-His), but not ionic copper, stimulated LHRH release, the magnitude of which was dependent on the dose of Cu-His. The chelator specificity for Cu complex action was such that Cu-His stimulated LHRH release 4.9-fold and Cu-Cys stimulated release 2.5-fold, whereas neither Cu-Thr, Cu-Gly-His-Lys, Cu-bovine serum albumin, nor ceruloplasmin stimulated LHRH release. Based on these results and those of others indicating that the concentration of copper in hypothalamic axonal terminals is 1-2 orders of magnitude greater than plasma, we propose that copper released in the vicinity of the LHRH neurons interacts with specific sites on the LHRH axonal terminals, which leads to release of the peptide.

Animals↗

PubMed1984

In vivo effect of staphylococcal enterotoxin A on peripheral blood lymphocytes.

Staphylococcal enterotoxin A (SEA) administration to monkeys produced an initial lymphocytic leukopenia lasting approximately 24 h. Lymphocytes isolated from blood circulation (PBL) during this stage had normal or decreased [3H]thymidine incorporating activity. After 48 h, however, a significant increase (five- to sixfold) in [3H]thymidine incorporating activity into PBL was apparent. The peak of incorporating activity (seven- to eightfold) was reached 3 to 4 days after SEA administration, followed by a gradual decline, reaching the baseline after 2 weeks. The increased levels of [3H] thymidine incorporation in PBL were concomitant with the conversion of lymphopenia into lymphocytosis, accompanied by the release of many immature cells into the circulation. Lymphocytes isolated 24 h after SEA administration in vivo did not respond to the mitogenic action of SEA in vitro. Lymphocytes isolated at later stages after SEA challenge were fully activated by toxin. From a series of studies, it was concluded that SEA administered to monkeys caused, during the initial 24 h, the removal of a great proportion of lymphocytes from the circulation, followed by the release of new immature cells with augmented DNA synthesis activity. The lymphocytic leukocytosis state declined gradually and reached normal levels between 3 and 4 weeks after the SEA challenge. The biological implications of the hematological changes occurring after SEA challenge in vivo are discussed.

Animals↗

PubMed1984

Evidence that copper-amino acid complexes are potent stimulators of the release of luteinizing hormone-releasing hormone from isolated hypothalamic granules.

Chelated copper has been previously shown to stimulate the release of LHRH from isolated hypothalamic granules. In this study, we evaluated the chelator specificity, the kinetic constants, and the characteristics of copper interaction with LHRH granules. LHRH granules were isolated from the median eminence area of adult male rats and then incubated in a buffered medium at 37 C. Release of LHRH into the incubation medium was assessed by RIA of LHRH remaining in the granules after incubation. It was found that CuHistidine (CuHis) as well as CuCysteine markedly stimulated LHRH release from the isolated granules, release being 56% and 63%, respectively, of the total LHRH content of granules incubated in buffer alone. In contrast, neither CuGly-His-Lys nor CuBSA stimulated LHRH release. The CuHis-stimulated release of LHRH was a saturable function of the concentration of CuHis. The Michaelis-Menten constants of this release process were estimated; the apparent Km for copper was found to be 4 microM, and the maximal velocity was 65% of the granule content of LHRH released in 5 min. In addition, we noted that CuHis-stimulated release of LHRH, assessed 6 min after CuHis, was completely abolished when dithiothreitol (DTT) was added immediately after CuHis, partially abolished when added 1 or 2 min after CuHis, and not affected at all when added 3 min after CuHis. This time course of DTT inhibition of LHRH release suggests that a period of 2-3 min of copper interaction with the granules is required for the 6-min manifestation of copper action. Furthermore, this DTT-inhibitable interaction of copper did not occur when granules were incubated at 4 C. In summary the findings that copper, chelated to putative circulating chelators, markedly stimulates LHRH release and that the apparent Km of 4 microM for copper in this process is within the concentration range for the physiological action of copper support the proposal that blood-borne copper can interact rapidly with the LHRH granule in an energy-requiring fashion and that, consequent to this interaction, LHRH release occurs.

Animals↗

PubMed1983

Aging-related reduced release of LH-releasing hormone from hypothalamic granules.

The effects of aging on the secretory function of hypothalamic granules containing luteinizing hormone releasing hormone (LHRH) were examined, using chelated copper as a test substance. Granules prepared from old and young female rats were each incubated with various concentrations of CuATP and the kinetic constants of LHRH release estimated. The LHRH content of the hypothalamus of old female rats (3.4 +/- 0.1 ng; mean +/- S.E.; n = 3 determinations) was lower than that of young female rats (4.7 +/- 0.1 ng; n = 3). CuATP-stimulated LHRH release (pg/hypothalamus) from granules isolated from old animals was significantly (p less than 0.001) lower than that from granules isolated from young animals; Vmax being 58.5 +/- 1.6 pg (n = 36) and 76.7 +/- 2.7 pg (n = 34) of LHRH released/min, respectively. In contrast, the fractional amount of CuATP-stimulated release of LHRH from granules isolated from old and young rats was similar; Vmax being 60 +/- 0.5% and 62 +/- 1% of granule LHRH released, respectively. In addition, the apparent Km for CuATP-stimulated release of LHRH from granules of old and young rats was also similar, 5.1 and 5.5 microM copper, respectively. These results are suggestive that aging does not alter the responsiveness of LHRH-containing granules to a secretion-stimulus such as copper but it does alter the capacity (amount) of peptide released.

Adenosine Triphosphate↗

PubMed1983

Two distinct events underlie the MgATP-stimulated release of luteinizing hormone releasing hormone from isolated hypothalamic granules.

We have previously demonstrated that MgATP stimulates the release of luteinizing hormone releasing hormone (LHRH) from isolated hypothalamic granules and that this stimulation has a requirement for monovalent ions. To assess the role of the monovalent ions in the release process, granules (isolated from hypothalami of adult male rats) were incubated in buffered medium containing MgATP in the presence or absence of KCl. When granules were incubated in the absence of KCl, MgATP did not lead to LHRH release. However, when KCl was added to the MgATP-treated granules, a rapid (1 min) release of LHRH occurred. Based on these results we propose that the process of MgATP-stimulated release of LHRH from isolated hypothalamic granules is comprised of two distinct events: a priming event and a release event. During the priming event, MgATP interacts with the LHRH granule which leads to a change in the granule such that the release event, initiated by monovalent ions, can occur.

Adenosine Triphosphate↗

PubMed1983

Copper-induced release of immunoreactive alpha-melanotropin from isolated hypothalamic granules.

Previously, we demonstrated that copper chelates stimulate the release of luteinizing hormone releasing hormone (LHRH) from isolated hypothalamic granules. To assess the generality of the copper-stimulated release process, we determined the effects of copper on the release of immunoreactive alpha-melanotropin (alpha-MSHi) from isolated granules. When granules were incubated with various copper complexes, CuATP stimulated alpha-MSHi release by 54 +/- 6% (mean +/- S.E.), Cu tartarate by 56 +/- 4%, CuBSA by 32 +/- 5% and Cu histidine by 29 +/- 2%. CuATP-stimulated alpha-MSHi release from granules incubated under N2 was 57% of that incubated under air. Furthermore, the reducing agent dithiothreitol (DTT) inhibited CuATP-stimulated alpha-MSHi release (p less than 0.01), whereas oxidized DTT did not do so. Pretreatment of granules with the thiol-blocking reagents iodoacetic acid or 5, 5'-dithiobis-(2-nitrobenzoic acid) inhibited CuATP-stimulated alpha-MSHi release by 52 +/- 3 and 38 +/- 4%, respectively. Thus, chelated copper, rather than ionic copper, is the active form of the metal and the action of copper involves the oxidation of thiols. These data are similar to those previously observed for the copper-stimulated release of LHRH. Hence, the effects of copper on the permeability of granule membranes may be a generalized phenomenon which underlies susceptibility of storage granules to the reduction-oxidation status of the cellular milieu.

Animals↗

PubMed1983

A possible role for copper-mediated oxidation of thiols in the regulation of the release of luteinizing hormone releasing hormone from isolated hypothalamic granules.

A role for copper in the release of luteinizing hormone releasing hormone (LHRH) from hypothalamic neurons has been previously proposed. To elucidate further the mechanism of action of copper, we addressed two questions: (a) what is the active form of copper that interacts with the LHRH granule (ionic or chelated)? and (b) is copper-stimulated LHRH release a result of an interaction of copper with thiol groups and, if so, does it require oxygen? Granules were isolated from hypothalami of adult male rats and were then incubated at 37 degrees C for 3-5 min in a buffered medium. When granules were incubated with various copper complexes, CuATP stimulated LHRH release by 45 +/- 4% (mean +/- SE), copper tartrate by 44 +/- 4%, CuBSA by 27 +/- 7%, and copper histidine by 16 +/- 6%. Neither CuEDTA nor CuCl2 stimulated LHRH release. CuATP-stimulated LHRH release from granules incubated under N2 was 50% of that incubated under air. Furthermore, the CuATP-stimulated release of LHRH was completely inhibited by dithiothreitol or glutathione (10(-3) M each), partially (40-50%) by iodoacetate or 5,5-dithiobis-(2-nitrobenzoic acid), and not at all by oxidized dithiothreitol. Thus, chelated copper, rather than ionic copper, is the active form of the metal, and the action of copper involves an oxidation reaction and granule thiol groups. The precise mechanism of action of copper, however, has yet to be elucidated. We propose that copper may affect LHRH release as follows: copper, bound to an intracellular chelator (protein, peptide, or amino acid), oxidizes thiols of the LHRH granule, leading to a change in granule-membrane permeability and hence to LHRH release.

Adenosine Triphosphate↗

PubMed1983

Acetylation of adrenocorticotropin and beta-endorphin by hypothalamic and pituitary acetyltransferases.

Acetylation of beta-endorphin- and adrenocorticotropin (ACTH)-related peptides was assessed in particulate fractions obtained from the neuro-intermediate lobe and anterior lobe of the pituitary gland and the hypothalamus of adult male rats. All three tissues acetylated ACTH(1-24), whereas only the neuro-intermediate lobe acetylated beta-endorphin and none of the tissues acetylated dynorphin. The apparent Km of the neuro-intermediate lobe enzyme for ACTH(1-24) and beta-endorphin was 1.6 and 22.3 microM, respectively; that of the anterior lobe and hypothalamic enzyme for ACTH(1-24) was 14.6 and 18.9 microM, respectively. N-acetylation appears to be the primary acetylation reaction of the neuro-intermediate lobe and hypothalamic enzyme. When the neuro-intermediate lobe enzyme was incubated in the presence of saturating concentrations of either ACTH(1-24) or ACTH(1-13)NH2 each mixed with beta-endorphin, acetylation of the peptide mixture equalled that seen in the presence of one peptide alone. These results are suggestive that the same neuro-intermediate lobe enzyme acetylates the N-terminal amino acid of both ACTH- and endorphin-related peptides and that this enzyme differs from the hypothalamic acetylating enzyme in its substrate specificity and apparent affinity for ACTH(1-24).

Acetylation↗

PubMed1982

A reduction in the concentration of immunoreactive corticotropin, melanotropin and lipotropin in the brain of the aging rat.

We have previously shown that the hypothalamic concentration of immunoreactive alpha-melanotropin (alpha-MSHi) is markedly lower in the aging female rat than in the young rat. The current view is that alpha-MSH is derived from corticotropin (ACTH), and ACTH, in turn, is derived from a large molecular-weight precursor (pro-opiocortin); pro-opiocortin also serves as the precursor to beta- and gamma-lipotropin (LPH). To ascertain if the age-related reduction in the concentration of alpha-MSHi may be a result of a decline in the production of pro-opiocortin, we determined the content of immunoreactive ACTH (ACTHi), alpha-MSH (alpha-MSHi), gamma-LPH (gamma-LPHi), and protein, in 3 regions of the brain of young (4 months) and old (26-28 months) female rats: the medial basal hypothalamus (MBH, the region containing the perikarya of the ACTH/MSH/LPH neurons), the preoptic anterior hypothalamus (POA), and the thalamus (regions containing axons of these neurons). The concentration of ACTHi, alpha-MSHi (mol/mg protein), or gamma-LPHi (U/mg protein) in the MBH of old rats was 30-50% of that in the MBH of young rats. Moreover, the concentration of ACTHi, alpha-MSHi or gamma-LPHi in the POA and thalamus of old rats was also lower than that in the POA and thalamus of young rats. Based on these findings, we propose that aging causes a reduction in the production of pro-opiocortin in the brain of the female rat and that such a change may be related to the altered function of the brain of the aged.

Adrenocorticotropic Hormone↗

PubMed1982

Molecular-weight profiles of immunoreactive corticotropin in the hypothalamus of the aging rat.

Extracts of the medial basal hypothalamus (MBH), the preoptic anterior hypothalamus (POA), or the pituitary gland of young (4-month-old) and old (18-month-old) female rats were fractionated on columns of Sephadex G-75 superfine. Five forms of immunoreactive corticotropin (ACTHi) were found in the MBH or POA: greater than 40K, 30-40K, 20-30K, 5.7K, and 4.5K. In contrast, 4 forms of ACTHi were found in the pituitary gland: 30-40K, 20-30K, 5.7K, and 4.5K. Thus, hypothalamic tissue contains a large form of ACTH (greater than 40K ACTH) which is not present in the pituitary gland. We tentatively identified greater than 40K ACTH as a large form of pro-opiocortin, 30-40K ACTH as pro-opiocortin, 20-30K ACTH as ACTH biosynthetic intermediate, 5.7K ACTH as glycosylated ACTH 1-39, and 4.5K ACTH as ACTH 1-39. The content of ACTHi in the MBH and POA of old rats was lower than that of young rats. Nevertheless, regardless of the age of the animals, the fractional amount of 30-40K ACTHi was high in the MBH (a region that includes the presumed site of biosynthesis of pro-opiocortin) compared to that in the POA (a region that is distant to the site of biosynthesis of pro-opiocortin). Moreover, the reduced fractional amount of 30-40K ACTHi in the POA was associated with an increased fractional amount of greater than 40K, 20-30K, 5.7K, and 4.5K ACTHi. These findings are consistent with a precursor-product relationship between the 30-40K ACTHi and 20-30K ACTHi, 5.7K ACTHi, and 4.5K ACTHi.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenocorticotropic Hormone↗

PubMed1982

MgATP-stimulated release of luteinizing hormone releasing hormone from isolated hypothalamic granules: evidence for a selective requirement for monovalent cations.

The purpose of this study was to characterize the kinetics and monovalent ion requirements of the MgATP-stimulated release of luteinizing hormone releasing hormone (LHRH) from isolated hypothalamic granules. LHRH granules were obtained from homogenates of hypothalami of adult male rats by differential centrifugation and then incubated in buffered media containing KCl. Under these conditions, the pH optimum for the MgATP-stimulated release of LHRH was between 7.5 and 8.0; the rate of release was linear for the first 5 min of incubation; and the magnitude of the release was a saturable function of KCl concentration. When the potassium in the incubation mixture was replaced with various monovalent cations (Li+, Na+, Cs+, choline, or tetraethylammonium), the magnitude of the MgATP-stimulated release of LHRH was inversely related to the size of the cation; release in the presence of Li+ was 12%, whereas release in the presence of tetraethylammonium was 0% of the total LHRH. When the chloride in the incubation mixture was replaced with various monovalent anions (Br-, I-, or isethionate), the magnitude of the MgATP-stimulated release of LHRH was not related to the size of the anion. Moreover, inclusion of anion transport inhibitors in the incubation mixture did not affect LHRH release, suggesting that anion transport is not required for the MgATP-stimulated release of LHRH. In summary, we suggest that the MgATP-stimulated release of LHRH from isolated hypothalamic granules involves an enzymatic event and that this release process does not have a selective requirement for monovalent anions, but has a selective requirement for monovalent cations.

Adenosine Triphosphate↗

PubMed1982

Comparison of the effects of ATP, Mg2+, and MgATP on the release of luteinizing hormone-releasing hormone from isolated hypothalamic granules.

The effect of ATP, Mg2+, or MgATP on the release of luteinizing hormone-releasing hormone (LH-RH) from hypothalamic granules was examined under in vitro conditions. Granules, isolated from adult make hypothalami, were incubated at 37 degrees C in a buffered (pH 7.8) medium containing 0.15 M-KCl. The addition of ATP to the incubation mixture did not stimulate the release of LH-RH. In contrast, the addition of MgATP stimulated the release of LH-RH, the release being 62% greater than control. The addition of Mg2+ to the incubated granules also stimulated the release of LH-RH. However, the magnitude of this Mg2+-stimulated release of LH-RH was significantly (P less than 0.01) lower than that of the MgATP-stimulated release, indicating that ATP stimulates LH-RH release in a Mg2+-dependent manner. As both MgATP and Mg2+ alone stimulated LH-RH release, we characterized further these two release processes by incubating the granules under one of the following conditions: incubation at 4 degrees C in a buffered medium containing 0.15 M-KCl or incubation at 37 degrees C in a medium that does not contain KCl. Under these two incubation conditions, the MgATP-stimulated release of LH-RH was not manifested, whereas the Mg2+-stimulated release of LH-RH was manifested. On the basis of these differences, we propose that two different processes can lead to the release of LH-RH from isolated hypothalamic granules: one process involves ATP and Mg2+ (MgATP) and another process involves Mg2+ alone.

Adenosine Triphosphate↗