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Biomedical subjects

R L Matteri

Publications and source records attributed to R L Matteri.

At least 55 records · Page 3Linked to original sources

Relationships of serum insulin-like growth factor I concentrations to growth, composition, and reproductive traits of swine.

Insulin-like growth factor I (IGF-I) is a peptide hormone that has been shown to be involved in metabolic regulation of growth and reproduction in livestock species. The objectives of this study were to quantify concentrations of IGF-I in growing pigs and determine whether IGF-I concentration can be used as a predictor of growth, composition, and reproductive traits. Forty male and 60 female pigs, divided equally between two locations, were weighed and bled at 3-wk intervals from 6 to 21 wk of age. At each sampling, two blood samples were collected via jugular venipuncture at an interval of at least 1 h. Serum was separated and IGF-I concentration determined via RIA. Pigs were weighed at each sampling date. Backfat and longissimus muscle area were measured with the use of B-mode ultrasound and adjusted to 100 kg. Age at puberty and first-parity litter size were measured on gilts. Effects of age, sex, location, and pig within sex x location on log-transformed IGF-I concentrations were determined by analyzing data as a split-plot. Performance traits were fitted to a model including the effects of IGF-I concentration, sex, location, and interactions. The IGF-I concentrations increased (P < .05) from 3 to 18 wk of age before dropping at 21 wk of age. Concentrations increased more rapidly in males than in females and differed significantly between sexes from 12 to 21 wk of age. Repeatability of IGF-I concentration was .29 +/- .02; IGF-I concentrations of samples collected at 6 wk were not correlated with those at later ages. Correlations between IGF-I concentrations of samples at later ages ranged from .27 to .51. Heritability of IGF-I concentration was .27 +/- .07. There was a tendency for weight to be affected by a sex x age interaction (P = .09). Weight of boars exceeded weight of gilts only at 21 wk (111.4 +/- 1.1 vs 107.1 +/- .8 kg). Regressions of weight on IGF-I concentrations were positive at all ages but greatest at 6 wk. The IGF-I concentration did not affect backfat thickness, longissimus area, percentage of lean, age at puberty, or litter size.

Aging↗

Relationship between in vitro somatotroph function and growth in three-week-old barrows.

The relationship between in vitro somatotroph function and growth was examined in piglets demonstrating a continuous range of growth characteristics. Twenty barrows were sacrificed at 3 weeks of age for the collection of pituitary tissue and blood. Pituitary cells from each animal were cultured and exposed to vehicle (culture medium); .1, 1, and 10 nM growth hormone-releasing hormone (GHRH); 2 mM 8-Br-cAMP (cAMP); 100 nM phorbol myristate acetate (PMA); and 59 mM KCl. All secretagogue treatments stimulated growth hormone (GH) secretion (p < .0001). Basal and stimulated GH secretion in culture, intracellular GH content (icGH), and serum insulin-like growth factor-1 (IGF-1) concentrations were all positively correlated with 3-week weight gain (p < .05). Concentrations of GH in the serum sample taken at sacrifice were not related to growth (p > .3). Intracellular GH content was correlated with in vitro GH secretion (p < .01) and serum IGF-1 concentrations (p < .001). Somatotroph function was contrasted in the 7 largest and 7 smallest piglets (large, 8.3 +/- .3 kg, n = 7; small, 4.5 +/- .2 kg, n = 7). Treatment with GHRH produced a dose-related increase in GH secretion in both experimental groups (p < .0001). No significant size x GHRH interaction was detected (p = .09), When contrasted with the small group, the large group demonstrated elevated GH secretion in culture (p < .01), icGH content (p < .001), and circulating IGF-1 (p < .001). The results of this study raise the possibility of a functional relationship between porcine somatotroph secretory activity and growth, mediated by IGF-1, which may be regulated by the quantity of GH available for release.

Animals↗

Somatotroph, lactotroph and thyrotroph function in three-week-old gilts reared in a hot or cool environment.

The influence of the thermal environment on the ability of the pituitary gland to secrete growth hormone (GH), prolactin (PRL), and thyroid-stimulating hormone (TSH) was examined in gilts reared under hot (H: 27-32 degrees C, 50-90% RH, n = 6) or cool (C: 20 degrees C, 50% RH, n = 6) conditions. Piglets were sacrificed at 3 wks of age. Pituitary cells from each animal were cultured and exposed to vehicle (culture medium); 1, 1, and 10 nM thyrotropin-releasing hormone (TRH); .1, 1, and 10 nM growth hormone-releasing hormone (GHRH); 59 mM KCl; 2 mM 8-Br-cAMP; and 100 nM phorbol myristate acetate. Rearing in the H, compared to the C, environment increased plasma PRL concentrations (p < .001), in vitro PRL secretion subsequent to all secretagogue treatments (p < .001), and cellular PRL content (p < .001). The stimulated release of TSH in culture was reduced (p < .001), but cellular TSH content was increased (p < .05) by exposure to the H environment. The total amount of TSH in culture (secreted + cellular) was not affected by thermal environment. The release of GH in vitro, cellular GH content, total GH in culture, and plasma GH concentrations were similar between H and C groups. The only dose-response curves that differed in slope between thermal groups were those produced by the TSH response to TRH (p < .001). The results of this study suggest that chronic exposure to a hot environment can 1) enhance PRL secretion by a mechanism which affects the quantity of releasable PRL rather than lactotroph sensitivity to secretagogues and 2) reduce TSH secretion by inhibiting thyrotroph secretory response to stimulation.

8-Bromo Cyclic Adenosine Monophosphate↗

Somatotroph and lactotroph function in relation to growth in six-week-old pigs reared in a hot or cool environment.

The influences of thermal environment and individual growth rate on somatotroph and lactotroph function were examined in 6-week-old barrows reared entirely in a hot (H: 27-32 degrees C, n = 8) or cool (C: 21 degrees C, n = 10) environment. Growth hormone (GH) and prolactin (PRL) cell contents and responses to growth hormone-releasing hormone (GHRH) or thyrotropin-releasing hormone (TRH) were evaluated in cultured pituitary cells from each animal. Plasma GH, PRL, and insulin-like growth factor-1 (IGF-1) concentrations also were monitored. Thermal environment did not affect in vitro GH secretion, cellular GH content, or plasma GH concentrations. Stimulated in vitro GH release (GHRH-basal) and plasma GH were inversely related to average daily gain (ADG, r = -.76, p < .005 and r = -.51, p < .05, respectively). Cellular GH content also declined as ADG increased (r = -.57, p < .05). Plasma IGF-1 concentrations were not affected by thermal environment and were not related to ADG. Pituitary cells from H animals secreted and contained more PRL than cells from C animals (p < .05). Plasma PRL values were correlated with ADG (r = .54, p < .05), but did not differ between thermal groups. Stimulated in vitro PRL (TRH-vehicle) secretion was positively related with ADG only in the H group (r = .97, p < .001). In contrast, cellular PRL content decreased with ADG in cells from the H barrows (r = -.8, p < .05). Lactotroph function was not related to growth in cells from C pigs. In summary, 1) heat enhanced PRL secretion and cell content; 2) growth and somatotroph function were inversely related; and 3) serum PRL and the PRL response to TRH in cells from H barrows were positively related to growth.

Animals↗

Luteinizing hormone synthesis in cultured fetal human pituitary cells exposed to gonadotropin-releasing hormone.

To investigate whether GnRH regulates LH synthesis during human development, pituitary cells from second trimester fetuses were incubated with [35S]methionine ([35S]met) and [3H]glucosamine ([3H]gln) for 48 h with 0, 10(-9), and 10(-7) mol/L GnRH. Immunoassayable (i) LH was measured in media and cellular lysates, and dual label scintillation analysis was used to quantitate incorporation of radiolabeled precursors into cells, trichloroacetic acid-precipitable proteins, and immunoprecipitated LH subjected to electrophoresis. Exposure of cells to GnRH did not affect cellular uptake or incorporation of precursors into proteins, but specifically increased total (secreted plus cellular) LH synthesis. Both GnRH concentrations significantly increased iLH release and enhanced secreted and cellular [3H]gln-LH. The secretion of [35S] met-LH was stimulated only by 10(-7) mol/L GnRH. The proportion of newly synthesized LH that was secreted and the 3H/35S ratio of secreted and cellular LH were uninfluenced by GnRH. Although basal LH synthesis was not sex dependent, total iLH content and GnRH-stimulated LH translation were greater in cells from females than in those from males. Therefore, GnRH regulates LH synthesis by second trimester fetal human gonadotrophs without influencing the proportion of total radiolabeled LH that is secreted. The existence of a sex difference in total iLH content and GnRH-stimulated LH translation is consistent with the sexual dimorphism in pituitary LH content occurring during human development.

Cells, Cultured↗

Association between surges of follicle-stimulating hormone and the emergence of follicular waves in heifers.

The effects of ablation of a dominant follicle and treatment with follicular fluid on circulating concentrations of follicle-stimulating hormone (FSH) were studied and the temporal relationships between surges of FSH and follicular waves were studied in heifers with two or three follicular waves/interovulatory interval. Cauterization of the dominant follicle on Day 3 or Day 5 (ovulation on Day 0) (six control and six treated heifers/day) resulted in a surge (P less than 0.05) in FSH beginning the day after cautery. The FSH surge prior to wave 2 (first post-treatment follicular wave) occurred 4 days (Day 3 cautery) and 2 days (Day 5 cautery) before the surge in control groups, corresponding to a 4-day and a 2-day advance in emergence of wave 2 compared with controls. It was concluded that the dominant follicle on Day 3 and Day 5 was associated with the suppression of circulating FSH concentrations. Heifers (n = 4/group) were untreated or treated intravenously with a proteinaceous fraction of bovine follicular fluid on Days 0-3, 3-6, or 6-11. Concentrations of FSH were suppressed (P less than 0.05) for the duration of treatment, regardless of the days of treatment. Cessation of treatment was followed within 1 day by the start of a surge in FSH. The FSH surge prior to wave 2 occurred 2 days earlier (treatment on Days 0-3), 1 day later (treatment on Days 3-6), and 6 days later (treatment on Days 6-11) than in controls, corresponding to an equivalent advance or delay, respectively, in the emergence of wave 2 compared with controls. The results suggest that the effects of exogenous follicular fluid on follicular development were mediated, in whole or in part, by altering plasma FSH concentrations. Control heifers combined for the two experiments were separated into those with 2-wave (n = 11) or 3-wave (n = 5) interovulatory intervals. Two-wave heifers had two FSH surges and 3-wave heifers had three apparent FSH surges during the interovulatory interval. Results of the cautery and follicular fluid experiments indicated that a surge in FSH necessarily preceded the emergence of a wave. The FSH surges in treated and control heifers began 2-4 days before the detectable (ultrasound) emergence of a follicular wave (follicles of 4 and 5 mm), peaked 1 or 2 days before emergence and began to decrease approximately when the follicles of a wave begin to diverge into a dominant follicle and subordinate follicles (follicles 6-7 mm).

Animals↗

Effect of progesterone on ovarian follicles, emergence of follicular waves and circulating follicle-stimulating hormone in heifers.

The hypothesis was tested that greater growth of the dominant follicle of wave 1 (first follicular wave of an interovulatory interval), compared with that of subsequent anovulatory waves, is due to lower circulating concentrations of progesterone during the growing phase of the follicle. Control heifers (n = 6) were compared with heifers (n = 6) treated with a decreasing dose of progesterone from day 0 to day 5 (ovulation = day 0). Maximum diameter (12.7 +/- 0.9 versus 15.3 +/- 0.7 mm) and mean diameter of the dominant follicle of wave 1, averaged over days, were smaller (P < 0.05) in the progesterone-treated than in the control group. Progesterone treatment did not suppress circulating follicle-stimulating hormone (FSH); but the second FSH surge was earlier, resulting in earlier emergence of wave 2 as indicated by a tendency (P < or = 0.1) for group x day interactions attributed to earlier detection of the dominant follicle and an earlier rise in the total number of follicles detected. The stated hypothesis was supported. We also tested the hypothesis that exposure to low circulating concentrations of progesterone at the end of the growing phase of the anovulatory dominant follicle of wave 1 results in continued growth and prolonged maintenance of the dominant follicle. Heifers (n = 6 per group) were given a luteolytic dose of prostaglandin F2 alpha (PGF2 alpha) on day 6 and treated with a low (30 mg day-1), physiological (150 mg day-1), or high (300 mg day-1) dose of progesterone on days 6 to 20. Continued periodic emergence of anovulatory follicular waves occurred (2.1 +/- 0.0 waves, 2.8 +/- 0.2 waves, 3.8 +/- 0.3 waves, respectively; P < 0.05) until treatment was stopped (interovulatory intervals: 26.2 +/- 1.0, 30.8 +/- 0.6 and 40.3 +/- 1.7 days, respectively; P < 0.05). Compared with the physiological dose group, the growth of the dominant follicle was inhibited to a lesser degree in the low-dose group since it grew for longer (P < 0.05) and to a larger diameter (P < 0.05), and persisted for longer (P < 0.05). Prolonged dominance of this oversized (> 20 mm) follicle was associated with delayed emergence of wave 2. The hypothesis was supported. Results also showed that the high dose of progesterone suppressed the dominant follicle more than the physiological dose when given during the growing phase, but not when given after the growing phase.(ABSTRACT TRUNCATED AT 400 WORDS)

Analysis of Variance↗

Regulation of the biopotency of primate luteinizing hormone by gonadotropin-releasing hormone in vitro and in vivo.

Gonadotropin biological/immunological (B/I) ratios have proven to be valuable indicators of the biopotencies of LH and FSH. Observations of rapidly changing LH B/I have been made which suggest the existence of a readily mobilized pool of highly bioactive pituitary gonadotropins. To test this hypothesis, we have examined the role of GnRH in the regulation of LH B/I in vivo and in vitro. The rhesus monkey was used as a model due to its many physiological similarities with the human. A rapid elevation in circulating LH B/I was observed following GnRH administration to male monkeys that was sustained for at least 2 h (15 min; p less than 0.05). The administration of 1 or 10 nM GnRH to cultured pituitary cells was found to significantly increase the B/I of secreted, but not intracellular, LH (p less than 0.05). In unstimulated controls, the B/I of intracellular LH was higher than that of secreted LH (p less than 0.05). These findings are consistent with the notion that a pool of highly active LH exists within the gonadotrophs in primates. One way that GnRH may regulate the bioactivity of circulating LH is by rapidly mobilizing this gonadotropin pool.

Animals↗

Microheterogeneity of equine follicle-stimulating hormone.

The present report demonstrates the microheterogeneity of equine follicle-stimulating hormone (eFSH), and its subunits, and characterizes eFSH isoforms with respect to receptor-binding and immunological properties. The isoelectric microheterogeneity of eFSH was demonstrated by the isolation of two highly purified hormone preparations (eFSH-A and eFSH-B) from different cation-exchange chromatography fractions (Sephadex SP-C50). Equine FSH-A eluted before eFSH-B, indicating a greater apparent negative charge. This apparent charge difference was substantiated by subsequent analysis by chromatofocusing. These two hormone preparations were then analyzed by FSH radioreceptor assay (RRA) and eFSH radioimmunoassay (RIA). The FSH receptor-binding/immunological activity (RRA/I) ratio of eFSH-A was significantly greater than that of eFSH-B (p less than 0.05). No significant differences between the two FSH preparations were detected by RIA. To study eFSH microheterogeneity in more detail, a standard eFSH preparation was analyzed by the technique of chromatofocusing. A variety of immunoreactive forms were observed in the range of pH 7 to 14, with a large amount of material eluting at less than pH 4.0. Chromatofocusing fractions corresponding to pI values of 6.6, 6.4, 5.8, 5.5, 5.2, 4.8, 4.6, 4.3, 4.1, and less than 4.0 were analyzed by RRA and RIA. The respective RRA/I ratio values were 0.35, 0.61, 0.82, 1.16, 1.23, 1.70, 1.73, 1.46, 1.09, and 1.55. Values that differ by more than 0.2 are significantly different (p less than 0.05, Student-Newman-Keuls multiple comparison). This is in agreement with the data obtained with eFSH-A and -B, which showed the more negatively charged preparation possessed the higher RRA/I activity ratio.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Characterization of a monoclonal antibody which detects luteinizing hormone from diverse mammalian species.

The present study describes the development and characterization of a monoclonal antibody (518B7) generated against bovine LH (bLH). Although 518B7 was extremely specific for LH, very low species specificity was observed. A RIA using this antibody and radioiodinated equine LH (eLH) showed good sensitivity for all mammalian LH preparations tested, with the exception of human LH (15%, relative to the eLH reference standard). Activities of most mammalian LH's ranged between approximately 50-200%. Much less activity was detected with reptilian LH (less than 1.5%). Amphibian and avian LH fractions were essentially inactive. The reactivities of LH alpha and beta subunits from a variety of mammals clearly showed that the antibody reacts with the beta subunit. Sensitive RIAs were also developed utilizing 125I-bovine and 125I-rat LH. Interestingly, all hormone preparations which showed sufficient reactivity for statistical analysis within the dose ranges used in the present study (0.01-1000 ng/tube) produced a displacement curve parallel to the reference standard. We have also validated the use of 518B7 in detecting LH in serum. Parallel dilution curves relative to purified LH reference standards were observed with equine and bovine serum samples and equine pituitary extract. High (average 94%) recoveries were also seen with bovine serum with known amounts of exogenously added bLH. Similar patterns of LH secretion were detected with a RIA based upon 125I-bLH and 518B7 and a previously described polyclonal antibody-based RIA in bovine serum samples during estrus. Thus, a monoclonal antibody for LH has been produced which can be used to develop sensitive and specific RIAs in many different mammalian species.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Characterization of equine luteinizing hormone by chromatofocusing.

Three equine luteinizing hormone (LH) preparations (eLH-A, -B, and -C) recently have been isolated in our laboratory and were shown to differ in average basicity (eLH-A greater than -B greater than -C). The present study further characterizes these preparations by chromatofocusing. Each of these preparations are comprised of a family of isohormones, with 5 major immunoreactive peaks in the pH range of 7 to 4 (approx. pIs = 6.6, 6.1, 5.7, 5.2, and 4.8), with varying amounts of material eluting to either side of the pH gradient. Although similar isoforms are seen in all three LH preparations, the relative proportions of different isoforms vary in a manner reflecting the average charge properties of eLH-A, -B, and -C. While eLH-A contains predominantly basic forms, eLH-C consists largely of acidic material, and eLH-B is composed mostly of isohormones with pIs intermediate to eLH-A and -C. Chromatofocusing of a crude extract from a single horse pituitary gland revealed isohormone peaks corresponding to those found in the highly purified LH preparations. Peak fractions of the various isoforms were used to generate a variety of activity ratios (LH bioactivity:LH radioimmunoassay (RIA), LH radioreceptorassay (RRA):LH RIA, LH bioactivity:LH RRA, follicle-stimulating hormone (FSH) RRA:LH RIA, and FSH RRA:LH RRA activity ratios). The LH bioactivity:LH receptor binding potency ratio showed a linear increase with increasing isohormone acidity (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Biological and immunological properties of zebra pituitary gonadotropins: comparison with horse and donkey gonadotropins.

Previous studies from this laboratory have described the properties of purified luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from horse and donkey anterior pituitary glands. The present study afforded the opportunity to further characterize these previously purified hormone preparations and to compare them with enriched gonadotropin fractions from zebra pituitary glands. Although a single LH and FSH fraction was usually obtained for each pool of pituitaries, two separate zebra LH and two donkey FSH preparations were generated. Purified hormone preparations from the horse were designated eLH and eFSH. Preparations zLH-A, zLH-B, and zFSH were obtained from zebra pituitaries, and fractions dLH, dFSH-A, and dFSH-B were prepared from donkey pituitary glands. These preparations were analyzed by LH and FSH radioimmunoassays (RIAs), radioreceptor assays (RRAs), LH bioassay, and chromatofocusing. Clear immunological differences were observed between equid gonadotropins. Homologous RIAs for eLH and eFSH did not cross-react similarly, or in a parallel fashion, with gonadotropins from the donkey and zebra. In contrast, RIAs capable of assessing LH or FSH in a wide number of species showed all equid gonadotropin preparations to have considerable activity and to produce parallel dilution curves. Relative to eLH (1.00), zLH-A was found to have higher LH bioactivity:LH RIA (2.50), LH RRA:LH RIA (1.42), and LH bioactivity: LH RRA (2.21) activity ratios. The dLH and zLH-B fractions only differed from eLH in LH RRA:LH RIA activity (0.69 and 0.62, respectively). Only LH from the horse possessed clear intrinsic FSH-receptor-binding activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Isolation and characterization of three forms of luteinizing hormone from the pituitary gland of the horse.

Three isoforms of equine luteinizing hormone (eLH-A, eLH-B and eLH-C) have been isolated from horse pituitary glands. Separation was achieved on the basis of charge heterogeneity by ion-exchange chromatography. These charge differences were apparent after final purification, as determined by electrophoretic mobility on polyacrylamide disc gels (RF = 0.14, 0.19 and 0.26 for eLH-A, -B and -C, respectively). Apparent size differences were also noted between the isohormones by gel filtration on Sephadex G-100. Ve/Vo ratios for eLH-A, -B and -C were 1.72, 1.54 and 1.47, respectively. All 3 isoforms were found to contain an equivalent amount of hexose (9.0-9.2%). Isohormones eLH-B and eLH-C, however, possess more sialic acid than eLH-A (6.6-6.7%, vs. 4.5%). The eLH-A and eLH-B preparations contain a similar amount of hexosamine, which is slightly lower than the amount of eLH-C (8.8-9.1% vs. 11.2%). No differences were noted between the isohormones by rat Leydig cell LH bioassay, equine testis LH radioreceptor assay (RRA) or calf testis follicle-stimulating hormone (FSH) RRA. Slight, but nonsignificant, variations were noted between preparations in an eLH radioimmunoassay (RIA). Although chemical variations were detected between the eLH isoforms, no significant differences were observed in in vitro biological and immunological activities. The differences detected in sialic acid content raises the possibility that differences in in vivo clearance rates may exist.

Animals↗

Adrenocorticotropin-induced changes in ovine pituitary gonadotropin secretion in vitro.

In vitro pituitary perifusion experiments were conducted to examine the effect of ACTH and related peptides on basal and GnRH-stimulated gonadotropin release. Treatments of 5 X 10(-7) M ACTH-(1-39), ACTH-(1-24), or ACTH-(18-39) were examined for their ability to influence basal gonadotropin secretion and the subsequent response to a 10(-9)- or 10(-8) M GnRH challenge. Administration of the 1-39 or 18-39 peptide sequences of ACTH similarly stimulated the release of LH and FSH (P less than 0.01). ACTH-(1-24) had no effect on basal gonadotropin secretion. Pretreatment with ACTH-(1-39) inhibited the LH and FSH responses to 10(-9) and 10(-8) M GnRH (P less than 0.05). Suppression of the LH response to 10(-8) M GnRH (P less than 0.05) and the FSH response to 10(-9) M GnRH (P less than 0.05) was observed after ACTH-(1-24) treatment. The administration of ACTH-(18-39) had no significant effect on GnRH-induced gonadotropin release. PRL concentrations were not affected by any of the ACTH peptides. Exposure to 10(-10) M GnRH or 5 X 10(-7) M synthetic ACTH-(1-39) produced an equivalent stimulation of LH secretion. GnRH pretreatment enhanced (P less than 0.05), while ACTH-(1-39) diminished (P less than 0.05), the subsequent response to GnRH. The GnRH receptor antagonist [D-pGlu1, D-Phe2, D-Trp3,6]GnRH attenuated the LH and FSH responses to GnRH and ACTH-(1-39) (P less than 0.05). The results obtained in this study indicate that certain portions of the ACTH molecule may affect gonadotropin secretion, perhaps by interacting with the GnRH receptor.

Adrenocorticotropic Hormone↗

Histamine and increased ovarian blood flow mediate LH-induced superovulation in the cyclic hamster.

Constant infusion of LH (400 micrograms NIH-S24) through an osmotic minipump inserted on Day 1 (oestrus) of the cycle in the hamster resulted in spontaneous superovulation (approximately equal to 29 ova) at the next expected oestrus, increased blood flow (P less than 0.001) to the ovary on Day 3, and slight depletion (0.1 greater than P greater than 0.05) of histamine in the ovary. Treatment with antihistamine (alpha-fluoromethylhistidine, an irreversible inhibitor of histidine decarboxylase, or cimetidine, an H2 blocker) by injections or by infusion using separate osmotic minipumps significantly (P less than 0.01) reduced the number of ova shed in the LH-treated hamsters. Infusion of LH with alpha-fluoromethylhistidine in the same osmotic minipump reduced the bioactivity of the LH. Infusion of antihistamine alone did not alter the normal number of ova shed. The results suggest that the LH-induced superovulation involves stimulation of histamine release; the histamine than may increase ovarian blood flow thus allowing more gonadotrophins to reach the ovary.

Animals↗

Deglycosylation of gonadotropins with an endoglycosidase.

A commercially available endoglycosidase (N-glycanase, Genzyme, Boston, Mass.) purified from Flavobacterium meningosepticum with a specificity for cleaving asparagine-linked carbohydrate moieties in glycoproteins was tested on several pituitary and chorionic gonadotropins as substrates. All intact hormones tested were resistant to the action of the enzyme as were all beta subunits from the respective gonadotropins. All alpha subunits, however, were susceptible to the enzyme as evidenced by a decrease in molecular size when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Preparative experiments with ovine luteinizing hormone subunit (oLH alpha) indicated that only 35-40% of the carbohydrate was removed after N-glycanase treatment, suggesting that perhaps only one of the two carbohydrate moieties was cleavable under the conditions employed. The enzyme-modified subunit (DG-oLH alpha) was able to recombine with untreated oLH beta. An in vitro steroidogenic bioassay (rat Leydig cell) showed that the recombinant (DG-oLH alpha-oLH beta) was about 22% as potent as the native oLH, but in a testicular membrane binding assay for LH, it was equal in potency to the native hormone in competing with the radioligand.

Animals↗

Stress or acute adrenocorticotrophin treatment suppresses LHRH-induced LH release in the ram.

This study examined the effects of varying treatment durations and doses of adrenocorticotrophin (ACTH) as well as restraint stress on the LH response to exogenous LHRH. Injection of 80 i.u. of a concentrated ACTH preparation at 11, 6, 3 or 1.5 h before LHRH administration was effective in suppressing the LH response. Injection of 40, but not 20 or 10, i.u. ACTH 3 h before an LHRH challenge inhibited the magnitude of the LH response, while cortisol values did not vary between ACTH doses. Injection of 200 micrograms of synthetic ACTH1-24 also resulted in a reduced LH response when given 3 h before LHRH. Restraint stress caused elevated corticosteroid levels and reduced LHRH responsiveness. The results of this study suggest that stress may cause an inhibition of pituitary gland ability to respond to LHRH by way of an hormonal component of the adrenocortical axis. A glucocorticoid-independent mechanism may be involved.

Adrenal Cortex Hormones↗