MEASUREMENT OF RAT LH WITH A DOUBLE-ANTIBODY SOLID-PHASE RADIOIMMUNOASSAY: EFFECT OF LH-RH AND OF TESTOSTERONE OENANTHATE IN CASTRATED ANIMALS

1975 ◽  
Vol 78 (3) ◽  
pp. 451-460 ◽  
Author(s):  
J. S. E. Dericks-Tan ◽  
Hans-Dieter Taubert
Keyword(s):  
Solid Phase ◽  
Test System ◽  
Subsequent Treatment ◽  
Male Rats ◽  
Ventral Prostate ◽  
Phase Method ◽  
Double Antibody ◽  
Serum Lh ◽  
Solid Phase Radioimmunoassay ◽  
Lh Rh

ABSTRACT A double-antibody solid-phase radioimmunoassay for the measurement of rat LH was described. The use of a solid-phase method for the second incubation resulted in a considerable saving of time as compared to the conventional double-antibody method. The sensitivity and accuracy of the method were not affected by this modification. It was found that there was a cross-reaction between the NIAMD-anti-rat-LH-serum-1 used and NIAMD-rat-FSH-RP-1. This indicates that the test system does not specifically measure rat LH but gonadotrophic activity. The RIA was tested under different experimental conditions. In male rats, the effect of castration and of subsequent treatment with testosterone oenanthate upon serum LH, and upon the weight of the seminal vesicles, the ventral prostate, and the levator ani, was examined. Serum LH rose rapidly after castration and reached 2 to 4 weeks after orchidectomy a maximum (8–9 fold increase). Subsequent treatment with testosterone oenanthate caused a rapid decrease of serum LH. The accessory sex organs showed reciprocal changes of weight. In oophorectomized rats whose pituitaries had been blocked with oestradiol monobenzoate and progesterone, a linear log-dose response of plasma LH was found after administration of various doses of synthetic LH-RH.

POTENCY ESTIMATES OF HUMAN GONADOTROPHINS BY A BIOASSAY AND THREE IMMUNOASSAY METHODS

1971 ◽  
Vol 67 (3) ◽  
pp. 417-433 ◽  
Author(s):  
C. Robyn ◽  
M. L'Hermite ◽  
P. Petrusz ◽  
E. Diczfalusy
Keyword(s):  
Biological Activity ◽  
Solid Phase ◽  
Haemagglutination Inhibition ◽  
Reproductive Organs ◽  
Male Rats ◽  
Equilibrium System ◽  
Antibody Technique ◽  
Double Antibody ◽  
Human Menopausal Gonadotrophin ◽  
Solid Phase Radioimmunoassay

ABSTRACT Human chorionic gonadotrophin (HCG), human menopausal gonadotrophin (HMG) and human hypophysial gonadotrophin (HHG) preparations were assayed by a bioassay and three immunoassay methods. The bioassay method was based on the increase in weight of the accessory reproductive organs of immature male rats. The immunoassays included haemagglutination inhibition, a radioimmunoassay using the double antibody technique and a solid phase radioimmunoassay using coated tubes. The same anti-HCG serum was employed in all immunoassays. When relatively crude HCG preparations (< 6000 IU/mg) were assayed, there was a highly significant (P < 0.001) correlation between the estimates obtained by all methods. However, when highly purified HCG preparations (> 6000 IU/mg) were assayed, the bioassay gave significantly higher potency estimates than the immunoassays. Furthermore, the estimates obtained by radioimmunoassays were significantly higher than those obtained by haemagglutination inhibition. However, the results obtained by the two radioimmunoassays showed a highly significant (P < 0.001) correlation. The regression coefficient of the line relating the logit transform of the response to loge, of the dose was significantly higher (b =−1.700±0.7322)) in assays employing the double antibody technique (a non-equilibrium system) than in solid phase (b =−0.945 ± 0.116) immunoassays (an equilibrium system). When highly purified HCG preparations were assayed by the double antibody technique, the regression coefficients were significantly higher than those obtained with the standard preparations. In relation to their biological activity, HMG preparations contained much less immunological activity, than HHG preparations. In addition, the regression lines obtained with HCG, HMG and HHG preparations showed significant deviations from parallelism when estimated by radioimmunoassays. The precision and sensitivity of the four assay methods increased in the following order: bioassay, haemagglutination inhibition, solid phase radioimmunoassay and radioimmunoassay involving the use of the double antibody technique. It is concluded that in relation to their biological activity, marked differences exist in the immunological activities of highly purified and relatively crude HCG preparations. In addition, the immunological activities of HCG, HMG and HHG preparations are dissimilar, when immunoassayed by the use of an anti-HCG serum.

EFFECT OF ACTINOMYCIN D ON THE PITUITARY RESPONSE TO LH-RH

1976 ◽  
Vol 81 (1) ◽  
pp. 73-81 ◽  
Author(s):  
Jesus A. Vilchez-Martinez ◽  
Akira Arimura ◽  
Andrew V. Schally
Keyword(s):  
Initial Phase ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Male Rats ◽  
Continuous Stimulation ◽  
Immature Male ◽  
Serum Lh ◽  
Pre Treatment ◽  
Lh Rh

ABSTRACT The effect of Actinomycin D (Act D) on the release of LH and FSH induced by LH-RH was investigated in rats. Immature male rats received an iv infusion over a period of 3–4 h or a quick iv injection of synthetic LH-RH. Infusion of LH-RH significantly increased serum LH and FSH levels at 1, 2, 3 and 4 h after the initiation of infusion. Pre-treatment with 100 μg/100 g b. w. Act D failed to affect the rise of serum LH and FSH levels 1 h after the infusion but significantly suppressed the response at 2, 3 and 4 h. The increase in serum LH and FSH levels after a quick injection of LH-RH was unaffected by pre-treatment with Act D whether the antibiotic was injected 1 or 2 h before LH-RH. The results suggest that the initial phase of the pituitary response to LH-RH does not require DNA-dependent RNA synthesis, whereas that in the later period does. RNA synthesis may be necessary only to maintain the increased secretion of both LH and FSH during a continuous stimulation with LH-RH.

Effect of short-term starvation on reproductive hormone gene expression, secretion and receptor levels in male rats

1989 ◽  
Vol 121 (3) ◽  
pp. 409-417 ◽  
Author(s):  
M. Bergendahl ◽  
A. Perheentupa ◽  
I. Huhtaniemi
Keyword(s):  
Male Rats ◽  
Ventral Prostate ◽  
Male Rat ◽  
Testicular Function ◽  
Mrna Levels ◽  
Short Term ◽  
Α Subunit ◽  
Gnrh Receptors ◽  
Androgen Synthesis ◽  
Serum Lh

ABSTRACT The effects of 4–6 days of food deprivation on the pituitary-testicular function of adult male rats were studied. Fasting decreased body weights on average by 23% (P<0·01) and those of seminal vesicles by 55% (P<0·01) in 4 days. No consistent changes were found in testicular and ventral prostate weights. The pituitary levels of gonadotrophin-releasing hormone (GnRH) receptors decreased by 50% (P<0·01). Serum and pituitary levels of LH, FSH and prolactin decreased by 25–50% (P<0·01 for all). Testicular and serum levels of testosterone decreased by 70–80%, testicular LH receptors by 26%, those of prolactin by 50% (P<0·01 for all), but those of FSH remained unaffected. Acute (2 h) stimulation by a GnRH agonist (buserelin, 10 μg/kg i.m.) resulted in similar LH, FSH and testosterone responses in the fasted and control animals, and human chorionic gonadotrophin (hCG) stimulation (30 IU/kg i.m.) in similar increases in testosterone. A 42% decrease was found in pituitary content of mRNA of the common α subunit (P<0·05), but the mRNAs of the LH- and FSH-β chains and prolactin were unaffected by fasting for 4 days. When the same mRNAs were measured after 6 days of fasting, the decrease of the mRNA of FSH-β also became significant (50%, P<0·01). In contrast, the mRNA of LH-β was increased twofold (P<0·01) at this time and serum LH levels were similar in control and starved animals. It is concluded that during short-term starvation of male rats: (1) the decrease in gonadotrophin and prolactin synthesis and secretion is first noticed on the level of translation (protein synthesis), and the mRNA levels of these hormones may respond more slowly to starvation, (2) decreased pituitary GnRH receptors indicate decreased GnRH release from the hypothalamus, (3)the gonadotrophin and prolactin loss results secondarily in decreased testicular androgen synthesis and LH and prolactin receptor levels, (4) no decrease occurs during starvation in acute gonadotrophin response to GnRH, or testicular testosterone response to hCG, (5) the primary response to starvation in male rat pituitary-testicular function is the loss of normal hypothalamic support of gonadotrophin and prolactin secretion, rather than direct nutritional effects on the pituitary and testis, and (6) when starvation is continued beyond 4 days, a recovery is seen in pituitary mRNA on the LH-β chain and in serum LH, most probably because the starvation-associated decrease serum testosterone is a more potent positive stimulus of LH synthesis than the direct hypothalamic-pituitary inhibition. Journal of Endocrinology (1989) 121, 409–417

HYPOTHALAMIC-PITUITARY-TESTICULAR SYSTEM FOLLOWING TESTICULAR X-IRRADIATION

1976 ◽  
Vol 83 (1) ◽  
pp. 190-200 ◽  
Author(s):  
H. L. Verjans ◽  
K. B. Eik-Nes
Keyword(s):  
Interstitial Cell ◽  
Serum Testosterone ◽  
Male Rats ◽  
Ventral Prostate ◽  
Testis Weight ◽  
X Ray ◽  
Serum Lh ◽  
Testicular Weight ◽  
Unknown Factor ◽  
X Irradiation

ABSTRACT Testes of adult, male rats were exposed to a total dose of 1500 R of X-irradiation. Testicular weight decreased from day 8 after X-ray treatment. This decrease was, however, preceded by an increment of the testis weight on day 4 following treatment. X-ray treatment of testes was associated with significant increases in serum FSH. Testicular irradiation had, however, no effect on ventral prostate and seminal vesicles weights. Serum testosterone increased only on day 1, 2 and 4 after irradiation, while serum LH levels tended to increase from day 8 post-irradiation. These changes were not significant, however, when compared with non-irradiated controls. At 7, 13 and 20 days following 1500 R of bilateral, testicular X-irradiation, the hypothalamic-pituitary unit was still capable of responding to exogenous gonadotrophin releasing factor. Serum FSH may in male rats be regulated at least partly by circulating steroids of testicular origin and partly by an unknown factor of non-interstitial cell nature.

scholarly journals A Rapid Solid-Phase Radioimmunoassay for Human Plasma Follicle-Stimulating Hormone

1980 ◽  
Vol 17 (1) ◽  
pp. 38-44
Author(s):  
AC Lovesey
Keyword(s):  
Human Plasma ◽  
High Precision ◽  
Reference Values ◽  
Plasma Levels ◽  
Solid Phase ◽  
Follicle Stimulating Hormone ◽  
Diagnosis And Management ◽  
Double Antibody ◽  
Solid Phase Radioimmunoassay ◽  
Stimulating Hormone

The measurement of plasma levels of human follicle-stimulating hormone (FSH) has proved to be of value for the study of the hypothalamic-hypophyseal-gonadal axis, greatly facilitating the diagnosis and management of problems relating to the menopause and infertility. In the present work a solid-phase radioimmunoassay for human FSH has been developed. This system is characterised by high precision, is economical, and is considerably faster to operate than conventional double antibody systems used in the hospital assay service. Reference values for plasma FSH in various endocrine states are recorded and discussed.

Basal levels of prolactin in goat blood measured throughout a 24-h period by a rapid double antibody–solid phase radioimmunoassay

1973 ◽  
Vol 40 (2) ◽  
pp. 235-245 ◽  
Author(s):  
I. C. Hart
Keyword(s):  
Solid Phase ◽  
Virgin Female ◽  
Double Antibody ◽  
Lactating Goats ◽  
Highly Sensitive ◽  
Solid Phase Radioimmunoassay ◽  
Circulating Levels

SummaryThe development of a rapid, highly sensitive double antibody–solid phase (DASP) radioimmunoassay for ovine/caprine prolactin is described. The assay was applied to the measurement of basal levels of prolactin in the blood of 4 anoestrous lactating female, 4 anoestrous virgin female and 4 castrated male goats. For several hours after milking there was a chronic gradually decreasing release of prolactin in the lactating goats, such that the circulating levels of prolactin were, for 14–16 h of the day, at a much higher level than those in the virgin female or castrated male goats. The basal levels of prolactin in the castrated male goats (87–200ng/ml) were somewhat higher than those in the virgin females (47–77ng/ml). The effects of feeding and stress on prolactin levels are discussed.

Double-antibody radioimmunoassay of thyroid microsomal antibody in serum.

1981 ◽  
Vol 27 (1) ◽  
pp. 39-42 ◽  
Author(s):  
V T Kung ◽  
P M Weber ◽  
L V dos Remedios
Keyword(s):  
Solid Phase ◽  
Sodium Deoxycholate ◽  
Published Data ◽  
Health Examination ◽  
Standard Curve ◽  
Double Antibody ◽  
Microsomal Antibody ◽  
Solid Phase Radioimmunoassay ◽  
Chloramine T ◽  
Antimicrosomal Antibody

Abstract We describe a double-antibody radioimmunoassay for antimicrosomal antibodies in serum. Antigen in centrifuged pellets of microsome is solubilized by treatment with sodium deoxycholate solution and radiolabeled (125I] by the Chloramine T method. A 10-microL aliquot of human serum or antimicrosomal antibody standard is incubated overnight at room temperature with 125I-labeled microsome. Bound radiolabeled microsome is separated by precipitation with goat antihuman IgG antiserum. The percentage of 125I-labeled microsome bound by each serum is calculated, and its concentration of antimicrosomal antibody interpolated from a standard curve. The prevalence of abnormal antimicrosomal antibody was 16% in a self-referred asymptomatic or not acutely ill group of patients having a periodic multiphasic health examination. Values for patients with certain thyroid diseases generally agreed with other published data. We conclude that our assay is sensitive, precise, and accurate, and is more efficient for routine measurement of antimicrosomal antibody than the currently used solid-phase radioimmunoassay.

scholarly journals Assay of Australia Antigen Employing Double-Antibody and Solid-Phase Radioimmunoassay Techniques

Kanzo ◽  
1974 ◽  
Vol 15 (7) ◽  
pp. 427-432
Author(s):  
Shinichi KAKUMU ◽  
Shozo ITO ◽  
Sumihiko OKUYAMA ◽  
Seikichi INADA ◽  
Katsuhiko TAGO
Keyword(s):  
Solid Phase ◽  
Australia Antigen ◽  
Double Antibody ◽  
Solid Phase Radioimmunoassay

THE INFLUENCE OF TIME, DOSE AND GONADAL STEROIDS ON LH-RH-INDUCED GONADOTROPHIN RELEASE IN RATS

1975 ◽  
Vol 78 (4) ◽  
pp. 695-704 ◽  
Author(s):  
Wolfgang Lotz
Keyword(s):  
Time Course ◽  
Time Lapse ◽  
Male Rats ◽  
Ovariectomized Rats ◽  
Response Curves ◽  
Intact Male ◽  
Maximal Increase ◽  
Serum Lh ◽  
Lh Rh ◽  
Lh Releasing Hormone

ABSTRACT Hypophyseal responses to LH-releasing hormone (LH-RH) were studied in ovariectomized rats, pre-treated once with 50 μg oestradiol-3-benzoate alone or in combination with 25 mg progesterone either 16, 24, 48 or 72 h before the administration of LH-RH. The highest serum gonadotrophin increase was detected in ovariectomized rats pre-treated with oestrogen 48 h before the LH-RH injection. The serum LH concentration change in intact male rats was markedly lower in terms of order of magnitude than in ovariectomized, oestradiol-3-benzoate, progesterone-treated rats ("O. Oe. P."-rats). These latter test animals showed the maximal increase of serum LH concentration, depending on the dose of LH-RH, after 10 to 25 min and that of FSH after 40 min. The dose-response curves for LH release in "O. Oe. P."-rats were linear for short (10 and 15 min) and sigmoid for longer time intervals (20, 30 and 40 min) between LH-RH injection and sacrifice. The possible reasons for the difference in both magnitude and time-lapse for maximal increase of serum LH and FSH concentration, the influence of steroid pre-treatment and the variation of the time-course of serum LH concentration, depending on the dose of LH-RH, are discussed.

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