scholarly journals Effect of single-chain ovine gonadotropins with dual activity on ovarian function in sheep

Reproduction ◽  
2014 ◽  
Vol 148 (2) ◽  
pp. 129-136 ◽  
Author(s):  
Heloisa M Rutigliano ◽  
Betty M Adams ◽  
Albina Jablonka-Shariff ◽  
Irving Boime ◽  
Thomas E Adams
Keyword(s):  
Ovarian Function ◽  
Single Injection ◽  
Corpora Lutea ◽  
Single Chain ◽  
Comparable Amount ◽  
Free Protein ◽  
Preovulatory Follicles ◽  
Ovulatory Follicles ◽  
The Impact ◽  
Follicle Maturation

We examined the half-life and biological activity of two single-chain proteins that combined portions of ovine FSH and LH. We proposed the hypothesis that these chimeric proteins would display LH and FSH activities and would promote follicle maturation in ewes. Estrus activity was synchronized using progestogen-impregnated vaginal pessaries. To negate the impact of endogenous LH and FSH, animals received serum-containing antibodies against GNRH 1 day before pessary removal (PR). At PR sheep (five animals per group) received a single injection (10 IU/kg, i.v.) of either the ovine-based (oFcLcα) gonadotropin analog, an ovine-based analog containing oLHβ truncated at the carboxyl terminus (oFcL(ΔT)cα), or a human-based gonadotropin analog (hFcLcα). Control animals received a comparable amount of gonadotropin-free protein. Ovulation was induced 3 days after PR using human chorionic gonadotropin (1000 IU, i.v.). Ovaries were collected 11 days after PR. Neither estradiol (E2) or progesterone (P4) production, development of preovulatory follicles or corpora lutea (CL) were noted in control animals receiving gonadotropin-free protein. Significant increase in the synthesis of E2 and P4 was noted in sheep receiving the dually active gonadotropin analogs. The number of CLs present 11 days after PR was significantly increased in sheep receiving the chimeric glycoproteins compared with control animals. The magnitude of the secretory and ovarian responses did not differ between hFcLcα and oFcLcα or between oFcLcα and oFcL(ΔT)cα. Immunoactivity of LH and FSH was low in control animals, but was significantly elevated in sheep receiving the gonadotropin analogs. In conclusion, ovine-based gonadotropin analogs are functionally active in sheep and a single injection is adequate to induce the development of multiple ovulatory follicles.

199 The effect of different doses of follicle-stimulating hormone during superovulation on ovarian function in dairy cattle

2019 ◽  
Vol 31 (1) ◽  
pp. 224
Author(s):  
K. Karl ◽  
F. Jimenez-Krassel ◽  
E. Gibbings ◽  
K. E. Latham ◽  
J. J. Ireland
Keyword(s):  
Ovarian Reserve ◽  
Ovarian Function ◽  
Prostaglandin F2α ◽  
Antral Follicle ◽  
Ovulation Rate ◽  
Corpora Lutea ◽  
Ovulatory Follicle ◽  
Study Results ◽  
Ovulatory Follicles ◽  
Higher Doses

Cattle with an unknown antral follicle count (AFC) during follicular waves respond to different FSH doses during superovulation in a curvilinear fashion with the highest doses decreasing or not increasing ovulatory follicle number, number of transferable embryos, and number of corpora lutea (CL). These observations support the hypothesis that heifers treated with different FSH doses during superovulation will reach a superovulation maximum (SOVmax) and that doses exceeding SOVmax are excessive, resulting in a decrease in ovarian function. To test this hypothesis, we used eight 12-month-old Holstein heifers with a low AFC (<10 follicles ≥3mm in diameter, index for small ovarian reserve). These 8 heifers were subjected to 4 different superovulations with a 21-day intervals between each superovulation. During each superovulation, which began on Day 1 of a synchronized oestrous cycle before selection of a dominant follicle, 2 of the 8 heifers were treated (twice-daily FSH injections×4 days) with 20, 40, 80, or 120mg of Folltropin-V (Vetoquinol, Lure, France). At the end of the study, the same 8 heifers had been superovulated 4 times with each of the 4 FSH doses. To enhance ovulation rate in response to hCG, each superovulated heifer was injected with 3 different injections of prostaglandin F2α (PGF2α) spaced 12h apart to induce luteolysis of the original CL. Two PGF2α injections were given after each FSH injection on the last day of FSH treatment and the last PGF2α injection was given after the hCG (2500IU) injection, which was 12h after the last FSH injection. Daily ultrasonography was used to measure AFC and number of ovulatory follicles and CL during the study. Results of t-test analysis showed that at time of hCG, AFC was greater (P>0.05) for heifers treated with the 40-mg (mean±s.e.m., 27±3.7) v. 20-mg (17.9±2.5) FSH dose, but the response to higher FSH doses (27.9±4.4; 27±3.3) did not differ from the response to the 40-mg dose. Number of ovulatory follicles was greater (P<0.05) for heifers treated with the 40-mg (17.5±2.3) v. 20-mg (10.1±1.4) FSH dose, but the response to higher doses (18.9±3.3; 17.3±2.4) did not differ from the response to the 40-mg dose. At Day 7 post-hCG, CL number was greater (P<0.05) for heifers treated with the 40-mg (13.8±2.9) v. 20-mg (7.4±1.4) dose, but response to higher doses (9.9±3.2; 9.1±2.3) did not differ from the response to the 40-mg dose. Ovulation rate (ovulatory follicle number divided by CL number) was greater (P<0.05) for heifers treated with the 40-mg (79±0.08%) v. 80-mg (52±0.09%) or 120-mg (53±0.09%) dose, but similar to that of heifers treated with 20-mg (73±0.09%) FSH dose. These results support the conclusion that the 40-mg FSH dose achieves SOVmax and FSH doses greater than SOVmax in cattle with a low AFC and small ovarian reserve are excessive and detrimental to ovarian function. This project was supported by Agriculture and Food Research Initiative Competitive Grant no. 2017-67015-26084 from the USDA National Institute of Food and Agriculture (Washington, DC).

scholarly journals Ovarian function in ewes after treatment with mifepristone early during the oestrous cycle

Reproduction ◽  
2003 ◽  
pp. 205-210 ◽  
Author(s):  
EM Paslay ◽  
U Salli ◽  
F Stormshak ◽  
Keyword(s):  
Ovarian Function ◽  
Venous Blood ◽  
Oestrous Cycle ◽  
Corpora Lutea ◽  
Blood Samples ◽  
Treatment Groups ◽  
Significant Difference ◽  
Preovulatory Follicles ◽  
Prostaglandin F ◽  
The Mean

The aim of this study was to determine whether endogenous progesterone regulates synthesis and secretion of luteal oxytocin. In Expt 1, mature ewes (n = 5 per group) were assigned randomly to control or mifepristone (RU486) treatment groups. Ewes were injected s.c. twice a day with vehicle or 10 mg RU486 on days 5-7 of the oestrous cycle (oestrus = day 0). On day 8, after an i.v. injection with prostaglandin F(2alpha) (250 microg cloprostenol), venous blood samples were collected at frequent intervals to determine plasma oxytocin concentrations. Plasma oxytocin concentrations of RU486-treated ewes were not significantly different from those of control ewes. In Expt 2, ewes were injected s.c. each day with vehicle or 175 mg RU486 on days 2-5 of the oestrous cycle followed by administration of prostaglandin F(2alpha) on day 6. Four of five RU486-treated ewes showed 'split-oestrus' (oestrous behaviour for 36 h and then again at 84-108 h after the onset of initial oestrus). There was no significant difference in mean plasma oxytocin or progesterone concentrations between treatment groups. The mean masses of mature corpora lutea from control and RU486-treated ewes on day 6 of the oestrous cycle did not differ significantly (394.8 +/- 28.8 versus 319.5 +/- 48.3 mg). RU486-treated ewes contained mature corpora lutea, new corpora lutea (two of four ewes) and preovulatory follicles (>or= 10 mm, two of four ewes). The average interoestrous interval for RU486-treated ewes was 9 days more than that for control animals (26.2 +/- 2.9 versus 17 +/- 0.5 days; P < 0.025).

THE »EARLY-ANDROGEN« SYNDROME; DIFFERENCES IN RESPONSE TO PRE-NATAL AND POST-NATAL ADMINISTRATION OF VARIOUS DOSES OF TESTOSTERONE PROPIONATE IN FEMALE AND MALE RATS

1964 ◽  
Vol 47 (1) ◽  
pp. 37-50 ◽  
Author(s):  
H. E. Swanson ◽  
J. J. van der Werff ten Bosch
Keyword(s):  
Critical Period ◽  
Ovarian Function ◽  
Testosterone Propionate ◽  
Single Injection ◽  
Male Rats ◽  
Male Rat ◽  
High Dose ◽  
Corpora Lutea ◽  
Female Rat ◽  
Time Of Administration

ABSTRACT The interaction between dose and time of administration of testosterone propionate (TP) on the development of sexual function was studied by giving a single dose of 5, 10, 50 or 500 μg TP to young rats of both sexes on the day of birth (day 1) or on day 2, 4 or 5. The effectiveness of androgen administration before birth was studied by giving a single injection of 2500 μg TP to pregnant rats on day 19 to 22 after conception. Pre-natal administration had no effect on the function of the ovaries of female offspring, although the dose was sufficient to cause masculinization of the external genitalia. The weight of the testes and accessories of the male offspring were not affected. The effects of post-natal TP administration on ovarian function varied with the dose and with the time of administration. Threshold doses (5 and 10 μg) were more effective the earlier they were given after birth. With these small doses, most of the rats had normal luteinized ovaries at 10 weeks and were able to bear and suckle normal litters. Some time later ovulations ceased so that at 21 weeks they were no longer fertile; at 27 weeks there were no more corpora lutea in the ovaries. In males, a dose of 50 μg of TP or more resulted in permanently reduced weight of testes, seminal vesicles and prostate. The earlier the treatment, the more marked was the depression of weight. From these results and others reported in the literature the following deductions can be made: (1) the critical period of brain sensitivity to physiological amounts of androgen probably lies between days 4 and 6 (day of birth counted as day 1); (2) a rough estimate of the amount of androgen secreted by the newborn male rat during the critical period would seem to be the equivalent of a single injection of 5–50 μg TP; (3) after the physiological critical period has elapsed a female rat can still be »masculinized« if a high dose of TP is given, up to a period of between 10–20 days after birth.

scholarly journals Subfertility with Defective Folliculogenesis in Female Mice Lacking Testicular Orphan Nuclear Receptor 4

2008 ◽  
Vol 22 (4) ◽  
pp. 858-867 ◽  
Author(s):  
Lu-Min Chen ◽  
Ruey-Sheng Wang ◽  
Yi-Fen Lee ◽  
Ning-Chun Liu ◽  
Yu-Jia Chang ◽  
...  
Keyword(s):  
Nuclear Receptor ◽  
Ovarian Function ◽  
Corpora Lutea ◽  
Orphan Nuclear Receptor ◽  
Estrous Cycles ◽  
Female Mice ◽  
Preovulatory Follicles ◽  
Direct Binding ◽  
Normal Spermatogenesis ◽  
Functional Analyses

Testicular orphan nuclear receptor 4 (TR4) plays essential roles for normal spermatogenesis in male mice. However, its roles in female fertility and ovarian function remain largely unknown. Here we found female mice lacking TR4 (TR4−/−) displayed subfertility and irregular estrous cycles. TR4−/− female mice ovaries were smaller with fewer or no preovulatory follicles and corpora lutea. After superovulation, TR4−/− female mice produced fewer oocytes, preovulatory follicles, and corpora lutea. In addition, more intensive granulosa apoptosis was found in TR4−/− ovaries. Functional analyses suggest that subfertility in TR4−/− female mice can be due to an ovarian defect with impaired folliculogenesis rather than a deficiency in pituitary gonadotropins. Molecular mechanism dissection of defective folliculogenesis found TR4 might induce LH receptor (LHR) gene expression via direct binding to its 5′ promoter. The consequence of reduced LHR expression in TR4−/− female mice might then result in reduced gonadal sex hormones via reduced expression of enzymes involved in steroidogenesis. Together, our results showed TR4 might play essential roles in normal folliculogenesis by influencing LHR signals. Modulation of TR4 expression and/or activation via its upstream signals or unidentified ligand(s) might allow us to develop small molecule(s) to control folliculogenesis.

The ovary and sexual maturation of the brain

Development ◽  
1967 ◽  
Vol 17 (1) ◽  
pp. 1-10
Author(s):  
W. N. Adams Smith
Keyword(s):  
Corpus Luteum ◽  
Direct Effect ◽  
Sexual Maturation ◽  
Ovarian Function ◽  
Testosterone Propionate ◽  
Single Injection ◽  
Female Rats ◽  
Corpora Lutea ◽  
Litter Mate ◽  
The Brain

Pfeiffer (1935, 1936) reported the induction of constant oestrus in female rats following the transplantation of testes from litter-mate males just after birth and noted that the ovaries of these animals did not contain corpora lutea. These changes remained after removal of the testis transplants. The same effects were obtained by Bradbury (1941) following the administration of multiple doses of testosterone propionate. Barraclough & Leathern (1954) found that a single injection of 1 mg of testosterone propionate at 5 days of age led to permanent sterility in female mice, with no corpus luteum formation in their ovaries. Similar results were obtained in rats by Barraclough (1961) with the administration of a single injection of 1·25 mg of testosterone propionate. This permanent change in ovarian function does not appear to be a direct effect upon the ovary (Bradbury, 1941).

scholarly journals Single-chain human gonadotropin analogs induce follicle development in sheep

2007 ◽  
Vol 196 (3) ◽  
pp. 593-600 ◽  
Author(s):  
Elyse P Lemke ◽  
Betty M Adams ◽  
Albina Jablonka-Shariff ◽  
Irving Boime ◽  
Thomas E Adams
Keyword(s):  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
Single Injection ◽  
Corpora Lutea ◽  
Follicle Development ◽  
Serum Concentrations ◽  
Single Chain ◽  
Ovarian Weight ◽  
Treatment Groups ◽  
Basal Serum

The biopotency of single-chain analogs of human hFSH, human chorionic gonadotropin (hCG), and a dually active gonadotropin construct (FcCGβα) was examined. Sheep (bwt=61.4±1.1 kg; n=6 ewes/treatment) received a single injection (5 IU/kg, i.v.) of the hFSH analog (Fcα), the hCG analog (CGβα), FcCGβα, or Fcα and CGβα. Control animals received conditioned media. Ovulation was induced 3 days after analog administration using hCG (1000 IU, i.v.). Basal serum concentrations of estradiol (E2) were maintained in control animals. Neither Fcα nor CGβα alone induced significant E2 production during the pre-hCG period. Conversely, serum concentrations of E2 were increased (P<0.05) 2 days after administration of FcCGβα or Fcα+ CGβα. Although P4 concentrations were maintained at basal levels in control animals, significant increase was noted in all other treatment groups during the post-hCG period. Final ovarian weight was significantly increased (P<0.05) in animals receiving Fcα, Fcα+ CGβα, or FcCGβα, but not CGβα alone. Most of the ovarian enlargement was attributed to the formation of corpora lutea. Collectively, these observations demonstrate that the single-chain analogs of the human gonadotropins are active in sheep. The construct with singular FSH activity supports follicle development but not E2 production. Conversely, the construct that incorporates β-domains from both CG and FSH has dual activity. The long-lived nature of the single-chain constructs suggests that these recombinant gonadotropins may be effective alternatives to pituitary- or placenta-derived gonadotropins in out-of-season breeding and/or superovulation protocols.

Neonatal testosterone exposure induces early development of follicular cysts followed by sympathetic ovarian hyperinnervation

2016 ◽  
Vol 28 (11) ◽  
pp. 1753 ◽  
Author(s):  
Gabriel Anesetti ◽  
Rebeca Chávez-Genaro
Keyword(s):  
Ovarian Function ◽  
Sympathetic Innervation ◽  
Temporal Association ◽  
Ovarian Activity ◽  
Corpora Lutea ◽  
Neonatal Rats ◽  
Ovarian Morphology ◽  
The Impact ◽  
Cyst Development

This study analysed the temporal association between ovarian cyst development induced by neonatal androgenisation and sympathetic innervation. Neonatal rats (postnatal Days 1 to 5) were treated with testosterone or dihydrotestosterone and the effects were evaluated at postnatal Days 20, 40, 90 or 180. Ovulation rate, number of cystic follicles and density of sympathetic fibres were analysed. The effects of surgical denervation or gonadotrophin stimulation were also assessed. Rats exposed to testosterone showed no oestrous cycle activity and did not ovulate, maintaining a polycystic ovarian morphology at all ages studied. Also, a significant increase in ovarian density of noradrenergic fibres was detected at postnatal Days 90 and 180. Sympathectomy was unable to re-establish ovarian activity; however, human chorionic gonadotrophin stimulation was enough to induce ovulation. The impact of dihydrotestosterone on ovarian function was less noticeable, showing the coexistence of corpora lutea and cystic structures without changes in sympathetic innervation. Our findings suggest that a remodelling of ovarian sympathetic innervation occurs as a response to modifications in the pattern of follicular growth induced by testosterone. A role of sympathetic innervation in the maintenance of the polycystic condition is suggested.

THE »EARLY-ANDROGEN« SYNDROME; ITS DEVELOPMENT AND THE RESPONSE TO HEMI-SPAYING

1964 ◽  
Vol 45 (1) ◽  
pp. 1-12 ◽  
Author(s):  
H. E. Swanson ◽  
J. J. van der Werff ten Bosch
Keyword(s):  
Sexual Maturity ◽  
Compensatory Growth ◽  
Testosterone Propionate ◽  
Single Injection ◽  
Weight Increase ◽  
Corpora Lutea ◽  
Transition Stage ◽  
Adult Females

ABSTRACT The »early-androgen« syndrome in the rat – i. e. anovulatory ovaries in adult females after a single injection of testosterone propionate (TP) within a week of birth – may not become apparent until some time after the attainment of sexual maturity. Large doses (50 or 100 μg) of TP were effective earlier than lower doses (5 or 10 μg). Rats which received 5 μg TP were ovulating at 10 weeks of age, mated but were infertile at 13 weeks of age, and were anovulatory at 21 weeks. In rats between 10 and 13 weeks old there was a marked fall in the number of corpora lutea in the ovaries of animals which had been given 5 μg TP. Hemi-spaying was followed by compensatory growth of the remaining ovary which consisted of corpora lutea in ovulating, and of follicles in anovulatory rats; little or no compensatory weight increase occurred in animals which seemed to be in the transition stage from the ovulatory to the anovulatory condition.

scholarly journals A rodent model of human dose-equivalent progestin-only implantable contraception

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Heather C. M. Allaway ◽  
Roger A. Pierson ◽  
Jesse Invik ◽  
Susan A. Bloomfield
Keyword(s):  
Repeated Measures ◽  
Ovarian Function ◽  
Virgin Female ◽  
Rodent Model ◽  
High Dose ◽  
Corpora Lutea ◽  
Average Dose ◽  
Estrus Cycle ◽  
Human Dose ◽  
Dose Dependent

Abstract Background Long-acting, reversible contraceptives (LARC; progestin only) are an increasingly common hormonal contraceptive choice in reproductive aged women looking to suppress ovarian function and menstrual cyclicity. The overall objective was to develop and validate a rodent model of implanted etonogestrel (ENG) LARC, at body size equivalent doses to the average dose received by women during each of the first 3 years of ENG subdermal rod LARC use. Methods Intact, virgin, female Sprague-Dawley rats (16-wk-old) were randomized to 1 of 4 groups (n = 8/group) of ENG LARC (high-0.30μg/d, medium-0.17μg/d, low-0.09μg/d, placebo-0.00μg/d) via a slow-release pellet implanted subcutaneously. Animals were monitored for 21 days before and 29 days following pellet implantation using vaginal smears, ultrasound biomicroscopy (UBM), saphenous blood draws, food consumption, and body weights. Data were analyzed by chi-square, non-parametric, univariate, and repeated measures 2-way ANOVA. Results Prior to pellet implantation there was no difference in time spent in estrus cycle phases among the treatment groups (p > 0.30). Following pellet implantation there was a dose-dependent impact on the time spent in diestrus and estrus (p < 0.05), with the high dose group spending more days in diestrus and fewer days in estrus. Prior to pellet insertion there was not an association between treatment group and estrus cycle classification (p = 0.57) but following pellet implantation there was a dose-dependent association with cycle classification (p < 0.02). Measurements from the UBM (ovarian volume, follicle count, corpora lutea count) indicate an alteration of ovarian function following pellet implantation. Conclusion Assessment of estrus cyclicity indicated a dose-response relationship in the shift to a larger number of acyclic rats and longer in duration spent in the diestrus phase. Therefore, each dose in this model mimics some of the changes observed in the ovaries of women using ENG LARC and provides an opportunity for investigating the impacts on non-reproductive tissues in the future.

scholarly journals Solubility assessment of single-chain antibody fragment against epithelial cell adhesion molecule extracellular domain in four Escherichia coli strains

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Fatemeh Sadat Javadian ◽  
Majid Basafa ◽  
Aidin Behravan ◽  
Atieh Hashemi
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Epithelial Cell ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Extracellular Domain ◽  
Epithelial Cell Adhesion Molecule ◽  
Single Chain ◽  
E Coli ◽  
The Impact

Abstract Background Overexpression of the EpCAM (epithelial cell adhesion molecule) in malignancies makes it an attractive target for passive immunotherapy in a wide range of carcinomas. In comparison with full-length antibodies, due to the small size, the scFvs (single-chain variable fragments) are more suitable for recombinant expression in E. coli (Escherichia coli). However, the proteins expressed in large amounts in E. coli tend to form inclusion bodies that need to be refolded which may result in poor recovery of bioactive proteins. Various engineered strains were shown to be able to alleviate the insolubility problem. Here, we studied the impact of four E. coli strains on the soluble level of anti-EpEX-scFv (anti-EpCAM extracellular domain-scFv) protein. Results Although results showed that the amount of soluble anti-EpEX-scFv obtained in BL21TM (DE3) (114.22 ± 3.47 mg/L) was significantly higher to those produced in the same condition in E. coli RosettaTM (DE3) (71.39 ± 0.31 mg/L), and OrigamiTM T7 (58.99 ± 0.44 mg/L) strains, it was not significantly different from that produced by E. coli SHuffleTM T7 (108.87 ± 2.71 mg/L). Furthermore, the highest volumetric productivity of protein reached 318.29 ± 26.38 mg/L in BL21TM (DE3). Conclusions Although BL21TM (DE3) can be a suitable strain for high-level production of anti-EpEX-scFv protein, due to higher solubility yield (about 55%), E. coli SHuffleTM T7 seems to be better candidate for soluble production of scfv compared to BL21TM (DE3) (solubility yield of about 30%).

Sign in / Sign up

Export Citation Format

Share Document