Combined treatment with oestradiol benzoate, d-cloprostenol and oxytocin permits cervical dilation and nonsurgical embryo recovery in ewes

The high variability of responses to multiple ovulation and embryo transfer (MOET) programs is related to several factors, including the ovarian population present at the beginning of hormonal protocols. Thus, our objective was to evaluate the effect of the presence or absence of corpus luteum (CL) in females presenting small antral follicles (SF) at the beginning of a progesterone (P4)-based oestrus synchronization protocol on superovulatory response and embryo yields. The study was conducted during September and October in northeast Brazil (03°40′26″S and 40°14′20″W) using 15 adult Santa Inês ewes, in which the ovarian population present were SF (≤3.75mm, n=5) or SF plus CL (SF+CL, n=10) at the beginning of P4-based oestrus synchronization protocol. The females received a new or reused intravaginal device of progesterone (CIDR Eazi-breed, Zoetis) on Day 0, which remained for 9 days. On Day 7, the pFSH (133mg, Folltropin V, Vetoquinol) treatment was begun, with 6 decreasing doses (25, 25, 15, 15, 10, and 10%) injected IM at 12-h intervals. On Day 9, two equal doses of D-cloprostenol were injected at a 12-h interval (37.5μg, Prolise, Agener União). Flunixin meglumine (24.9 mg; Banamine, MSD Animal Health) was administered IM on Days 13 to 15. All ewes showed oestrus and were mated with fertile rams. On Day 16 non-surgical embryo recovery (NSER) was performed after cervical dilation using D-cloprostenol and oestradiol benzoate at 16h and ocytocin at 20min before. Transrectal B-mode ultrasound evaluations (Z5 Vet, Mindray) were performed on Day 0 and just before NSER to evaluate the ovarian population present. Data are showed in mean±s.d. and compared by t-test for independent samples (P<0.05). The number of CL present on NSER day (21.5±4.4 and 4.0±8.5) and the number of viable embryos (11.1±14.9 and 4.0±8.5) differed (P<0.05) between SF+CL ewes and SF ewes, respectively. However, the number of luteinizing unovulated follicles present on NSER day (4.5±2.5 and 12.4±7.8) and the number of recovered structures (i.e. embryos plus unfertilized eggs; 14.2±15.6 and 4.2±8.2) did not differ (P>0.05) between SF+CL ewes and SF ewes, respectively. In conclusion, the presence of a CL at the beginning of the oestrous synchronization protocol used for superovulation treatment affects responses to the MOET program. Financial support for this study was provided by Embrapa (02.13.06.026.00.02 and 02.13.06.026.00.04) and FAPEMIG (PPM 00201-17).

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FURTHER STUDIES ON THE INFLUENCE OF OESTROGENS ON ANDROGEN DEPENDENT FRUCTOSE FORMATION IN SEX ACCESSORY ORGANS

Acta Endocrinologica ◽

10.1530/acta.0.0530455 ◽

1966 ◽

Vol 53 (3) ◽

pp. 455-461 ◽

Cited By ~ 4

Author(s):

John A. Thomas ◽

Edward T. Knych

Keyword(s):

Combined Treatment ◽

Seminal Vesicles ◽

Synergistic Action ◽

Initial Injection ◽

Oestradiol Benzoate ◽

Anterior Prostate

ABSTRACT Testosterone alone had a stimulating effect on the fructose formation in castrate mice, but the combined treatment of testosterone and oestrogen caused significant reductions in anterior prostate levels of this carbohydrate. Oestrogens were more effective in counteracting the action of testosterone when injected early rather than late after castration. Ethynyl oestradiol and oestradiol benzoate were more effective in counteracting testosterone than other oestrogens studied. There was a greater reduction in fructose levels when lower doses of injected testosterone were simultaneously administered with oestrogen(s). In the seminal vesicles a synergistic action between testosterone and various oestrogens on fructose levels was commonly observed, though antagonism is also evident. Increasing the period of time between castration and initial injection appeared to enhance the synergistic actions of the two hormones.

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TESTOSTERONE AND OESTRADIOL SUPPRESSION OF LH AND FSH IN ADULT MALE RATS: DURATION OF CASTRATION, DURATION OF TREATMENT AND COMBINED TREATMENT

Acta Endocrinologica ◽

10.1530/acta.0.0730011 ◽

1973 ◽

Vol 73 (1) ◽

pp. 11-21 ◽

Cited By ~ 29

Author(s):

R. S. Swerdloff ◽

P. C. Walsh

Keyword(s):

Adult Male ◽

Testosterone Propionate ◽

Combined Treatment ◽

Male Rats ◽

Low Doses ◽

Duration Of Treatment ◽

Serum Lh ◽

Oestradiol Benzoate ◽

Hypothalamic Pituitary Axis ◽

Androgen Effect

ABSTRACT The effects of androgens and oestrogens on serum LH and FSH in castrated rats were evaluated with regard to the modifying influences of duration of castration, duration of treatment and combined oestrogen-androgen effect. Serum LH was not greatly influenced by these variables. In contrast, serum FSH was shown to be more resistant to suppression by both steroids after at least five days of castration, requiring a longer duration of treatment to be suppressed to intact levels. Combined treatment of submaximally suppressive doses of testosterone propionate and oestradiol benzoate resulted in no additive effect on lowering serum FSH. Low doses of both androgens and oestrogens resulted in elevated levels of serum LH and FSH, suggesting that the adult male hypothalamic-pituitary axis may be responsive to positive feedback. In all studies, testosterone preferentially suppressed serum LH as compared to serum FSH. In contrast, oestradiol administration produced parallel inhibition of both LH and FSH. It is emphasized that neither oestrogen nor androgen alone, nor in combination, resulted in preferential inhibition of serum FSH over LH.

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EFFECTS OF DIHYDROTESTOSTERONE AND OESTRADIOL ON SEXUAL DIFFERENTIATION IN MALE RATS

Journal of Endocrinology ◽

10.1677/joe.0.0840397 ◽

1980 ◽

Vol 84 (3) ◽

pp. 397-407 ◽

Cited By ~ 27

Author(s):

P. VAN DER SCHOOT

Keyword(s):

Sexual Differentiation ◽

Testosterone Propionate ◽

Neonatal Period ◽

Combined Treatment ◽

Female Rats ◽

Male Rats ◽

Normal Male ◽

Oestradiol Benzoate ◽

The Brain ◽

Copulatory Behaviour

Adult male rats which had been castrated at birth and treated with the non-aromatizable androgen dihydrotestosterone propionate (DHTP) showed incomplete copulatory behaviour. When tested with oestrous female rats during treatment with testosterone propionate (TP) they readily mounted these females and showed frequent penile intromissions but rarely ejaculated. In a long series of observations the proportion of ejaculating rats in tests of 30 min did not exceed 50%. Neonatally castrated rats treated with DHTP during infancy thus seemed to be capable of ejaculation in adulthood during treatment with TP, but the threshold for the occurrence of the ejaculatory reflex seemed to be higher than in normal male rats. By replacing treatment in adulthood with TP by a combined treatment with DHTP and oestradiol benzoate (OB), the frequency of ejaculation was not increased. It was concluded that the incomplete copulatory behaviour was not due to reduced efficiency of aromatization of androgen within the brain of these rats. The addition of OB to DHTP during the neonatal period of treatment enhanced the frequency of ejaculation in adulthood. The combined treatment of 0·1 mg DHTP on days 1, 3 and 5 with 0·01 mg OB on day 1 made adult copulatory behaviour during treatment with TP indistinguishable from that of rats castrated on day 10 or rats castrated at birth and treated with TP during infancy. It was concluded that the masculine organization of systems and structures involved in the display of male copulatory behaviour occurs under the influence of both non-aromatizable androgen and oestrogen, oestrogen being most likely the substance required to 'organize' the central nervous aspects of the regulation of this behaviour. The absence neonatally of nonaromatizable androgen and/or oestrogen results in specific deficiencies in adult copulatory behaviour as compared with the behaviour of normal male rats.

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Effect of clomiphene citrate on the in vitro release of LH and FSH by the pituitary gland of the long-term ovariectomized rat pretreated with LRH or with LRH and oestradiol benzoate

Acta Endocrinologica ◽

10.1530/acta.0.1130035 ◽

1986 ◽

Vol 113 (1) ◽

pp. 35-41

Author(s):

G. A. Schuiling ◽

H. Moes ◽

T. R. Koiter

Keyword(s):

Clomiphene Citrate ◽

Combined Treatment ◽

In Vitro Release ◽

Pituitary Gonadotropin ◽

Ovx Rats ◽

Oestradiol Benzoate ◽

Pituitary Glands ◽

Pre Treatment

Abstract. The effect of a combined in vivo pre-treatment with luteinizing hormone-releasing hormone (LRH) and either oestradiol benzoate (OB), clomiphene (-citrate) or OB plus clomiphene on the autonomous and the supramaximally LRH-stimulated in vitro secretion of LH and FSH by pituitary glands of long-term ovariectomized (OVX) rats was studied using a hemi-pituitary perifusion system. The concentration of LRH in the perifusion medium was 1 μg/ml. Pre-treatment with LRH during 5 days was effected by means of sc implanted Alzet® osmotic minipumps; control rats received a piece of silastic with the dimensions of a minipump. OB, 3 μg/injection, clomiphene 100 μg/injection or solvent were given on days 2 and 4 (day of perifusion: day 5). In rats not pre-treated with LRH neither OB, nor clomiphene changed the content of the pituitary gonadotropin stores. There was only a small but significant positive effect of the combined treatment with OB and clomiphene on the pituitary FSH content. LRH (partly) depleted the gonadotropin stores. This effect of LRH was potentiated by OB, but not by clomiphene. Clomiphene prevented the depletion-potentiating effect of OB. OB raised the LRH-stimulated secretion of LH and FSH as well as the autonomous secretion of LH. Clomiphene raised the LRH-stimulated (not the autonomous) secretion of LH and FSH. OB plus clomiphene had the same effect as OB alone. Clomiphene also raised the LRH-stimulated secretion of LH and FSH after pre-treatment with LRH, but OB did not do so: LRH prevented the stimulatory effect of OB but not of clomiphene. OB plus clomiphene had the same effect as OB alone. The absence of a stimulatory effect of OB on the LRH-stimulated secretion of LH and FSH in the LRH-pretreated rat appeared to be due to the very low gonadotropin content of the pituitary glands after pre-treatment with LRH and OB: the effect of OB on the LRH-responsiveness proper (i.e. release of LH and FSH as related to the pituitary LH and FSH content) remained stimulatory. Also clomiphene enhanced the LRH-responsiveness proper, but this drug cannot potentiate the gonadotropin stores-depleting effect of LRH. These results demonstrate that clomiphene exclusively 'behaves' like an oestrogen-agonist, able to enhance the LRH-stimulated gonadotropin secretion. Also in the LRH-pre-treated rat clomiphene acts like an oestrogen-agonist, but unlike oestradiol clomiphene cannot potentiate the LRH-induced depletion of the pituitary gonadotropin stores. Therefore, it can also raise the LRH-stimulated secretion of LH and FSH in the LRH-pre-treated OVX rat.

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Comparison of the intravenous and intravaginal route of oxytocin administration for cervical dilation protocol and non‐surgical embryo recovery in oestrous‐induced Santa Inês ewes

Reproduction in Domestic Animals ◽

10.1111/rda.13499 ◽

2019 ◽

Vol 54 (9) ◽

pp. 1230-1235 ◽

Cited By ~ 4

Author(s):

Lucia Prellwitz ◽

Fabiana Nunes Zambrini ◽

José Domingos Guimarães ◽

Marco Antonio Paula de Sousa ◽

Maria Emília Franco Oliveira ◽

...

Keyword(s):

Cervical Dilation ◽

Embryo Recovery ◽

Santa Inês

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EFFECTS OF CASTRATION AND TESTOSTERONE, DIHYDROTESTOSTERONE OR OESTRADIOL REPLACEMENT TREATMENT IN NEONATAL RATS ON MOUNTING BEHAVIOUR IN THE ADULT

Journal of Endocrinology ◽

10.1677/joe.0.0760251 ◽

1978 ◽

Vol 76 (2) ◽

pp. 251-260 ◽

Cited By ~ 27

Author(s):

P. SÖDERSTEN ◽

S. HANSEN

Keyword(s):

Testosterone Propionate ◽

Combined Treatment ◽

Daily Treatment ◽

Glans Penis ◽

High Dose ◽

Neonatal Treatment ◽

Replacement Treatment ◽

Oestradiol Benzoate ◽

Lordosis Behaviour ◽

The Brain

Castration of rats on the day of birth abolished the capacity to ejaculate and reduced the capacity to show intromissions in response to testosterone propionate (TP) treatment in adults. Day 10 castrated rats treated daily with oil or day 0 castrated rats treated daily with testosterone benzoate (TB) during the first 10 days of life achieved intromissions and ejaculation after TP treatment in adulthood. Treatment of day 0 castrated rats with a high dose of TB during the first 10 days of life enhanced their capacity to ejaculate in response to TP treatment in adulthood to a level above that of day 10 castrated rats given oil in infancy and similarly treated with TP as adults. Castration on the day of birth greatly reduced the increase in penis weight and the development of cornified papillae on the glans penis which were seen in day 10 castrated rats after TP treatment in adulthood. These peripheral effects of neonatal testicular secretions are reversed by neonatal treatment of day 0 castrated rats with TB. Daily treatment of day 0 castrated rats with dihydrotestosterone benzoate (DHTB) during the first 10 days of life facilitated the increase in weight of the penis and the development of cornified papillae on the glans penis but did not enhance the capacity to ejaculate in response to TP treatment in adulthood. Daily treatment of day 0 castrated rats with oestradiol benzoate (OB) during the first 10 days of life facilitated ejaculation without increasing penis sensitivity to TP in adulthood. Combined treatment of the neonate with OB and DHTB was no more effective in facilitating ejaculation in the adult than was OB alone. Neonatal treatment with OB was considerably more potent than neonatal treatment with TB in enhancing ejaculatory behaviour in adulthood. It is suggested that both the inhibition of the development of lordosis behaviour and the facilitation of the development of mounting behaviour by testicular secretions in newborn rats may be dependent upon, but variously sensitive to, the amount of oestradiol formed in the brain from testosterone in the blood during the first 10 days of life.

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Anti-oestrogenic effects of tamoxifen on mammary gland and hypophysis in female rats

Acta Endocrinologica ◽

10.1530/acta.0.1050360 ◽

1984 ◽

Vol 105 (3) ◽

pp. 360-370 ◽

Cited By ~ 12

Author(s):

Stephan Götze ◽

Yukishige Nishino ◽

Friedmund Neumann

Keyword(s):

Mammary Gland ◽

Combined Treatment ◽

Secretory Activity ◽

Female Rats ◽

Ovariectomized Rats ◽

Immunoperoxidase Staining ◽

Immunoperoxidase Technique ◽

Basal Secretion ◽

Whole Mount ◽

Oestradiol Benzoate

Abstract. Adult ovariectomized rats were treated for 14 days with oestradiol benzoate (E2B) 15 μg/kg/d and oestradiol benzoate 15 μg/kg/d + progesterone (PRO) 15 mg/kg/d for induction of mammary gland parenchymal stimulation. Histological examination and whole mount preparation demonstrated that ductal growth in the mammary gland after E2B treatment was completely antagonized by tamoxifen (TAM) 0.5 mg/kg/d. Parallel DNA concentrations in the mammary gland were decreased to control levels by TAM 0.5, 5 and 15 mg/kg/d. E2B-induced hyperprolactinaemia in the forenoon (basal secretion was equally reduced by TAM 0.5, 5 and 15 mg/kg/d). In the afternoon, when prolactin (Prl) secretion is at its maximum, TAM 0.5 mg/kg/d turned out to be ineffective to abolish Prl surge, but TAM 5 and 15 mg/kg/d reduced serum Prl concentrations in a doserelated manner. Immunoperoxidase staining of Prl cells in the pars distalis of the hypophysis indicated that adaptive hypertrophy and signs of hypersecretion after E2B were abolished by TAM 5 mg/kg/d. Luteotrophic cells clearly showed cellular atrophy, regression and secretory inactivity. Maximal tubulo-alveolar mammary parenchymal stimulation in rats treated with E2B-PRO was slightly inhibited by TAM 0.5 mg/kg/d. Histology showed a small disseminated parenchymal islet. DNA concentrations only were partially decreased by the anti-oestrogen though serum Prl concentrations were found to be completely decreased to control levels. Secretory activity of Prl cells was reduced by TAM 0.5 mg/kg/d. In E2B-PRO treated rats lisuride had poor inhibitory activity on Prl levels and none on DNA concentrations in the mammary gland. Combined treatment with TAM and lisuride significantly decreased DNA concentration in the mammary gland compared to animals which received E2B-PRO. Also Prl levels were at a minimum. Histology performed on the mammary gland showed only slight tubulo- but no tubulo-alveolar activation. Luteotrophic cells in the pituitary gland stained by the immunoperoxidase technique appeared regressive, shrunken and atrophied.

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Blockade of LH and FSH secretion by LH-releasing hormone, by the LH-releasing hormone analogue, buserelin, and by combined treatment with LH-releasing hormone and oestradiol benzoate

Journal of Endocrinology ◽

10.1677/joe.0.1030301 ◽

1984 ◽

Vol 103 (3) ◽

pp. 301-309 ◽

Cited By ~ 6

Author(s):

G. A. Schuiling ◽

N. Pols-Valkhof ◽

G. C. J. van der Schaaf-Verdonk ◽

T. R. Koiter

Keyword(s):

Half Life ◽

Combined Treatment ◽

Plasma Concentrations ◽

Blocking Agent ◽

Releasing Hormone ◽

Oestradiol Benzoate ◽

Pituitary Glands ◽

Lh Rh ◽

Lh Releasing Hormone ◽

Dose Levels

ABSTRACT The LH and FSH release-stimulating (experiment 1) and -blocking (experiment 2) effects of LH-releasing hormone (LHRH) and of the LHRH analogue d-Ser(But)6-des-Gly10-LHRH-ethylamide (buserelin), as well as the effect of combined treatment with LH RH and oestradiol benzoate (OB; experiment 3) on the 'supra-maximally' LHRH-stimulated release of LH and FSH were studied in rats ovariectomized for 2 weeks. Pretreatment with LHRH (250 or 500 ng/h) or buserelin (250 ng/h) for 6 days was effected by means of subcutaneously implanted Alzet osmotic minipumps; control rats received a 'sham pump', i.e. a piece of silicone elastomer with the dimensions of a minipump. Oestradiol benzoate (3 μg/injection) or solvent was injected subcutaneously 75 and 27 h before the induction of LH/FSH responses. Experiment 1 revealed that after infusion of LHRH and buserelin, both at the rate of 1 μg/h, plasma LHRH concentrations were established which were about twice as low as the plasma buserelin concentrations. This might suggest that buserelin has a longer half-life than LHRH. As an LH and FSH release-stimulating substance, however, it appeared that buserelin was about as effective as LHRH. Experiment 2, however, suggested that as an LH/FSH release-blocking agent buserelin was much more effective than LHRH. In addition, after buserelin pretreatment the pituitary glands contained much less LH and FSH than after LHRH pretreatment at both dose levels used. However, this may also (at least partly) be due to the fact that buserelin has a longer half-life so that after infusion of buserelin and LHRH at the same rate the plasma concentrations of buserelin are higher than those of LHRH; after buserelin infusion the pituitary gland is therefore stimulated at a higher intensity. Experiment 3 showed that in OB-injected, sham-implanted rats the LHRH-stimulated secretion of LH and FSH was significantly higher than in the oil-injected, sham-implanted rats. In the LHRH-pretreated rats (LHRH: 250 ng/h for 6 days), however, the already depressed LHRH-stimulated secretion of LH and FSH was still further depressed by OB treatment. These latter results suggest that the increase of the pituitary LHRH responsiveness during exposure to oestrogen requires a reduction of the LHRH stimulation, which is normally caused by the negative feedback of oestrogen on the hypothalamus. J. Endocr. (1984) 103, 301–309

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MAINTENANCE OF MALE SEXUAL BEHAVIOUR BY COMBINED TREATMENT WITH OESTROGEN AND DIHYDROTESTOSTERONE IN CD-1 MICE

Journal of Endocrinology ◽

10.1677/joe.0.0660257 ◽

1975 ◽

Vol 66 (2) ◽

pp. 257-262 ◽

Cited By ~ 32

Author(s):

C. J. WALLIS ◽

W. G. LUTTGE

Keyword(s):

Adult Male ◽

Sexual Behaviour ◽

Mating Behaviour ◽

Combined Treatment ◽

Complete Loss ◽

Male Mating ◽

Intact Control ◽

Male Sexual Behaviour ◽

Oestradiol Benzoate

SUMMARY After castration, adult male sexually experienced CD-1 strain mice were treated with dihydrotestosterone (200 μg/day, DHT), oestradiol benzoate (1 μg/day, OB) or DHT (200 μg/ day) plus OB (1 μg/day). Oestradiol benzoate and the combined treatment DHT+OB maintained male sexual behaviour at levels comparable to a group of intact control mice, while DHT maintained behaviour at a lower level. Halving all hormone dosages resulted in a decrease in the number of OB-treated animals mating and a complete loss of mating in the DHT-treated animals. The decrease in dosage did not result in any change in the behaviour of the mice receiving both hormones. Adrenalectomy was found to have no effect on the mating behaviour of the OB-treated and untreated animals, but it did reduce the number of DHT+OB-treated animals mating. Thus, both OB alone and combined DHT+OB treatment can maintain male mating behaviour in castrated CD-1 strain mice and these effects do not appear to be due to the effects of oestradiol on the adrenal.

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