Implantation Of Estradiol-17 Beta Research Articles

Kisspeptin Neurons in the Ovine Arcuate Nucleus and Preoptic Area Are Involved in the Preovulatory Luteinizing Hormone Surge

Kisspeptin is the product of the Kiss1 gene that regulates GnRH secretion. In sheep, Kiss1 mRNA-expressing cells are found in the preoptic area (POA) and arcuate nucleus (ARC), and expression is up-regulated in the caudal ARC during the periovulatory period. We hypothesized that kisspeptin neurons in the ARC are activated by estradiol-17beta prior to the preovulatory LH surge. Ovariectomized ewes were treated as follows: 1) estradiol-17beta implants (sc 2 wk) to cause tonic negative feedback; 2) vehicle (no estrogen negative or positive feedback); or 3) positive feedback/GnRH surge-inducing injection of estradiol-17beta (50 microg iv). For groups 2 and 3, brains were collected 1 h after treatment and kisspeptin/Fos immunoreactivity was examined. In the caudal and mid-ARC, the percentage of kisspeptin cells that were Fos immunoreactive increased after acute estradiol treatment (group 3) over that seen in the other two groups. Kisspeptin/Fos colocalization was also quantified in ewes during the luteal and late-follicular phase of the estrous cycle, showing a trend toward an increase in colocalization in the late-follicular phase. Kisspeptin/Fos colocalization was similar in the POA across groups in both experiments. Analysis of Kiss1 mRNA by in situ hybridization revealed an increase in expression during the late-follicular phase in the caudal ARC and POA. These data suggest kisspeptin neurons located in the caudal extent of the ARC are involved in generating the positive feedback preovulatory GnRH/LH surge in the ewe, but there may also be a role for Kiss1-expressing cells in the POA.

Suppression of Follicle Wave Emergence in Cyclic Ewes by Supraphysiologic Concentrations of Estradiol-17beta and Induction with a Physiologic Dose of Exogenous Ovine Follicle-Stimulating Hormone1

Follicle waves are preceded by follicle-stimulating hormone (FSH) peaks in ewes. The purpose of the present study was to see whether estradiol implant treatment would block FSH peaks to create a model in which the effect of the timing and mode of FSH peaks could be studied by ovine FSH (oFSH) injection. In Experiment 1, 10 ewes received estradiol-17beta implants on Day 4 after ovulation (Day 0, day of ovulation); five ewes received large implants, and five ewes received small implants. Five control ewes received empty implants. In Experiment 2, 12 ewes received large implants on Day 4. On Day 9, six ewes received oFSH twice, 8 h apart (0.5 microg/kg; s.c.). Implants were left in place for 10 days in both experiments. In both studies, ovarian ultrasonography and blood sampling was done daily. In Experiment 1, estradiol concentrations were significantly higher in ewes with large implants (10.4 +/- 0.7 pg/ml) compared with controls (3.9 +/- 0.7 pg/ml) and ewes with small implants (5.4 +/- 0.7 pg/ml; P < 0.001). A significant reduction was found in mean FSH peak concentration (31%; P < 0.05) and FSH peak amplitude (45%; P < 0.05) in ewes with large implants compared with controls. Mean and basal FSH concentrations were unaffected by the large implants. The large implants halted follicle-wave emergence between Day 0 and 8 after implant insertion. The small follicle pool (2-3 mm in diameter) was unaffected by the large implants. When oFSH was injected into ewes with large implants, a follicle wave emerged 1.5 +/- 0.5 days after injection; however, in ewes given saline alone, a follicle wave emerged 4.8 +/- 0.8 days after injection (P < 0.01). We concluded that truncation of FSH peaks by estradiol implants prevented follicle-wave emergence, but injection of physiologic concentrations of oFSH reinitiated follicle-wave emergence.

Quantification of prolactin (PRL) and PRL receptor messenger RNA in gilthead seabream (Sparus aurata) after treatment with estradiol-17beta.

Prolactin (PRL) in fish is considered to be an osmoregulatory hormone, although some studies suggest that it may influence the production of steroid hormones in the gonads. The objective of the present study was to establish if PRL is involved in reproduction of the gilthead seabream-a protandrous hermaphrodite. Adult and juvenile gilthead seabream received implants of estradiol-17beta (E(2)) for 1 wk during the breeding season, and the mRNA expressions of PRL and PRL receptor (sbPRLR) were determined. Northern blot analysis revealed a single pituitary PRL transcript, the expression of which was significantly reduced by E(2) treatment in adults but significantly increased in juvenile fish. In adult gonads, four sbPRLR transcripts of 1.1, 1.3, 1.9, and 2.8 kilobases were observed. A competitive reverse transcription-polymerase chain reaction was developed and used to determine how E(2) treatment alters expression of the gonadal sbPRLR gene. Seabream PRLR was detectable in all samples analyzed by this assay. Levels of sbPRLR mRNA increased significantly (50-fold) after E(2) treatment in adults, but a 24-fold decrease was measured in juveniles. Immunohistochemistry using specific polyclonal antibodies raised against an oligopeptide from the extracellular domain of sbPRLR detected the receptor in spermatogonia and oocytes. Taken together, the preceding results suggest that in the seabream, PRL may act on both testis and ovary via its receptor and that the stage of maturity influences this process. The full characterization and relative importance of the different transcripts of sbPRLR in eliciting the action of PRL in the gonads remain to be elucidated.

The later administration of progesterone more rapidly activates dormant mouse mammary tumor cells initiated by 3'-methyl-4-dimethylaminoazobenzene.

Implantation of progesterone at 1 month of age induced the development of mammary tumors in female C57BL/6 x DS-F1 mice that had been treated with 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) neonatally, and that had undergone ovariectomy and received implants of estradiol-17beta (E2) pellets at 1 month of age, and the incidence of mammary tumors became 100% at 15 months of age. On the other hand, no mammary tumors developed in these mice with implants of E2 pellets alone. Implantation of progesterone alone also induced no mammary tumors in mice that had been treated with 3'-Me-DAB neonatally, and had undergone ovariectomy at 1 month of age. Implantation of progesterone at 4, 6, 8, and 10 months of age also caused the prompt development of mammary tumors as implantation of progesterone at 1 month of age. When ages at which the incidence became 50% were estimated on curves of the incidences, these ages on implantation of progesterone at 1, 4, 6, 8, and 10 were about 11, 13, 14, 14, and 14 months of age. These results suggest that progesterone together with estrogen promotes the development of mammary tumors induced by 3'-Me-DAB, and that the later progesterone is administered, the more rapidly it activates dormant mammary tumor cells initiated by 3'-Me-DAB.

Suppression by oestrogen of hepatocellular tumourigenesis induced in mice by 3'-methyl-4-dimethylaminoazobenzene.

Treatment of female C57BL/6 x DS-F1 mice with 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) neonatally resulted in the development of adenomatous nodules and glucose-6-phosphatase (G-6-Pase) deficient foci at 8 and 6 months of age, respectively. Ovariectomy of these mice at 1 month of age hastened the development and increased the incidences of these lesions. Subcutaneous implantation of estradiol-17 beta (E2) with ovariectomy at 1 month of age markedly decreased the incidences of adenomatous nodules and G-6-Pase deficient foci at 10 or 12 months of age, but subcutaneous implantation of progesterone did not reduce their incidences. Subcutaneous implantation of E2 into ovariectomised mice at 6 months of age resulted in significant decreases in the incidences of adenomatous nodules and G-6-Pase deficient foci at 10 months of age, but implantation of E2 into the spleen of ovariectomised mice of the same age had no effect on their incidences. The present results suggest that E2 suppresses the development of adenomatous nodules and G-6-Pase deficient foci induced in the mouse liver by 3'-Me-DAB by actions on tissues other than the liver.

Growth and endocrine responses of cattle to implantation of estradiol-17 beta during continuous or discontinuous grazing of high- and low-endophyte-infected tall fescue

Forty-eight Angus x Hereford steers (initial BW = 336 +/- 8.3 kg) were used in a 56-d study to evaluate growth and endocrine responses to continuous or discontinuous grazing of high-endophyte-infected Kentucky-31 (K; > 57% infestation rate) or low-endophyte-infected Johnstone tall fescue (J; < 1% infestation rate) and implantation with 0 or 24 mg/steer of estradiol-17 beta (E2; Compudose). Steers were allotted by weight to eight 3-ha paddocks (four paddocks of each fescue variety) with six steers per paddock. Two paddocks of each variety were grazed continuously (KK and JJ), whereas steers on the remaining four paddocks were rotated every 14 d from K to J (KJ) or from J to K (JK). Three steers in each paddock were implanted with E2 on d 0. The study extended from May 25, 1988 to July 20, 1988 with steers exposed to potential heat-stress conditions for 52 d of the 56-d study. Body weights were obtained on d 0, 28, and 56, and blood samples were taken on d 28 and 56. Overall ADG, serum prolactin, and serum alkaline phosphatase activity were greater (P < .05) in JJ than in KK steers. Rotation from K to J did not increase overall ADG, prolactin, insulin-like growth factor I (IGF-I), or alkaline phosphatase activity compared with the continuously grazed KK, whereas JK steers had lower (P < .10) ADG, prolactin, and alkaline phosphatase activity than JJ steers. Estradiol-17 beta increased (P < .10) IGF-I in JJ, KJ, and JK steers but not in KK steers.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of Estrogen Administration on Endogenous and Luteinizing Hormone-Releasing-Hormone-Induced Luteinizing Hormone Secretion and Follicular Development in the Lactating Sow1

The objectives of this study were to investigate whether estradiol treatment during lactation modifies 1) the patterns of endogenous LH, FSH, and prolactin (PRL) release; 2) the sensitivity of the pituitary to exogenous injections of LHRH; and 3) the responsiveness of the ovarian follicles to gonadotropin. Plasma LH, FSH, and PRL were determined in samples taken repeatedly from 18 sows on Days 24-27 of lactation. Ovaries were then recovered, and follicular development was assessed by measuring the follicular diameter (FFD) and follicular fluid estradiol-17 beta concentration (FFE) of the ten largest follicles dissected from each ovary. Sows were randomly allocated to one of four treatments: 1) Group C (4 sows) received no treatment; 2) Group LHRH (5 sows) received 800 ng of LHRH every 2 h throughout the sampling period; 3) Group E2 (4 sows) received subcutaneous implants containing estradiol-17 beta 24 h after start of sampling; 4) Group LHRH + E2 (5 sows) were administered a combination of LHRH and estradiol-17 beta implants. Between-animal variability for plasma LH, FSH, and PRL was considerable. LH concentration and LH pulse frequency increased (p less than 0.05) after LHRH treatment in the LHRH and LHRH + E2 groups; however, an acute inhibition of LH secretion was observed in the latter group immediately after estradiol implant application. In the absence of LHRH treatment, estradiol caused chronic inhibition of LH secretion. Follicular development was greater in the LHRH and LHRH + E2 groups compared to the C and E2 groups (p less than 0.05 for both FFD and FFE).(ABSTRACT TRUNCATED AT 250 WORDS)

Control of the Bovine Estrous Cycle with Melengestrol Acetate (MGA): A Review

Expanded use of artificial insemination in the beef cattle industry depends on successful application of treatments designed to synchronize estrus. Regulation of estrous cycles is associated with control of the corpus luteum (CL), whose life span and secretory activity are subject to trophic and lytic mechanisms. The advantages of melengestrol acetate (MGA) in estrous synchronization incorporate ease of administration, lower cost relative to other estrous synchronization products, and potential for use to induce estrus in prepubertal heifers. Treatments first designed to synchronize estrous cycles of normally cycling heifers by feeding MGA were imposed daily for 14 to 18 d at levels of .5 to 1 mg. The minimal daily effective dose required to inhibit ovulation was .42 mg. Longer feeding periods of MGA were associated with low fertility at the first synchronized estrus, but at the second estrus, conception was normal. Low fertility at the synchronized estrus resulted in development of alternative treatment practices, which combined feeding of MGA with injections or implants of estradiol-17 beta, estradiol cypionate, luteinizing hormone, human chorionic gonadotropin, pregnant mare serum gonadotropin, or oxytocin. Estrus was synchronized after MGA and estradiol-17 beta or estradiol cypionate treatments, but fertility was low. Short-term feeding of MGA (5 to 7 d) combined with prostaglandin F2 alpha or its analogs (PGF) on the last day of MGA reduced fertility at the synchronized estrus. The reduced conception at first service occurred in animals that began treatment after d 12 of the estrous cycle. However, feeding MGA for 14 d and then injecting PGF 17 d later avoided problems with reduced conception. Fertility of animals after this treatment was similar to that of contemporaries synchronized with Syncro-Mate-B. However, the length of the treatment period creates a need for increased management and may extend management beyond practical limits. Further research is warranted to address problems associated with reduced fertility after short-term treatment with MGA.

Luteinizing hormone secretion in ovariectomized gilts: effects of age, reproductive state and estrogen replacement.

Crossbred gilts were ovariectomized (OVX) at 120, 150, 180 and 210 d of age to determine whether various characteristics of luteinizing hormone (LH) concentrations are influenced by age and reproductive state (prepuberal vs postpuberal). All 120-d-old gilts were prepuberal and all 210-d-old animals were postpuberal, whereas gilts 150 and 180 d old included both prepuberal and postpuberal animals. Blood was collected at 15-min intervals for 2 h, 2 d before OVX (d -2), and 2 (d +2), 8 (d +8) and 14 (d +14) d after OVX. Mean LH concentrations for prepuberal gilts were similar among age groups (P greater than .05) on d -2 and +2; however, LH increased (P greater than .05) from d -2 to +2. No change in LH secretion was found in postpuberal gilts during these two periods. After OVX, LH increased from d +2 to +14 in both prepuberal and postpuberal gilts in all age groups. In postpuberal gilts, LH increased linearly (P less than .05) between d +2 and +14; rate of increase accelerated with advancing age (P less than .01). In prepuberal gilts, LH increased in a nonlinear manner, but it did not increase between d +2 and +8. The increase observed in prepuberal and postpuberal gilts after OVX resulted primarily from an increase in magnitude of peak concentrations of LH. Implants of estradiol-17 beta (E2) were used to determine whether the postovariectomy increase in LH is affected differently by E2 in prepuberal and postpuberal gilts during advancing ages.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of ovarian secretions and dopamine on secretion of luteinizing hormone and prolactin in ewes.

In the present study, effects of dopamine (DA) and ovarian secretions on release of luteinizing hormone (LH) and prolactin (PRL) were examined using two methods of withdrawal of progesterone. Eight to 10 d after estrus, 36 ewes were assigned to one of six treatments: 1) lutectomy; 2) lutectomy and DA; 3) ovariectomy; 4) ovariectomy and DA; 5) ovariectomy and estradiol-17 beta or 6) ovariectomy, estradiol-17 beta and DA. Estradiol-17 beta was administered by Silastic implants inserted during surgery. Immediately after surgery, ewes were infused iv with DA (6.6 micrograms X kg-1 X min-1) or saline for 48 h. Infusion of DA did not affect secretion of LH after surgical removal of the corpus luteum. In lutectomized ewes, concentrations of LH in serum increased between 0 and 12 h from .5 to 3.3 ng/ml, declined between 12 and 16 h and then plateaued at 1.1 to 1.5 ng/ml. In contrast, concentrations of LH in ovariectomized ewes increased between 0 and 12 h and then remained at 3 to 5 ng/ml. Surges of LH occurred between 40 and 50 h postsurgery in eight of 12 lutectomized ewes, 11 of 12 ovariectomized ewes with implants of estradiol-17 beta and zero of 12 ovariectomized ewes. The magnitude of the surge of LH was higher in lutectomized ewes than in ovariectomized ewes with implants of estradiol-17 beta. Infusion of DA inhibited release of PRL, but there was no effect of method of progesterone withdrawal. Lutectomized ewes, but not ovariectomized ewes with implants of estradiol-17 beta, had a surge of PRL associated with the surge of LH.(ABSTRACT TRUNCATED AT 250 WORDS)