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PubMed1984

Reduction of sperm transport in ewes by superovulation treatments.

Oestrus was synchronized in ewes by (a) withdrawal of an intravaginal progestagen sponge after 12 days or (b) injection of PGF-2 alpha on Day 11 of the oestrous cycle. In addition, ewes were treated with (a) no hormone, (b) 1500 i.u. PMSG 48 h before sponge removal or PG injection, or (c) 24 mg porcine pituitary FSH in multiple injections commencing 48 h before sponge removal or PG injection, in a 2 X 3 factorial design. Ewes were inseminated with 0.2 ml fresh undiluted semen into the neck of the cervix 48 h after sponge removal or PG injection. Normally cyclic ewes were similarly inseminated within 12 h of observed standing oestrus. At 24 h after insemination one uterine horn and one oviduct were flushed for recovery of spermatozoa and ova. When compared with naturally cyclic ewes, PG synchronization resulted in a marked reduction in the numbers of spermatozoa recovered (P less than 0.05), and sponge synchronization led to a small, non-significant, reduction. Within the synchronized ewes, PMSG and FSH resulted in an equivalent superovulatory response, but there was a marked reduction in sperm recovery when compared with unstimulated animals (P less than 0.01), with the greatest reduction attributable to PMSG treatment. Spermatozoa were recovered from fewer ewes treated with PMSG than with FSH (P less than 0.05). Despite the observed impairment of sperm transport, a high fertilization rate was observed within each group and there were no differences between treatments.

Animals↗
PubMed2004

A simple ultrasound test to predict the superstimulatory response in cattle.

We tested the hypotheses that: (1) the superstimulatory response is related to the intrinsic number of follicles recruited into a follicular wave; and (2) the number of follicles recruited into a wave is correlated to the number of follicles recruited into the successive wave. A positive correlation will form the basis of a test for predicting the superstimulatory response. Cows (n = 141) were treated with estradiol and progesterone to synchronize follicular wave emergence (first synchronization) and ranked according to the number of follicles > or =2mm at wave emergence to select the upper and lower 10% of the herd. Follicular wave emergence was synchronized again in the high-end (n = 16) and low-end (n = 20) groups (second synchronization), and cows were treated with FSH twice daily for 3 days. High-end cows had a greater number of follicles (P < 0.001) than low-end cows at the time of wave emergence after both the first and second synchronizations in the 2-3 and 4-6mm categories. The numbers of 2-3 and 4-6mm follicles at wave emergence after the first and second synchronizations were positively correlated (P < 0.001; r = 0.77 and 0.71, respectively). Endogenous FSH peak at the time of wave emergence was higher in the low-end group than in the high-end group. Superstimulatory treatment resulted in more than double the number of follicles (P < 0.003) in the 5-7mm and > or =8mm categories in the high-end group than in the low-end group (16.8 +/- 2.2 versus 8.1 +/- 0.9 and 22.7 +/- 4.1 versus 9.7 +/- 1.6, respectively). The number of follicles > or =5 and > or =8mm at the end of superstimulation was positively correlated (P < 0.001) with the total number of follicles > or =2mm at the time of wave emergence after both the first (r = 0.64 and 0.54, respectively) and second ( r = 0.65 and 0.5, respectively) synchronizations. Based on the results of this study, the superstimulatory response can be predicted by the number of follicles > or =2mm at wave emergence. For practical purposes, practitioners can expect the number of follicles > or =5mm after ovarian superstimulation to be approximately 71% of the number of follicles > or =2mm at the time of wave emergence. Results validated the proposed simple ultrasound-based test for predicting the superstimulatory response of individual cows.

Animals↗
PubMed2006

The application of reproductive technologies to natural populations of red deer.

Over the past decade, there has been increasing interest in the application of reproductive technology to the conservation and management of natural populations of deer. The application of assisted reproduction technologies within natural population of deer is in its infancy. However, its future potential is enormous, particularly in relation to genetic management or conservation. This paper reviews the present state of such technologies for a wild subspecies of red deer, the Iberian red deer (Cervus elaphus hispanicus), by discussing the major components of oestrous synchronization, semen collection/cryopreservation and insemination techniques. In addition, findings made during the course of studies on natural populations have enormous potential for the understanding of novel reproductive mechanism that may not be uncovered by livestock or human studies. A summary of these results are also reviewed here.

Animals↗
PubMed1977

[Histological findings on the uterus of gilts following synchronized ovulation].

Histological studies were undertaken with the view to testing uterus structure and function of gilts following synchronised ovulation by means of different PMSG doses. All proband groups received 500 I.U. HCG. All histomorphological, histochemical, and histometric checks revealed 500 I.U. PMSG to be too low a dosage, whereas the optimum amount was found to be between 750 and 1,000 I.U. PMSG. The uterine glands of all treated animals in all three groups were less favourably developed than those of the untreated controls. The best morphologico-histochemical pattern was observed following administration of 750 I.U. PMSG.

Animals↗
PubMed1981

Mating behaviour of a bull and its relationship with serum LH levels in a group of oestrous cows.

A PGF-2 alpha analogue (cloprostenol) was injected into 9 cows to synchronize oestrus. The cows were placed with or without a bull in a free-stall in groups of 3 at 46 h after the injection. Sexual behaviour was observed and serum LH concentrations were measured during the next 34 h. The bull ejaculated with the cows in regular sequence and did not return to cows after an ejaculatory series was completed. The mating behaviour of the bull was closely related to the LH surge of the cows.

Animals↗
PubMed1999

Synchronization of oestrous cycles in sable antelope.

Techniques for manipulating the oestrous cycle of sable antelope, Hippotragus niger, were evaluated in a captive population of 24 females maintained at the Smithsonian Institution's Conservation and Research Center in Front Royal, VA, USA. A secondary objective was to demonstrate the effectiveness of fecal steroid monitoring techniques as a non-invasive method of tracking experimental manipulations. Controlled Internal Drug Releasing (CIDR) devices designed for cattle (type B, reduced in length by 5 cm to fit the sable antelope's smaller reproductive tract) were more effective than CIDR devices designed for goats (type G) at delivering progesterone into circulation, and maintained serum progesterone at levels up to 86.1+/-7.8% of normal luteal concentrations in females whose spontaneous ovarian activity had been inhibited with melengestrol acetate. Serum progesterone and fecal progestagen measurements were highly correlated (P<0.05). Synchronization treatments of prostaglandin (PG) F2alpha alone and in combination with modified CIDR-B devices (12-day insertion interval) were both effective in inducing synchronized ovulation, however the PGF2alpha/modified CIDR-B treatment resulted in more precise synchrony and a shorter latency to ovulation than did PGF2alpha alone. In a separate experiment to characterize the temporal relationship between synchronization treatment, behavioral oestrus and ovulation, onset of behavioral oestrus occurred 34.1+/-5.7 h following PGF2alpha/modified CIDR-B treatment. Mean duration of the induced oestrus was 24.9+/-4.3 h. The first detectable rise in fecal progestagens occurred 5.1+/-1.0 and 4.1+/-1.0 days following PGF2alpha/modified CIDR-B treatment in groups of females housed with and without an adult male, respectively, indicating that the presence of a male did not accelerate the onset of the induced cycle.

Administration, Intravaginal↗
PubMed1996

Oestrous synchronization, semen preservation and artificial insemination in the Mohor gazelle (Gazella dama mhorr) for the establishment of a genome resource bank programme.

Gazella dama mhorr is an endangered species with an extant population of about 190 animals distributed between several zoos. Semen was collected by electro-ejaculation from 12 adult males, and cryopreserved in TEST-yolk diluent containing 6% glycerol. The effects of the concentration of egg yolk (5%, 10% and 20%) and the presence or absence of sodium triethanolamine lauryl sulfate (equex) on sperm motion and acrosomal integrity after thawing were examined. Increasing concentrations of egg yolk resulted in more acrosomal damage and poorer motility after thawing. The presence or absence of equex had no effect on either parameter. The frozen spermatozoa were used in an insemination trial, in which 13 females were treated with intravaginal progesterone-releasing devices to synchronize oestrus. Seven females were inseminated with frozen-thawed semen 48 h after removal of the devices, and six were inseminated after 60 h. Three females in the first group and one in the second group became pregnant. However, only one pregnancy (from the 48-h group) was carried to term. The study demonstrated the feasibility of applying artificial insemination in this species, but revealed that a number of outstanding technical problems remain to be solved.

Acrosome↗
PubMed1999

Ovarian and hormonal responses of cows to treatment with an analogue of gonadotrophin releasing hormone and prostaglandin F2 alpha.

Blood samples were taken from 11 cows and their ovaries were scanned by ultrasound at least daily. Around day 5 of an induced cycle, they were injected with 10 micrograms buserelin, an analogue of gonadotrophin releasing hormone, and on day 12 they received 0.5 mg cloprostenol, an analogue of prostaglandin F2 alpha (PGF2 alpha). Two days later six of the cows (the treated group) received a second injection of 10 micrograms buserelin, but the remaining five received no further treatment (control group). The dominant, that is, the largest follicle in each cow disappeared after the first buserelin injection and was replaced by a new one which grew synchronously in all the cows until after the treatment with PGF2 alpha. Ovulation occurred significantly earlier after PGF2 alpha in the treated group than in the control group (72 to 96 hours v 96 to 120 hours; P < 0.05). Plasma progesterone concentrations then increased more rapidly in the treated group than in the control group and were significantly higher on days 3 and 4 after ovulation (P < 0.05).

Animals↗
PubMed2000

Synchronization of ovulation in crossbred dairy heifers using gonadotrophin-releasing hormone agonist, prostaglandin F2alpha and human chorionic gonadotrophin or estradiol benzoate.

Girolando (Gir x Holstein) is a very common dairy breed in Brazil because it combines the rusticity of Gir (Bos indicus) with the high milk yield of Holstein (Bos taurus). The ovarian follicular dynamics and hormonal treatments for synchronization of ovulation and timed artificial insemination were studied in Girolando heifers. The injection of a gonadotrophin-releasing hormone (GnRH) agonist was followed 6 or 7 days (d) later by prostaglandin F2alpha (PGF2alpha). Twenty-four hours after PGF2alpha injection either human chorionic gonadotropin (hCG, GPh-d6 and GPh-d7 groups) or estradiol benzoate (EB, GPE-d6 and GPE-d7 groups) was administered to synchronize ovulation and consequently allow timed artificial insemination (AI) 24 and 30 h after hCG and EB injection, respectively. Follicular dynamics in Girolando heifers was characterized by the predominance of three follicular waves (71.4%) with sizes of dominant follicles (10-13 mm) and corpus luteum (approximately 20 mm) similar to those for Bos indicus cattle. In the GnRH-PGF-hCG protocol, hCG administration induced earlier ovulation (67.4 h, P<0.01) compared to the control group (GnRH-PGF) and a better synchronization of ovulation, since most of it occurred within a period of 12 to 17 h. Pregnancy rate after timed AI was 42.8 (3/7, GPh-d6) to 50% (7/14, GPh-d7). In contrast, estradiol benzoate (GnRH-PGF-EB protocol) synchronized ovulation of only 5 of 11 heifers from the GPE-d7 group and of none (0/7) from the GPE-d6 group, which led to low pregnancy rates after timed AI (27.3 and 0%, respectively). However, since a small number of Girolando heifers was used to determine pregnancy rates in the present study, pregnancy rates should be confirmed with a larger number of animals.

Analysis of Variance↗
PubMed2005

Increased cervical electrical activity during oestrus in progestagen treated ewes: Possible role in sperm transport.

The aim of this investigation was to characterize the pattern of cervical myoelectrical activity (EMG) in the sheep, during the periovulatory period, after synchronization of oestrus with progestagen and eCG. EMG was measured with a computerised modular system in five ewes previously fitted with a pair of monopolar Teflon needle electrodes in the muscle layers of the cervix. Each ewe was submitted to oestrus synchronization treatment with intravaginal progestagen sponge during 12 days, and the administration of 500 IU of eCG at the withdrawal of sponge. EMG was recorded in each animal during 19 h, starting 44 h after withdrawal of sponge. The number and duration of events were determined every hour during the experiment. Two distinct event durations were identified: one lasting less than 200 s and another lasting between 300 and 500 s. The two types of events analysed (less than 200 s and lasting between 300 and 500 s) had a similar pattern during the period of observation although they were not in synchrony. For events lasting less than 200 s, activity increased between 48 and 50 h after sponge withdrawal, with the peak of activity being observed between 51 and 53 h. For events of 300-500 s duration, the peak of activity was observed between 48 and 50h after sponge withdrawal and activity was maintained until 51-53 h. The increase in cervical motility observed in progestagen-eCG treated ewes is in keeping with the increase in cervical activity observed by others in natural cycling animals, and suggests that exogenous hormones used in synchronization protocols had no deleterious action on cervical motility during periovulatory period. The enhanced activity of cervical muscle layer found around the time of mating and/or AI suggests it may play an important role as a regulatory mechanism of sperm transport. Taking advantage of the cervical responsiveness to various drugs, experimental modulation of cervical activity could be used to facilitate cervical sperm transport and consequent improve of fertility after cervical AI.

Animals↗
PubMed1978

[Induction of labor and its synchronization in hogs].

The authors describe the synchronization of farrowing by the induction of parturition with a prostaglandin F2alpha analogue (Cloprostenol). The time sequence of the beginning of parturitions was recorded in 147 sows after administration of 175 microgram Cloprostenol. 93.7% of the parturitions take place between the 14th and 39th hour after administration, the peak time being between the 24th and 27th hour (31.2%). The schedule of the farrowing days is described as a prerequisite for a qualitative change in the organization of work in large pig herds.

Animals↗
PubMed1978

[Biotechnical increase of litter size in early pregnant sows using steroid application. 1. The effect of a single application of hydroxyprogesterone capronate on fertility response in ovulation-synchronized gilts and old sows].

One single intramuscular injection of hydropyrogesteroncapronate was given in an experiment to 199 gilts and 239 old sows on the 13th or 15th or 17th day after insemination to test various fertility performance parameters. Doses were 50 mg, 75 mg or 100 mg. Positive effects on litter size were established from the majority of nine compared experimental variants. The number of live-born piglets was increased up to two. Several groups of gilts had higher mean litter weights. More studies are necessary to prepare an adequate therapeutic regime by which to reduce loss of embryos in swine.

Age Factors↗
PubMed1982

Effects of creep feeding and monensin on reproductive performance and lactation of beef heifers.

A 23 factorial arrangement of treatments was utilized to determine the effect of breed, creep feeding and monensin on subsequent reproductive performance and lactation of 32 primiparous heifers. One-half of each breed (Angus and Hereford) group had access to creep feed (2.67 Mcal metabolizable energy/kg) while nursing their dams. Approximately 40 d before breeding through 120 d of lactation, all heifers were fed a suboptimal energy diet and 50% of each breed and creep group received 200 mg monensin/head daily. Estrus was synchronized with a progestogen. Volatile fatty acids (VFA) were determined periodically throughout the monensin-feeding portion of the experiment. Twenty-four hour milk production, percentage butterfat and percentage solids-not-fat were determined at 60 and 120 d postpartum. Weaning weights (adjusted to 205 d and for age of dam) of the original heifers were heavier (P less than .05) for those that were creep fed compared with those not creep fed (219 vs 202 kg). Monensin-supplemented females gained significantly more weight from the initiation of treatment to immediately postcalving and gave birth to heavier calves (P less than .05) even though they received comparable amounts of dietary energy as those that did not receive monensin. The energy stressed, monensin-supplemented first-calf heifers exhibited a shorter postpartum interval (P less than .05) to first estrus than did those that did not receive monensin (55.7 vs 69.1 d, respectively). First-calf heifers that had been creep fed while nursing their dams had a lower (P less than .05) daily milk yield at 120 d post-partum than those that did not have access to creep feed. No detrimental effects were observed due to long-term monensin supplementation.

Animals↗
PubMedDate not supplied

The value of progesterone, oestradiol benzoate and cloprostenol in controlling the timing of oestrus and ovulation in dairy cows and allowing successful fixed-time insemination.

The relative merits of three hormone treatments of dairy cows: (1) intravaginally administered progesterone and oestradiol benzoate; (2) intravaginally administered progesterone and injected cloprostenol; and (3) injected cloprostenol; begun 35-75 days after calving and designed to synchronize oestrus and ovulation and allow successful artificial insemination (AI) at fixed times, have been assessed utilizing information from progesterone concentrations in milk. From this it was concluded that 89% of the cows had ovulated one to three times between calving and the beginning of treatment. Treatment (2) was more effective than (1) in synchronizing ovulation. This was due to the fact that when treatments began early in the ovulation cycle, the requirement for a rapidly effective luteolytic agent was provided by cloprostenol but not by oestradiol benzoate. Treatment (2) was also more effective than (3) in synchronizing ovulation. This is interpreted as meaning that progesterone treatment for 12 days had a beneficial effect in restoring normal cyclic ovarian function in the cows after calving. Whilst cloprostenol administered alone did not have this beneficial effect, there is no evidence that it had a detrimental effect. Based on all cows in treatment groups, the proportion that became pregnant to the fixed-time AI was significantly greater after treatment (2) than after (1), but when based on numbers of cows with synchronized ovulation, there were no significant differences among treatments in the proportions becoming pregnant. The progesterone/cloprostenol treatment had a disadvantage in that when begun during the 11-22 day period of the ovulation cycle, so resulting in a long, total period of suppression of ovulation (mean, 32.1 days), fertility to the fixed-time AI was poor despite effective synchronization of ovulation. Ovulation cycles immediately following the failed, fixed-time AI were normal, both in length and in maximum, luteal-phase progesterone concentration and indicated normal corpus luteum function. Thus the infertility could be ascribed neither to poor timing of AI nor to gross degeneration of follicles prior to their synchronized ovulation following the prolonged suppression of ovulation. The 12-day progesterone treatments when given to anovulatory cows gave, within 5.5 h of their beginning, a concentration of progesterone in milk that was not significantly different from the maximum reached. This concentration declined during the 12 days of the treatment but remained above pretreatment level until 5.5 h after treatment withdrawal; the maximum reached was about half that in normal ovulation cycles.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

Exogenous progesterone and progestins as used in estrous synchrony regimens do not mimic the corpus luteum in regulation of luteinizing hormone and 17 beta-estradiol in circulation of cows.

Our working hypothesis was that the low concentrations of progesterone (P4) and synthetic progestins administered in hormonal regimens to control estrous cycles of cows would have similar effects on secretion of LH and 17 beta-estradiol (E2). In addition, we hypothesized that concentrations of exogenous P4 typical of the midluteal phase of the estrous cycle and the corpus luteum (CL) would have similar effects on LH and E2, and the effects would be different from those of synthetic progestins and low concentrations of P4. Cows (n = 29) were randomly assigned to one of five treatment groups: 1) one Progesterone Releasing Intravaginal Device (1PRID; n = 6); 2) two PRIDs (2PRID; n = 6); 3) norgestomet, as in Syncro-Mate-B regimen (SMB; n = 6); 4) melengestrol acetate (MGA; 0.5 mg/day; n = 5); and 5) control (CONT; n = 6). Treatments were administered for 9 days (Day 0 = initiation of treatment). All cows from 1PRID, 2PRID, SMB, and MGA groups were injected with prostaglandin F2 alpha (PGF2 alpha) on Days 2 and 5 of the treatment period to regress CL. Cows in the 1PRID and SMB groups were also administered exogenous estrogen according to the respective estrous synchronization protocol for these products. Daily blood samples were collected from Day 0 to 35 to determine concentrations of P4. On Day 8, blood samples were collected at 15-min intervals for 24 h to determine pattern of LH secretion. On Day 9, all treatments ceased and cows in the CONT group received injections of PGF2 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

Factors affecting superovulation in heifers treated with PMSG.

In this study we determined 1) if the immunoneutralization of PMSG affected the ovulatory response, the number of large follicles and embryo yield compared with that of PMSG alone or pFSH, and 2) whether the stage of the estrous cycle at which PMSG was injected affected the ovulatory response and yield of embryos in superovulated heifers. Estrus was synchronized in 99 (Experiment 1) and 71 (Experiment 2) heifers using prostaglandin F2alpha (PG) analogue, cloprostenol, given 11 d apart in replicate experiments over 2 yr. In Experiments 1 and 2, heifers were randomly allocated to 1 of 3 treatments (initiated at mid-cycle): Treatment 1--24 mg of pFSH (Folltropin) given twice daily for 4 d; Treatment 2--a single injection of 2000 IU PMSG; Treatment 3--2000 IU PMSG followed by 2000 IU of Neutra-PMSG at the time of first insemination. In Experiment 3, 116 heifers were given 2000 IU PMSG on Day 2 (n = 28), Day 3 (n = 27), Day 10 (n = 41) or Day 16 (n = 20) of the estrous cycle. The PG was given at 48 h (500 microg cloprostenol) and 60 h (250 microg cloprostenol) after the first gonadotropin treatment. Heifers were inseminated twice during estrus, and embryos were recovered on Day 7, following slaughter and graded for quality. The numbers of ovulations and large follicles (> or =10 mm) were also counted. There was no effect of treatment on ovulation rate in Experiment 1, but in Experiment 2 it was greater (P < 0.002) in heifers given PMSG (14.7 +/- 1.5) than pFSH (7.5 +/- 1.4) or PMSG-neutra-PMSG (8.7 +/- 1.5). The number of large follicles was higher following PMSG than pFSH treatment in Experiment 1, and it was higher (P < 0.004) in heifers given PMSG (5.5 +/- 0.8) than pFSH (1.12 +/- 0.7) or PMSG-neutra-PMSG (2.7 +/- 0.8) in Experiment 2. The use of Neutra-PMSG did not affect the numbers of embryos recovered or numbers of Grade 1 or 2 embryos, but it did decrease the number of Grade 3 embryos in both experiments. In Experiment 3, the ovulation rate decreased (P < 0.004) when PMSG was given on Day 3 (5.7 +/- 1.46) of the cycle rather than on Day 2 (12.3 +/- 1.64), Day 10 (13.4 +/- 1.45) or Day 16 (12.5 +/- 1.87). There was no effect of day of treatment on the numbers of large follicles. The mean numbers of embryos recovered were lower (P < 0.01) in heifers treated on Day 3 (2.1 +/- 0.67) than on Day 2 (6.8 +/- 1.0), Day 10 (6.4 +/- 0.86) or Day 16 (7.8 +/- 1.87). It is concluded that Neutra-PMSG given to heifers treated with PMSG did not improve embryo yield or quality and that treatment with PMSG early in the cycle can result in acceptable embryo yields provided sufficient time elapses between treatment and luteolysis.

Journal Article↗
PubMed1983

Heat synchronization in dairy cows.

Dairy cows were heat synchronized with two injections of cloprostenol (PG1/PG2) with an interval of 13 days and inseminated 72 hours after PG2. The total pregnancy rate obtained was 42% The probability of achieving luteal phase at PG2, the probability of undergoing luteolysis after PG2 and the probability of achieving pregnancy were all equal whether the cows were in luteal phase, follicular phase, had follicular cysts or small ovaries at PG1. Close correlations were found between the signs of heat at the insemination and the interval from calving to PG1 to the pregnancy rates.

Animals↗
PubMed2006

In vivo effect of growth hormone on the expression of connexin-43 in bovine ovarian follicles.

This study assessed the in vivo effects of recombinant growth hormone (rGH) administration on the expression of connexin-43 (Cx43) in bovine ovarian follicles. Two independent experiments were carried out using either estrous unsynchronized or synchronized multiparous Aberdeen Angus cows. rGH-treated animals were inoculated with a single dose of hormone (500 mg, intramuscular) while control animals were inoculated with hormone diluent. Five and 14 days after treatment (Experiments 1 and 2, respectively), ovarian Cx43 and apoptosis expression were assessed using immunohistochemistry. In both experiments primary, secondary, and tertiary follicles from rGH-treated and control groups distinctly expressed Cx43 protein. Primordial and atretic follicles were Cx43-negative. Interestingly, the number of Cx43 dots per granulosa cell did not show significant variation at different folliculogenesis stages neither in the rGH-treated nor in the control group. In unsynchronized animals, Cx43-positive follicles per total number of follicles ratio showed an interaction between stage of folliculogenesis and treatment due to significant differences between treatment groups in the early secondary follicle stage. In synchronized animals, there were significant differences between treatment groups and folliculogenesis stage. In both experiments, atretic follicles showed apoptosis-related DNA-fragmentation as determined by terminal uridin nick end labeling (TUNEL) assay. Tertiary follicles presented moderate TUNEL staining. Our results show significant increment in the number of ovarian follicles expressing the gap junction subunit Cx43 after in vivo rGH treatment. Therefore, we conclude that growth hormone can modulate in vivo gap junction assembly at early stages of folliculogenesis.

Animals↗