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Temporary exposure of ovine embryos to an advanced uterine environment does not affect fetal weight but alters fetal muscle development.

Embryo transfer techniques may result in fetuses that are heavier at birth and that have been described as highly muscled. The aim of this study was to investigate myogenesis in lambs derived from embryo transfer. Embryos were transferred at Day 3 (estrus = Day 0) to a 3 days-advanced uterine environment, maintained there for 3 days, recovered, and then returned to a synchronous (Day 6) uterus; these fetuses comprised the asynchronous group. Control animals were created by synchronous embryo recovery and single transfer at Day 3. Asynchronous transfer did not affect fetal weight or curved crown-rump length between 46 and 135 days of gestation. No differences were detected between groups at Days 110-135 with respect to muscle mass or protein, RNA, and DNA content. However, total muscle fiber number was significantly increased in plantaris muscles from the asynchronous groups at Day 110 and Day 125, suggestive of prolonged hyperplasia. In addition, the levels of Myf 5 protein and the secondary-to-primary fiber ratio were altered in plantaris muscle from the asynchronous group. The growth data are in contrast to previously reported findings. The results show that fetal myogenesis can be altered by very early events in embryogenesis and suggest that any inferences made solely on the basis of fetal or muscle weight may be fallacious.

Animals↗

A large-scale program in laparoscopic intrauterine insemination with frozen-thawed semen in Australian Merino sheep in Argentine Patagonia.

This report shows the results of a large-scale laparoscopic intrauterine insemination program on a flock of Australian Merino sheep in Argentine Patagonia. The study was carried out on a total of 1824 ewes (3-to-7-yr-old) and 480 ewe hoggets (19-20 months old) on 2 farms in the southeastern region of Santa Cruz Province, in April and May 1996. The animals, divided into 15 groups, were synchronized with vaginal sponges containing 60 mg medroxyprogesterone acetate for 14 d and injected with 200 IU PMSG upon sponge removal. Estrus was screened every 12 h by means of vasectomized marker rams. The animals were inseminated laparoscopically by the intrauterine route using 2 schemes: 1) at a fixed time (12 h) after estrus detection, or 2) at a fixed time (60 h) after sponge removal irrespective of estrus. Pregnancy was determined at 30 d by transrectal ultrasound imaging. The results showed that 1) the onset of estrus occurs most often between 24 and 48 h after sponge removal, 2) ewe hoggets undergo estrus significantly earlier than sexually mature ewes, 3) in those animals showing estrus, there appears to be no relationship between fertility (as assessed by pregnancy outcome) and time of estrus, 4) there is a significant association between the percentage of estrus occurrence and pregnancy rate, 5) fertility is significantly higher in ewes than in hoggets, 6) for practical purposes insemination at a fixed time after the onset of estrus has no advantage over that of to insemination at a fixed time after sponge removal. It is concluded that large-scale laparoscopic intrauterine insemination can be successfully applied in Australian Merino ewes and ewe hoggets in low-productivity areas such as that of Argentine Patagonia and that estrus detection is unnecessary when insemination is performed at 60 h after sponge removal.

Journal Article↗

Estrogen-triggered delays in mammary gland gene expression during the estrous cycle: evidence for a novel timing system.

During the estrous cycle and beginning in estrus, the mammary gland undergoes pregnancy-like development that depends on transcriptional regulation by the estrogen and progesterone receptors (ER, PR) and Pax-2 as well as the action of the growth factors Wnt-4 and RANKL. In this report, we first describe the decay and delayed expression of ERalpha, PR, and Pax-2 proteins as well as depression of Wnt-4 and RANKL mRNA coincident with the strong estrogen surge in proestrus. In time-course studies using ovari-ectomized mice, a single estrogen injection replicated these delays and caused an 18 h delay in Wnt-4 expression. Molecular time-delay systems are at the core of cellular cycles, most notably the circadian clock, and depend on proteasome degradation of transcriptional regulators that exhibit dedicated timing functions. The cytoplasmic dynamics of these regulators govern delay duration through negative transcription/translation feedback loops. A proteasome inhibitor, PS-341, blocked estrogen-stimulated ERalpha, PR, and Pax-2 decay and proteasome chymotryptic activity, assayed using a fluorogenic substrate, was elevated in proestrus correlating with the depletion of the transcription factors. The 18-h delay in Wnt-4 induction corresponded to the turnover time of Pax-2 protein in the cytoplasm and was eliminated in Pax-2 knockout mammary tissue, demonstrating that Pax-2 has a unique timing function. The patterns of estrogen-triggered ERalpha, PR, and Pax-2 turnover were consistent with a negative transcriptional feedback. Retarding the expression of ERalpha, PR, and Pax-2 may optimize preparations for pregnancy by coordinating expression of critical receptors and transcription factors with rising estrogen and progesterone levels in estrus. The estrogen surge in proestrus has no defined mammotropic function. This study provides the first evidence that it is a synchronizing signal triggering proteasome-dependent turnover of mammary gland ERalpha, PR, and Pax-2. We hypothesize that the delays reflect a previously unrecognized timing system, which is present in all ovarian target tissues.

Animals↗

Metabolic hormone secretion and FSH-induced superovulatory responses of beef heifers fed dietary fat supplements containing predominantly saturated or polyunsaturated fatty acids.

Ovulatory responses following FSH treatment were examined in beef heifers fed dietary fat supplements expected to produce differential effects on serum insulin concentrations and follicular recruitment patterns. Twenty-one heifers (n = 7/group) exhibiting regular estrous cycles were assigned randomly to either a control diet or to 1 of 2 fat-supplemented diets consisting of soybean oil (polyunsaturated fatty acids) or animal tallow (saturated fatty acids). The diets were formulated to be isoenergetic and isonitrogenous, and were fed until ovariectomy between experimental Days 35 and 45. Experimental Day 1 was defined for each heifer as the first day all of the treatment diet was consumed. After 20 d of diet consumption, estrous cycles were synchronized with prostaglandin F(2alpha) (PGF(2alpha)), and ovarian follicle populations were monitored via transrectal ultrasound for 4 d. Four days after estrus, the dominant follicle was aspirated and heifers were treated with FSH-P to induce superovulation. Ovulation rate was determined at ovariectomy 5 d after the superovulatory estrus (experimental Days 35 to 45). Both soybean oil and animal tallow diets increased (P < 0.05) the number of medium-sized follicles and increased (P < 0.02) serum concentrations of GH relative to the control diet. The soybean oil diet also increased (P < 0.001) serum concentrations of insulin on Days 14, 28, and 5 d after the superovulatory estrus. However, the number of ovulations following FSH treatment did not differ due to diet. Procedures employed in the current study were ineffective in recruiting the increased number of medium-sized follicles into the superovulatory pool.

Journal Article↗

Do Norway rats (Rattus norvegicus) synchronize their estrous cycles?

Estrous synchrony was tested using 10 pairs of sibling female rats (Rattus norvegicus). A Monte Carlo bootstrap simulation was used to construct random control groups to avoid previous statistical errors and to test for significance when there are irregular cycles. The 10 pairs of females did not exhibit estrous synchrony. The effect of cycle irregularity on the limits of synchrony was analyzed using an equation that the related degree of cycle regularity to the degree of synchrony. This equation significantly predicted the limits of synchrony: cycle irregularity limits both the maximum and minimum degree of synchrony that can occur between two females. Finally, simulations of the expected levels of synchrony in groups of five rats were compared to the original study on estrous synchrony. The simulations indicated that the results of the original study were consistent with chance levels of synchrony. It is concluded that there is no evidence that Norway rats synchronize their estrous cycles. Evolutionary implications are discussed.

Algorithms↗

Breeding capacity, behavior and fertility of bulls with Brahman genetic influence during synchronized breeding of beef females.

The objective of this experiment was to evaluate the feasibility of using bulls of Brahman-derived breeds for synchronized breeding of females treated with Syncro-Mate-B (SMB). Suckled, postpartum cows (n = 261) and virgin heifers (n = 227) were given the standard SMB treatment. Calves were removed from cows for 48 h beginning at implant removal, and individual bulls were placed in pens with 15 to 20 females (x = 1:15.7 ) for 48 h beginning at implant removal. Bulls (n = 31) were 18 to 36 mo of age and had scored satisfactorily on a breeding soundness evaluation (BSE). The mean +/- SEM percentage of females in estrus, percentage of estrous females serviced and percentage of total females serviced were 77.2 +/- 2.4, 72.9 +/- 2.9 and 55.7 +/- 3%, respectively. Total average services per bull was 23.6 +/- 1.6; however, average number of individuals serviced was only 8.6 +/- 0.42. Mean +/- SEM percentage of females conceiving of those serviced, percentage conceiving of those in estrus and percentage conceiving of total females available were 57.3 +/- 3.7, 40.6 +/- 2.6 and 32.6 +/- 2.5%, respectively. Mean BSE scores of eight Simbrah bulls tested immediately before and 5 d after synchronized breeding did not differ (P > 0.05). These data suggest that the conception rate of Brahman-influenced bulls servicing SMB-synchronized cows is within the normal range. However, the total number of individual females serviced and the total pregnancy rate under the protocol employed was low. This occurred because all estrous females were not inseminated, and some bulls exhibited unpredicted low fertility.

Journal Article↗