Effects of dose and route of administration of cloprostenol on luteolysis,estrus and ovulation in beef heifers

Four experiments were conducted (with crossbred beef heifers) to determine the effects of dose and route of administration of cloprostenol on luteolysis, estrus and ovulation. In Experiment 1, 19 heifers with a CL > or = 17 mm in diameter were randomly allocated to receive cloprostenol as follows: 100 microg s.c., 250 microg s.c., or 500 microg i.m. Heifers given 100 microg s.c. had a longer (P<0.03) interval (120.0 h+/-10.7 h; mean+/-S.E.M.) from treatment to ovulation than those given either 250 microg s.c. or 500 microg i.m. (92.0 h+/-7.4 h and 84.0 h+/-8.2 h, respectively). In Experiment 2, 28 heifers were given porcine LH (pLH), followed in 7 days by cloprostenol (same doses and routes as in Experiment 1), and a second dose of pLH 48 h after cloprostenol. Luteolysis occurred in all heifers, and no difference was detected among treatment groups in the interval from cloprostenol treatment to ovulation (mean, 101 h; P<0.9). In Experiment 3, 38 heifers at random stages of the estrous cycle (but with plasma progesterone concentrations > or =1.0 ng/ml) received 500 or 125 microg cloprostenol by either i.m. or s.c. injection (2/2 factorial design). There was no difference (P<0.4) among groups in the proportions of heifers that were detected in estrus or that ovulated. However, the interval from cloprostenol treatment to estrus was shorter (P<0.02) in the group that received 500 microg i.m. (58.5h) than in the other three groups (500 microg s.c., 75.0 h; 125 microg i.m., 78.0 h; and 125 microg s.c., 82.3h). In Experiment 4, 36 heifers were treated (as in Experiment 3) on Day 7 after ovulation. The proportions of heifers detected in estrus and ovulating after 125 microg s.c. (33 and 44%, respectively) or 125 microg i.m. (55 and 55%) were lower (P<0.05) than in those that received 500 microg s.c. (100 and 100%), but not different from those receiving 500 microg i.m. (78 and 89%, respectively). Overall, ovulation was detected in 9/18 heifers given 125 microg and 17/18 heifers given 500 microg of cloprostenol, on Day 7 (P<0.01) and was detected in 17/20 heifers given 125 microg and 18/18 heifers given 500 microg of cloprostenol, at random stages of the estrous cycle (P>0.05). Although there was no significant difference in luteolytic efficacy between i.m. and s.c. injections of the recommended dose (500 microg) of cloprostenol, variability in responsiveness to a reduced dose depended upon CL sensitivity, therefore, reduced doses cannot be recommended for routine use.     

Induction of parturition, progesterone secretion, and delivery of placenta in beef heifers given relaxin with cloprostenol or dexamethasone

Sixty primiparous beef heifers from a crossbreeding study were used to examine the effects of inducing parturition with relaxin (3,000 U/mg) combined with cloprostenol (500 micrograms, i.m., n = 30) or dexamethasone (20 mg, i.m., n = 30) at Day 273, 10 +/- 1 days before expected parturition (Day 283). Heifers were assigned at random within cloprostenol and dexamethasone groups to receive relaxin (1 mg, n = 5/treatment), i.m. or in the cervical os (OS), at 0 h (the same time as cloprostenol and dexamethasone) or 24 h later. Eleven and six first-calving heifers and sixteen and nine second-calving cows also received cloprostenol + relaxin and cloprostenol + phosphate-buffered saline, respectively. Radioimmunoassay of daily plasma samples indicated an abrupt decrease in progesterone with time (p less than 0.001), from 7.5 +/- 0.50 to 1.0 +/- 0.30 ng/ml (mean +/- SE) within 48 h for all groups. The mean rate of progesterone decrease (ng/ml in 24 h) was accelerated (p less than 0.01) in relaxin-treated heifers (5.3 +/- 0.36), in contrast to dexamethasone- and cloprostenol-treated control heifers (2.8 +/- 0.40). Relaxin combined with cloprostenol or dexamethasone shortened the calving period in these heifers by reducing the interval between treatment and calving (33 vs. 56 h; p less than 0.01). The incidence and duration of retained placenta were reduced by 22 vs. 75% and 14 vs. 34 h for relaxin combined with cloprostenol or dexamethasone as compared with cloprostenol- or dexamethasone-treated controls, respectively (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)     

Failure of the LH-releasing hormone agonist, deslorelin, to prevent development of a persistent follicle in heifers synchronized with norgestomet

The use of exogenous progestagens for estrus synchronization in cattle can result in a persistent dominant follicle which is associated with reduced fertility. We examined whether the LHRH agonist, deslorelin, would prevent the formation of a persistent follicle in heifers synchronized with norgestomet. The estrous cycles of heifers were synchronized with cloprostenol, and on Day 7 of the ensuing cycle the heifers received one of the following treatments for 10 d: Group C (n = 5), untreated control; Group N (n = 6), injection of a luteolytic dose of cloprostenol on Days 7 and 8 and implant of norgestomet from Day 7 to Day 17 (i.e. typical 10-day norgestomet implant period); Group D (n = 6), injection of cloprostenol on Days 7 and 8 and implants of deslorelin from Day 7 to Day 17; Group ND (n = 6), injections of cloprostenol and both norgestomet and deslorelin implants as above. Follicle growth was monitored using ultrasonography. Group-N heifers showed continued follicle growth and had larger follicles on Day 17 of the cycle than Group-C heifers (16.8 +/- 1.6 and 10.4 +/- 1.6 mm). Follicle growth for Group-D and ND heifers was similar and variable, and seemed to depend on follicle status at the initiation of treatment. Heifers with follicles of 5 to 10 mm (n = 9) in diameter either showed no follicle growth (2 9 ) or developed large follicles (7 9 ), while heifers with follicles approximately 12 mm (n = 3) in diameter showed follicle atresia with no further significant growth. On Day 17, size of the largest follicle was similar for Group-ND (14.3 +/- 2.9) and Group-D (16.8 +/- 1.6) heifers. Heifers in Group N showed estrous behavior 1.8 +/- 0.2 d after treatment, whereas heifers in Groups D and ND did not show estrus for 2 to 4 wk. The results show that combined treatment with progestagen and an LHRH agonist does not consistently prevent the development of a persistent dominant follicle and that return to estrus can be delayed after treatment with an LHRH agonist.     

Single appointment insemination for heifers after prostaglandin or progestin synchronization of estrus

Pregnancy rates to a single appointment insemination were compared in seven groups of beef or dairy heifers following estrus synchronization with the prostaglandin F(2a) product, cloprostenol, versus the progestín product, Syncro-Mate-B. For cloprostenol synchronization, two injections of cloprostenol were given 11 d apart, with insemination occurring at 61+/-1 h after the second injection. The Syncro-Mate-B treatment consisted of a norgestomet/estradiol injection and a norgestomet implant on Day 0, followed by insemination at 49+/-1 h after implant removal on Day 9. Treatments were coordinated so all heifers in each group were housed together at the time of and for 48 h before a common insemination time so that the treatment received by individual heifers was not known. Overall pregnancy rates did not differ between treatment groups: 42% for 111 heifers treated with cloprostenol and 38% for 108 heifers treated with Syncro-Mate-B. Four blood samples per heifer taken during each replicate showed that two groups (n = 67) had many noncyclic heifers, while five groups (n = 152) had very few. Differences in pregnancy rates between these categories existed for both estrus synchronization methods, 18 versus 53% for cloprostenol and 21 versus 45% for Syncro-Mate-B. Thus the two methods were equally effective for cyclic heifers and equally ineffective for noncyclic heifers.     

Synchronization of estrus in beef cows and heifers with fenprostalene, cloprostenol sodium, and prostaglandin F2 alpha

The effects of fenprostalene, cloprostenol sodium and prostaglandin F(2) alpha (PGF(2alpha)) on estrus, conception rate, pregnancy rate, and the interval from Day 1 of the breeding season to calving were studied on 135 purebred Angus cows and heifers. The cows and heifers were randomly allotted within age to the three estrus synchronization treatments and a control group. The calving percentages (for cows and heifers combined) that resulted from artificial insemination (AI) were 32.3, 31.4, 43.6, and 51.1% for the control, fenprostalene, cloprostenol sodium, and PGF(2alpha) groups, respectively. The calving percentage during the AI period by ages of dam at breeding were 54.2% for yearling heifers, 30.5% for two-year-olds, 47.6% for three-year-olds, and 26.1% for four-year-old or older cows. The percentage of cows and heifers detected in estrus and the percentage that conceived after the first injection for control, fenprostalene, cloprostenol sodium, and PGF(2alpha) groups were 51.6 and 22.3%, 59.3 and 32.1%, 76.8 and 44.1%, and 66.6 and 50.2%, respectively. The intervals from Day 1 of the breeding season to calving and from Day 1 of the calving season within each treatment to the birth of each calf were control, 285.9 and 23.8 d; fenprostalene, 283.6 and 13.4 d; cloprostenol sodium, 285.5 and 6.5 d; and PGF(2alpha), 284.0 and 11.1 d.     

Short synchronization system for estrus cycles in dairy heifers: a preliminary report

In previous studies we demonstrated that the administration of a luteolytic dose of cloprostenol to dairy cows in luteal phase, followed by hCG plus estradiol benzoate (EB) 12 h later, led to successful timed AI 48 h after the initiation of treatment. This article reports two consecutive studies. In Study 1 we determined the pregnancy rate of dairy heifers in luteal phase (established by palpation per rectum) treated with cloprostenol followed by 250 IU of hCG plus 1 mg of EB 12 h later, and inseminated 48 h after cloprostenol injection. Study 2 was designed to evaluate the efficiency this synchronization protocol, irrespective of the estrus stage of the animals. In Study 1, 1272 Friesian heifers aged 14 to 16 months with a palpable corpus luteum received 500 mcg cloprostenol. Heifers were then synchronized either according to the hCG plus EB protocol (hCG-EB, n=637), or by a second dose of cloprostenol 11 d later (PG, n=636). Animals in this last group served as controls and were inseminated 72 and 96 h after the second cloprostenol injection. The pregnancy rate was significantly higher (P<0.0001) in the hCG-EB group (59.5%, 379/637) than in PG (44.8%, 285/636). In Study 2, 135 contemporary heifers (with no corresponding information on estrus stage) were subjected to the same protocol as those in the hCG-EB group of Study 1. These animals were classified in retrospect according to estrus stage established by plasma progesterone concentration. Pregnancy rates were 66.7% (24/36), 51% (25/49) and 58% (29/50) for animals in the follicular, early/late luteal, and mid-luteal phase, respectively. The total pregnancy rate was 57.8% (78/135). These findings indicate an improved pregnancy rate for heifers subjected to single insemination after cloprostenol/hCG/EB synchronization, compared to double insemination after synchronization by 2 cloprostenol injections 11 d apart. The cloprostenol/hCG/EB protocol did result in acceptable pregnancy rates after timed AI of dairy heifers regardless of their estrus cycle phase.     

Timing of the onset and duration of ovulation in superovulated beef heifers

Thirty-two beef heifers were induced to superovulate by the administration of follicle stimulating hormone-porcine (FSH-P). All heifers received 32 mg FSH-P (total dose) which was injected twice daily in decreasing amounts for 4 d commencing on Days 8 to 10 of the estrous cycle. Cloprostenol was administered at 60 and 72 h after the first injection of FSH-P. Heifers were observed for estrus every 6 h and were slaughtered at known times between 48 to 100 h after the first cloprostenol treatment. The populations of ovulated and nonovulated follicles in the ovaries were quantified immediately after slaughter. Blood samples were taken at 2-h intervals from six heifers from 24 h after cloprostenol treatment until slaughter and the plasma was assayed for luteinizing hormone (LH) concentrations. The interval from cloprostenol injection to the onset of estrus was 41.3 +/- 1.25 h (n = 20). The interval from cloprostenol injection to the preovulatory peak of LH was 43.3 +/- 1.69 h (n = 6). No ovulations were observed in animals slaughtered prior to 64.5 h after cloprostenol (n = 12). After 64.5 h, ovulation had commenced in all animals except in one animal slaughtered at 65.5 h. The ovulation rate varied from 4 to 50 ovulations. Approximately 80% of large follicles (> 10 mm diameter) had ovulated within 12 h of the onset of ovulation. Onset of ovulation was followed by a dramatic decrease in the number of large follicles (> 10 mm) and an increase in the number of small follicles (相似文献   

Prolactin and epidermal growth factor regulation of the proliferation,morphogenesis, and functional differentiation of normal rat mammary epithelial cells in three dimensional primary culture

The epithelial cell-specific effects of prolactin and epidermal growth factor (EGF) on the development of normal rat mammary epithelial cells (MEC) were evaluated using a three dimensional primary culture model developed in our laboratory. Non-milk-producing MEC were isolated as spherical end bud-like mammary epithelial organoids (MEO) from pubescent virgin female rats. The cultured MEO developed into elaborate multilobular and lobuloductal alveolar organoids composed of cytologically and functionally differentiated MEC. Prolactin (0.01–10 μg/ml) and EGF (1–100 ng/ml) were each required for induction of cell growth, extensive alveolar, as well as multilobular branching morphogenesis, and casein accumulation. MEO cultured without prolactin for 14 days remained sensitive to the mitogenic, morphogenic, and lactogenic effects of prolactin upon subsequent exposure. Similarly, cells cultured in the absence of EGF remained sensitive to the mitogenic and lactogenic effects of EGF, but were less responsive to its morphogenic effects when it was added on day 14 of a 21-day culture period. If exposure to prolactin was terminated after the first week, the magnitude of the mitogenic and lactogenic effects, but not the morphogenic response was decreased. Removal of EGF on day 7 also reduced the mitogenic response, but did not have any effect on the magnitude of the lactogenic or morphogenic responses. These studies demonstrate that physiologically relevant development of normal MEC can be induced in culture and that this model system can be used to study the mechanisms by which prolactin and EGF regulate the complex developmental pathways operative in the mammary gland. © 1995 Wiley-Liss, Inc.     

Induction of abortion in feedlot heifers with cloprostenol (ICI 80,996)

Abortion was induced in two groups of feedlot heifers with cloprostenol. Response varied with the stage of gestation and dose of cloprostenol. Results of this limited trial indicate that cloprostenol is a safe and effective abortifacient in feedlot heifers at a dose of 250 μg to day 120 of gestation and at a dose of 500 μg from day 121 to day 150.     

Reproductive characteristics of cyclic beef heifers treated with the prostaglandin analog luprostiol

One hundred and twenty crossbred Angus heifers, after exhibiting a 17- to 23-d estrous cycle, were placed into six groups of 20 heifers each and administered 2 ml i.m. propylene glycol containing either 0 (controls), 3.75, 7.5, 15.0 or 30.0 mg of luprostiol, or saline containing 0.5 mg cloprostenol (Groups 1 through 6, respectively). Heifers were observed for estrus every 6 h and all treatments were given 6.5 to 8.0 d after heifers were observed in standing estrus. Blood samples were collected after treatments from 10 heifers in each groups. Blood serum was assayed for progesterone. The synchronization period was considered to be 120 h after administration of luprostiol or cloprostenol. There were 0, 16, 17, 18, 20 and 18 heifers observed in estrus during the synchronization period in Groups 1 through 6, respectively. Progesterone concentrations in blood serum dropped below 1 ng/ml in 0, 8, 9, 10, 10 and 10 of the heifers from which blood samples had been taken in the six groups. All heifers observed in estrus were artificially inseminated. During the synchronization period, 0, 12, 14, 15, 16 and 10 heifers conceived in Groups 1 through 6, respectively. The interval from injection to estrus for the 89 heifers that exhibited estrus in the synchronization period averaged 49.0 h and was not different among the luprostiol and cloprostenol treated groups. Control heifers returned to estrus an average of 13.2 d after the treatment. The number of heifers that conceived at first insemination, regardless of when estrus occurred, was 16, 15, 16, 16, 16 and 12, and the total number that conceived at the first and second inseminations was 18, 18, 17, 19, 19 and 16 for Groups 1 through 6, respectively. Based on serum progesterone concentration and/or interval from treatment to estrus, 15 and 30 mg of luprostiol effectively regressed corpora lutea (100%) when administered between 6.5 and 8.0 d after estrus, and the estrous response and conception rate for these two groups equalled or exceeded that of the control and cloprostenol groups.     

Control of estrus in dairy cows with a synthetic analogue of prostaglandin F2 alpha

Trials were carried out on 1184 dairy cows calved at least six weeks before treatment and 255 heifers to determine effectiveness of the prostaglandin analogue, cloprostenol to control estrus. In trial 1, following two injections of cloprostenol given 12 days apart, there was no difference in calving rate following AI either at 72 and 96 hr after treatment (71 163 ) or at a detected estrus (53 118 ) compared to control cows bred at estrus (54 110 ). In trial 2, treated cows were injected once after 5 to 7 days of estrous detection and AI. The calving rate following AI either at 72 and 96 hr after cloprostenol (46 100 ) or at a detected estrus (39 71 ) was similar to that in control cows bred at estrus (45 86 ). In trial 3, cows were bred at a detected estrus after the first cloprostenol injection. Twelve days after this injection, cows not bred were given a second injection and bred 72 and 96 hr later. The calving rate in treated cows bred at estrus after the first injection (66 138 ) was similar to calving rate in controls (55 95 ). However, calving rate in cows given a second injection and bred 72 and 96 hr later was significantly (P angle 0.05) lower (30 98 ). Similar results were obtained in heifers, except calving rate in trial 3 after the second cloprostenol injection was not reduced.     

Effects of an LHRH antagonist on gonadotrophin and oestradiol secretion, follicular development, oestrus and ovulation in Holstein heifers

Mature cyclic Holstein heifers were given a luteolytic dose of cloprostenol followed by two i.v. injections, 12 h apart, of various doses of [Ac-D-Nal1, D-p-Cl-Phe2, D-Trp3, D-Arg6, D-Ala10]-LHRH, beginning either at the time of first observation of behavioural oestrus, or 48 h after the cloprostenol injection. When treatment began at the first observation of oestrus, the time of ovulation, as determined by ultrasonic echography, was significantly delayed by total doses of 0.8 mg or more of the antagonist. When given at 48 and 60 h after cloprostenol injection, a total dose of 1.5 mg of the antagonist significantly delayed the growth of the ovulating follicle, the onset of oestrus, the preovulatory surges of oestradiol, LH and FSH, and ovulation. It is concluded that the LHRH antagonist can effectively suppress endogenous LH secretion and may therefore be useful in the study of follicular development, ovulation, and other events in the oestrous cycle of the cow.     

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.     

Estrus induction with prostaglandin F2alpha, cloprostenol or fenprostalene during the normal estrous cycle, superovulation and after embryo collection

Holstein heifers used as embryo donors were treated with three luteolytic agents (PGF2alpha, cloprostenol, fenprostalene) during the normal estrous cycle, superovulation or after embryo collection to determine the interval from treatment to estrus. A similar return-to-estrus interval was observed for each luteolytic agent among the three groups of heifers. Nevertheless, after embryo collection, fenprostalene had a tendency to induce the longest delays (p = 0.08). This tendency is supported by a higher proportion of delayed luteolysis and more heifers showing estrus later than 11 d post treatment. Also, during normal estrous cycles, 5/10 and 0/8 fenprostalene- and cloprostenol-treated heifers, respectively, showed progesterone concentrations higher than 1 ng/mL 48 h after treatment. Regardless of the luteolytic agent used, estrus was induced earlier (P < 0.005) during superovulation than when heifers were treated between Days 9 to 16 of the normal estrous cycle or after embryo collection. However, the return-to-estrus interval was similar between heifers treated during superovulation and those treated between Days 6 to 8 of the normal estrous cycle. After embryo collection, intervals before the return to estrus increased with the number of Corpora lutea (CL) palpated except in the nonresponding group (0 to 1 CL), which returned to estrus later than the low responding group (2 to 4 CL).     

Overexpression of Mos, Ras, Src, and Fos inhibits mouse mammary epithelial cell differentiation.

Mammary epithelial cells terminally differentiate in response to lactogenic hormones. We present evidence that oncoprotein overexpression is incompatible with this hormone-inducible differentiation and results in striking cellular morphological changes. In mammary epithelial cells in culture, lactogenic hormones (glucocorticoid and prolactin) activated a transfected beta-casein promoter and endogenous beta-casein gene expression. This response to lactogenic hormone treatment was paralleled by a decrease in cellular AP-1 DNA-binding activity. Expression of the mos, ras, or src (but not myc) oncogene blocked the activation of the beta-casein promoter induced by the lactogenic hormones and was associated with the maintenance of high levels of AP-1. Mos expression also increased c-fos and c-jun mRNA levels. Overexpression of Fos and Jun from transiently transfected constructs resulted in a functional inhibition of the glucocorticoid receptor in these mouse mammary epithelial cells. This finding clearly suggests that glucocorticoid receptor inhibition arising from oncogene expression will contribute to the block in hormonally induced mammary epithelial cell differentiation. Expression of Src resulted in the loss of the normal organization and morphological phenotype of mammary epithelial cells in the epithelial/fibroblastic line IM-2. Activation of a conditional c-fos/estrogen receptor gene encoding an estrogen-dependent Fos/estrogen receptor fusion protein also morphologically transformed mammary epithelial cells and inhibited initiation of mammary epithelial differentiation-associated expression of the beta-casein and WDNM 1 genes. In response to estrogen treatment, the cells displayed a high level of AP-1 DNA-binding activity. Our results demonstrate that high cellular AP-1 levels contribute to blocking the ability of mammary epithelial cells in culture to respond to lactogenic hormones. This and other studies indicate that the oncogene products Mos, Ras, and Src exert their effects, at least in part, by stimulating cellular Fos and probably cellular Jun activity.     

Effect of photoperiod on LH, FSH and prolactin patterns in ovariectomized oestradiol-treated heifers

Angus and Angus crossbred heifers were ovariectomized, treated with oestradiol implants and randomly assigned to the natural photoperiod of fall to spring for 43 degrees N latitude or extra light simulating the photoperiod of spring to fall. Weekly blood samples were taken for 6 months (fall to spring equinox). All heifers were cannulated every 4 weeks and blood samples were taken for 4 h at 15-min intervals. Sera were assayed for LH, FSH, prolactin and oestradiol. In samples taken weekly, serum LH and FSH concentrations were higher while serum prolactin was lower in heifers exposed to natural photoperiod. There was a photoperiod X time interaction for both FSH and prolactin with concentrations diverging as photoperiod diverged. Circulating concentrations of oestradiol were not different between groups. In samples taken every 4 weeks at 15-min intervals, baseline concentrations of LH and FSH and LH pulse amplitude were higher while prolactin pulse frequency was lower in heifers exposed to natural photoperiod. There was a photoperiod X time interaction for each of these pulsatile characteristics. The correlation between LH and prolactin concentrations estimated from the 15-min samples differed between the two photoperiod treatment groups. The pooled correlation coefficient (r) was -0.12 under natural photoperiod and +0.50 under extra light. There was also a photoperiod X time interaction with negative correlations occurring when photoperiod was decreasing and positive correlations occurring when photoperiod was increasing. These results support the hypothesis that photoperiod alters serum concentrations of LH, FSH and prolactin in cattle.     

Effects of amiloride on the induction of DNA synthesis and casein gene expression in rabbit mammary explants

Amiloride, an inhibitor of Na+/H+ exchange, was added at various concentrations to the culture medium of rabbit mammary explants. In the concentration range 100-250 microM, amiloride progessively inhibited 14C-thymidine incorporation induced by insulin, EGF or prolactin. Up to 250 microM, amiloride, which did not inhibit basal protein synthesis, was not cytotoxic, but it reduced basal DNA synthesis at the highest concentration. Addition of amiloride to the culture medium of mammary explants also strongly inhibited the induction of casein synthesis and casein mRNA accumulation by prolactin. The inhibition by amiloride is therefore not specific of the mitogenic action of prolactin since this drug also prevented its lactogenic action. The data reported here describe a new inhibitory action of amiloride on the transmission of the lactogenic signals.     

Neither duration of progesterone insert nor initial GnRH treatment affected pregnancy per timed-insemination in dairy heifers subjected to a Co-synch protocol

Our objectives were to: 1) compare response to cloprostenol, synchrony of ovulation, and pregnancy per timed-AI (P/TAI) in a 5 d versus a 7 d Co-synch + PRID protocol (Experiment 1); and 2) investigate whether the initial GnRH is necessary to achieve acceptable P/TAI in a 5 d Co-synch + PRID protocol (Experiment 2) in dairy heifers. In Experiment 1, 64 Holstein heifers, 15 to 17 mo, were assigned by age to receive 100 μg of GnRH and a PRID for 5 or 7 d (PRID5 and PRID7, respectively). At PRID removal 500 μg of cloprostenol (PGF) was given i.m. Heifers received the second GnRH treatment concurrently with TAI at 72 (PRID5) or 56 (PRID7) h after PRID removal. Transrectal ultrasonography monitored ovarian dynamics, ovulation synchrony, and pregnancy status (28 and 45 d after TAI). Plasma progesterone concentrations were determined at PRID removal and TAI. Five of seven heifers that ovulated before TAI became pregnant, and only two heifers did not respond to PGF treatment in the PRID5 group. Five PRID5 and 2 PRID7 heifers failed to ovulate after the second GnRH. However, P/TAI did not differ between PRID5 (59.4%) and PRID7 (58.1%). Overall ovulation response to first GnRH treatment was only 31.7%, and a larger proportion of heifers that did not ovulate became pregnant (65.1 versus 45.0%). In Experiment 2, 56 Holstein heifers, assigned as in Experiment 1, were subjected to a PRID5 protocol with (PRID5G) or without (PRID5NoG) GnRH at PRID insertion; all heifers were TAI 72 h after PRID removal. Transrectal ultrasonography and progesterone determinations were performed as in Experiment 1. Pregnancy per TAI did not differ whether or not heifers received GnRH at PRID insertion (67.9 versus 71.4%). Consistent with our previous findings, seven of nine heifers that ovulated before TAI became pregnant, and only two heifers did not respond to PGF treatment. Combining both experiments, length of proestrus but not ovulatory follicle diameter was identified as a significant predictor of probability of pregnancy 28 d after TAI, with a maximum predicted probability of 80.1% when the length of proestrus was 3 d. In summary, a PRID5 protocol resulted in comparable P/TAI to a PRID7 protocol. Most of the heifers that ovulated before TAI in the PRID5, PRID5G, and PRID5NoG protocols became pregnant. More than one PGF or a GnRH treatment at PRID insertion in a 5 d Co-synch + PRID protocol was not required to achieve acceptable P/TAI in dairy heifers.     

Superovulation of beef heifers with Pergonal (HMG): A dose response trial

A study was designed to establish a dose-response curve for Pergonal (Human Menopausal Gonadotrophin) and to compare its efficacy in inducing superovulation with commercial FSH-P. A recognized treatment schedule for HMG of two ampoules at 0, 12, 24 and 36 hours and one ampoule at 48, 60, 72, 84, 96 and 108 hours was considered to be our 100% effective dose level. Fifty mature cycling cross-bred beef heifers were superovulated on day 10 +/- 1 of their cycle. Treatment groups were HMG I (200% dose), HMG II (100% dose), HMG III )50% dose), HMG IV (25% dose) and FSH-P (total dose 32 mg). All groups received 500 ug of cloprostenol 72 hours after initiation of treatments. The heifers were observed for onset of estrus and inseminated at 12, 24 and 36 hours. All heifers were slaughtered on day 7 post-estrus and their reproductive tracts removed for processing. All heifers were bled once daily for progesterone estimation and four times daily for two days beginning 24 hours after cloprostenol injection, for luteinizing hormone and estradiol-17beta estimations. A dose response to HMG was demonstrated for number of corpora lutea and all classes of ova/embryos. HMG II (100% dose) closely approximated the optimum dose for superovulation. There was no significant difference between the HMG II group and the FSH-P group for mean number of transferable embryos. The 200% HMG dose did not increase the numbers of ovulations or ova recovered but did decrease the numbers of fertilized and transferable ova.