Concentrations of Prolactin, LH, Testosterone, TSH and Thyroxine in Normospermic Dogs of Different Breeds

Concentrations of prolactin (PRL), LH, testosterone (T), TSH and thyroxine (T₄) were determined before and at 20, 120 and 180 min after a single iv injection of thyrotropin-releasing hormone (TRH) in eight Beagles, eight Fox Terriers, six Labrador Retrievers and five Great Danes that were normospermic. Mean basal PRL concentrations were lower in the Fox Terriers compared with the Great Danes (p < 0.05). Mean LH concentrations were higher in the Fox Terriers than in the Beagles, and T was lower in the Fox Terriers at some times but not others (p < 0.05). Thyroid Stimulating Hormone (TSH) concentrations did not differ among breeds, while mean basal T₄ values were lower in Fox Terriers compared with Labrador Retrievers and Great Danes (p < 0.05). Stimulation of T₄ secretion 120 and 180 min after iv TRH injection was most pronounced in the Beagles and less in the Fox Terriers (p < 0.05). The results of the present study indicate that potential breed differences in circulating concentrations of PRL, LH, T, TSH and T₄ in male dogs with apparently normal fertility can be encountered, but further studies are needed to determine whether the observed differences are typical features of these breeds, reflect subsets of dogs within breeds, or are in part because of possible uncontrolled parameters such as sample timing, ambient photoperiod, housing conditions or diet.     

Plasma Hormone Levels and Semen Quality in Male Cats during Non-Breeding and Breeding Seasons

Female cats are known to be seasonal breeders and male cats annual breeders. Despite this, there are limited data on the influence of breeding season (BS) on hormone concentration and semen quality in the male cat. This study compared plasma concentrations of LH and testosterone (T), and semen quality during the non-breeding season (NBS) and BS in five male cats subject to natural hours of daylight but a constant environmental temperature. Plasma LH and T concentrations were higher during the BS in 2/35 and 3/5 cats, respectively, although when comparing both hormones combined, values were higher during the BS than the NBS in all cats (p < 0.01). There were no significant differences in the percentage of abnormal sperm between the cats. Overall, semen quality was superior during the BS with larger semen volume in 2/5, sperm motility in 2/5 and sperm viability in 3/5 cats. Although there was a clear seasonal effect on hormone secretion and semen quality, during the NBS all cats were likely to have been fertile.     

Effect of an Injectable Cabergoline Formulation on Serum Prolactin (PRL) and Milk Secretion in Early Postpartum Beagle Bitches

This study was conducted in order to evaluate effects on prolactin (PRL) concentration and mammary milk secretion of an injectable cabergoline formulation administered to five lactating Beagle bitches during early postpartum (PP). Bitches were bled twice daily (from PP day 3 to PP day 12) and then daily (from PP day 13 to PP day 16) to assay serum PRL. On PP day 6, a subcutaneous (SC) injection of 0.1 ml/kg of placebo was administered. On PP day 9, a SC 0.1 ml/kg dose of injectable cabergoline was administered. All bitches were checked for milk production, using a clinical scoring in order to quantify milk expression from each teat. A circadian variation of serum PRL was evident during the 6 days of pre-treatment monitoring. The day after cabergoline injection, an 80% decrease of PRL serum concentration was observed (p < 0.05). The circadian oscillatory pattern of PRL secretion disappeared after administration of cabergoline, and PRL values remained significantly lower than in the previous days for the first 60 h following treatment (p < 0.001). Milk production was drastically reduced when comparing pre-treatment to post-treatment scores (p < 0.001). A single dose of injectable cabergoline caused a significant reduction in serum PRL concentration and a significant reduction in milk flow. The injectable formulation of cabergoline appeared to be safe and well tolerated.     

The effects of equine somatotropin on pituitary and testicular function in the stallion during the nonbreeding season

An experiment was conducted to determine the effects of equine somatotropin on the reproductive axis of the stallion during the nonbreeding season. Adult stallions were treated with equine somatotropin (20 μg/kg body weight [BW]; n = 5) or saline (n = 4) daily for 21 days starting in January. During the last week of treatment, stallions were subjected to low- and high-dose injections of luteinizing hormone (LH), as well as low- and high-dose injections of gonadotropin-releasing hormone (GnRH) and thyrotropin-releasing hormone (TRH). Two months after the onset of somatotropin treatment, semen was collected from all stallions every other day for 14 days. Treatment with equine somatotropin increased (P < .001) daily IGF-1 concentrations but had no effect (P > .1) on concentrations of LH, follicle-stimulating hormone (FSH), or testosterone. The testosterone responses to injections of LH were similar (P > .1) between treatments. Likewise, the LH, FSH, prolactin, and testosterone responses to the injections of GnRH/TRH were similar (P > .1) between groups. At seminal collections, stallions treated with somatotropin exhibited greater volumes of gel-free semen (P < .01) and gel (P < .05) and had decreased time until ejaculation (P < .05). In conclusion, somatotropin treatment for 21 days may alter the long-term accessory gland contribution to seminal volume but does not appear to alter pituitary gonadotrope function or testicular testosterone secretion.     

Effects of dopamine, norepinephrine and serotonin on secretion of luteinizing hormone, follicle-stimulating hormone and prolactin in ovariectomized, pituitary stalk-transected ewes

Two experiments were conducted in ovariectomized, pituitary stalk-transected ewes to determine if dopamine (DA), norepinephrine (NE) or serotonin (5-HT) alter secretion of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL). In experiment 1, ewes were infused (iv) with saline (control), DA (66 micrograms/kg/min), NE (6.6 micrograms/kg/min) or 5-HT (6.6 micrograms/kg/min). Treatments did not alter pulse frequency, but 5-HT increased (P less than .05) amplitude of pulses of LH and mean concentrations of LH, DA and NE were without effect on basal secretion of LH. DA but not NE or 5-HT decreased (P less than .05) the release of LH in response to gonadotropin hormone-releasing hormone (GnRH, 25 micrograms, im). Concentrations of FSH were not affected by treatments. Secretion of PRL was reduced (P less than .05) by treatment with DA and NE but not 5-HT. Each amine reduced (P less than .05) the release of PRL in response to thyrotropin-releasing hormone (TRH; 3 micrograms, im). In experiment 2, ewes were given DA at doses of 0, 0.66, 6.6 or 66.0 micrograms/kg/min, iv. No dose altered basal LH, but each dose reduced (P less than .05) basal and TRH-induced release of PRL. Key findings from these studies include direct pituitary action for: (1) 5-HT enhanced basal secretion of LH, (2) suppression of GnRH-induced secretion of LH by DA. (3) DA and NE inhibition of PRL secretion, and (4) DA, NE and 5-HT inhibition of release of PRL in response to TRH.     

Effect of hCG in the Presence of hCG Antibodies on the Follicle, Hormone Concentrations, and Oocyte in Mares

Follicle blood flow, follicular-fluid and plasma hormone concentrations, and oocyte quality were studied 30 h after an ovulation-inducing hCG treatment when the pre-ovulatory follicle was 32 mm. Mares were grouped as positive (n = 16) and negative (n = 44) for hCG antibodies before the experimental hCG treatment. Percentage of the follicle wall with blood flow signals was less (p < 0.05) in the antibody positive group than in the negative group. The concentrations of follicular-fluid oestradiol and free IGF1, and plasma oestradiol were greater (p < 0.001), and follicular-fluid progesterone (p < 0.001) and plasma LH (p < 0.02) were less in the antibody-positive group than in the negative group. For recovered oocytes at 30 h (n = 37), the antibody-positive group had fewer (p < 0.001) mature (MII) oocytes than the antibody-negative group. Results were attributable to highly effective neutralization of the hCG in the antibody-positive group.     

Evidence for endogenous opioid modulation of serum luteinizing hormone and prolactin in the steer

Opioid modulation of LH and prolactin (PRL) concentrations in Angus steers was investigated. In Exp. 1, morphine sulfate (M) was administered at either 1, 2 or 3 mg/kg BW (n = 4) as an i.v. injection. Blood samples were obtained at 15-min intervals for 4 h pre- and post-treatment for serum hormone analyses. Mean serum LH concentration and number of LH secretory pulses decreased (P less than .1) for 2 h after M (4.1 to nadir of 2.4 ng/ml, and .33 vs. .21 pulses/h; pre- vs post-treatment). Luteinizing hormone pulse amplitude decreased (P less than .01; 7.3 vs 2.6 ng/ml; pre- vs post-treatment) during the 2 h following M. Prolactin concentrations increased 126.6%, 170.6% and 187.6% following 1, 2 and 3 mg M/kg BW, respectively (P less than .05, 1 vs 2; P less than .01, 1 vs 3). In Exp. 2, either saline solution (S, n = 6) or M (.31 mg/kg BW, i.v. injection followed by .15 mg/(kg.h) infusion; n = 6) was given for 7 h. Concentration of LH was unaffected. Response of LH to naloxone was determined in Exp. 3. Blood samples were obtained for 2 h pre- and post-administration of either naloxone (1 mg/kg BW, i.v. injection; n = 5) or S (n = 5). Response of LH at 15, 30 and 45 min posttreatment was greater (P less than .05) in naloxone- compared with S-treated steers. In summary, M had no significant effect on serum LH concentration or LH pulse frequency, but it decreased pulse amplitude and increased serum PRL concentrations. In contrast, naloxone increased LH secretion. These observations taken together indicate a physiological role for opioid modulation of LH and PRL secretion in the steer.     

Reproductive characteristics of stallions during the breeding and non-breeding season in a tropical region

The objective of this study was to investigate reproductive characteristics of stallions at a tropical zone in the breeding and non-breeding seasons. The following parameters were assessed: testicular volume; semen quality; and serum concentrations of LH, FSH, and testosterone; in addition to the percentages of germ cells and proportions of germ cells/Sertoli cells by testicular cytology in stallions. Semen was collected from eight adult stallions twice a week during a 12-week period in both seasons (6?weeks before and 6?weeks after the summer and winter solstices). Jugular blood samples were collected periodically for hormone analysis by radioimmunoassay during the same periods. Testicular measures and cytological samples were taken at the end of each period. Mean concentration of testosterone was significantly higher (P?=?0.04) during the breeding season and the proportion of Sertoli cells/100 germ cells in cytological smears was significantly lower during the breeding season (P?=?0.0001). Effects of season were not significant either for testicular volume or for any semen parameter (P?>?0.05). Seasonal changes in the mean concentrations of LH and FSH were not observed (P?>?0.05). There were also no significant differences in the mean percentages of germ cell types between both seasons (P?>?0.05). Lack of seasonal differences in the testicular volume and semen parameters of tropical stallions are probably due to the small variation in duration of natural light between the observed periods, slightly under 3?h.     

Development of One vs Multiple Ovulatory Follicles and Associated Systemic Hormone Concentrations in Mares

Ablation of follicles ≥  6 mm in diameter and treatment with PGF2α 10 days after ovulation were used to induce the development of ovulatory waves. Comparisons were made between induced waves with one (33 waves, 72%) and multiple (13 waves, 28%) ovulatory follicles. Diameter deviation was defined as the separation of follicles into dominant and subordinate categories. Multiple ovulatory follicles were preceded by more (p < 0.001) follicles ≥ 20 mm at the beginning of deviation, higher LH preceding deviation (approached significance, p < 0.08), lower (p < 0.05) concentrations of FSH on the day of deviation and thereafter, and higher (p < 0.0003) oestradiol by 2 days after deviation. During the peri-ovulatory period, systemic hormone concentrations for waves with multiple ovulations involved higher oestradiol before ovulation (approached significance, p < 0.07), lower FSH (p < 0.04) before and after ovulation, and both higher progesterone (p < 0.05) and lower LH (p < 0.05) beginning the day after ovulation. Results indicated that by the beginning of deviation there were more follicles ≥  20 mm and subsequently greater oestradiol production in waves that led to the development of multiple ovulatory follicles, and during the peri-ovulatory period differences between one and multiple ovulations were consistent with the negative effects of the ovarian hormones on the gonadotropins.     

Effect of Antioxidants During Bovine In Vitro Fertilization Procedures on Spermatozoa and Embryo Development

Increased amounts of reactive oxygen species (ROS) during in vitro fertilization (IVF) may cause cytotoxic damage to gametes, whereas small amounts of ROS favour sperm capacitation. The aim of this study was to investigate the effect of antioxidants [50 μ m β-mercaptoethanol (β-ME) and 50 μ m cysteamine (Cyst)] or a pro-oxidant (5 m m buthionine sulfoximine) on the quality and penetrability of spermatozoa into bovine oocytes and on the subsequent embryo development and quality when added during IVF. Sperm quality, evaluated by the integrity of plasma and acrosomal membranes, and mitochondrial function, was diminished (p < 0.05) after 4-h culture in the presence of antioxidants. Oocyte penetration rates were similar between treatments (p > 0.05), but antioxidants adversely affected the normal pronuclear formation rates (p < 0.05). The incidence of polyspermy was high for β-ME (p < 0.05). No differences were observed in cleavage rates between treatments (p > 0.05). However, the developmental rate to the blastocyst stage was adversely affected by Cyst treatment (p < 0.05). The quality of embryos that reached the blastocyst stage, evaluated by total, inner cell mass (ICM) and trophectoderm cell numbers and ICM/total cell ratio was unaffected (p > 0.05) by treatments. The results indicate that ROS play a role in the fertilizing capacity in bovine spermatozoa, as well as in the interaction between the spermatozoa and the oocytes. It can be concluded that supplementation with antioxidants during IVF procedures impairs sperm quality, normal pronuclear formation and embryo development to the blastocyst stage.     

Relationships among LH, FSH and prolactin secretion, storage and response to secretagogue and hypothalamic GnRH content in ovariectomized pony mares administered testosterone, dihydrotestosterone, estradiol, progesterone, dexamethasone or follicular fluid

Thirty-five ovariectomized pony mares were used to study the relationships among luteinizing hormone (LH), follicle stimulating hormone (FSH) and prolactin (PRL) concentrations in blood (secretion), in pituitary (storage) and in blood after secretagogue administration, as well as the content of gonadotropin releasing hormone (GnRH) in hypothalamic areas, under various conditions of steroidal and nonsteroidal treatment. Five mares each were treated daily for 21 d with vegetable shortening (controls), testosterone (T; 150 micrograms/kg of body weight, BW), dihydrotestosterone (DHT; 150 micrograms/kg BW), estradiol (E2; 35 micrograms/kg BW), progesterone (P4; 500 micrograms/kg BW), dexamethasone (DEX; 125 micrograms/kg BW) or charcoal-stripped equine follicular fluid (FF; 10 ml). Secretagogue injections (GnRH and thyrotropin releasing hormone, TRH, at 1 and 4 micrograms/kg of BW, respectively) were given one d prior to treatment and again after 15 d of treatment. Relative to controls, treatment with T, DHT and DEX reduced (P less than .05) LH secretion, storage and response to exogenous GnRH, whereas treatment with E2 increased (P less than .05) these same characteristics. Treatment with P4 reduced (P less than .05) only LH secretion. Treatment with T, DHT, E2 and DEX reduced (P less than .05) FSH secretion, whereas treatment with P4 increased (P less than .05) it and FF had no effect (P greater than .1). All treatments increased (P less than .05) FSH storage, whereas only treatment with T and DHT increased (P less than .05) the FSH response to exogenous GnRH. Other than a brief increase (P less than .05) in PRL secretion in mares treated with E2, secretion of PRL did not differ (P greater than .1) among groups. Only treatment with E2 increased (P less than .01) PRL storage, yet treatment with T or DHT (but not E2) increased (P less than .05) the PRL response to exogenous TRH. Content of GnRH in the body and pre-optic area of the hypothalamus was not affected (P greater than .1) by treatment, whereas treatment with T, E2 and DEX increased (P less than .1) GnRH content in the median eminence. For LH, secretion, storage and response to exogenous GnRH were all highly correlated (r greater than or equal to .77; P less than .01). For FSH, only storage and response to exogenous GnRH were related (r = .62; P less than .01). PRL characteristics were not significantly related to one another. Moreover, the amount of GnRH in the median eminence was not related (P greater than .1) to any LH or FSH characteristic.     

Comparison of Two Doses of the GnRH Antagonist, Acyline, for Pregnancy Termination in Bitches

Gonadotropin-releasing hormone (GnRH) antagonists are particularly useful when a rapid inhibitory effect on the gonadal axis is required. The aim of this study was to test the efficacy and clinical safety of a low and high dose of the third generation GnRH antagonist, acyline, on pregnancy termination in female dogs. The effect of the antagonist on the progesterone (P₄) serum concentration was also described. Twenty-one mid-pregnant bitches were randomly assigned to a single subcutaneous (SC) dose of a placebo (PLACE; n = 7), a low (ACY-L; 110 μg/kg; n = 6) or high (ACY-H; 330 μg/kg; n = 8) dose of acyline. The animals were followed up for 15 days. All ACY treated but no placebo-treated animals terminated their pregnancy by abortion (p < 0.01). The ACY-L and ACY-H groups interrupted their pregnancy 7 ± 1.9 and 6.4 ± 1.3   days after treatment, respectively (p = 0.7). A significant interaction between treatment and day was found (p < 0.01) for P₄ serum concentrations when PLACE was compared with both ACY groups. No difference was found for this hormone between both ACY groups (p > 0.05) where P₄ diminished throughout the study. The decreasing rate varied among animals and was closely related to the time of abortion when P₄ reached basal concentrations. In PLACE animals, gestation progressed normally and P₄ did not change throughout the study (p > 0.05). None of the bitches presented side effects. It was concluded that acyline safely terminated mid-pregnancy by permanently decreasing P₄ serum concentrations.     

Effect of furazolidone and nitrofurazone on egg production, on plasma luteinising hormone and on prolactin concentrations in turkeys

1. Furazolidone or nitrofurazone were given orally to laying turkeys at doses of 7.5, 15 or 30 mg/kg for 7 d. Plasma concentrations of luteinising hormone (LH), prolactin (PRL) and egg production were measured before, during and after treatment. 2. Both drugs produced dose-dependent decreases in LH concentration which were statistically significant at doses of 15 and 30 mg/kg. Plasma PRL concentration was significantly increased in birds receiving 15 or 30 mg/kg of nitrofurazone, and tended to increase in the other treated groups, but this was not statistically significant. 3. Egg production was lowered in a dose-dependent manner by both drugs. However, nitrofurazone appeared to be more potent in reducing egg production than furazolidone. 4. Birds given 15 mg/kg of either drug were injected intramuscularly with luteinising hormone releasing hormone (LHRH) at a dose of 5 micrograms/kg and blood was collected immediately before and 30 min after LHRH administration. 5. Nitrofurazone significantly reduced the rise in LH induced by LHRH. Seven days after withdrawing the drug, the LHRH-induced LH release was not significantly different when compared to that in the control group or that seen on day 7 of treatment.     

Endocrine changes in sows exposed to elevated ambient temperature during lactation

Seven sows were placed into one of two environmental chambers at 22 C, 5 d prior to farrowing. On day 9 of lactation, one chamber was changed to 30 C (n = 4) and the other remained at 22 C (n = 3). On days 24 and 25, blood samples were collected every 15 min for 9 hr and 7 hr, respectively. On day 24, thyrotropin releasing hormone (TRH) and gonadotropin releasing hormone (GnRH) were injected iv at hour 8. On day 25 naloxone (NAL) was administered iv at hour 4 followed 2 hr later by iv injection of TRH and GnRH. Milk yield and litter weights were similar but backfat thickness (BF) was greater in 22 C sows (P less than .05) compared to 30 C sows. Luteinizing hormone (LH) pulse frequency was greater (P less than .003) and LH pulse amplitude was less (P less than .03) in 22 C sows. LH concentrations after GnRH were similar on day 24 but on day 25, LH concentrations after GnRH were greater (P less than .05) for 30 C sows. Prolactin (PRL) concentrations were similar on days 24 and 25 for both groups. However, PRL response to TRH was greater (P less than .05) on both days 24 and 25 in 30 C sows. Growth hormone (GH) concentrations, and the GH response to TRH, were greater (P less than .0001) in 30 C sows. Cortisol concentrations, and the response to NAL, were less (P less than .03) in 30 C sows. NAL failed to alter LH secretion but decreased (P less than .05) PRL secretion in both groups of sows. However, GH response to NAL was greater (P less than .05) in 30 C sows. Therefore, sows exposed to elevated ambient temperature during lactation exhibited altered endocrine function.     

Interaction between salsolinol (SAL) and thyrotropin-releasing hormone (TRH) or dopamine (DA) on the secretion of prolactin in ruminants

We have recently demonstrated that salsolinol (SAL), a dopamine (DA)-derived compound, is present in the posterior pituitary gland and is able to stimulate the release of prolactin (PRL) in ruminants. The aim of the present study was to clarify the effect that the interaction of SAL with thyrotropin-releasing hormone (TRH) or DA has on the secretion of PRL in ruminants. A single intravenous (i.v.) injection of SAL (5mg/kg body weight (b.w.)), TRH (1microg/kg b.w.), and SAL plus TRH significantly stimulated the release of PRL in goats (P<0.05). The cumulative response curve (area under the curve: AUC) during 120min was 1.53 and 1.47 times greater after the injection of SAL plus TRH than either SAL or TRH alone, respectively (P<0.05). A single i.v. injection of sulpiride (a DA receptor antagonist, 0.1mg/kg b.w.), sulpiride plus SAL (5mg/kg b.w.), and sulpiride plus TRH (1microg/kg b.w.) significantly stimulated the release of PRL in goats (P<0.05). The AUC of PRL during 120min was 2.12 and 1.78 times greater after the injection of sulpiride plus TRH than either sulpiride alone or sulpiride plus SAL, respectively (P<0.05). In cultured bovine anterior pituitary (AP) cells, SAL (10(-6)M), TRH (10(-8)M), and SAL plus TRH significantly increased the release of PRL (P<0.05), but the additive effect of SAL and TRH detected in vivo was not observed in vitro. In contrast, DA (10(-6)M) inhibited the TRH-, as well as SAL-induced PRL release in vitro. All together, these results clearly show that SAL can stimulate the release of PRL in ruminants. Furthermore, they also demonstrate that the additive effect of SAL and TRH on the release of PRL detected in vivo may not be mediated at the level of the AP, but that DA can overcome their releasing activity both in vivo and in vitro, confirming the dominant role of DA in the inhibitory regulation of PRL secretion in ruminants.     

Effect of supplementation with the reduced form of coenzyme Q10 on semen quality and antioxidant status in dogs with poor semen quality: Three case studies

Oxidative stress owing to an imbalance between reactive oxygen species and antioxidants, such as coenzyme Q10 (CoQ10), is a major contributor to male infertility. We investigated the effects of the reduced form of CoQ10 (ubiquinol) supplementation on semen quality in dogs with poor semen quality. Three dogs received 100 mg of ubiquinol orally once daily for 12 weeks. Semen quality, serum testosterone, and seminal plasma superoxide dismutase (SOD) activity were examined at 2-week intervals from 2 weeks before ubiquinol supplementation to 4 weeks after the treatment. Ubiquinol improved sperm motility, reduced morphologically abnormal sperm, and increased seminal plasma SOD activity; however, it had no effect on testosterone level, semen volume, and sperm number. Ubiquinol supplementation could be used as a non-endocrine therapy for infertile dogs.     

Libido, hormone concentrations in blood plasma and semen characteristics in Holstein bulls

Relationships among bull libido, serum hormone concentrations and semen characteristics were studied using 18 Holstein bulls that were 4 to 5 yr old. The hormones studied included testosterone, estradiol (E2), prolactin (PRL), LH and cortisol. Two ejaculates were collected three times per week from each bull during a 5-wk trial. During the last week of the trial, on a day semen was not collected, blood was collected from indwelling catheters every 15 min for 6 h to determine the hormonal profiles of each bull. On the following day, blood was sampled every 10 min before and after the time of semen collection. Libido factors were quantified, and semen volumes and sperm concentrations were recorded. The libido factors included reaction time to first service, latency time between the first and second semen collections, and duration of time the bull mounted the teaser prior to the first (TM1) and second (TM2) semen collection. Average reaction and latency times were correlated (r = .524; P = .026), as were TM1 and TM2 (r = .597; P = .015). Latency times were correlated with average TM2 (r = .669; P = .003). Average PRL concentrations were correlated with average latency times (r = .467; P = .05). Low libido bulls tended to have higher E2:testosterone ratios than did high libido bulls. Both PRL and cortisol concentrations increased at semen collection.     

Effect of Progesterone and Ionomycin on Domestic Cat Sperm Motility Patterns and Acrosome Reaction

This study was aimed at assessing the changes in sperm motion patterns and the percentage of acrosome reaction (AR) in domestic cat semen after treatment with either ionomycin or progesterone (P₄). Ten ejaculates were collected from five tomcats using an artificial vagina, and were diluted, centrifuged and resuspended in a capacitation medium. Samples were evaluated and divided into seven equal aliquots and, after 2 h at 25°C, were incubated for 30 min at 38°C in 5% CO₂ and then analyzed. Computer-assisted sperm analysis and a combination of three fluorescent probes were used to assess sperm plasma, acrosomal membrane integrity and mitochondrial transmembrane potential. Thirty minutes after the start of incubation, P₄ was added (10 μg/ml) to the P1 group. Groups P2 and P3 were supplemented with P₄ (10 and 20 μg/ml, respectively) only after 2 h of incubation, and groups I1 and I2 were supplemented with ionomycin (4 and 8 μ m , respectively) 2 h after incubation. Group E was supplemented with ethanol (0.6%) at 2 h after incubation and group C received no supplementation. Ionomycin and P₄ treatments led to a hyperactivation-like sperm motion and an increase (p < 0.05) in the percentage of AR. Although a higher (p < 0.05) percentage of AR was obtained in group I2 when compared with all P₄ groups, a decrease (p < 0.05) in total and progressive motility was observed in I2 group. As I1 group was similar to I2 to induce AR without diminishing sperm motility, we can conclude that ionomycin at 4 μ m seems to be more suitable to trigger AR in domestic cat sperm.     

Bromocriptine inhibits salsolinol‐induced prolactin release in male goats

The secretion of prolactin (PRL) is under the dominant and tonic inhibitory control of dopamine (DA); however, we have recently found that salsolinol (SAL), an endogenous DA‐derived compound, strongly stimulated the release of PRL in ruminants. The aim of the present study was to clarify the inhibitory effect of DA on the SAL‐induced release of PRL in ruminants. The experiments were performed from late June to early July. Male goats were given a single intravenous (i.v.) injection of SAL (5 mg/kg body weight (BW)), a DA receptor antagonist (sulpiride, 0.1 mg/kg BW), or thyrotropin‐releasing hormone (TRH, 1 µg/kg BW) before and after treatment with a DA receptor agonist (bromocriptine), and the effect of DA on SAL‐induced PRL release was compared to that on sulpiride‐ or TRH‐induced release. Bromocriptine completely inhibited the SAL‐induced release of PRL (P < 0.05), and the area under the response curve (AUC) for a 120‐min period after the treatment with bromocriptine was 1/28 of that for before the treatment (P < 0.05). Bromocriptine also completely inhibited the sulpiride‐induced release (P < 0.05). The AUC post‐treatment was 1/17 that of pre‐treatment with bromocriptine (P < 0.05). Bromocriptine also inhibited the TRH‐induced release (P < 0.05), though not completely. The AUC post‐treatment was 1/3.8 that of pre‐treatment (P < 0.05). These results indicate that DA inhibits the SAL‐induced release of PRL in male goats, and suggest that SAL and DA are involved in regulating the secretion of PRL. They also suggest that in terms of the regulatory process for the secretion of PRL, SAL resembles sulpiride but differs from TRH.     

Assessment of a combined anterior pituitary function test in beagle dogs: Rapid sequential intravenous administration of four hypothalamic releasing hormones

A combined anterior pituitary (CAP) function test was assessed in eight healthy male beagle dogs. The CAP test consisted of sequential 30-second intravenous administrations of four hypothalamic releasing hormones in the following order and doses: 1 μg of corticotropin-releasing hormone (CRH)/kg, 1 μg of growth hormone-releasing hormone (GHRH)/kg, 10 μg of gonadotropinreleasing hormone (GnRH)/kg, and 10 μg of thyrotropin-releasing hormone (TRH)/kg. Plasma samples were assayed for adrenocorticotropin, cortisol, GH, luteinizing hormone (LH), and prolactin (PRL) at multiple times for 120 min after injection. Each releasing hormone was also administered separately in the same dose to the same eight dogs in order to investigate any interactions between the releasing hormones in the combined function test.Compared with separate administration, the combined administration of these four hypothalamic releasing hormones caused no apparent inhibition or synergism with respect to the responses to CRH, GHRH, and TRH. The combined administration of these four hypothalamic releasing hormones caused a 50% attenuation in LH response compared with the LH response to single GnRH administration. The side effects of the combined test were confined to restlessness and nausea in three dogs, which disappeared within minutes after the administration of the releasing hormones. It is concluded that with the rapid sequential administration of four hypothalamic releasing hormones (CRH, GHRH, GnRH, and TRH), the adenohypophyseal responses are similar to those occurring with the single administration of these secretagogues, with the exception of the LH response, which is lower in the CAP test than after single GnRH administration.