Effect of prostaglandin F2 alpha on the adenylate cyclase and phosphodiesterase activity of ovine corpora lutea

The objective of the present study was to determine the temporal relationships among luteal adenylate cyclase activity, luteal phosphodiesterase activity, luteal progesterone concentration and plasma progesterone concentration during prostaglandin F2 alpha (PGF2 alpha)-induced luteolysis in ewes. Corpora lutea (CL) were removed from cycling ewes on d 9 (d 0 = first day of estrus) at 0, 2, 4, 6, 12 and 24 h (seven to eight ewes/group) after PGF2 alpha administration (im). Jugular blood samples were collected at the time of enucleation of CL and analyzed for progesterone. Plasma and luteal progesterone concentrations were decreased (P less than .05) by 4 and 12 h after PGF2 alpha injection, respectively. Basal adenylate cyclase, luteinizing hormone (LH)-activated adenylate cyclase, guanylylimidodiphosphate [Gpp(NH)p]-activated adenylate cyclase and LH plus Gpp(NH)p-activated adenylate cyclase activities were decreased (P less than .05) by 2 h after PGF2 alpha injection. The decrease in adenylate cyclase activity paralleled the decrease in plasma progesterone concentration over time. Luteinizing hormone stimulated (P less than .05) adenylate cyclase activity relative to basal activity at 0, 2, 12 and 24 h post-PGF2 alpha; whereas, Gpp(NH)p stimulated (P less than .01) adenylate cyclase activity relative to basal activity at each time point. In contrast to the decrease in adenylate cyclase activity, phosphodiesterase activity was increased (P less than .05) at 2 and 4 h post-PGF2 alpha. These results suggest that a decrease in adenylate cyclase activity coupled with an increase in phosphodiesterase activity may decrease the intracellular adenosine 3',5' cyclic monophosphate (cAMP) concentration.(ABSTRACT TRUNCATED AT 250 WORDS)     

Semen Characteristics and Plasma Levels of Testosterone after Bilateral Vasectomy in Bucks

The effects of bilateral vasectomy on the seminal characteristics were assessed in six bucks of the Canarian breed. In addition, we tried to establish the effects of vasectomy on the plasmatic concentrations of testosterone and the libido of the bucks. Semen samples were collected once a week from 8 weeks before to 16 weeks after vasectomy; blood samples were collected prior to vasectomy, and then at once and 1 week after vasectomy and every 2 weeks from the week 4 to the end of the experiment. One week after the vasectomy, ejaculated spermatozoa were non‐motile and the percentage of live spermatozoa was below 5% in all vasectomized males; in addition, the total number of cells/ejaculate was 3100 × 10⁶ and 30 × 10⁶ spermatozoa in the control and vasectomized males, respectively. Our results suggest that the vasectomized males may be used as oestrus detectors, without risks of accidental fecundating, only 1 week after vasectomy. Before vasectomy, no significant differences were observed in plasma levels of testosterone between the vasectomized and control males (5.4 ± 1.2 and 3.9 ± 1.4 ng/ml, respectively); from 4 to 12 weeks after vasectomy, a marked decrease in the testosterone concentration in all males (vasectomized and control bucks) was observed. From 12 weeks after vasectomy until the end of the experiment, four of the vasectomized males and the control males recovered their normal libido. The results suggest that vasectomy did not exert a remarkable effect on the steroidogenic functionality of the testicle.     

Clinical and Pathological Findings in Testis, Epididymis, Deferens Duct and Prostate following Vasectomy in a Dog

We report the case of a bilateral and multilocular spermatocele and sperm granuloma in a dog that was vasectomized 5 years before. Clinical examination revealed scrotal dermatitis and benign prostatic hyperplasia. Orchiectomy was performed, and gross and histological examination showed testicular degeneration associated with epididymal sperm granuloma. In relation to this case, the literature about long-term effects of vasectomy in dogs has been reviewed. On the basis of these results, a preventive sonogram and physical assessment in prostate and other reproductive structures before vasectomy is recommended.     

Inhibition of macrophage adenylate cyclase by the alpha-methylene-gamma-lactone moiety of sesquiterpene lactones from forage plants

Inhibition of murine macrophage adenylate cyclase activity by sesquiterpene lactones isolated from toxic forage plants was highly correlated with the presence of the alpha-methylene-gamma-lactone moiety on the molecule (ie, hymenovin and helenalin). Tenulin, a sesquiterpene lactone which does not contain this reactive moiety, caused minimal inhibition of the enzyme. Reaction of the alpha-methylene-gamma-lactone moiety of hymenovin and helenalin with cysteine decreased the number of reactive moieties available to alkylate the enzyme, thus decreasing the inhibition of adenylate cyclase by these 2 sesquiterpene lactones. As the reaction time available for the reduction by cysteine of the alpha-methylene-gamma-lactone moiety decreases, the amount of adenylate cyclase inhibition increases. Stimulation of the hymenovin- or helenalin-inhibited adenylate cyclase by prostaglandin E1 or E2 or by sodium fluoride did not reverse the inhibition of the enzyme, but did stimulate the undamaged adenylate cyclase in the sesquiterpene lactone treatment groups to the same degree as in the nontreated control. These data indicate that sesquiterpene lactones containing an alpha-methylene-gamma-lactone moiety are potent inhibitors of macrophage adenylate cyclase activity. This moiety may have a significant role in the toxicity of some sesquiterpene lactones in poisonous plants when ingested by livestock.     

猪脂肪沉积的品系差异及其cAMP的调控作用

体重、健康状况相近的８月龄混系和丹系长白肥育猪１６头，空腹采血测定ｃＡＭＰ、胰岛素、激素敏感脂酶（ＨＳＬ）、游离脂肪酸（ＦＦＡ）和甘油三酯；屠宰后测定胴体指标，并采背部脂肪和肝脏组织，分析了ｃＡＭＰ和腺苷酸环化酶（ＡＣ）活性以及脂肪组织中脂肪酸组成．结果表明，与混系相比，丹系瘦肉率、眼肌面积和后腿比分别高９．４４％（Ｐ＜０．０１）、９．６７％和４．００％，背膘厚、脂肪率和板油比分别低７．９７％、１８．１０％（Ｐ＜０．０５）和１４．２５％。血浆ｃＡＭＰ、ＨＳＬ和ＦＦＡ分别比混系高３２．８５％（Ｐ＜０．０５）、７．４１％和１２３．４１％；甘油三酯和胰岛素则分别低１２．２３％和１８．０６％．丹系脂肪组织中总脂，１４碳和１６碳脂肪酸分别高１０．８５％、３９．６６％（Ｐ＜０．０１）和７．３１％，而１８碳脂肪酸则低４．３１％；丹系肝和脂肪组织中ｃＡＭＰ和ＡＣ显著高于混系。结果证明，猪脂肪沉积的品系差异与肝、脂肪组织及血中ｃＡＭＰ的水平有关，脂肪沉积的ｃＡＭＰ调控具有明显的品系差异。     

Effect of flushing the vasa deferentia at the time of vasectomy on the rate of clearance of spermatozoa from the ejaculates of dogs and cats

Flushing the vasa deferentia (ductus deferentes) at the time of vasectomy reduced to zero the number of intact spermatozoa by postvasectomy day 6 in the dog and by postvasectomy day 7 in the cat and shortened the time from vasectomy to azoospermia in the dog, but not in the cat. The fluid used to flush the vasa deferentia was not eliminated through the penile urethra, but flowed into the urinary bladder, indicating that the least resistant pathway for the exit of vasal content in the anesthetized dog and cat is toward the urinary bladder. Both control and treated dogs and cats had spermatozoa in the urine obtained by cystocentesis immediately after ejaculation or ejaculation and flushing of the vasa deferentia. Flushing the vasa deferentia at the time of vasectomy is easy to do, safe, and can be used in clinical practice to decrease the time from vasectomy to the safe utilization of dogs and cats as teasers. The procedure has potential application to males of other species.     

Changes and interrelationships among luteal LH receptors, adenylate cyclase activity and phosphodiesterase activity during the bovine estrous cycle

Changes and interrelationships among the cellular mechanisms that may regulate luteal progesterone synthesis during the bovine estrous cycle were studied. Corpora lutea (CL) were enucleated from 30 cows via a transvaginal incision on d 4, 7, 10, 13, 16 and 19 following estrus (estrus = d 0). Mean corpus luteum weight, and luteal progesterone and plasma progesterone concentrations increased (P less than .05) from d 4 to 7 or 10, and declined following luteal regression (d 19). Unoccupied LH receptor (UOR) concentrations increased (P less than .05) more than fourfold from d 4 (38 fmol/mg protein) to d 16 (173 fmol/mg protein) before declining (P less than .05) by d 19 (30 fmol/mg protein). The dissociation constant for UOR concentrations increased (P less than .05) during the estrous cycle. Concentrations of occupied LH receptors (OR) were less (P less than .05) on d 7 than other days; however, the total number of OR/CL increased fourfold from d 4 (47 fmol/CL) to d 10 (221 fmol/CL) and remained similar thereafter. Basal adenylate cyclase, LH-activated adenylate cyclase, and Gpp(NH)p-activated adenylate cyclase activities were greatest (P less than .05) on d 7 to 16 as compared with d 4 and 19. Luteinizing hormone stimulated (P less than .05) adenylate cyclase activity relative to basal activity on d 7, 10, 13, and 16; whereas, Gpp(NH)p stimulated (P less than .05) adenylate cyclase activity relative to basal activity at each period. Phosphodiesterase was 46% greater on d 19 as compared with d 4.(ABSTRACT TRUNCATED AT 250 WORDS)     

Progesterone stimulation by LH involves the phospholipase-C pathway in bovine luteal cells

Luteinizing hormone (LH)-stimulated steroidogenesis in luteal cells is known to be mediated through the activation of cyclic AMP (cAMP)-dependent protein kinase, and to be also modulated by calcium-dependent mechanisms. In the present study, we tested the hypothesis that LH stimulates progesterone (P4) production in bovine luteal cells through activation of phospholipase (PL) C by using a cell culture system. Bovine mid-luteal cells (Days 8-12 of the estrous cycle) were cultured for 24 h and then exposed to a PLC inhibitor (U-73122; 10 microM) with or without LH (10 ng/ml) for 4 h. U-73122 blocked LH-stimulated P4 production without affecting cAMP accumulation. Moreover, exposure of luteal cells to PLC increased P4 production in a dose-dependent manner. These results support the hypothesis that the luteotropic action of LH in bovine luteal cells is mediated not only by activation of adenylate cyclase but also by activation of PLC.     

Cyclic AMP Signalling During Mammalian Sperm Capacitation – Still Largely Terra Incognita

Cyclic AMP is known to play a major role in intracellular signalling during mammalian sperm capacitation. However, despite much research, many of the molecular details of cyclic AMP's involvement remain obscure. In this review, I discuss the following aspects, presenting some original data as illustration where relevant. With respect to cyclic AMP synthesis, uncertainties exist as to the number of forms of adenylyl cyclase that are present in the spermatozoon, whether they are cytosolic or bound to subcellular structures, and which physiological effectors they respond to (e.g. bicarbonate, Ca²⁺, or receptor‐coupled G‐proteins). While net intracellular levels of cyclic AMP in spermatozoa depend upon the relative activities of adenylyl cyclase and phosphodiesterase, there are wide between‐sample variations within species, both in basal levels and in levels attained after activation of the cyclase (e.g. after sperm treatment with bicarbonate). Moreover, minor changes in bulk cyclic AMP levels can result in large changes in cyclic AMP‐dependent functions. Finally, while cyclic AMP levels respond very rapidly to sperm treatment by effectors such as bicarbonate and Ca²⁺ (key components of capacitating media), there are big discrepancies between the rates of functional response. For example, enhancement of motility and collapse of phospholipid asymmetry take place within a few minutes, whereas more than 1 h of exposure to capacitating conditions is needed for cyclic AMP‐dependent protein tyrosine phosphorylation to become detectable or for the sperm population to attain a capacitated state.     

Unilateral and Bilateral Vasectomy in the Dog: Alkaline Phosphatase as an Indicator of Tubular Patency

Contents The objective of this research was to use a model of unilateral and bilateral occlusion of the ductus deferens in the dog to study the use of alkaline phosphatase (AP) as an indicator of tubular patency. Seven healthy cross bred dogs weighing 10–15 kg BW with normal spermiogram and AP concentrations in semen were used. From each dog, three semen samples were obtained before (intact) and after right (unilateral) and left (bilateral) vasectomy. The AP concentrations were measured in duplicates by a colorimetric method in each of the three fractions (first, second (sperm-rich), third) of each ejaculate. In addition, a macroscopic and microscopic evaluation of each ejaculate was carried out to assure its quality. Data were analysed by least squares analysis of variance using SAS^®. In intact and unilateral vasectomized dogs, 96.6% of AP measured in semen corresponded to the second sperm-rich fraction whereas 1.53 and 1.83% corresponded to the first and third fractions respectively. Total AP concentrations (first and second and third fraction) in vasectomized dogs were lower than in intact animals (19.857 vs 2284.431 ± 4.347 UAL; p < 0.001). AP concentrations were much lower in bilateral than in unilateral vasectomized dogs (142 vs 39.572 ± 4.347 UL, p < 0.001). In summary, AP concentrations in semen can be used as an early indicator of unilateral or bilateral lack of patency of the epididymal and deferent ducts in the dog.     

Effects of Vasectomy on Seminal Plasma Alkaline Phosphatase in Male Alpacas (Vicugña pacos)

Azoospermia is a common finding in male alpacas which present for infertility. The challenge is to differentiate azoospermia of testicular origin from non‐testicular origin. In several species, alkaline phosphatase (AP) concentrations in seminal plasma have been used as a diagnostic marker of contributions of the testis and epididymis to the ejaculate. The purpose of this study was to determine whether AP assay could differentiate testicular from non‐testicular azoospermia in male alpacas. An experimental model of bilateral outflow obstruction (pre‐scrotal vasectomy) was used in 22 male alpacas, aged 2–9 years. No reproductive history was available. Animals were submitted for electroejaculation (EE) under general anaesthesia and vasectomy performed. Five weeks later, animals were submitted for EE. Vasectomy was not successful in one animal, which was removed from analysis. AP levels were compared in seminal plasma in the pre‐ and post‐vasectomy samples. The mean ± SEM concentration of AP in pre‐vasectomy seminal plasma was 504.29 ± 166.45 U/l (range 10–2910); the post‐vasectomy levels were 252.48 ± 81.77 U/l (range 0–1640; p = 0.06). In 71.4% of animals, AP levels decreased, varying from 18% to 100% reduction. Results of this study suggest that AP is not produced exclusively by the testis and epididymis in alpacas and that AP assay is not a valid diagnostic test for determination of origin of azoospermia; the gold standard for diagnosis of origin of azoospermia remains testicular biopsy.     

The effect of cholera toxin and heat labile and heat stable Escherichia coli enterotoxin on cyclic AMP concentrations in small intestinal mucosa of pig and rabbit.

The effect of cholera toxin, heat labile and heat stable Escherichia coli enterotoxin on mucosal cyclic AMP concentrations was determined on the proximal jejunum of weanling pigs and young rabbits. Ligated loops were injected with solutions containing no enterotoxin for control and either cholera toxin, heat labile or heat stable E. coli enterotoxin. The loops were drained after either two, four or six hours incubation at which time accumulated fluid was recorded and mucosal samples removed for determination of cyclic AMP concentration. In the rabbit, cholera toxin and heat labile, but not heat stable E. coli enterotoxin stimulated intestinal secretion while in the pig all three enterotoxins induced net fluid accumulation. Cholera toxin and heat labile, but not heat stable E. coli enterotoxin elevated rabbit mucosal cyclic AMP concentrations. In the pig these enterotoxins had no significant effect on mucosal cyclic AMP concentrations. The results are inconsistent with the hypothesis that the adenyl cyclase system is an essential step for enterotoxin induced intestinal secretion. The activation of intestinal adenyl cyclase by bacterial enterotoxins may only be an associated and not a necessary event for the stimulation of intestinal secretion.     

Surgical and chemical vasectomy in the cat

Ejaculates of surgically vasectomized cats had spermatozoa as long as 49 days after vasectomy, indicating that spermatozoa in the ejaculate from intact cats originated from the epididymides and vasa deferentia. Intraepididymal injections of an aqueous solution of 4.5% chlorhexidine digluconate into the caudae of the epididymides induced a lasting oligospermia or azoospermia in 7 of 8 cats. Of these 7 cats, 4 were azoospermic and 1 cat had no intact spermatozoa in his ejaculates 140 days after treatment. The method of chemical vasectomy by intraepididymal injection of sclerosing agents appears to be safe and may be suitable for large-scale sterilization programs for controlling the growth of the feline population.     

Cyclic nucleotides in hamster retina

The concentrations of cyclic 3':5'-adenosine monophosphate (cyclic AMP) and cyclic 3':5'-guanosine monophosphate (cyclic GMP) were determined in white light- and dim red light-adapted golden Syrian hamster retinas. Retinas from animals in dim red light had cyclic AMP concentrations of 9.29 +/- 2.94 pmol/mg of protein and cyclic GMP concentrations of 110.62 +/- 32.98 pmol/mg of protein. After white light adaptation, retinal cyclic AMP and cyclic GMP concentrations were reduced to 74% and 45% of the previous values, respectively. In another experiment with white light-adapted animals, the sex or method of immobilization of the animals (cervical dislocation vs sodium pentobarbital) had no significant effect on cyclic nucleotide values.     

Disappearance of spermatoza from the ejaculates of vasectomized dogs.

Ejaculates of bilaterally vasectomized dogs contained spermatozoa as long as 21 days after vasectomy, indicating that spermatozoa in the canine ejaculate originated from both the epididymides and the vasa deferentia, not from the epididymides alone.     

Effect of bilateral vasectomy on testosterone levels in the seminal and blood plasmas of boars]

In two sexually mature boars subjected to bilateral vasectomy the content of testosterone was followed in the semen and blood plasma prior to the operation and in the course of 26 days after the operation. In the third boar only the operation without actual vasectomy as the control was made to eliminate the possible effect of narcosis and of the surgical stress. Bilateral vasectomy induced an average decrease in the initial levels of testosterone in the semen plasma of the first and second animal by 63.24% and 47.67% respectively, and 47.17% and 56.46% respectively in the blood plasma. In the control boar no analogous post-operation drop of testosterone concentration in the semen plasma was observed, the content of testosterone in the blood plasma decreased over the followed period on an average only by 14.38%. The results imply that bilateral vasectomy in boars results, in the first followed month after the surgical procedure, in a decrease of semen and plasma testosterone, which proves that the operation under investigation affects testicular incretion.     

垂体腺苷酸环化酶激活多肽(PACAP)的研究概况

垂体腺苷酸环化酶激活多肽(PACAP)属于促胰液素胰高血糖素/VIP家族的新成员,最初发现其对大鼠垂体细胞腺苷酸环化酶(AC)具有高激活作用。随着研究的深入,发现其具有多种生物功能,文中就其影响动物对营养物质的消化,动物的繁殖性能,动物的能量平衡和动物的免疫系统等做简要概述。     

Effect of cohabitation with white-faced ewes on estrous activity of Hampshire and Suffolk ewes exposed to rams in June

Two groups of 24 Hampshire and 26 Suffolk purebred ewes each were used to study effects of cohabitation with cyclic white-faced (WF) ewes on estrous activity in June. Ewes lambed in January, February and March and had been isolated from mature rams since the previous fall breeding. From June 1 to July 2, treated (T) ewes were exposed to vasectomized rams and to 65 WF ewes; control (C) ewes were exposed only to vasectomized rams. Ovulation was assessed with biweekly serum progesterone assays; crayon marks were used to detect estrus. Daily observations of ram behavior were conducted to assess sexual activity of rams joined with T and C ewes. Cohabitation with WF ewes increased (P less than .01) ovulation percentages from 46% in C (42% for Hampshires and 50% for Suffolks) to 76% in T ewes (79% for Hampshires and 73% for Suffolks). Mating percentage also was increased (P less than .05) by cohabitation with WF ewes from 14% for C ewes to 30% for T ewes. Rams with T + WF ewes spent more (P less than .05) time checking ewes for estrus than did rams with C ewes. Hence, cohabitation with cycling WF ewes increased ovulation and mating percentages. Many acyclic T ewes first ovulated after 10 or more days of teasing, possibly due to increased ram contact in the presence of WF ewes.     

Differential effects of phosphodiesterase inhibitors on platelet activating factor (PAF)- and adenosine diphosphate (ADP)-induced equine platelet aggregation

Compounds that activate adenylate cyclase, increasing intracellular cyclic adenosine monophosphate (cAMP), inhibit equine platelet aggregation. Cyclic AMP is broken down by phosphodiesterase (PDE) and, in the present study, the effects of theophylline, a nonselective PDE inhibitor, and selective inhibitors of PDE isoenzymes PDE3, PDE4 and PDE5, on equine platelet aggregation in response to platelet activating factor (PAF) and adenosine diphosphate (ADP) have been examined. Theophylline and the PDE3 inhibitors, trequinsin and quazinone, inhibited both PAF and ADP-induced aggregation in a concentration dependent manner. The inhibition of PAF-induced aggregation was, however, significantly greater than that of the response to ADP. The inhibitory effects of theophylline and the PDE3 inhibitors on ADP- but not PAF-, induced aggregation were reversed by addition of the calcium ionophore, A23187. Rolipram and zaprinast, inhibitors of PDE4 and PDE5, respectively, had no effect on either PAF- or ADP-induced aggregation. These results demonstrate that inhibition of aggregation caused by PAF or ADP can be achieved by selective inhibition of PDE3 but suggest that there may be agonist-specific differences in the intracellular signalling pathways that regulate equine platelet aggregation.