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1.
Participation of phosphofructokinase,malate dehydrogenase and isocitrate dehydrogenase in capacitation and acrosome reaction of boar spermatozoa 
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下载免费PDF全文 E Breininger D Dubois VE Pereyra PC Rodriguez MM Satorre PD Cetica 《Reproduction in domestic animals》2017,52(5):731-740
The aim of this work was to determine the enzymatic activity of phosphofructokinase (PFK), malate dehydrogenase (MDH) and isocitrate dehydrogenase (IDH) in boar spermatozoa and study their participation in bicarbonate‐induced capacitation and follicular fluid‐induced acrosome reaction. Enzymatic activity of these enzymes was determined spectrophotometrically in extracts of boar spermatozoa. Sperm suspensions were incubated in the presence of bicarbonate (40 mM), a well‐known capacitation inducer, or follicular fluid (30%), as an acrosome reaction inducer, and different concentrations of oxoglutarate, oxalomalate and hydroxymalonate, inhibitors of PFK, IDH and MDH, respectively. Capacitation percentages were determined by the fluorescence technique of chlortetracycline (CTC), and true acrosome reaction was determined by trypan blue and differential–interferential contrast, optical microscopy. The activity of PFK in boar spermatozoa enzymatic extracts was 1.70 ± 0.19 U/1010 spermatozoa, the activity of NAD‐ and NADP‐dependent IDH was 0.111 ± 0.005 U/1010 and 2.22 ± 0.14 U/1010 spermatozoa, respectively, and the activity of MDH was 4.24 ± 0.38 U/1010 spermatozoa. The addition of the specific inhibitors of these enzymes prevented sperm capacitation and decreased sperm motility during capacitation and inhibited the acrosome reaction (AR), without affecting the sperm motility during this process. Our results demonstrate the participation of PFK, IDH and MDH in bicarbonate‐induced capacitation and follicular fluid‐induced acrosome reaction in boar spermatozoa, contributing to elucidate the mechanisms that produce energy necessary for these processes in porcine spermatozoa. 相似文献
2.
The effects of a diet containing a high proportion of rapeseed meal on the activity of certain plasma enzymes were studied in laying birds. The enzymes studied were alkaline phosphatase (AP), aspartate transaminase (AST), L-gamma-glutamyl-transferase (gamma-GT), isocitrate dehydrogenase (ICDH), leucine arylamidase (LAP), malate dehydrogenase (MDH) and sorbitol dehydrogenase (SDH). No notable differences were observed between the plasma AP, LAP or SDH activities of the birds given the rapeseed meal and the birds receiving a soyabean meal control diet throughout the experiment. However, the plasma AST and gamma-GT activities of the treated birds showed slight elevations while their plasma ICDH and MDH activities showed more marked elevations, which are indicative of liver damage, in response to the diet. Macroscopic observations of the livers of the birds at the end of the experiment were in fairly good accord with the elevation in plasma ICDH and MDH activities noted for the individual birds. 相似文献
3.
From the distribution of enzymes, the alveoli of a uropygial gland may be divided into inner and outer zones ; the former being the region near the cavity, and the latter the peripheral region of the gland.
In the inner zone, there appeared strong activities of 17β‐hydroxy‐steroid dehydrogenase (17β‐OH‐SDH), 30α‐OH‐SDH, glucose‐6‐phosphate dehydrogenase (G‐6‐PDH), lactate dehydrogenase (LDH), NAD‐dependent malate dehydrogenase (MDH) and acid phosphatase (Acid Pase) weak activities of succinate dehydrogenase (SuDH), NADP‐dependent MDH, monoamine oxidase (MAO) and non‐specific esterase (Etase), but no activity of β‐hydroxybutyrate dehydrogenase (β‐HbDH).
In the outer zone, there appeared strong activities of LDH and MDH (NAD), moderate activities of 17β‐OH‐SDH, 3α‐OH‐SDH, G‐6‐PDH and β‐HbDH, weak activities of SuDH, MDH (NADP) and MAO, but no activities of Etase and Acid Pase.
The results were negative concerning ?5‐3β‐OH‐SDH, glutamate dehydrogenase, a‐glycerophosphate dehydrogenase, phosphorylase and alkaline phosphatase in both zones. From these results, it was concluded that there must be functional differences between the zones, and that only a part of the metabolic pathways of steroids is being performed. 相似文献
4.
奶牛精子膜的分离及纯化 总被引：2，自引：0，他引：2
本实验采用高压爆破匀浆法将精子膜与精子分离。分离后的精子膜粗品采用蔗糖密度梯离心法进行纯化。经电镜观察，膜制品均一性很好。经酶学分析，膜特异性酶－碱性磷酸酯酶在膜制品中含量很高。 相似文献
5.
Koonjaenak S Chanatinart V Ekwall H Rodriguez-Martinez H 《Journal of veterinary medicine. A, Physiology, pathology, clinical medicine》2007,54(4):169-178
The appearance and incidence of sperm abnormalities was studied in 115 ejaculates, collected periodically over 1 year covering all seasons from five mature, healthy swamp buffalo (Bubalus bubalis) bulls reared under tropical conditions and serving as the current source of semen for artificial insemination (AI) in Thailand. Light microscopy of stained smears was used to investigate sperm head shape morphology, while unstained wet smears were used to examine other sperm abnormalities. The most commonly found morphological aberrations were pear-shaped spermatozoa, knobbed acrosomes, proximal cytoplasmic droplets, simple bent tails and coiled tails under the head, whose ultrastructure (scanning electron microscopy) corresponded to what has been found in other species of bovidae, including varieties of buffalo. The mean prevalence (as least squares mean +/- SEM) of sperm abnormalities was low (below 15%), corresponding to healthy spermiograms. The younger bulls (<10 years old, n = 3) had less abnormalities than the older ones (10.1 +/- 0.6% versus 14.1 +/- 0.8%, P < 0.001, n = 2), including abnormalities of sperm head shape (1.1 +/- 0.3% versus 3.6 +/- 0.3, P < 0.001), acrosome defects with knobbed acrosomes (1.1 +/- 0.2% versus 1.2 +/- 0.3%, P < 0.001), spermatozoa with proximal cytoplasmic droplets (2.7 +/- 0.1% versus 1.4 +/- 0.2%, P < 0.001), defective mid-pieces (0.2 +/- 0.1% versus 0.3 +/- 0.1%) and abnormal sperm tails (3.1 +/- 0.3% versus 5.7 +/- 0.4%, P < 0.001). The within-bull effect of the year solely affected the incidence of pear-shaped spermatozoa while the incidences of abnormal contour, variable size of sperm head shapes, abnormal mid-piece and simple bent tail among bulls were affected by ejaculate (week of collection). Interaction between age and ejaculate affected only the prevalence of spermatozoa with proximal cytoplasmic droplets. In conclusion, the types of defects encountered were similar to those found in other bovidae, with a very low prevalence over the year the AI sires were followed through. 相似文献
6.
Use of Glutamine and Low Density Lipoproteins Isolated from Egg Yolk to Improve Buck Semen Freezing 总被引：1，自引：0，他引：1
MZ Ali Al Ahmad G Chatagnon L Amirat-Briand M Moussa D Tainturier M Anton F Fieni 《Reproduction in domestic animals》2008,43(4):429-436
To improve the results obtained with a reference cryopreservation extender (control extender: Triladyl® + 20% (v/v) egg yolk + 6.4% (v/v) glycerol) for freezing caprine semen, glutamine was added to 18 split ejaculates at concentrations of 0, 20, 40, 80 and 120 m m (experiment 1). In experiment 2, glutamine was added to 18 split ejaculates at concentrations of 20, 25, 30, 35 and 40 m m . In the third experiment, the egg yolk was replaced with the low-density lipoprotein (LDL) fraction of egg yolk. The quality of frozen then thawed spermatozoa in each extender was compared using computer-assisted semen analysis. In experiment 1, glutamine at concentrations of 20 m m and 40 m m significantly improved sperm motility compared with the control extender. However, at 120 m m , a significant decrease in motility and velocity was observed. In experiment 2, motility, curvilinear velocity and amplitude of lateral head displacement (ALH) were improved in glutamine at 25 m m compared with the control. In experiment 3, 8% LDL and 25 m m glutamine significantly improved sperm motility, straight line velocity and ALH. In the fourth experiment, the quality of the previously defined freezing extender (Triladyl® + 8% (v/v) LDL + 25 m m glutamine + 6.4% (v/v) glycerol) was tested by comparing acrosome, tail membrane, plasma membrane and DNA integrity in 18 split ejaculates of frozen then thawed spermatozoa with spermatozoa that had been frozen then thawed in the control extender, and with spermatozoa from fresh, unfrozen sperm. The percentage of spermatozoa with intact acrosomes and tail membranes was significantly higher with the newly defined extender than that observed with the control extender. There was no significant difference in the percentage of spermatozoa with intact DNA between the frozen and fresh semen. 相似文献
7.
Tornquist SJ Dodson L Lanning DV 《Veterinary clinical pathology / American Society for Veterinary Clinical Pathology》2000,29(1):16-18
Abstract: Serum and heparinized plasma samples were collected from 11 adult, clinically healthy llamas. Aliquots were assayed for sorbitol dehydrogenase (SDH) activity after storage at room temperature (20°C), 4°C, or −20°C for defined time intervals up to 1 week postcollection. Sorbitol dehydrogenase activity in all samples was within reference intervals for our laboratory. No difference was found between serum and plasma SDH activity when measured immediately (within 1 hour) after collection. Sorbitol dehydrogenase activity decreased to 79% of initial activity by 24 hours in serum stored at room temperature; plasma had 94% of initial SDH activity under the same conditions. Sorbitol dehydrogenase activity was stable in both plasma and serum stored for up to 1 week at 4°C or −20°C. With the exception of serum stored at 20°C for > 8 hours, in vitro stability of llama SDH was adequate for its use in diagnostic testing. 相似文献
8.
哺乳动物精子经过附睾成熟后才能获得运动及受精的能力,为解释水牛精子在附睾中的成熟过程,本研究选用性成熟期的沼泽型水牛附睾,利用乙烯吡咯烷酮包裹的硅胶微小颗粒（Percoll）梯度离心纯化分别提取附睾头、体和尾部精子,应用计算机辅助精子分析系统（CASA）检测精子活力,透射电镜观察附睾不同部位精子的超微结构,对精子进行荧光标记后,利用流式细胞仪和荧光显微镜观察检测不同部位精子质膜完整率、线粒体鞘膜电位和顶体差异。结果表明,Percoll分离得到附睾头、体和尾3部位精子的纯度达95%,不同部位精子活力分别为8.35%、20.21%和65.60%;附睾不同部位精子都存在着结构完整的精子以及相同的畸形类型,附睾尾部精子线粒体鞘高膜电位比率最高,精子质膜完整率从附睾头部到尾部逐渐升高,精子顶体完整率从附睾头部到尾部逐渐升高。本研究直观地展示了水牛附睾不同部位精子特征以及差异,为研究水牛精子成熟机理提供理论依据。 相似文献
9.
Rota A Manuali E Caire S Appino S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2008,70(1):123-125
This paper describes the case of a 2-year-old English Bulldog with severe teratozoospermia that consisted mainly of "Dag-like" defects, which is strong coiling of the tail. Although libido, semen volume and sperm concentration were normal, sperm motility was 5.0%, and 93.3% of spermatozoa exhibited morphological abnormalities affecting the tail. Transmission electron microscopic examination of the spermatozoa revealed strong folding, coiling and fracture of sperm midpieces and tails, axonemal defects and the presence of swollen and unevenly distributed mitochondria. Taking into account the dog's history and examinations and the constantly high percentage of abnormal spermatozoa over time, the defect was considered to be genetic in origin. 相似文献
10.
Barth AD 《The Canadian veterinary journal. La revue veterinaire canadienne》1989,30(8):656-662
The prevalence of abaxial sperm tails, accessory tails and double tails, was determined from semen evaluation records of 600 western Canadian range bulls and 449 bulls in Canadian artificial insemination centers. Spermatozoa with abaxial tails were produced by 10.5% of bulls, however, only 0.48% produced sperm with greater than 50% abaxial tails, and 0.86% of the 1049 bulls produced sperm with accessory and double tails. Three experiments were done to determine the importance of abaxial sperm tails to fertility. In experiments 1 and 2, frozen semen with 88%, 50% and 0% abaxial tails was used to inseminate synchronized heifers, which were allowed to develop pregnancy, and superovulated heifers, which were slaughtered prior to embryo recovery seven days after breeding. In experiment 3, a bull which produced 100% abaxial sperm was used in a competitive mating situation with three control bulls in a herd of 118 cows. The combined results of these experiments indicate that spermatozoa with abaxial tail attachment fertilize ova at a normal rate and are not associated with any increase in embryonic death. 相似文献
11.
S Dhandayuthapani M P Balasubramanian K Nellaiappan K Ramalingam 《Veterinary parasitology》1983,12(1):65-69
Isozyme pattern of lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) of Gastrothylax crumenifer from sheep, goat and buffalo was studied using polyacrylamide gel electrophoresis. LDH of G. crumenifer from buffalo, goat and sheep consists of four fractions, three fractions and two fractions, respectively. The parasite from buffalo shows two fractions of MDH, whereas those from goats or sheep show only a single fraction. The significance of these results is discussed. 相似文献
12.
对猪体内发育30，40，60，80，95日猪囊尾蚴囊壁的已糖激酶（HK），丙酮酸激酶（PK），乳酸脱氢酶（LDH），苹果酸脱氢酶（MDH），葡萄糖－6－磷酸脱氢酶（G6PDH）活性和葡萄糖（GLc)，乳酸（LAC）含量进行测定，结果表明，HK，PK，MDH活性和GLc含量在60，80，95日高于或显著高于30，40日（P＞0．05，P＜0．05），说明随虫体的生长发育糖代谢逐渐增加。 相似文献
13.
Washizu T Azakami D Bonkobara M Washizu M Arai T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(6):615-616
Alterations in the activities of enzymes related to energy metabolism in canine lymphoma cells were investigated. Cytosolic pyruvate kinase (PK) and mitochondrial malate dehydrogenase (MDH) activities in lymphoma cells were significantly higher than those in lymphocytes obtained from lymph nodes of healthy dogs, whereas cytosolic lactate dehydrogenase (LDH) activity was significantly lower in lymphoma cells. The cytosolic M/L ratio (MDH activity/LDH activity), which is considered to be a good indicator of energy metabolism related to glucose utilization in animal tissues, was significantly higher in lymphoma cells than in the normal lymphocytes. 相似文献
14.
Magori E Nakamura M Inoue A Tanaka A Sasaki N Fukuda H Mizutani H Sako T Kimura N Arai T 《Research in veterinary science》2005,78(1):39-44
The activities of the enzymes in the malate-aspartate shuttle were measured in peripheral leucocytes of spontaneous type 1 diabetic dogs and cats treated with insulin injections. In the diabetic dogs and cats, fasting plasma glucose concentrations were three- or fourfold greater than the control levels in spite of insulin injections and the activities of cytosolic malate dehydrogenase (MDH), one of pivotal enzymes in the malate-aspartate shuttle, were remarkably lower than the controls. Depressed expression of cytosolic MDH mRNA was confirmed by RT-PCR analysis in the diabetic animals. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity (M / L ratio) in leucocytes of the diabetic animals was significantly lower than that of normal control animals. The smaller M / L ratio appeared to reflect depression of energy metabolism in the diabetic animals. Intrinsically lower and further decreased MDH activities may be factors that induce insulin resistance observed in diabetic cats. 相似文献
15.
Two matched case-control studies were performed at an abattoir with a capacity of 780 pigs per hour, each study using the approximately 7000 pigs slaughtered on one day. In the first study, the severity of tail biting and pneumonia were recorded in pigs with bitten or intact tails. In the second study, the tail score, sex, and the presence of pleuritis, externally visible abscesses and trimming were recorded in pigs with bitten or intact tails. In study 1, there was no significant association between the tail score and the percentage of lung tissue affected by lesions typical of enzootic pneumonia, but there was a significant association between the severity of tail biting and the prevalence of lungs with abscesses and/or pleuritic lesions (P<0.0001). In study 2, there were significant associations between the severity of tail biting, and the prevalence of external carcase abscesses and carcase trimming; the carcases of castrated males had evidence of tail biting more frequently than the carcases of females (P<0.05). 相似文献
16.
Bhattacharyy HK Bujarbaruah KM Goswami BK Biswas RK Khan I 《Journal of the South African Veterinary Association》2010,81(2):80-81
Genital organs of 10 healthy, adult Mithun bulls (6-8 years old) that were slaughtered at the dwellings of tribal people for meat were collected. Immediately after collection, spermatozoa from 3 different regions of the epididymis, i.e. the head, body and tail, were obtained to study morphological changes of the spermatozoa during passage through these regions. The prevalence of proximal cytoplasmic droplets significantly decreased from the head to the tail of the epididymis. Conversely, the percentage of distal cytoplasmic droplets increased significantly from the head to the tail region. The incidence of tailless heads rose significantly from head to body and then reduced significantly in the tail region. The percentage of total head abnormalities did, however, not change markedly, but total mid-piece and tail abnormalities differed significantly between the three epididymal regions. 相似文献
17.
Washizu T Nakamura M Izawa N Suzuki E Tsuruno S Washizu M Nakajo S Arai T 《Veterinary research communications》2002,26(5):341-346
The activities of malate dehydrogenase (MDH) and lactate dehydrogenase (LDH) and the pattern of the isoenzymes of LDH were determined in the peripheral blood leukocytes of dogs, rabbits and cats. Rabbits had significantly higher plasma glucose concentrations than dogs or cats. Feline leukocytes showed higher LDH and lower MDH activities than canine or rabbit leukocytes. The M/L ratio, defined as the MDH activity divided by the LDH activity in cytosolic fractions, was considered to be a good indicator with which to evaluate the metabolic state in animal tissues. The M/L ratio was highest in canine and lowest in feline leukocytes. LDH-2 and LDH-3 isoenzymes were dominant in canine leukocytes. LDH-1 and LDH-2 were dominant in rabbit leukocytes, whereas LDH-5 was dominant in feline leukocytes. It was evident that there were significant differences in energy metabolism between the leukocytes of dogs, rabbits and cats. 相似文献
18.
In the endodermal cells of the yolk sac membrane of chicken embryos incubated for 12 to 18 d, the activity of glucose‐6‐phosphatase (G‐6‐Pase) was high, the activity of acid phosphatase (Acid Pase) was moderate, the activities of lactate dehydrogenase (LDH) and of glyceraldehyde‐3‐phosphate dehydrogenase (G‐3‐PDH) were low, and succinate dehydrogenase (SDH) and alkaline phosphatase (Alk Pase) activities were not detected. On the day before hatching, however, SDH became evident with low activity, and the activities of Acid Pase, LDH and G‐3‐PDH increased and remained higher until the day after. Five days after hatching, there were marked decreases in the activities of all the enzymes except SDH. Alk Pase was not detected at any time. The function of the yolk sac endo‐derm in the absorption of yolk is discussed in the light of these observations. 相似文献
19.
Kimura N Yoshimura I Sako T Inoue A Tadami K Arai T 《Veterinary research communications》2005,29(1):19-26
Glucose, triglyceride, cholesterol and immunoreactive insulin (IRI) concentrations, some enzyme activities in plasma, and activities of enzymes related to energy metabolism in peripheral leukocytes were measured in fattening Japanese Black Wagyu x Holstein steers fed on different diets at 8, 12, 16, 20 and 24 months of age. The plasma IRI concentrations at 20 and 24 months of age were significantly higher than those at 8 months of age. Activities of hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PD), aspartate aminotransferase (AST), and malate dehydrogenase (MDH) in cytosolic fractions, and glutamate dehydrogenase (GLDH), MDH and AST in mitochondrial fractions in peripheral leukocytes of steers at 24 months of age were significantly higher than those at 8 months. Increasing plasma insulin concentration was considered to induce acceleration of glucose utilization in leukocytes of fattening steers. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity in leukocytes increased significantly in the fattening process and was considered to be a useful indicator for evaluating changes in energy metabolism in steers. 相似文献
20.
The Hereditary 'Short Tail' Sperm Defect - A New Reproductive Problem in Yorkshire Boars 总被引：2，自引：0，他引：2
M Andersson Oat Peltoniemi A Makinen A Sukura H Rodriguez-Martinez 《Reproduction in domestic animals》2000,35(2):59-63
This study describes a new sperm defect in Yorkshire boars. The length of the sperm tail is markedly reduced, resulting in a total immotility in all spermatozoa. At transmission electron microscopy level, the morphology of the midpiece microtubular components area is seriously affected. This boar sperm defect differs from the ‘tail stump’ defect observed in bulls, the tails being longer in most spermatozoa than those found in affected bulls. Therefore, the term ‘short tail’ sperm defect is more adequate. The authors observed the first case in 1987. In 1998, this defect became a noteworthy reproductive problem, when it was observed in nine boars intended for breeding. In one litter, three littermates were affected with the ‘short tail’ sperm defect. At the present time the authors believe that the defect is recessively inherited. 相似文献