Participation of phosphofructokinase,malate dehydrogenase and isocitrate dehydrogenase in capacitation and acrosome reaction of boar spermatozoa          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   

Protein Composition of Seminal Plasma in Fractionated Stallion Ejaculates     

AM Kareskoski  MM Rivera del Alamo  K Güvenc  T Reilas  JJ Calvete  H Rodriguez‐Martinez  M Andersson  T Katila 《Reproduction in domestic animals》2011,46(1):e79-e84

Seminal plasma (SP) contains several types of compounds derived from the epididymides and accessory glands. The aim of this study was to examine the protein composition of different ejaculate fractions. Trial I: fractionated ejaculates were collected from two normal and two subfertile stallions. Samples containing pre‐sperm fluid and the first sperm‐rich jets (HIGH‐1), the main sperm‐rich portion (HIGH‐2), the jets with low sperm concentrations (LOW), and a combined whole‐ejaculate (WE) sample was centrifuged, and the SP was filtered and frozen. A part of each SP sample was stored (5°C, 24 h) with spermatozoa from HIGH‐2 and skim milk extender. Sperm motility was evaluated after storage in extender mixed with the stallion’s own SP or SP from one of the other stallions (sperm from a normal stallion stored in SP from a subfertile stallion and vice versa). Protein composition was analysed using reverse‐phase liquid chromatography (RP‐HPLC), N‐terminal sequencing and mass spectrometry. The area‐under‐the‐curve (AUC) was used for quantitative comparison of proteins within fractions. Trial II: semen samples were collected from seven stallions. Fractions with the highest (HIGH) and lowest (LOW) sperm concentrations and WE samples were examined using SDS‐PAGE and densitometry. No significant differences emerged between fractions in the AUC‐values of the Horse Seminal Protein‐1 (HSP‐1) and HSP‐2 peaks, or the peak containing HSP‐3 and HSP‐4 (HSP‐3/4). Levels of HSP‐1, HSP‐2 and HSP‐3/4 were not significantly correlated with total sperm motility, progressive sperm motility or average path velocity after storage. Significant differences between ejaculate fractions in the amount of different protein groups present in SP were not found in Trial I; but in Trial II, the proteins in the 60–70 kDa range were more abundant in LOW than in HIGH and WE, indicating that this band contained proteins derived mainly from the seminal vesicles, which produce most of the SP in LOW.  相似文献   

Oesophageal sarcocystosis observed at slaughter provides a reliable and efficient proximate measure of Toxoplasma gondii seroprevalence in sheep     

PL Taggart  MM McAllister  D Rutley  CGB Caraguel 《Australian veterinary journal》2020,98(7):305-311

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Fusion of Boar Sperm with Nanoliposomes Prepared from Synthetic Phospholipids          下载免费PDF全文

VR Kasimanickam  MM Buhr 《Reproduction in domestic animals》2016,51(4):461-466

Liposomes are artificial membrane vesicles that can be used to test and model the functions and interactions of various biological membranes, or as a carrier system to deliver biologically active substances into the cells, or to incorporate lipids into the plasma membrane of target cells to modify membrane structure–function relationships. Sperm plasma membrane undergoes lipid modification during maturation in epididymis and during capacitation in the female reproductive tract to facilitate fertilization. Natural variation in the amounts and composition of lipids in the sperm plasma membrane may also contribute to the species‐specific sperm sensitivities to handling and storage conditions. Boar sperm are notoriously susceptible to membrane damage and are resistant to compositional alteration by artificial liposomes. This study used flow cytometry to demonstrate stable incorporation of nanoliposomes prepared from a complex mixture of various phospholipids (phosphatidylcholine : phosphatidylethanolamine : sphingomyelin : phosphatidylserine : phosphatidylinositol) with high fusion efficiency. Over 90% of sperm rapidly took up fluorescently labelled liposomes and retained the lipids for at least 60 min, in a significant time‐ and concentration‐dependent manner. This unique fusion efficacy could be used to alter sperm plasma membrane composition and hence membrane‐based functional responses.  相似文献   

Intranasal administration of midazolam in blue‐and‐yellow macaws (Ara araruana): evaluation of sedative effects          下载免费PDF全文

Débora PH Schäffer  Ana Cláudia S Raposo  Fernanda A Libório  Renata MM Silva  Nayone LLC Araujo  Arianne P Oriá 《Veterinary anaesthesia and analgesia》2016,43(4):459-460

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Relation between respiratory activity and sperm parameters in boar spermatozoa cryopreserved with alpha‐tocopherol and selected by Sephadex          下载免费PDF全文

MM Satorre  E Breininger  PD Cetica  M Córdoba 《Reproduction in domestic animals》2018,53(4):979-985

Our aim was to evaluate the effect of Sephadex filtration on respiratory activity of porcine spermatozoa and its relation with quality and functional sperm parameters. Samples were evaluated regarding oxygen uptake and sperm parameters: motility, plasma and acrosome membrane integrity, capacitation and acrosome reaction induction in vitro, plasma membrane functionality, determined by the hypo‐osmotic swelling test (HOST), and lipid peroxidation assessed by thiobarbituric acid assay. Sephadex filtration improved all routine quality parameters (motility, plasma and acrosome membrane integrity) and functional parameters (HOST, in vitro capacitation and true acrosome reaction levels) and produced a significant decrease in cryocapacitation and lipid peroxidation. Oxygen uptake increased in Sephadex samples (41 ± 7%) respect to single washing. Oxygen addition of carbonyl‐cyanide‐m‐chlorophenylhydrazone (CCCP) confirmed mitochondrial coupling in washed and Sephadex samples; showing an increase of 2.6 and 4.2 times for oxygen consumption in single washing and Sephadex ones, respectively. The increase in oxygen uptake with succinate addition with respect to basal oxygen uptake was significantly lower in Sephadex samples (63 ± 25%) than in the washed ones (183 ± 35%). Sephadex samples showed higher mitochondrial activity measured by oxygen consumption and improved quality and functional parameters. Our study recommends this protocol due to the fact that this filtration method removes dead or damaged spermatozoa allowing to obtain cryopreserved boar spermatozoa with optimized fertilizing capacity.  相似文献   

Conversion of Cortisone to Cortisol and Prostaglandin F2α Production by the Reproductive Tract of Cows at the Late Luteal Stage In Vivo     

HT Duong  DJ Skarzynski  KK Piotrowska‐Tomala  MM Bah  K Jankowska  P Warmowski  K Łukasik  K Okuda  TJ Acosta 《Reproduction in domestic animals》2012,47(6):939-945

Previous in vitro studies demonstrated that bovine endometrium has the capacity to convert inactive cortisone to biologically active cortisol (Cr) and that Cr inhibits cytokine‐stimulated prostaglandin F_2α (PGF) production. This study was carried out to test the hypothesis that bovine reproductive tract has the capacity to convert cortisone to Cr in vivo and to evaluate the effects of intravaginal application of exogenous cortisone on uterine PGF secretion during the late luteal stage. The temporal relationships between PGF and Cr levels in uterine plasma were also determined. Catheters were inserted into jugular vein (JV), uterine vein (UV), vena cava caudalis (VCC) and aorta abdominalis (AA) of six cows on Day 15 of the oestrous cycle (ovulation = Day 0) for frequent blood collection. On Day 16, the cows were divided randomly into two groups and infused intravaginally with vaseline gel (10 ml; control; n = 3) or cortisone dissolved in vaseline gel (100 mg; n = 3). Blood samples were collected at ?2, ?1, ?0.5, 0, 0.5, 1, 1.5, 2, 3, 4, 5 and 6 h after treatments (0 h). Intravaginal application of cortisone increased plasma concentrations of Cr between 0.5 and 1.5 h in UV, at 0.5 h in VCC, at 1 h in JV and at 1.5 h in AA. The plasma concentrations of PGF in UV and of PGF metabolite in JV were greater at 0.5 and 1 h in the cortisone‐treated animals than in control animals. The levels of PGF in UV blood plasma decreased after Cr reached its highest levels. The overall findings suggest that the female reproductive tract has the capacity to convert cortisone to Cr in vivo. Based on the temporal changes of PGF and Cr levels in the uterine plasma, a biphasic response in PGF secretion was found to be associated to the Cr increase induced by the cortisone treatment at the late luteal stage in non‐pregnant cows.  相似文献   

Effects of Matrix Filtration of Low-Quality Boar Semen Doses on Sperm Quality     

E Bussalleu  E Pinart  MM Rivera  M Briz  S Sancho  M Yeste  I Casas  A Fàbrega  T Rigau  JE Rodriguez-Gil  S Bonet 《Reproduction in domestic animals》2009,44(3):499-503

The aim of this work was to develop a method to enhance the sperm parameters of ejaculates with low sperm quality from Piétrain boars. Seminal doses were filtered through columns of DEAE Sephadex (length 2.5 ± 0.5 cm), CM Sephadex (length 5 ± 0.5 cm), glass wool (length 2 ± 0.5 cm) or glass bead (length 10 ± 0.5 cm), with an exit flow rate of 1 ml/40 s in all cases. For each male, 10 ml of the sperm cell-rich fraction diluted at 1 : 6 were filtered. Sperm quality was assessed before and after filtration. Sperm morphology, sperm motility and sperm concentration were determined using the computer program sca ^® 2002 Production, and sperm viability was evaluated by fluorescence multistaining. Osmotic resistance test and hyperosmotic resistance test were used to determine the osmotic resistance of spermatozoa, whereas l -lactate production estimated the metabolic activity. Results showed a decrease of sperm concentration and osmotic resistance of spermatozoa after filtration in the four matrixes. However, an increase in the frequency of viable spermatozoa with intact acrosome after filtration in glass bead columns and an increase of morphologically normal spermatozoa after filtration in Sephadex CM-50, glass wool and glass bead columns were observed. Despite the decrease in the frequency of progressive motile spermatozoa, l -lactate production and mitochondrial sheath integrity maintained constant after filtration. Our findings indicate that column filtration is an effective method to enhance the sperm quality by selecting viable and morphologically normal spermatozoa without altering DNA, plasma membrane, mitochondrial sheath integrity or inducing premature acrosome reaction.  相似文献   

Clinical and Subclinical Endometritis in the Mare: Both Threats to Fertility     

MM LeBlanc  RC Causey 《Reproduction in domestic animals》2009,44(S3):10-22

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Relation of growth hormone response to growth hormone-releasing hormone to estimation of milk production via deuterium oxide dilution in beef cattle     

Auchtung TL  Baer DJ  Erdman RA  Barao SM  Dahl GE 《Journal of animal science》2002,80(5):1270-1274

Current methods of estimating milk production in beef cows can be time-consuming, labor-intensive, and subject to high variability. The weigh-suckle-weigh (WSW) method requires repeated separation of offspring from their dams. Machine milking requires that animals be acclimated to the equipment prior to the estimation. The objective of Exp. 1 was to validate a deuterium oxide (D2O) dilution method of estimating milk production in cattle. In Exp. 1, Holstein calves (n = 5) averaging 29+/-2 d of age and 52.6+/-2.5 kg (+/- SE) were used as the model. Blood was collected for baseline D2O measurements followed by an injection of 300 mg D2O/kg BW. Syringes were weighed before and after the injection to gravimetrically determine the dose. Another blood sample was collected after D2O was allowed to equilibrate with body water for 2 h, and on each of the next five consecutive days, prior to feeding. Actual milk intake was measured by disappearance (i.e., amount of milk replacer offered to the calf minus the amount refused). Deuterium oxide in plasma was measured by mass spectrometry and milk intake was computed from the disappearance curve of D2O in blood plasma for each calf. Accumulated milk intake estimated by D2O dilution was highly correlated (y = 0.9x + 0.6; R2 = 0.99; P < 0.001) with actual milk intake. The objectives of Exp. 2 were to determine whether 1) D2O dilution was comparable to a standard measure of milk production in beef heifers and 2) growth hormone (GH) response to GH-releasing hormone (GHRH) in heifers at weaning is predictive of subsequent milk production. Deuterium oxide dilution and WSW were compared using 14 first-calf Angus heifers and their calves. Deuterium oxide dilution was used to estimate milk production of 40 first-calf Angus heifers that had been challenged with GHRH at weaning. Results indicate that the D2O dilution method is correlated (R2 = 0.89; P = 0.04) to the WSW estimation of milk production. Growth hormone response to GHRH in weanling heifers is positively related (R2 = 0.22; P = 0.03) to their subsequent milk production. Deuterium oxide dilution in calves offers an additional approach to the estimation of milk production of the dam in typical beef cattle production settings.  相似文献