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1.
Polymorphonuclear leucocytes (PMN) were isolated from milk and blood of healthy cows, and the generation of reactive oxygen by the two cell populations was compared by measuring chemiluminescence (CL) after stimulation with zymosan. The ratio of milk to blood PMN CL was relatively constant in a given animal, but varied widely between different cows, ranging from 0.3 to 1.3.The relative contributions of various oxygen species to CL was studied by measuring quenching using different oxygen scavengers. While the relative contributions of H2O2, ?O2 and ′O2 seemed to be similar in both milk and blood PMN, the OH· radical was clearly more prominent in PMN isolated from milk than from blood. In addition, blood PMN CL was more dependent on the presence of glucose in the reaction medium than milk PMN CL. Furthermore, the CL response to phorbol myristate acetate, to the Ca ionophore A23187 and to Sendai virus was different in the two cell types. The results suggest that CL generation in milk PMN differs from that in blood PMN in quantitative as well as qualitative aspects. 相似文献
2.
The aim of the present study was to determine the mechanism of cytosolic calcium ion concentration [Ca2+]i elevation in chicken and rat phagocytes stimulated with phorbol myristate acetate (PMA), leukotriene B4 (LTB4), formyl‐methionyl‐leucyl‐phenylananine (fMLP) and Saccaromyces cerevisiae culture supernatant (SCS). Pretreatment with EGTA completely suppressed the PMA‐induced [Ca2+]i elevation in rat and chicken phagocytes, suggesting that all the [Ca2+]i elevation induced in the PMA‐stimulated rat and chicken phagocytes was attributable to the influx of extracellular Ca2+. On the other hand, the elevation of LTB4‐, FMLP‐ and SCS‐induced [Ca2+]i was only partially suppressed by ethyleneglycol‐bis (β‐aminoethyl)‐N,N,N′,N′‐tetraacetic acid ethylene (EGTA) pretreatment of phagocytes. The results indicated that two pathways of [Ca2+]i elevation, recruitment from the intracellular Ca2+ store and influx of extracellular Ca2+, are involved in the [Ca2+]i elevation of LTB4‐, fMLP‐ and SCS‐stimulated phagocytes. In fMLP‐stimulated rat neutrophils, [Ca2+]i elevation showed a two‐phase pattern in which the time lag between the first and second phase was approximately 1 min. The EGTA treatment of the fMLP‐stimulated cells induced a reduction of the first phase level and a disappearance of the second phase. The reason for the special influence of EGTA observed in fMLP‐stimulated cells is unknown, but the disappearance of the second phase of the [Ca2+]i may be elicited by the EGTA‐induced decrease of the first phase [Ca2+]i elevation that depends on IP3 and diacylglycerol induced by fMLP. 相似文献
3.
The effects of acellular milk on the activity of the microbicidal cationic enzymes of the polymorphonuclear cells of goats were studied in an attempt to explain the phenomenon by which PMN functions fail in mastitis. Assays were undertaken on the myeloperoxidase, lysozyme and elastase activities in a polymorphonuclear cell (PMN) lysate, both in the presence and absence of acellular milk from homologous species. There was a significant decrease (p<0.05) in the activity of lysozyme, myeloperoxidase and elastase in the presence of acellular milk. Superoxide and H2O2 production following activation of caprine PMNs by lipopolysaccharide (LPS) was significantly reduced (p<0.05) in the presence of acellular milk. Thus, the microbicidal function of PMNs is significantly impaired in the presence of acellular milk and this may contribute to the development of mastitis in dairy animals. 相似文献
4.
Phagocytosis and Related Phenomena in Polymorphonuclear Leukocytes from Cow's Milk 总被引:1,自引:1,他引:0
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There were fewer efficient phagocytes among leukocytes collected from artificially irritated mammary glands than among the leukocytes from blood of the same animals. The milk polymorphonuclear (PMN) leukocytes adhered poorly to a column of siliconised glass beads when compared with the blood cells. However, investigations of the O2 uptake and CO2 production of the milk PMN leukocytes revealed that these cells appeared to utilize metabolic pathways similar to those used by human and guinea pig PMN leukocytes during phagocytosis. These pathways are associated with degranulation and the production of H2O2 following particle ingestion. It is therefore suggested that the milk PMN leukocytes appear not to have lost the ability to produce this bactericidal substance. 相似文献
5.
Yingxiao Fu Jianhong Gu Yi Wang Yan Yuan Xuezhong Liu Jianchun Bian Zong-Ping Liu 《Journal of veterinary science (Suw?n-si, Korea)》2015,16(2):151-156
The purpose of this study was to determine whether the Ca2+ signaling pathway is involved in the ability of osteoprotegerin (OPG) to inhibit osteoclast differentiation and maturation. RAW264.7 cells were incubated with macrophage colony-stimulating factor (M-CSF) + receptor activator of nuclear factor-κB ligand (RANKL) to stimulate osteoclastogenesis and then treated with different concentrations of OPG, an inhibitor of osteoclast differentiation. The intracellular Ca2+ concentration [Ca2+]i and phosphorylation of Ca2+/calmodulin-dependent protein kinase II (CaMKII) in the different treatment groups were measured by flow cytometry and Western blotting, respectively. The results confirmed that M-CSF + RANKL significantly increased [Ca2+]i and CaMKII phosphorylation in osteoclasts (p < 0.01), and that these effects were subsequently decreased by OPG treatment. Exposure to specific inhibitors of the Ca2+ signaling pathway revealed that these changes varied between the different OPG treatment groups. Findings from the present study indicated that the Ca2+ signaling pathway is involved in both the regulation of osteoclastogenesis as well as inhibition of osteoclast differentiation and activation by OPG. 相似文献
6.
In this study, the effect of reproductive hormones and substances with hormonal activity on the oxidative burst activity of blood polymorphonuclear leucocytes (PMN) high yielding dairy cows was evaluated. Different concentrations of: progesterone, oestradiol 17β, FSH, LH, GnRH, cortisol and PGF2α were incubated in vitro for 4 h with PMN of seven high milk yielding cows, during the period of anoestrous postpartum. Controls were run in parallel in which each hormone was replaced by its solvent. After incubation with hormones the competence of PMN to generate H2O2 was monitored by flow cytometry. A down‐regulation on the oxidative burst activity of PMA‐stimulated PMN was observed when cells were incubated with progesterone. Significant (p ≤ 0.001) differences between control and progesterone incubated cells were observed from 6.56 μg/ml. The same predisposition was observed when PMNs were incubated with cortisol. Besides for all concentrations employed, a decrease in the burst activity was observed, only beyond 0.19 mg/ml, statistical differences between the results obtained by the control and the cortisol incubated cells were obtained. Concerning oestradiol 17β, an increase on H2O2‐production was observed when PMN were incubated with 15 pg/ml and 45 pg/ml of this steroid (p ≤ 0.05), followed by a depression of the cell’s activity when unphysiological concentrations were employed. Significant (p ≤ 0.05) differences between the obtained with the control and oestradiol 17β incubated cells were observed only in the highest concentration of oestradiol. No statistical differences were observed in the metabolic burst activity of PMN incubated with FSH, GnRH and LH when compared with the results obtained by the control. 相似文献
7.
Shang-Jin Kim In-Gook Cho Hyung-Sub Kang Jin-Shang Kim 《Journal of veterinary science (Suw?n-si, Korea)》2006,7(1):31-36
A change in pH can alter the intracellular concentration of electrolytes such as intracellular Ca2+ and Na+ ([Na+]i) that are important for the cardiac function. For the determination of the role of pH in the cardiac magnesium homeostasis, the intracellular Mg2+ concentration ([Mg2+]i), membrane potential and contraction in the papillary muscle of guinea pigs using ion-selective electrodes changing extracellular pH ([pH]o) or intracellular pH ([pH]i) were measured in this study. A high CO2-induced low [pH]o causes a significant increase in the [Mg2+]i and [Na+]i, which was accompanied by a decrease in the membrane potential and twitch force. The high [pH]o had the opposite effect. These effects were reversible in both the beating and quiescent muscles. The low [pH]o-induced increase in [Mg2+]i occurred in the absence of [Mg2+]o. The [Mg2+]i was increased by the low [pH]i induced by propionate. The [Mg2+]i was increased by the low [pH]i induced by NH4Cl-prepulse and decreased by the recovery of [pH]i induced by the removal of NH4Cl. These results suggest that the pH can modulate [Mg2+]i with a reverse relationship in heart, probably by affecting the intracellular Mg2+ homeostasis, but not by Mg2+ transport across the sarcolemma. 相似文献
8.
Ducusin RJ Nishimura M Sarashina T Uzuka Y Tanabe S Otani M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(4):535-539
To determine the effects of ozone on the phagocytosis of bovine polymorphonuclear leukocytes (PMNs), ozone gas was administered in vitro on the blood and milk of healthy lactating cows, cows with acute mastitis, and cows with milk fever. In the blood of healthy dairy cattle, although there was no significant effect of ozone gas on the viability of the leukocytes, phagocytosis of PMNs significantly decreased. In contrast, ozone gas administration in vitro significantly increased phagocytosis of PMNs from the blood of cows with acute mastitis and milk fever, and from mastitic milk. These findings showed that ozone administration in vitro has positive and negative effects on bovine PMN phagocytosis, depending on the health status of the animal. 相似文献
9.
Glycogen content was determined quantitatively by the Anthrone reagent method in leukocytes obtained from blood and milk of five cows. Distribution of glycogen in leukocytes was studied by microscopic examination of slides stained by Periodic acid-Schiff (PAS) reaction. Blood glucose concentrations were investigated in these animals by standard procedures. In two of five cows both blood glucose levels and blood leukocyte glycogen levels on the same day were determined for six consecutive days. One hundred and two blood leukocyte samples from five cows had a mean glycogen content of 1.32 ± 0.04 (S.E.) mg/109 WBC, and 6.11 ± 0.17 (S.E.) mg/109 PMNs. Leukocyte preparations from 80 samples of milk comprising 97 to 98% PMNs contained 3.81 ± 0.18 (S.E.) mg glycogen/109 milk leukocytes. In PAS preparations of blood and milk leukocytes glycogen was found almost exclusively in PMNs. Glycogen granules, present frequently in PMNs and occasionally in monocytes and large lymphocytes from blood, were not observed in those from milk. The glycogen level in milk leukocytes was significantly lower (P = <0.01) than that of the blood PMNs in every cow, and the overall mean difference between levels for milk leukocytes and blood PMNs was highly significant (P = <0.001). Mean blood glucose concentration in the five cows was 44.46 ± 0.66 (S.E.) mg%. There was no significant relationship between blood glucose and blood leukocyte glycogen levels in the five corresponding cows; nor between blood glucose and blood PMN glycogen levels on the same day in either of two cows investigated. Leukocyte preparations from milk samples obtained on the second day following intramammary infusion of endotoxin consistently contained markedly less glycogen than the leukocyte preparations from first day post-infusion samples.
These tended to level off and became intermediate between first and second day levels. It is postulated that the poor phagocytic competence of leukocytes from bovine mammary glands compared to their counterparts in blood observed by various workers may be due partially to low energy reserves in these cells.
相似文献10.
Apoptosis and necrosis of blood and milk polymorphonuclear leukocytes in early and midlactating healthy cows. 总被引:4,自引:0,他引:4
Increased milk somatic cell counts (SCC) are used as an indicator for bovine mastitis. During mastitis, polymorphonuclear leukocytes (PMN) become the predominant cell type. Shortly after parturition, the severity of mastitis is increased and several PMN functions are downregulated. Apoptotic and necrotic processes of PMN could influence SCC and PMN functions. In this study, the percentages of apoptotic and necrotic PMN in blood and milk from early and midlactating healthy cows were compared. Apoptosis and necrosis of PMN were quantified using a dual-color flow cytometric procedure with fluorescein labeled annexin-V (green) and propidium iodide (red). Using this technique three different subpopulations of bovine PMN could be detected: apoptotic cells (high intensive green fluorescence), necrotic cells (high intensive green and high intensive red fluorescence) and viable cells (low intensive green and low intensive red fluorescence). Following a 4 h incubation of blood from both groups of cows at 37 degrees C to induce apoptosis, the mean percentage of apoptotic blood PMN was significantly higher (P < 0.01) in early lactating cows (15.1%, n = 9) compared with midlactating cows (5.3%, n = 10). The mean percentage of necrotic PMN remained lower than 5% in all cows. In contrast to blood, no significant difference was found between the percentage of apoptotic PMN in milk from early (41.2%, n = 7) and midlactating cows (34.0%, n = 8). The percentage of necrotic PMN in milk from early lactating cows (25.9%, n = 7) was significantly higher than that in midlactating cows (14.2%, n = 8) (P < 0.05). Higher percentages of apoptotic as well as necrotic PMN were consistently found in milk compared to blood in all cows. From these results, it can be concluded that spontaneously induced apoptosis was higher in blood PMN from early lactating cows than in blood PMN from midlactating cows. The higher percentage of necrotic milk PMN in early lactating cows than in midlactating cows could be explained by the induction of secondary necrosis. 相似文献
11.
Tetsuji OKAZAKI Shuji YOSHIDA Hisanori TESHIMA Masayuki SHIMADA 《Animal Science Journal》2011,82(3):412-419
In our previous study, seminal plasma effectively suppressed the induction of sperm to capacitation‐like status and acrosome loss during the thawing process. However, because boar seminal plasma is contaminated with various kinds of bacteria and/or viruses, it is necessary to develop a thawing solution without animal‐derived materials. In this study, we focused on the increase of intracellular Ca2+ ([Ca2+]i) in sperm after thawing and the negative effects of sperm qualities. After thawing, the fluorescent intensity of [Ca2+]i indicator, Fluo‐3/AM, and the level of phosphorylated tyrosine residue of protein were increased in the sperm. Next, we investigated whether the addition of Ca2+ chelators (ethylenediaminetetraacetic acid (EDTA) and ethyleneglycoltetraacetic acid (EGTA)) improved post‐thawed sperm motility. When the frozen–thawed sperm were treated with 6 mmol/L EDTA + 6 mmol/L EGTA, sperm motility was significantly increased as compared with control (6 mmol/L EDTA alone) at all incubation periods (P < 0.05). The combinational treatment significantly suppressed the elevation of [Ca2+]i and the tyrosine phosphorylation, which improved the acrosomal status and fertilizing ability in vitro. Furthermore, when the thawing method was applied for artificial insemination, the fertilization rate was significantly higher than control (P < 0.05, 33% vs. 82%). Thus, we conclude that the addition of EDTA + EGTA to thawing solution is a beneficial tool for artificial insemination using frozen–thawed boar sperm. 相似文献
12.
Sylvia J. Bedford-Guaus Lori A. McPartlin Dickson D. Varner 《Journal of Equine Veterinary Science》2012
In all mammalian species studied thus far, fertilization results in a series of intracellular Ca2+ ([Ca2+]i) increases, referred to as oscillations, responsible for driving oocyte activation and embryonic development. Current evidence supports the notion that sperm-borne phospholipase C zeta (PLCZ) is responsible for the initiation of these [Ca2+]i oscillations. Although this appears to be a highly conserved mechanism for oocyte activation, differences in PLCZ sequence, activity, and expression do exist among different species. Herein, we summarize the information supporting PLCZ as the oocyte-activating factor in mammals and present our current knowledge regarding the characterization of this protein in the horse. The equine sequence yielded a protein of high relative [Ca2+]i-releasing activity. Equine PLCZ was expressed over the head region overlying the acrosome, equatorial segment, connecting piece between the head and midpiece, and on the principal piece of the flagellum of stallion sperm. Equine PLCZ expressed both over the head and tail sperm regions was catalytically active, with the latter representing a characteristic unique to the horse. We also present preliminary data in subfertile stallions displaying PLCZ expression defects, although further research is required to establish a clear association between these defects and fertility problems in the horse. In summary, the information presented raises the questions of whether equine PLCZ could play diverse roles in sperm physiology and/or become a marker for the evaluation of stallion fertility, both of which are worthy of further investigation. 相似文献
13.
J. -L. Riond N. Kocabagli U. E. Spichiger M. Wanner 《Veterinary research communications》1995,19(3):195-203
Ion-selective electrodes have recently been designed for determining the ionized concentration of magnesium (Mg2+) in serum. This development may allow new insights into some metabolic diseases of cattle. For this report, the concentrations of Mg2+, total magnesium (Mgtot), ionized calcium (Ca2+), total calcium (Catot), and inorganic phosphate (Pi) were determined in sera from seventeen 3-to 16-year-old Brown Swiss and crossed Simmental/Red Holstein cows during the periparturient period. In each animal, a transient increase of Mg2+ and Mgtot serum concentrations was observed in association with the transient decrease in serum concentrations of Ca2+, Catot and Pi after parturition. On average, throughout the study, the serum Mg2+ concentrations were 68.5% of those of Mgtot, whereas the serum Ca2+ concentrations were 52% of those of Catot. The possible mechanisms involved in the transient increase of Mg2+ and Mgtot serum concentrations are discussed.Abbreviations Ca2+
ionized calcium
- Catot
total calcium
- Mg2+
ionized magnesium
- Mgtot
total magnesium
- Pi
inorganic phosphate
- PTH
parathyroid hormone
- PTHrP
parathyroid homrone related protein 相似文献
14.
《Domestic animal endocrinology》1997,14(2):99-107
The effects of amino acids on growth hormone (GH) release and cytosolic calcium concentration ([Ca2+]i) were investigated in caprine anterior pituitary cells cultured for 3 d in Dulbecco modified Eagle medium. The addition of an amino acid mixture consisting of seven nonessential amino acids (NEAA: l-Asp, Gly, l-Ala, l-Ser, l-Pro, l-Asn, and l-Glu; concentration of each 12.5–200 μmol/l) in the medium significantly raised GH release from the cultured cells in a concentration-dependent manner with the maximum release at 200 μmol/l NEAA. Although an addition of l-Asp (0.1–100 μmol/l) caused a significant rise in GH release in a concentration-dependent manner, neither the individual amino acids contained in NEAA except l-Asp nor others (l-Leu, l-Phe, l-Gln, l-Met, and l-Arg) caused a rise in GH release when added alone to the medium. The rise in GH release induced by NEAA (200 μmol/l) and GH-releasing hormone (GHRH, 10 nmol/l) was significantly reduced by the addition of EGTA (l.8 mmol/l) and nifedipine (1 μmol/l) to the medium, respectively. The addition of NEAA (200 μmol/l) caused a rapid and transient [Ca2+]i increase, followed thereafter by a steady increase. The prior addition of nifedipine (1 μmol/l), which itself significantly reduced the basal [Ca2+]i, completely abolished the response induced by NEAA or GHRH. From these findings, we conclude that: 1) NEAA raises GH release and [Ca2+]i in cultured caprine anterior pituitary cells, and 2) Ca2+ influx from the medium may be responsible for the cellular action of NEAA. 相似文献
15.
Hannah Y. Y. Lee James D. Morton Julie Sanderson Roy Bickerstaffe Lucinda J. G. Robertson 《Veterinary ophthalmology》2008,11(6):347-355
Objective To investigate biochemical changes accompanying Ca2+-induced lens opacification and the possible role of calpain activation in opacification within an ovine lens culture system. Methods Sheep lenses were cultured in minimal media. Lens opacification was induced by exposure to the Ca2+ ionophore, ionomycin, and graded by digital image analysis. Cell viability was estimated by the release of lactate dehydrogenase into the culture medium. Opaque lenses were fixed and stained for a microscopic view of the lens structural changes. Ionic changes in the lens were measured by atomic absorption spectroscopy. Calpain activation was determined by zymography on casein gels and proteolysis was investigated by SDS–polyacrylamide gel electrophoresis (SDS–PAGE), two-dimensional gel electrophoresis (2DE) and Western blotting. The calpain inhibitor, SJA6017, was used to investigate the involvement of calpains in lens opacification. Results Treatment of cultured ovine lenses with ionomycin increased total lens Ca2+ concentration and caused the cortical region of the lens to become opaque. Addition of the Ca2+ chelator, EGTA, inhibited the ionomycin-induced changes. Progress of opacification correlated with the death of lens cells and lens swelling in differentiating fiber cells. Autolysis of calpain 2, following ionomycin treatment, suggested activation of this protease. 2DE revealed that the ionomycin did not result in substantial proteolysis of the crystallins. However, Western blotting revealed significant breakdown of the cytoskeletal proteins, spectrin and vimentin. The pattern of the breakdown products was consistent with calpain proteolytic activity. SJA6017 retarded the cortical opacity induced by Ca2+-overload in the ovine lens. Conclusion The ovine lens with Ca2+-induced opacification by ionomycin is associated with calpain activation and the subsequent proteolysis of cytoskeletal proteins. These events could be initial factors contributing to cell death and the loss of lens transparency which occurs in this ovine model of cataractogenesis. The ovine model supports the hypothesis that cytoskeletal proteins and Ca2+ homeostasis play an important role in maintaining lens transparency. 相似文献
16.
Shusei Mizushima Tomohiro Sasanami Tamao Ono Norio Kansaku Asato Kuroiwa 《The Journal of Poultry Science》2022,59(2):175
We previously reported that egg activation in Japanese quail is driven by two distinct types of intracellular Ca2+ ([Ca2+]i): transient elevations in [Ca2+]i induced by phospholipase Czeta 1 (PLCZ1) and long-lasting spiral-like Ca2+ oscillations by citrate synthase (CS) and aconitate hydratase 2 (ACO2). Although the blockade of inositol 1,4,5-trisphosphate receptors (ITPRs) before microinjections of PLCZ1, CS, and ACO2 cRNAs only prevented transient increases in [Ca2+]i, a microinjection of an agonist of ryanodine receptors (RYRs) induced spiral-like Ca2+ oscillations, indicating the involvement of both ITPRs and RYRs in these events. In this study, we investigated the isoforms of ITPRs and RYRs responsible for the expression of the two types of [Ca2+]i increases. RT-PCR and western blot analyses revealed that ITPR1, ITPR3, and RYR3 were expressed in ovulated eggs. These proteins were degraded 3 h after the microinjection of PLCZ1, CS, and ACO2 cRNAs, which is the time at which egg activation was complete. However, degradation of ITPR1 and ITPR3, but not RYR3, was initiated 30 min after a single injection of PLCZ1 cRNA, corresponding to the time of the initial Ca2+ wave termination. In contrast, RYR3 degradation was observed 3 h after the microinjection of CS and ACO2 cRNAs. These results indicate that ITPRs and RYR3 differentially mediate in creases in [Ca2+]i during egg activation in Japanese quail, and that downregulation of ITPRs and RYR3-mediated events terminate the initial Ca2+ wave and spiral-like Ca2+ oscillations, respectively. 相似文献
17.
Yan Sai Junfeng Chen Feng Ye Yuanpeng Zhao Zhongmin Zou Jia Cao Zhaojun Dong 《Journal of toxicologic pathology》2013,26(2):149-157
Rotenone is an inhibitor of mitochondrial complex I that produces a model of Parkinson’s
disease (PD), in which neurons undergo dopamine release dysfunction and other features. In
neurons, exocytosis is one of the processes associated with dopamine release and is
dependent on Ca2+ dynamic changes of the cell. In the present study, we have
investigated the exocytosis of dopamine and the involvement of Ca2+ in dopamine
release in PC12 cells administrated with rotenone. Results demonstrated that rotenone led
to an elevation of intracellular Ca2+ through Ca2+ influx by opening
of the voltage-gated Ca2+ channel and influenced the soluble N-ethylmaleimide
attachment protein receptor (SNARE) proteins expression (including syntaxin,
vesicle-associated membrane protein 2 (VAMP2) and synaptosome-associated
protein 25 (SNAP-25)); pretreatment with a blocker of L-type voltage-activated
Ca2+ channels (nifedipine) decreased the intracellular dopamine levels and
ROS formation, increased the cell viability and enhanced the neurite outgrowth and
exocytosis of synaptic vesicles. These results indicated that the involvement of
intracellular Ca2+ was one of the factors resulting in suppression of dopamine
release suppression in PC12 cells intoxicated with rotenone, which was associated with the
rotenone-induced dopamine neurotoxicity. 相似文献
18.
The effect of Brucella on the generation of microbicidal reactive oxygen and nitrogen metabolites by bovine peripheral polymorphonuclear cells (PMNs) was investigated. The PMNs were recovered from the peripheral blood of control calves and experimental calves previously vaccinated against brucellosis. Significantly larger quantities of NO and H2O2 were generated by PMNs from control and experimental calves following activation by heat-killed whole cells or outer membrane protein of Brucella abortus than by non-activated cells (p<0.05–0.01). In contrast, generation of H2O2 and NO decreased when PMNs were exposed to the lipopolysaccharide of Brucella. However, the generation of H2O2 and NO by activated PMNs from the control and experimental calves did not differ significantly. 相似文献
19.
Cytoskeletal and mitochondrial properties of bovine oocytes obtained by Ovum Pick‐Up: the effects of follicle stimulation and in vitro maturation
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Tamás Somfai Satoko Matoba Yasushi Inaba Michiko Nakai Kei Imai Takashi Nagai Masaya Geshi 《Animal Science Journal》2015,86(12):970-980
Follicle stimulation by follicular stimulating hormone (FSH) is known to improve developmental competence of bovine oocytes obtained by Ovum Pick‐Up (OPU); however, the exact factors in oocytes affected by this treatment have remained unclear. We compared in vitro matured (IVM) oocytes obtained at the immature stage from cows by OPU either without or with stimulation with FSH (non‐stimulated and stimulated OPU, respectively) to those obtained by superstimulation and in vivo maturation in terms of cytoskeleton morphology, mitochondrial distribution, intracellular adenosine triphosphate (ATP) content and H2O2 levels at the metaphase‐II stage and intracellular Ca2+ levels after in vitro fertilization (IVF). Confocal microscopy after immunostaining revealed reduced size of the meiotic spindle, associated with increased tendencies of microfilament degradation and insufficient mitochondrial re‐distribution in non‐stimulated OPU‐derived IVM oocytes compared with those collected by stimulated OPU, which in turn resembled in vivo matured oocytes. However, there was no difference in mitochondrial functions between oocytes obtained by stimulated or non‐stimulated OPU in terms of ATP content, cytoplasmic H2O2 levels, base Ca2+ levels and the frequencies and amplitudes of Ca2+ oscillations after IVF. Larger size of metaphase spindles in oocytes obtained by stimulated OPU may reflect and potentially contribute to their high developmental competence. 相似文献
20.
Objective: To determine whether in healthy horses and those with colic, exposure of peritoneal fluid to room air affects values obtained on biochemical analysis. Study Design: Prospective study. Animals: Adult horses with a primary complaint of acute abdominal pain (n=29) and 12 healthy horses. Methods: Peritoneal fluid was aseptically collected under aerobic and anaerobic conditions. After collection, pH, PCO2, PO2, HCO3?, Na+, ionized Ca2+, K+, lactate, and glucose were immediately measured using a commercial blood gas analyzer. Biochemical variables were compared between aerobically and anaerobically obtained samples using a paired t‐test. Results: In healthy horses, peritoneal fluid samples collected under anaerobic conditions had higher PCO2 and ionized Ca2+ and lower PO2, HCO3?, and pH compared with samples exposed to air. No differences were observed for K+, Na+, glucose, and lactate. In horses with colic, samples collected anaerobically had higher PCO2, ionized Ca2+, Na+, and glucose and lower PO2, HCO3?, and pH value compared with samples exposed to air. No differences were observed for K+ and lactate. Conclusion: Exposure of peritoneal fluid to room air had a significant effect on pH, PCO2, PO2, and variables associated or dependent on changes in pH such as HCO3? and ionized Ca2+. Interpretation of biochemical analysis of peritoneal fluid may be influenced by sample collection method. 相似文献