Glycosidase determination in bovine oviducal fluid at the follicular and luteal phases of the oestrous cycle
Glycosidase determination in bovine oviducal fluid at the follicular and luteal phases of the oestrous cycle
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DOI：
10.1071/rd08113
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发表时间：
2008-01-01
影响因子：
1.9
通讯作者：
Romar, Raquel
中科院分区：
文献类型：
--
作者：
Carrasco, Luis Cesar;Coy, Pilar;Romar, Raquel
配子识别和精子与输卵管的结合是碳水化合物介导的过程，几种糖苷酶被认为在其中起作用，尽管这还没有被明确地证明。输卵管液是受精和早期胚胎发育的生物环境，但输卵管液中的酶组成在很大程度上是未知的。本研究的目的是测定牛输卵管液中的糖苷酶活性和蛋白质含量，以及每条输卵管收集的液量。根据卵巢黄体形态，将屠宰场获得的输卵管分为卵泡期和黄体期。抽吸输卵管液，离心法测定输卵管液量。然后将样品冷冻，直到化验。以4-甲基伞形花环为底物，在pH值为7.2的条件下对7种糖苷酶的活性进行筛选。结果表明，BoF在周期的两个阶段都具有α-L岩藻糖苷酶、β-N-乙酰-氨基葡萄糖苷酶、β-D-半乳糖苷酶、α-D-甘露糖苷酶和β-N-乙酰半乳糖苷酶活性，后两种酶在卵泡期的比活力较高。在发情周期的两个阶段，pH为7.2时，BoF均未检测到N-乙酰神经氨酸酶和α-D-半乳糖苷酶活性，但在pH为4.4时，这两种糖苷酶均有活性。在周期的两个阶段，蛋白质浓度和收集的BoF体积没有差异。这些发现表明，输卵管液表现出糖苷酶活性，在整个发情周期中有特定的变化，表明这些酶在碳水化合物介导的事件中发挥作用。
Gamete recognition and binding of spermatozoa to the oviduct are carbohydrate-mediated processes in which several glycosidases are thought to have a role, although this has not been demonstrated unequivocally. Oviducal fluid is the biological milieu in which fertilisation and early embryo development take place, but the enzyme composition of oviducal fluid is largely unknown. The aim of the present study was to determine glycosidase activity and protein content in bovine oviducal fluid (bOF) and the volume of fluid collected per oviduct. Oviducts obtained from a slaughterhouse were classified as either in the follicular or luteal phase on the basis of ovarian luteal morphology. Oviducal fluid was aspirated, centrifuged and the volume determined. Samples were then frozen until assay. Substrates conjugated to 4-methylumbelliferyl were used to screen for the activity of seven glycosidases at pH 7.2. The results indicate that bOF has alpha-l-fucosidase, beta-N-acetyl-glucosaminidase, beta-D-galactosidase, alpha-D-mannosidase and beta-N-acetyl-galactosaminidase activity during both phases of the cycle, with the specific activity of the latter two enzymes being higher during the follicular phase. There was no N-acetyl-neuraminidase or alpha-D-galactosidase activity detected in bOF at either phase of the oestrous cycle at pH 7.2, but activity for both glycosidases was detected at pH 4.4. There were no differences in protein concentration or the volume of bOF collected between the two phases of the cycle. These findings indicate that oviducal fluid exhibits glycosidase activity, with specific variations throughout the oestrous cycle, suggesting that these enzymes play a role in carbohydrate-mediated events.