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游离氨基酸在虹鳟和鲤鱼精液中的作用。

The role of free amino acids in semen of rainbow trout Oncorhynchus mykiss and carp Cyprinus carpio.

机构信息

Department for Organismic Biology, University of Salzburg, Hellbrunnerstrasse 34, 5020 Salzburg, Austria.

出版信息

J Fish Biol. 2009 Sep;75(4):816-33. doi: 10.1111/j.1095-8649.2009.02317.x.

Abstract

The present study investigated (1) the free amino acid (FAA) composition in semen of rainbow trout Oncorhynchus mykiss and carp Cyprinus carpio, (2) enzyme systems involved in amino acid metabolism and (3) the effect of amino acids on sperm viability under in vitro storage conditions. In the seminal plasma of O. mykiss, the main FAAs were arginine, glutamic acid, isoleucine, leucine, methionine and proline, in spermatozoa cysteine, arginine and methionine. In the seminal plasma of C. carpio, the main FAAs were alanine, arginine, cysteine, glutamic acid, histidine, leucine, lysine, methionine and proline, in spermatozoa arginine, glutamic acid, histidine, leucine and lysine. When spermatozoa were incubated for 48 h together with the seminal plasma, the quantitative amino acid pattern changed in both species indicating their metabolism. In spermatozoa and seminal plasma of O. mykiss and C. carpio, the following enzymes were found to be related to amino acid metabolism: transaminases (specific for alanine, aspartate, isoleucine and leucine), decarboxylases (specific for valine and lysine), glutamate dehydrogenase and alpha-keto acid dehydrogenases (substrates: 3-methyl-2-oxovaleric acid and 4-methyl-2-oxovalerate). These data demonstrate that amino acid catabolism by transamination, decarboxylation and oxidative deamination can occur in semen of the two species. Also activity of methionine sulphoxide reductase was detected, an enzyme which reduces methionine sulphoxide to methionine. This reaction plays an important role in antioxidant defence. To determine the effect of FAAs on the sperm viability, C. carpio and O. mykiss spermatozoa were incubated in sperm motility inhibiting saline solution containing different amino acids. Methionine had a positive effect on the sperm viability in both species. Taken together this result with the in vivo occurrence of methionine and of methionine reductase in semen, it can be assumed that this amino acid plays an important role in antioxidant defence. Also isoleucine in O. mykiss and leucine in C. carpio had a positive effect on sperm viability. As seminal plasma and spermatozoa of the two species exhibit enzyme activities to catabolize leucine and isoleucine, they might serve as additional energy resources especially during prolonged incubation and storage periods.

摘要

本研究调查了(1)虹鳟 Oncorhynchus mykiss 和鲤鱼 Cyprinus carpio 精液中的游离氨基酸(FAA)组成,(2)参与氨基酸代谢的酶系统,(3)在体外储存条件下氨基酸对精子活力的影响。在 O. mykiss 的精液中,主要的 FAAs 是精氨酸、谷氨酸、异亮氨酸、亮氨酸、蛋氨酸和脯氨酸,在精子中是半胱氨酸、精氨酸和蛋氨酸。在 C. carpio 的精液中,主要的 FAAs 是丙氨酸、精氨酸、半胱氨酸、谷氨酸、组氨酸、亮氨酸、赖氨酸、蛋氨酸和脯氨酸,在精子中是精氨酸、谷氨酸、组氨酸、亮氨酸和赖氨酸。当精子与精液一起孵育 48 小时时,两种物种的定量氨基酸图谱发生变化,表明它们在代谢。在 O. mykiss 和 C. carpio 的精子和精液中,发现以下酶与氨基酸代谢有关:转氨酶(特异性为丙氨酸、天冬氨酸、异亮氨酸和亮氨酸)、脱羧酶(特异性为缬氨酸和赖氨酸)、谷氨酸脱氢酶和α-酮酸脱氢酶(底物:3-甲基-2-氧代戊酸和 4-甲基-2-氧代戊酸)。这些数据表明,两种物种的精液中可以通过转氨基、脱羧和氧化脱氨作用进行氨基酸分解代谢。还检测到了蛋氨酸亚砜还原酶的活性,该酶将蛋氨酸亚砜还原为蛋氨酸。该反应在抗氧化防御中起着重要作用。为了确定 FAAs 对精子活力的影响,将 C. carpio 和 O. mykiss 精子在含有不同氨基酸的精子运动抑制盐溶液中孵育。蛋氨酸对两种物种的精子活力都有积极影响。综合这一结果以及体内蛋氨酸和蛋氨酸还原酶在精液中的存在,可以假设这种氨基酸在抗氧化防御中起着重要作用。O. mykiss 中的异亮氨酸和 C. carpio 中的亮氨酸对精子活力也有积极影响。由于这两种物种的精液都具有分解亮氨酸和异亮氨酸的酶活性,因此它们可能是额外的能量来源,特别是在长时间孵育和储存期间。

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