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陈春娜,黄颖颖,陈先均,龙治海,杜军.2015.达氏鲟精子的主要生物学特性.动物学杂志,50(1):75-87.
达氏鲟精子的主要生物学特性
Biological characteristics of Acipenser dabryanus sperm
投稿时间:2013-10-23  修订日期:2015-01-04
DOI:DOI: 10.13859/j.cjz.201501010
中文关键词:  达氏鲟  精子  生物学特性  
英文关键词:Acipenser  dabryanus sperm  motility spermatozoa  structure seminal  plasma
基金项目:四川省财政基因工程青年(达氏鲟精子生理生态特性研究(2012QNJJ-016)
作者单位E-mail
陈春娜 四川省农业科学院水产研究所 hc_1981_2001@sina.com 
黄颖颖 四川省农业科学院水产研究所  
陈先均 四川省农业科学院水产研究所  
龙治海 四川省农业科学院水产研究所  
杜军 四川省农业科学院水产研究所 dujun9100@126.com 
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中文摘要:
      达氏鲟(Acipenser dabryanus)属淡水定居性鲟鱼类,为我国特有种,主要分布在长江上游干流及金沙江下游。长期人为的过度捕捞及其生存环境的持续污染和水利工程的影响,使得达氏鲟自然种群资源遭到严重破坏,其配子质量的下降己经成为限制其规模化人工繁殖成功的关键因素之一,因此为解决达氏鲟规模化人工繁殖过程中存在的关键性技术点,作者从达氏鲟精液基本特征、精浆元素组成、不同水体及Na , K 对达氏鲟精子活力的影响、精子超微结构方面入手,对达氏鲟精子的生理生态特性进行了研究。结果显示,达氏鲟精子平均密度为1.52×109 个/ml; 精浆元素以Na 含量最高,其次是K ,之后为Ca2 、Mg2 、Cu2 、Zn2 ,其中Na 、 K 、Zn2 在达氏鲟精浆中的含量有极显著性差异(P<0.01),Ca2 、Cu2 、Mg2 差异不明显; 精子在江水中的活力最高; 在Na 浓度为20 mmol/L时,精子活力最高,精子快速运动时间(FT)和寿命(LT)分别为(66.7±7.1)s和(177.0±14.9)s,达氏鲟精子对K 浓度变化较为敏感,在K 浓度为0.05 mmol/L时,精子FT和LT最长,分别为(109.0±16.1)s和(189.3±12.4)s,超过0.05 mmol/L后精子FT和LT急速下降,当K 浓度达到0.5 mmol/L以上时,精子活力立即受到抑制;达氏鲟精子细胞核长(5.67±0.20)μm,鞭毛长(63.16±2.79)μm, 全长为(70.35±2.92)μm。
英文摘要:
      Acipenser dabryanus lives only in the Yangtze River and Chin-sha River, and its natural populations have been damaged severely by long-term over-exploitation and increasing pollution. In 1988, the fish was listed as class Ⅰ state protected animal. It is immediately required to develop conservation measures for its populations, which requires both management and understanding of its biogeographical and biological chracteristics. Beacuase of the decline of natural spawning colony resource, there is litter knowledge about the sperm physio-ecological characteristics. Healthy male A.dabryanus individuals which had been cultured by our institute were selected and their spermatozoa were collected. The density of spermatozoa was determined by counting using a haemocytometer after 1 000× dilution of samples with 0.65% NaCl extender, and the results from three replicates were averaged. Milt concentration was enumerated by using milt collected into standard centrifuge tubes and centrifuged at 4 000 r/min for 30 min. The sperm cell volume percentage in the total volume of semen was calculated from 5 replicates. Semen pH was measured with a precision test paper. Ionic composition of seminal plasma was measured by the atomic absorption spectrophotometry method with the use of a AA_220 spectrometer (Varian USA). The effects of different aqueous solution on the sperm motility were measured by using different sources of test water, and Na , K sensitivity of sperm motility was assessed by activating sperm with 20, 40, 60, 80, 100, 120 mmol/L NaCL and 0.01, 0.05, 0.10, 0.50, 1.00, 1.50 mmol/L KCl, respectively. For testing the motility of spermatozoa, 1 μl of milt was mixed directly with 30 μl of the experimental solution, placed on a glass slide and examined under 400× magnification. The ultrastructure of spermatozoa was observed by using SEM (scanning electron microscopy) and TEM (transmission electron microscopy). All statistical analyses were conducted with SPSS STATISTICS, Version 17.0. For multiple comparisons between groups with LSD method, statistical significance was declared at P < 0.05. Results were presented as mean ± standard deviation (SD). The results showed that the sperm density of A.dabryanus was 0.63 × 109 to 2.73 × 109/ml, with the average of 1.52 × 109/ml, the concentration of sperm was 3.18% to 12.98%, with the average of 7.67%, and the pH value was between 7.5 and 7.9 (Table 2). The ions composition in seminal plasma of A.dabryanus included Na , K , Ca2 , Mg2 , Cu2 , Zn2 , and Na was the main ion in seminal plasma, followed by K , Mg2 , Ca2 , Cu2 , Zn2 . There was significant difference in the concentration of Na , K , and Zn2 (P < 0.01), but no significant difference in Ca2 , Cu2 , Mg2 (Table 3). In A. dabryanus, the Na , K concentrations were lower than in teleostean fish (Table 4).The sperm had the highest motility in river water, and the fast movement time (FT) and lifetime (LT) were 142 s and 373 s, respectively (Fig. 1). The activity of A. dabryanus sperm in NaCl solution reached the highest when its concentration was 20 mmol/L, with the FT and LT being 66.7 ± 7.1 s and 177.0 ± 14.9 s, respectively (Fig. 2a). When the concentration of K was 0.05 mmol/L, the motility of sperm was the highest, while the FT and LT were the longest, which were 109.0 ± 16.1 s and 189.3 ± 12.4 s, respectively (Fig. 2b). The average length of the nucleus was 5.67 ± 0.20 μm (Table 5). Three intertwining endonuclear canals traversed the nucleus longitudinally from the acrosomal end to the basal nuclear fossa region. There were three mitochondria in the midpiece. The flagellum (63.16 ± 2.79 μm in length), originating from the centriolar apparatus, had a typical 9 2 eukaryotic flagellar organization (PlateⅠ). Some A. dabryanus sperm characteristics are reported here, but further investigations are needed in order to improve our knowledge of reproductive biology of this species and to establish gamete management methods.
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