摘除眼柄对河南华溪蟹蜕皮激素与几丁质酶含量和β-NAGase活性的影响  

Effects of eyestalk ablation on the ecdysone and chitinase levels and N-acetyl-β-D-glocosaminidase activity of the freshwater crab( Sinopotamon henanense)

作　　者：裴思慧 落继先[1] 邹恩民 孙敏[1] 刘娜[1] 井维鑫[1] 王兰[1] 

机构地区：[1]山西大学生命科学学院,山西太原030006 [2]美国尼古拉斯州立大学生物科学系,路易斯安那州帝博多市70310

出　　处：《水产学报》2014年第5期677-683,共7页Journal of Fisheries of China

基　　金：国家自然科学基金(31272319);高等学校博士学科点专项科研基金(20111401110010);中国博士后科学基金(2012M510773);山西省教育厅专项(20111001)

摘　　要：为探索摘除眼柄对河南华溪蟹蜕皮机制的影响,采用灼伤挤压法摘除眼柄后,运用ELISA技术检测了河南华溪蟹血淋巴中蜕皮激素的含量和表皮组织中几丁质酶的含量,同时用比色法测定了表皮中N-乙酰-β-D-氨基葡萄糖苷酶(β-NAGase)的活性。结果显示,摘除眼柄后河南华溪蟹血淋巴中蜕皮激素的含量出现先升高后下降的总趋势,在摘除眼柄96 h时,血淋巴中蜕皮激素含量上升到最大值[♂:(23.25±4.56)ng/L;♀:(35.75±7.15)ng/L]。表皮中几丁质酶的含量在摘除眼柄后亦出现先升高后下降的变化趋势,在摘除眼柄48 h达到最大[♂:(16.29±3.91)ng/L;♀:(30.49±5.28)ng/L],到120 h仍显著高于对照组。表皮中N-乙酰-β-D-氨基葡萄糖苷酶(β-NAGase)的活性在摘除眼柄后持续升高,在144 h时达到最大[♂:(400.44±21.00)U/g;♀:(216.94±23.97)U/g],后逐渐下降,但依旧高于对照组。研究推测,摘除眼柄通过增加蜕皮激素含量、几丁质酶的含量和β-NAGase的活性来促进河南华溪蟹蜕皮。实验将为进一步研究甲壳动物蜕皮生长的调控机制提供新的理论基础。Molting is an essential stage during the growth of crustacean,which is coordinated by ecdysone secreted by Y-organ,chitinase synthesized by the epidermis; while negatively regulated by molting-inhibiting hormone secreted by X-organ sinus-gland complex at the bottom of eyestalk. However,the specific mechanisms of the molting triggered by eyestalk ablation are still unknown. In this study,two eyestalks were ablated and the ecdysone level in Sinopotamon henanense haemolymph and chitinase level in the epidermis were measured by ELISA,and N-acetyl-β-D-glocosaminidase（ β-NAGase） activity in the epidermis was measured by colormetry. Crabs were divided into one control group and seven eyestalk ablation groups respectively（ from 24 hours to 168 hours）. Our results showed that the level of ecdysone in haemolymph showed an increase at first and then decrease,with the maximum value at 96 h. [♂ ：（ 23. 25 ± 4. 56） ng / L; ♀：（ 35. 75 ± 7. 15） ng /L,P 0. 01]. This tendency was accompanied by similar changes as that of chitinase levels in the epidermis. After the two eyestalks were ablated for 48 h to 120 h,chitinase level increased significantly（ compared with the control group,P 0. 05）. Activity of β-NAGase increased continuously by the eyestalk ablation and reached the maximum value at 144 h[♂ ：（ 400. 44 ± 21. 00） U / g; ♀：（ 216. 94 ±23. 97） U /g,P 0. 05]. Then the activity of β-NAGase decreased gradually,but still higher than that of control group. The results imply that eyestalk ablation may affect the molting process by increasing the levels of ecdysone in the haemolymph,chitinase in the epidermis and the activity of β-NAGase. This study will provide the theoretical basis for the further study on the physiological regulation of crustacean molting and growth.

关 键 词：河南华溪蟹 蜕皮 蜕皮激素 几丁质酶 N-乙酰-Β-D-氨基葡萄糖苷酶 

分 类 号：Q175[生物学—水生生物学] S917.4[生物学—普通生物学]