The Application Of Mitochondria And DNA Integrity In Sperm Quality Assessment Of Eriocheir Sinensis

1. Study on sperm mitochondria of Eriocheir sinensisThree methods which were SDH activity, specific fluorescent probe (Rhl23) and mitochondrial 16S rDNA amplification, were used to detect the sperm mitochondria of Eriocheir sinensis, and three tissues which were muscle, branchia and testis were served as control. SDH activity in testis was only 28.06 U/mgprot which was significant lower than that in muscle (114.55U/mgprot) and in branchia (93.72 U/mgprot), while it was too low to be detected in sperm. Strong fluorescence was detected in testis by Rh123 but nothing in sperm. Primers used in PCR were designed based on the sequence of mitochondrial 16S rDNA (AJ250642, AF312718) and beta-actin (AY910691) of Eriocheir sinensis which had been deposited in Genebank. PCR analysis using total DNA from muscle, branchia, testis and sperm indicated that the two genes exhibit different amplifications in four tissues/cell. Beta-actin amplification was detected in high levels in all four tissues/cell and all patterns were the same. 16S rDNA gene amplification was detected in high levels in muscle and branchia, appreciable low in testis and very low in sperm. The PCR products of 16srDA were sequenced and aligned with the sequence of 16S rDNA from Genebank (AJ250642, AF312718), and the alignment result showed no difference between them. All the results suggest that there is mitochondria in Eriocheir sinensis sperm but much lower than that in testis and other tissues ,and this maybe the reason why SDH activity and Rhl23 as well as common histology and ultrastructure research can not detect the existence of sperm mitochondria.2. Research on Eriocheir sinensis sperm DNA damage by HgC₂ and CdCl₂ using single cell gel electrophoresisTo study the effects of mercuric chloride and cadmium chloride on DNA damage of Eriocheir sinensis sperm in vitrojsolated Eriocheir sinensis sperm were exposed to mercuric chloride and cadmium chloride respective concentration:0,0.01,0.05,0.1,1, 5mmol/L for one hour.The proportion of DNA damage and length of DNA...