| The autolysis of sea cucumber restricted its development of aquaculture and industry processing.Thus,the mechanism of sea cucumber autolysis should be further researched.Physical and chemical stimulation can lead to sea cucumber autolysis.Our previous studies found that the apoptosis appeared in the process of sea cucumber autolysis.Therefore,the apoptosis research can contribute to the mechanism of sea cucumber autolysis.This study could significance to explain the mechanism of sea cucumbers autolysis.The immunohistochemistry in situ labeling of cytochrome C during the sea cucumber autolysis was studied in this paper.Western blot was performed to detect the migration of cytochrome C in sea cucumber gut cytoplasm and mitochondrial.The presence of caspase-9 in cavity cell cytoplasm of sea cucumber was also detected.TUNEL method was used to label the apoptotic DNA fragments in sea cucumber body wall during the autolysis processing.Immunohistochemistry experimental results indicated that the cytochrome C staining particles in fresh sea cucumber body wall is larger than the samples that autolysis activated.With time lapse,the cytochrome C staining particles gradually become smaller and spread to the surroundings.Sea cucumber cytochrome C in intestines cytoplasm and intestines mitochondria were separated and marked by western blot.The molecular weight of sea cucumber cytochrome C is about 15KDa.With autolysis time lapsed,the concentration of cytochrome C in cytoplasmic increased,but it was decreased in mitochondria.The cytochrome C and caspase-9 in body cavity fluid cytoplasmic was marked by western blot.TUNEL method was used to label DNA fragments in situ of sea cucumber body wall after it was induced autolysis by UV irradiation.The nuclear DNA in sea cucumber body wall was gradually to fragment,with autolysis activated.These results proved the existence of apoptosis during sea cucumber autolysis and mitochondrial apoptotic pathway may involve during sea cucumber autolysis. |
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