Abstract:
Australian redclaw crayfish,
Cherax quadricarinatus,belongs to Decapoda order,and Parastacidae family.Because it grows rapidly and can reach commercial size in a few months at optimal conditions,it has been one of the most important economy shrimps in China.However,the physiological mechanisms of redclaw crayfish have not been well studied,especially the basic information of key nutrients and the related enzymes or proteins are missing.
Cathepsin L(
CatL)is an important enzyme related to digesting process in crustaceans,however,its molecular structure and correlation with dietary nutrients have not been investigated.In the present study,in order to obtain insights into the regulatory effects of key dietary nutrients on expression of
CatL gene,the
CatL gene of
C.quadricarinatus was cloned first using RACE-PCR methods.The tissue-specific
CatL gene expression was also performed,and the
C.quadricarinatus were fed with artificial diets containing different levels of vitamin C(0,100,200,400,800,1 600,3 200 mg/kg)for 8 weeks,and the expression of
CqCatL mRNA in different tissues and different groups were examined.The
CatL cDNA was isolated from the hepatopancreas of
C.quadricarinatus using RACE-PCR methods,which was designated
CqCatL(Genbank accession number:KJ913663).The full-length cDNA of
CqCatL obtained from the present study was 1 810 bp in length with an open reading frame of 1 026 bp,encoding a putative peptide of 341 amino acids,and the predicted molecular mass and isoelectric point were 37.63 ku and 5.17.By alignment,the amino acid sequence of
CqCatL of
C.quadricarinatus showed high homology with those of other animals.For example,the deduced amino acids sequence of
C.quadricarinatus CqCatL shared 79% homology with
Penaeus monodon,and 74%,59%,59%,58% and 58% with
Macrobrachium nipponense,Eriocheir sinensis,Metapenaeus ensis,Metapenaeus ensis and Litopenaeus vannamei,respectively.This suggested that
CqCatL was highly conservative.The tissue-specific expression pattern study indicated that
CqCatL mRNA was widely expressed in hemocytes,gill,hepatopancreas,muscle,ovary,intestine,stomach,heart,brain and antennal gland,indicating
CqCatL may play important roles in many tissues of the
C.quadricarinatus.The highest expression was observed in hepatopancreas,and the second was seen in hemocytes,while only very low expressions of
CqCatL mRNA were found in intestine and antennal gland.After feeding with seven levels of vitamin C,the mRNA expression of
CqCatL was the highest in the 400 mg/kg dietary vitamin C group and significantly higher than the other groups.The results showed that
CqCatL might play an important role in the physiological process of
C.quadricarinatus,and its gene expression and functions were likely regulated by nutrients,such as vitamin C.