A novel terminal ampullae peptide is involved in the proteolytic activity of sperm in the prawn, Macrobrachium rosenbergii.
As the distal part of the crustacean male reproductive tract, terminal ampullae play important roles in sperm development and storage of mature spermatophores. In the present study, the novel gene terminal ampullae peptide (TAP) was cloned from terminal ampullae of the prawn, Macrobrachium rosenbergii. The cDNA sequence consists of 768 nucleotides, with an open-reading frame of 264 nucleotides which encodes a putative 88-amino acid precursor protein with a 17-amino acid residue signal peptide. Western blotting and immunohistochemical analysis revealed that TAP was distributed on terminal ampullae and sperm, and its expression was related to gonad development. To elucidate the functional role of TAP in vivo, we disrupted the TAP gene by RNA interference (RNAi) and evaluated the effect on fertility and several sperm parameters. Although there was no difference in fertility between RNAi-induced prawns and controls, RNAi treatment decreased the sperm gelatinolytic activity and blocked proteolytic activity on the vitelline coat. These data provide evidence that TAP participates in regulating sperm proteolytic activity, and performs a crucial role in sperm maturation and degradation of the vitelline coat during fertilization.
Ma
Wen-Ming
WM
College of Life Sciences, Institute of Cell Biology and Genetics, Zhejiang University, Zijingang Campus, Hangzhou, Zhejiang 310058, People's Republic of China.
Qian
Ye-Qing
YQ
Wang
Mo-Ran
MR
Yang
Fan
F
Yang
Wei-Jun
WJ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D005260:Female; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D033541:Palaemonidae / Q000378:metabolism* / Q000502:physiology; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D013094:Spermatozoa / Q000201:enzymology*
D012333:RNA, Messenger; D010447:Peptide Hydrolases