Sperm motility is one of the most important determinants of male fertility. Elucidating the different expression proteins of nomal and asthenospermia spermatozoa should advance our understanding of the underlying molecular mechanisms. Tcte1 null mouse sperm reduced motility and failed to display the forward progressive motile. With tandem mass tag (TMT) labeling and mass spectrometry analysis， 3697 proteins were quantified from Widetype and Tcte1 deficient mouse spermatozoa. Of these proteins, 447 were found to be different expression proteins (DEPs), including 398 proteins down regulated (DDEPs, 89%) and only 49 proteins up regulated.