Identification and characterization of a novel splice variant of the PLCζ1 gene in bull testis tissues

Abstract

ABSTRACT. Phospholipase C zeta 1 (PLCζ1), which transcribes a key protein, has an important function in oocyte activation and embryo development because PLCζ1 can trigger a series of intracellular Ca 2+ oscillations in mammals. In this study, a novel splice variant in the testis tissues of adult and fetal Chinese Holstein bulls was characterized by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing analysis. The novel splice variant PLCζ1-sv1 was derived from the PLCζ1 complete transcript (PLCζ1-complete) by alternative splicing; the alternative splicing pattern exhibited alternative 5'-splice sites. The full-length transcript, PLCζ1-complete, is the main transcript found in fetal and adult cow testis tissue. Quantitative real-time PCR (qPCR) analysis demonstrated that the expression levels of the PLCζ1-complete transcript were significantly higher than those of the PLCζ1-sv1 splice variant in bovine testis tissues. PLCζ1 protein sequencing analysis showed that the amino acids at positions 453 to 457 were deleted in PLCζ1-sv1, thereby terminating transcription prematurely. In summary, this study provided information to elucidate the structure and function of the bovine PLCζ1 gene.

Phospholipase C Zeta 1 (PLCZ1): The Function and Potential for Fertility Assessment and In Vitro Embryo Production in Cattle and Horses

“…Bovine PLCZ1 is a 70–72 kDa protein compound of 634 amino acids and has a high homology with porcine PLCZ1 (83.0%) and lesser homology with equine PLCZ1 (79.4%) [ 13 , 68 , 105 , 148 ]. PLCZ1 is located in the equatorial and postacrosomal regions of bull sperm ( Figure 2 B,C); however, nonspecific immunoreactivity has also been detected in the acrosomal region and tail [ 13 , 63 , 64 ].…”

“…Also in Chinese Holstein bulls, a novel splice variant of PLCZ1 was identified in testis tissue. The protein sequencing analysis revealed amino acid deletions that resulted in altered transcription of the Y and C2 domains, impairing catalytic and lipid binding functions [ 148 ]. These findings confirm the existence of genotype and, consequently, phenotype diversity for PLCZ1 in bulls.…”