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초록
영어
During mammalian fertilization, germ cell-specific hyaluronidases, such as sperm adhesion molecule 1 (SPAM1) and hyaluronoglucosaminidase 5 (Hyal5), are important for the dispersal of the cumulus mass. In this study, we demonstrated that bull Hyal5 is a single copy gene on chromosome 4 that is expressed specifically in the testis. In addition, we expressed recombinant bull SPAM1 and Hyal5 in human embryonic kidney 293T cells and showed that these enzymes possessed hyaluronidase activity. We also demonstrated that a polyclonal antibody against bull sperm hyaluronidase inhibits sperm-egg interactions in an in vitro fertilization (IVF) assay. Our results suggested that bull Hyal5 may have a critical role in bull fertilization.
목차
ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Total RNA Extraction and Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)
Southern Blot Analysis
Antibody
Preparation of Protein Extracts
Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis(SDS-PAGE) and Western Blot Analysis
Expression of Bovine PH-20 and Hyal5
Zymography
Dispersal of Cumulus Cells In Vitro
In Vitro Maturation (IVM) and In Vitro Fertilization(IVF)
Statistical Analysis
RESULTS
Expression of PH-20 and Hyal5
Sequence Analysis of Hyal5 and SPAM1
SPAM1 and Hyal5 are Single Copy Genes in the Bovine Genome
Importance of Sperm Hyaluronidase in Sperm-Oocyte Interaction
DISCUSSION
REFERENCES
INTRODUCTION
MATERIALS AND METHODS
Total RNA Extraction and Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)
Southern Blot Analysis
Antibody
Preparation of Protein Extracts
Sodium Dodecyl Sulfate-polyacrylamide Gel Electrophoresis(SDS-PAGE) and Western Blot Analysis
Expression of Bovine PH-20 and Hyal5
Zymography
Dispersal of Cumulus Cells In Vitro
In Vitro Maturation (IVM) and In Vitro Fertilization(IVF)
Statistical Analysis
RESULTS
Expression of PH-20 and Hyal5
Sequence Analysis of Hyal5 and SPAM1
SPAM1 and Hyal5 are Single Copy Genes in the Bovine Genome
Importance of Sperm Hyaluronidase in Sperm-Oocyte Interaction
DISCUSSION
REFERENCES
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참고문헌
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