title: Characterization of pig sperm hyaluronidase and improvement of the digestibility of cumulus cell mass by recombinant pSPAM1 hyaluronidase in an in vitro fertilization assay.
abstract: Although sperm hyaluronidase is thought to play an important role in mammalian fertilization, the molecular function underlying these steps remains largely unknown. In mouse models, sperm-specific SPAM1 and HYAL5 hyaluronidase are believed to function in both sperm penetration of the cumulus matrix and sperm-ZP binding. However, gene-targeting studies for SPAM1 or HYAL5 show that hyaluronidases are not essential for fertilization, despite the fact that exogenous hyaluronidase can disrupt the cumulus matrix. Therefore, to evaluate whether sperm hyaluronidase is essential for mammalian fertilization, it is necessary to generate HYAL5/SPAM1 double-knockout mice. However, generating double-knockout mice is very difficult because these two genes exist on the same chromosome. Recently, investigators have begun to employ the pig model system to study human disease due to its similarities to human anatomy and physiology. In this study, we confirmed that pig SPAM1 exists as a single copy gene on chromosome 18 and is specifically expressed in the testis. In addition, we expressed recombinant pig SPAM1 in human embryonic kidney 293 cells and showed that these enzymes possess hyaluronidase activity. We also demonstrated that a polyclonal antibody against pig sperm hyaluronidase inhibits sperm-egg interactions in an in vitro fertilization (IVF) assay. Our results suggest that pig SPAM1 may play a critical role in pig fertilization and that recombinant SPAM1 can disperse the oocyte-cumulus complex in an IVF assay.
authors.affiliation: College of Pharmacy, Catholic University of Daegu, Gyeong-buk 712-702, Republic of Korea.
authors.forename: Sungwon
authors.initials: S
authors.lastname: Yoon
authors.affiliation: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chung-buk 363-883, Republic of Korea.
authors.forename: Kyu-Tae
authors.initials: KT
authors.lastname: Chang
authors.affiliation: College of Pharmacy, Catholic University of Daegu, Gyeong-buk 712-702, Republic of Korea.
authors.forename: Hongsang
authors.initials: H
authors.lastname: Cho
authors.affiliation: College of Pharmacy, Catholic University of Daegu, Gyeong-buk 712-702, Republic of Korea.
authors.forename: Jisang
authors.initials: J
authors.lastname: Moon
authors.affiliation: College of Applied Life Sciences, The Research Institute for Subtropical Agriculture and Biotechnology, Jeju National University, Jeju 690-756, Republic of Korea.
authors.forename: Ju-Sung
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Department of Biotechnology, College of Engineering, Daegu University, Gyeong-buk 712-714, Republic of Korea.
authors.forename: Sung-Hun
authors.initials: SH
authors.lastname: Min
authors.affiliation: Department of Biotechnology, College of Engineering, Daegu University, Gyeong-buk 712-714, Republic of Korea.
authors.forename: Deog-Bon
authors.initials: DB
authors.lastname: Koo
authors.affiliation: National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Chung-buk 363-883, Republic of Korea.
authors.forename: Sang-Rae
authors.initials: SR
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Sang-Hyun
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA.
authors.forename: Ki-Eun
authors.initials: KE
authors.lastname: Park
authors.affiliation: Department of Digital Media, Graduate School, Ewha Womans University, Seoul 120-750, Republic of Korea.
authors.forename: Young Il
authors.initials: YI
authors.lastname: Park
authors.affiliation: College of Pharmacy, Catholic University of Daegu, Gyeong-buk 712-702, Republic of Korea. Electronic address: ekyune@cu.ac.kr.
authors.forename: Ekyune
authors.initials: E
authors.lastname: Kim
mesh_terms: D000818:Animals; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism*; D054885:Cumulus Cells / Q000378:metabolism*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D016678:Genome; D057809:HEK293 Cells; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000235:genetics / Q000378:metabolism*; D008297:Male; D010063:Ovum / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism*; D013094:Spermatozoa / Q000201:enzymology* / Q000378:metabolism; D013552:Swine / Q000502:physiology*
chemical_list: D015815:Cell Adhesion Molecules; D011994:Recombinant Proteins; D006821:Hyaluronoglucosaminidase; C066852:hyaluronidase PH-20
keywords: Cumulus–oocyte complex; Fertilization; Pig SPAM1; Recombinant protein; Sperm hyaluronidase; Sperm–egg interaction