title: Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices.
abstract: Mammalian sperm motility has traditionally been analyzed to determine fertility using computer-assisted semen analysis (CASA) systems. To develop low-cost and robust male fertility diagnostics, we created a paper-based MTT assay and used it to estimate motile sperm concentration. When porcine sperm motility was inhibited using sperm enzyme inhibitors for sperm enzymes related to mitochondrial activity and glycolysis, we simultaneously recorded sperm motility and enzymatic reactivity using a portable motility analysis system (iSperm) and a paper-based MTT assay, respectively. When using our paper-based MTT-assay, we calculated the area mean value signal intensity (AMV) to evaluate enzymatic reactivity. Both sperm motility and AMV decreased following treatment with iodoacetamide (IODO) and 3-bromopyruvic acid (3BP), both of which are inhibitors of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a correlation between recorded motility using iSperm and AMV from our paper-based assay (P < 0.05), suggesting that a sperm-related enzymatic reaction is involved in sperm motility. Under this protocol, MTT reduction was coupled with catalysis of GAPDH and was promoted by electron transfer from NADH. Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.
authors.affiliation: Department of Biomedical Engineering, Faculty of Engineering, Okayama University of Science, 1-1 Ridai-Cho Kita-Ku, Okayama, 700-0005, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Matsuura
authors.affiliation: Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan.
authors.forename: Han-Wei
authors.initials: HW
authors.lastname: Huang
authors.affiliation: Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan.
authors.forename: Ming-Cheng
authors.initials: MC
authors.lastname: Chen
authors.affiliation: Department of Urology, Chang Gung Memorial Hospital and Chang Gung University, Taoyuan 33302, Taiwan.
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan.
authors.forename: Chao-Min
authors.initials: CM
authors.lastname: Cheng
mesh_terms: D000255:Adenosine Triphosphate; D000818:Animals; D015415:Biomarkers; D004305:Dose-Response Relationship, Drug; D005986:Glyceraldehyde 3-Phosphate / Q000378:metabolism; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000378:metabolism; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D055101:Semen Analysis / Q000379:methods; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D013552:Swine
chemical_list: D015415:Biomarkers; D005986:Glyceraldehyde 3-Phosphate; D000255:Adenosine Triphosphate; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)