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Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices

View Article: PubMed Central - PubMed

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.

No MeSH data available.


Relationship between motility and difference between AMV in MTT assay (P < 0.05).Data points (N) in this graph are 20.
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f4: Relationship between motility and difference between AMV in MTT assay (P < 0.05).Data points (N) in this graph are 20.

Mentions: Figure 4 shows the correlation between sperm motility and AMV in our MTT assay. Data points in Fig. 4 are chosen from averages of different treatment times detailed in Figs 2 and 3. The R2 value from this correlation analysis was 0.805 and a significant correlation was found (P < 0.05), suggesting a relationship between these two indicators and a coupling of enzymatic reaction detected by MTT assay with sperm motility. According to literature regarding the relationship between motility and farrowing rate, subfertility cut-off value is 70% motility, and subfertility AMV threshold must be approximately 90% of the original sample based on the regression line parameter calculation42. When comparing motility analyses using iSperm and MTT assay results, enzymatic activity and MTT reduction would be higher in semen with greater motility, and semen with higher motility produce more intense (more deeply purple) colorimetric signals.


Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices
Relationship between motility and difference between AMV in MTT assay (P < 0.05).Data points (N) in this graph are 20.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5384208&req=5

f4: Relationship between motility and difference between AMV in MTT assay (P < 0.05).Data points (N) in this graph are 20.
Mentions: Figure 4 shows the correlation between sperm motility and AMV in our MTT assay. Data points in Fig. 4 are chosen from averages of different treatment times detailed in Figs 2 and 3. The R2 value from this correlation analysis was 0.805 and a significant correlation was found (P < 0.05), suggesting a relationship between these two indicators and a coupling of enzymatic reaction detected by MTT assay with sperm motility. According to literature regarding the relationship between motility and farrowing rate, subfertility cut-off value is 70% motility, and subfertility AMV threshold must be approximately 90% of the original sample based on the regression line parameter calculation42. When comparing motility analyses using iSperm and MTT assay results, enzymatic activity and MTT reduction would be higher in semen with greater motility, and semen with higher motility produce more intense (more deeply purple) colorimetric signals.

View Article: PubMed Central - PubMed

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&thinsp;&lt;&thinsp;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.

No MeSH data available.