Limits...
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.


Inhibition of sperm motility recorded using iSperm and AMV of paper-based MTT assay using different inhibitors.We used (A) iSperm to test sperm motility and used (B) our paper-based device to calculate AMV. Error bars are standard deviation (SD) of three samples in each point (N = 3). Asterisks show the difference with the value of control group without inhibitor (P < 0.05). Inhibitor concentration of Iodo, 3NPA, 3BP and DOG were 2, 8, 0.2, and 8 mM, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC5384208&req=5

f2: Inhibition of sperm motility recorded using iSperm and AMV of paper-based MTT assay using different inhibitors.We used (A) iSperm to test sperm motility and used (B) our paper-based device to calculate AMV. Error bars are standard deviation (SD) of three samples in each point (N = 3). Asterisks show the difference with the value of control group without inhibitor (P < 0.05). Inhibitor concentration of Iodo, 3NPA, 3BP and DOG were 2, 8, 0.2, and 8 mM, respectively.

Mentions: Figure 2A shows inhibition of sperm motility using inhibitors of mitochondrial function and glycolysis. After mixing sperm with inhibitor (0 min), sperm motility of the original and the inhibitor-treated semen were approximately 90% and were quite similar, because inhibition of sperm motility had not begun. After 20, 40, and 60 min of inhibitor treatment, IODO and 3BP-treated sperm motility significantly decreased (P < 0.05). Sperm motility values from 3-NPA- and DOG-treated samples were similar to those of the control group. Although we increased concentration of the inhibitors to 8 mM, the motility was approximately 90% and almost the same as control sample motility. Treatment of porcine sperm with IODO and 3BP significantly reduced sperm motility after 20 min of incubation (P < 0.05).


Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices
Inhibition of sperm motility recorded using iSperm and AMV of paper-based MTT assay using different inhibitors.We used (A) iSperm to test sperm motility and used (B) our paper-based device to calculate AMV. Error bars are standard deviation (SD) of three samples in each point (N = 3). Asterisks show the difference with the value of control group without inhibitor (P < 0.05). Inhibitor concentration of Iodo, 3NPA, 3BP and DOG were 2, 8, 0.2, and 8 mM, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Inhibition of sperm motility recorded using iSperm and AMV of paper-based MTT assay using different inhibitors.We used (A) iSperm to test sperm motility and used (B) our paper-based device to calculate AMV. Error bars are standard deviation (SD) of three samples in each point (N = 3). Asterisks show the difference with the value of control group without inhibitor (P < 0.05). Inhibitor concentration of Iodo, 3NPA, 3BP and DOG were 2, 8, 0.2, and 8 mM, respectively.
Mentions: Figure 2A shows inhibition of sperm motility using inhibitors of mitochondrial function and glycolysis. After mixing sperm with inhibitor (0 min), sperm motility of the original and the inhibitor-treated semen were approximately 90% and were quite similar, because inhibition of sperm motility had not begun. After 20, 40, and 60 min of inhibitor treatment, IODO and 3BP-treated sperm motility significantly decreased (P < 0.05). Sperm motility values from 3-NPA- and DOG-treated samples were similar to those of the control group. Although we increased concentration of the inhibitors to 8 mM, the motility was approximately 90% and almost the same as control sample motility. Treatment of porcine sperm with IODO and 3BP significantly reduced sperm motility after 20 min of incubation (P < 0.05).

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.