Sperm Transcripts in Irpl

ObjectiveRecurrent pregnancy loss affects approximately 3-5% percent of couples attempting to have a child. Despite extensive evaluation majority of couple experiencing recurrent pregnancy loss (RPL), have no identifiable factors (iRPL). The aim of this study was to analyze the sperm transcripts level in cases of iRPL.DesignThe present study was set up to examine the level of relationship between sperm transcripts (PRM2, WNT5A, PSG1, ZNF582, HSP90) and iRPL. Fifty couples who experienced RPL and 30 fertile controls were enrolled in the study. Semen samples were collected by masturbation from after 4 days of abstinence. Semen analysis was performed as per WHO (1999) guidelines.Materials and MethodsTotal RNA was isolated from 1×107 spermatozoa using RNeasy mini kit. RNA was reverse-transcribed into complementary DNA using random hexamer priming and Revert AidTM M-MuLv reverse transcriptase. Quantitative analysis of each gene was performed by CFX96 real Time System (Bio-Rad, USA) using SsoFastTM EvaGreen Supermix (Bio-Rad Laboratories, CA). The relative quantification of target genes normalized to β-actin was calculated by 2ΔCt method. The relative gene expression levels between control and patient subjects were compared by student- t test. If the P value was ≤0.05, the difference was considered to be statistically significant.ResultsiRPL and control groups did not show any significant differences in the sperm parameters. There was a significant difference between fertile and iRPL group for PSG1, PRM2 ZNF582 and HSP90. However, no significant difference was found for WNT5A between fertile and iRPL patients.ConclusionThese findings, suggests that sperm PSG1, PRM2 ZNF582 and HSP90 mRNA (but not WNT5A) in the egg after fertilization, lead to the conclusion that these transcripts may have a functional role during early embryo development and/or implantation. However, its functional implication may require further validation. ObjectiveRecurrent pregnancy loss affects approximately 3-5% percent of couples attempting to have a child. Despite extensive evaluation majority of couple experiencing recurrent pregnancy loss (RPL), have no identifiable factors (iRPL). The aim of this study was to analyze the sperm transcripts level in cases of iRPL. Recurrent pregnancy loss affects approximately 3-5% percent of couples attempting to have a child. Despite extensive evaluation majority of couple experiencing recurrent pregnancy loss (RPL), have no identifiable factors (iRPL). The aim of this study was to analyze the sperm transcripts level in cases of iRPL. DesignThe present study was set up to examine the level of relationship between sperm transcripts (PRM2, WNT5A, PSG1, ZNF582, HSP90) and iRPL. Fifty couples who experienced RPL and 30 fertile controls were enrolled in the study. Semen samples were collected by masturbation from after 4 days of abstinence. Semen analysis was performed as per WHO (1999) guidelines. The present study was set up to examine the level of relationship between sperm transcripts (PRM2, WNT5A, PSG1, ZNF582, HSP90) and iRPL. Fifty couples who experienced RPL and 30 fertile controls were enrolled in the study. Semen samples were collected by masturbation from after 4 days of abstinence. Semen analysis was performed as per WHO (1999) guidelines. Materials and MethodsTotal RNA was isolated from 1×107 spermatozoa using RNeasy mini kit. RNA was reverse-transcribed into complementary DNA using random hexamer priming and Revert AidTM M-MuLv reverse transcriptase. Quantitative analysis of each gene was performed by CFX96 real Time System (Bio-Rad, USA) using SsoFastTM EvaGreen Supermix (Bio-Rad Laboratories, CA). The relative quantification of target genes normalized to β-actin was calculated by 2ΔCt method. The relative gene expression levels between control and patient subjects were compared by student- t test. If the P value was ≤0.05, the difference was considered to be statistically significant. Total RNA was isolated from 1×107 spermatozoa using RNeasy mini kit. RNA was reverse-transcribed into complementary DNA using random hexamer priming and Revert AidTM M-MuLv reverse transcriptase. Quantitative analysis of each gene was performed by CFX96 real Time System (Bio-Rad, USA) using SsoFastTM EvaGreen Supermix (Bio-Rad Laboratories, CA). The relative quantification of target genes normalized to β-actin was calculated by 2ΔCt method. The relative gene expression levels between control and patient subjects were compared by student- t test. If the P value was ≤0.05, the difference was considered to be statistically significant. ResultsiRPL and control groups did not show any significant differences in the sperm parameters. There was a significant difference between fertile and iRPL group for PSG1, PRM2 ZNF582 and HSP90. However, no significant difference was found for WNT5A between fertile and iRPL patients. iRPL and control groups did not show any significant differences in the sperm parameters. There was a significant difference between fertile and iRPL group for PSG1, PRM2 ZNF582 and HSP90. However, no significant difference was found for WNT5A between fertile and iRPL patients. ConclusionThese findings, suggests that sperm PSG1, PRM2 ZNF582 and HSP90 mRNA (but not WNT5A) in the egg after fertilization, lead to the conclusion that these transcripts may have a functional role during early embryo development and/or implantation. However, its functional implication may require further validation. These findings, suggests that sperm PSG1, PRM2 ZNF582 and HSP90 mRNA (but not WNT5A) in the egg after fertilization, lead to the conclusion that these transcripts may have a functional role during early embryo development and/or implantation. However, its functional implication may require further validation.