Quantification of sperm specific mRNA transcripts (PRM1, PRM2, and TNP2) in teratozoospermia and normozoospermia: New correlations between mRNA content and morphology of sperm

Abstract

Sperm mRNAs could be used as a predictor of fertilization capacity since the transcriptional profile of a gamete is critical for the production of viable human sperm. The aim of this study was to determine if PRM1, PRM2, and TNP2 transcripts in spermatozoa from normozoospermic and teratozoospermic men correlate with sperm morphology and/or assisted-reproduction outcomes. Human ejaculates were collected from 138 men referred to an infertility clinic, and were separated in two groups, teratozoospermic (n =72) and normozoospermic (n =66), based on World Health Organization criteria (2010). Chromomycin A3 and analine blue staining were used to evaluate protamination and chromatin integrity, respectively. Quantitative reverse-transcriptase PCR was performed for PRM1, PRM2, and TNP2. This analysis revealed significantly higher PRM1 and PRM2 mRNA copy numbers in normozoospermic versus teratozoospermic samples (P < 0.001). In contrast, TNP2 transcript abundance was significantly higher in teratozoospermic versus normozoospermic samples (P < 0.001) and positively correlated with sperm-head defects (P < 0.05). Sperm-tail defects negatively correlated (P < 0.05) with both PRM1 and PRM2 transcripts in normozoospermic samples. No significant differences were observed between the two groups when comparing transcript levels to the outcome of intracytoplasmic sperm injection cycles (P > 0.05), and a normal PRM1/PRM2 mRNA ratio (∼1) was observed in more than 70% of successful cycles. Thus, the quantity of PRM1, PRM2, and TNP2 transcripts and the PRM1/PRM2 mRNA ratio affect spermiogenesis, sperm morphology, and the function of mature human sperm. These mRNAs could therefore be used as biomarkers for the diagnosis of male infertility.

Testis Transcriptome Modulation in Klinefelter Patients with Hypospermatogenesis

“…FHL5 is a component of the CREM pathway regulating germ cells meiosis16, while PRM1 and PRM2 encode the proteins involved in the replacement of histones with protamines during spermiogenesis. It has been recently reported that the altered amount of PRM1 and PRM2 transcripts influences the whole spermiogenesis process and the sperm morphology, thereby affecting the functions of mature sperm1718. The expression levels of PRM1 and PRM2 genes are reported to be indicators of male infertility18.…”

“…It has been recently reported that the altered amount of PRM1 and PRM2 transcripts influences the whole spermiogenesis process and the sperm morphology, thereby affecting the functions of mature sperm1718. The expression levels of PRM1 and PRM2 genes are reported to be indicators of male infertility18.…”

The impact of sperm protamine deficiency and sperm DNA damage on human male fertility: a systematic review and meta-analysis

“…A total of nine studies with 633 patients and 463 controls in the first part, which fulfilled all of the inclusion criteria, were considered for the analysis. Five of the studies were prospective (de Mateo et al, 2009;Garcia-Peiro et al, 2011;Rogenhofer et al, 2013;Ni et al, 2014;Savadi-Shiraz et al, 2015) and four were retrospective (Mengual et al, 2003;Aoki et al, 2005aAoki et al, , 2006aSteger et al, 2008). Four studies used real-time RT-qPCR (mRNA) (Steger et al, 2008;Rogenhofer et al, 2013;Ni et al, 2014;Savadi-Shiraz et al, 2015) and five studies employed PAGE (protein) (Mengual et al, 2003;Aoki et al, 2005aAoki et al, , 2006ade Mateo et al, 2009;Garcia-Peiro et al, 2011) to calculate the protamine ratio.…”

“…Four studies used real-time RT-qPCR (mRNA) (Steger et al, 2008;Rogenhofer et al, 2013;Ni et al, 2014;Savadi-Shiraz et al, 2015) and five studies employed PAGE (protein) (Mengual et al, 2003;Aoki et al, 2005aAoki et al, , 2006ade Mateo et al, 2009;Garcia-Peiro et al, 2011) to calculate the protamine ratio. When comparing the differences in the protamine ratio between experimental and control groups, two studies compared normozoospermic healthy donors with subfertile patients (Steger et al, 2008;Savadi-Shiraz et al, 2015), five studies compared proven fertile donors with subfertile patients (Mengual et al, 2003;Aoki et al, 2005aAoki et al, , 2006ade Mateo et al, 2009;Garcia-Peiro et al, 2011), one study compared groups of men contributing to successful fertilization with those involved in fertilization failure after IVF and ICSI treatment (Rogenhofer et al, 2013), and one study compared spontaneous pregnant couples with non-pregnant couples (Ni et al, 2014). In the second part, the total number of studies included in the review was 12 comprising of 845 participants.…”

Lycium barbarum polysaccharide protects against ethanol‐induced spermiotoxicity and testicular degeneration in Immp2l +/− mice

“…Lack of TNP2 leads to mouse teratozoospermia (Adham et al, 2001). Research has indicated that the expression of TNP2 was obviously increased in teratozoospermic compared with normozoospermic groups ( p < .001) and it was associated with sperm‐head defects ( p < .05; Savadi‐Shiraz et al, 2015). TNP2 mRNAs can be used as biomarkers to diagnose male infertility.…”