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Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
Molecular study of human sperm RNA: Ropporin and CABYR in asthenozoospermia
M Pelloni1, D Paoli2, M Majoli1
1Laboratory of Seminology-Sperm Bank "Loredana Gandini", Department of Experimental Medicine, University of Rome "La Sapienza", Viale del Policlinico 155, 00161, Rome, Italy.
Background:
Sperm motility is an essential aspect of human fertility. Sperm contain an abundance of transcripts, thought to be remnants of mRNA, which comprise a genetic fingerprint and can be considered a historic record of gene expression during spermatogenesis. The aberrant expression of numerous genes has been found to contribute to impaired sperm motility; these include ROPN1 (rhophilin associated tail protein 1), which encodes a component of the fibrous sheath of the mammalian sperm flagella, and CABYR (calcium-binding tyrosine-(Y)-phosphorylation-regulated protein), which plays an important role in calcium activation and modulation. The aim of this study was to investigate ROPN1 and CABYR gene co-expression in asthenozoospermic semen samples in comparison with normozoospermic samples.
Methods:
We studied 120 semen samples (60 normozoospermic and 60 asthenozoospermic) from Caucasian patients attending our centre for an andrological check-up. Total RNA was extracted from purified spermatozoa with RNeasy mini kit. ROPN1 and CABYR mRNA expression was analysed using RT-qPCR. Continuous variables were described as means ± standard deviations.
Results:
ROPN1 and CABYR mRNA were simultaneously downregulated in asthenozoospermic in comparison with normozoospermic samples. There was also a positive correlation between total progressive motility and ROPN1 and CABYR gene expression and between total motile sperm number and ROPN1 and CABYR gene expression.
Conclusions:
The results demonstrated downregulation of both ROPN1 and CABYR in asthenozoospermic samples and importantly, a positive correlation between the expression of the two genes, suggesting that ROPN1 and CABYR co-expression is a prerequisite for normal flagellar function and sperm motility.
Insights
Sperm motility is crucial for fertility. This study found that reduced expression of ROPN1 and CABYR genes is linked to asthenozoospermia, suggesting their co-expression is vital for sperm function.
Area of Science:
- Reproductive Biology
- Sperm Physiology
- Molecular Genetics
Background:
- Sperm motility is critical for male fertility.
- Sperm mRNA transcripts offer insights into spermatogenesis.
- Aberrant gene expression, including ROPN1 and CABYR, can impair sperm motility.
Purpose of the Study:
- To investigate the co-expression of ROPN1 and CABYR genes in asthenozoospermic semen.
- To compare gene expression patterns between normozoospermic and asthenozoospermic samples.
Main Methods:
- Analysis of 120 semen samples (60 normozoospermic, 60 asthenozoospermic).
- RNA extraction from purified spermatozoa.
- Quantitative real-time PCR (RT-qPCR) to analyze ROPN1 and CABYR mRNA expression.
Main Results:
- Simultaneous downregulation of ROPN1 and CABYR mRNA in asthenozoospermic samples compared to normozoospermic ones.
- Positive correlation between total progressive motility and ROPN1/CABYR expression.
- Positive correlation between total motile sperm number and ROPN1/CABYR expression.
Conclusions:
- ROPN1 and CABYR genes are downregulated in asthenozoospermia.
- Co-expression of ROPN1 and CABYR is positively correlated.
- Co-expression of ROPN1 and CABYR appears necessary for normal sperm motility and flagellar function.
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