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Characterizing the Impact of Dysregulated Micrornas on CRISP3 Isoforms in Male Infertility
Delnya Gholami1,2, Farzane Amirmahani3, Reza Salman Yazdi4
1Personalized Medicine and Genometabolomics Research Center, Hope Generation Foundation, Tehran, Iran.
Abstract:
microRNAs (miRNAs) have a serious and dynamic function in spermatogenesis. These molecules have been recognized as crucial parts of the control of gene activity, and their involvement in the regulation of target genes has been extensively studied. This research aimed to determine the expression of CRISP3 and miR-493-5p, miR-204-5p, and miR-182-5p in the seminal plasma fluid and spermatozoa and to examine the relationship between CRISP3 and the mentioned miRNAs in 57 infertile men with Asthenozoospermia (AZ) (n = 19), Teratoasthenozoospermia (TAZ) (n = 19), and Normozoospermia (NZ) (n = 19). The selection of these three miRNAs, miR-493-5p, miR-204-5p, and miR-182-5p, was conducted using computational prediction algorithms. These miRNAs were nominated as CRISP3-associated miRNAs that can target CRISP3. We performed the quantitative real-time polymerase chain reaction (qRT-PCR) method to determine the levels of the studied miRNA expression. In the following stage, the expression of two protein isoforms of CRISP3, targeted by these miRNAs, was quantified using western blotting. The results demonstrate significant differences in the levels of miR-182-5p, miR-204-5p, miR-493-5p, and CRISP3 isoforms among the patient groups. In TAZ individuals, miR-182-5p and miR-204-5p expression decreased, while miR-493-5p expression increased compared to the control samples. Additionally, significant differences were observed in the expression levels of unglycosylated and glycosylated CRISP3 isoforms between the AZ and NZ groups. Correlation analysis revealed associations between miRNA expression and the expression of CRISP3 isoforms in the patient groups. Additionally, there were correlations between the expression of CRISP3 isoforms and sperm motility and morphology. These results offer valuable insights into the underlying molecular processes associated with male infertility.
Insights
microRNAs (miRNAs) and CRISP3 protein levels differ in infertile men. Specific miRNA changes correlate with CRISP3 expression and sperm quality, offering insights into male infertility causes.
Area of Science:
- Reproductive Biology
- Molecular Endocrinology
- Spermatogenesis Research
Background:
- microRNAs (miRNAs) play a critical role in regulating gene expression during spermatogenesis.
- CRISP3 is a protein implicated in sperm function and male fertility.
Purpose of the Study:
- To investigate the expression of CRISP3 and specific miRNAs (miR-493-5p, miR-204-5p, miR-182-5p) in seminal fluid and spermatozoa of infertile men.
- To examine the relationship between CRISP3 and these miRNAs in different infertility groups.
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) for miRNA expression analysis.
- Western blotting to quantify CRISP3 protein isoforms.
- Analysis of seminal plasma and spermatozoa from infertile men (Asthenozoospermia, Teratoasthenozoospermia) and normozoospermic controls.
Main Results:
- Significant differences in miR-182-5p, miR-204-5p, miR-493-5p, and CRISP3 isoform levels were observed across patient groups.
- Teratoasthenozoospermia patients showed decreased miR-182-5p and miR-204-5p, and increased miR-493-5p.
- Correlations were found between miRNA expression, CRISP3 isoforms, and sperm parameters (motility, morphology).
Conclusions:
- Aberrant expression of specific miRNAs and CRISP3 isoforms is associated with male infertility.
- These molecular changes may contribute to impaired sperm function and morphology.
- Findings provide potential biomarkers and therapeutic targets for male infertility.
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