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High quality RNA in semen and sperm: isolation, analysis and potential application in clinical testing.

ABSTRACT: PURPOSE:Male infertility is a complex health condition. To our knowledge there are no molecular biomarkers of male infertility. Sperm RNA is a potential biomarker for detecting sperm abnormalities and viability at infertility clinics. However, RNA use is hindered by its inconsistent quantity, quality, multiple cell types in semen and condensed sperm structure. MATERIALS AND METHODS:We tested the usefulness of high quality RNA isolated from mature sperm and whole semen by our protocol, which reduces RNA degradation by maintaining semen and protocol components at 37 C and decreasing processing time. We isolated RNA from 83 whole semen samples, 18 samples of motile sperm prepared by the swim-up protocol and 18 of sperm prepared by the standard Percoll gradient method. RESULTS:Electrophoretic and spectral analysis of RNA revealed high quality 18S and 28S rRNAs in 71 of 83 whole semen samples (86%) and 15 of 18 mature sperm swim-up samples (83%). However, high quality RNA was isolated from only 7 of 18 Percoll gradient sperm samples (39%). Interestingly quantitative reverse transcriptase-polymerase chain reaction analysis of 4 somatic and 10 germ cell markers showed that whole semen and swim-up samples had similar RNA profiles. RNA sequencing revealed that most encoded proteins were involved in mature sperm function, regulation of DNA replication, transcription, translation, cell cycle and embryo development. CONCLUSIONS:We believe that semen and sperm specific RNAs are highly informative biomarkers for germ cell stages and somatic cell contribution. Therefore, these RNAs could be valuable diagnostic indicators of sperm survival, fertilization and early embryogenesis, and could serve as a predictor of the in vitro fertilization prognosis.

SUBMITTER: Georgiadis AP

PROVIDER: S-EPMC4382362 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

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High quality RNA in semen and sperm: isolation, analysis and potential application in clinical testing.

Georgiadis Andrew P AP Kishore Archana A Zorrilla Michelle M Jaffe Thomas M TM Sanfilippo Joseph S JS Volk Etta E Rajkovic Aleksandar A Yatsenko Alexander N AN

The Journal of urology 20140801 1

<h4>Purpose</h4>Male infertility is a complex health condition. To our knowledge there are no molecular biomarkers of male infertility. Sperm RNA is a potential biomarker for detecting sperm abnormalities and viability at infertility clinics. However, RNA use is hindered by its inconsistent quantity, quality, multiple cell types in semen and condensed sperm structure.<h4>Materials and methods</h4>We tested the usefulness of high quality RNA isolated from mature sperm and whole semen by our proto ...[more]

PMID: 25088949

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Project description:BackgroundDespite the generally accepted World Health Organization guidelines on semen analysis, an individual's results can display significant variation when performed across time or in different laboratories. Semen parameters are in fact highly variable measures that can differ significantly between various analyses. Numerous researchers have discovered a wide range of semen parameters within each individual male, but only a few studies included the analysis of semen parameters variability in patients with infertility. The aim of this study was to evaluate the inter- and intra-individual variability of semen parameters in men of reproductive age with normozoospermia and those with oligozoospermia.MethodsFive hundred and thirteen who provided ≥ 2 semen samples (798 samples in total) using an at-home mail-in kit over a period of about 2 years were enrolled in the study. Semen samples collection using Give Legacy at-home mail-in semen collection kit; semen analysis at a CLIA-certified laboratory.ResultsThe degree of intra-subject variation across all semen parameters was lower in men with normozoospermia compared to men with oligozoospermia. Men with normozoospermia furthermore demonstrated a level of intra-subject variation that was lower than inter-subject variation across all measured parameters. No association was observed between intra-subject coefficients of variation in any of the semen parameters, including sperm concentration, sperm count, motile sperm count, total motility, progressive motility, the percentage of sperm with normal morphology, and the age, duration of abstinence, and BMI of the men.ConclusionThe results of this observational study confirm the significant variability in semen parameters in men with normozoospermia and oligozoospermia, as measured from at-home semen collection kit samples. This further underscore the importance of securing multiple samples for analysis to provide a robust assessment of male fertility.

Project description:Study questionWhat is the relationship between abnormal BMI and semen quality?Summary answerUnderweight was significantly associated with lower sperm concentration, total sperm number and total motile sperm count, while overweight was significantly associated with lower semen volume, total sperm number and total motile sperm count.What is known alreadyAbnormal BMI has been associated with lower semen quality, but the results remain somewhat controversial. In addition, most previous studies have focused on the influence of obesity or overweight on semen quality, and evidence on the association between underweight and semen quality is rare.Study design, size, durationThis research was an observational study investigating 3966 sperm donors from a large sperm bank in Wuhan city, China. These donors passed the screening for sperm donation and underwent 29 949 semen examinations between 1 January 2013 and 9 April 2018.Participants/materials, setting, methodsBMI was categorized into four groups: underweight (<18.5 kg/m2), normal weight (18.5-24.9 kg/m2), overweight (25-29.9 kg/m2) and obese (≥30 kg/m2). Semen volume, sperm concentration, total sperm number, total motility, progressive motility and total motile sperm count were determined by trained clinical technicians. Linear mixed models were used to conduct dose-response analyses between BMI and semen quality parameters.Main results and the role of chanceUnderweight was significantly associated with a 3.0% (95% CI: 0.1%, 5.8%), 6.7% (1.9%, 11.3%) and 7.4% (2.2%, 12.4%) reduction in sperm concentration, total sperm number and total motile sperm count, respectively. Overweight was significantly associated with a 4.2% (1.6%, 6.8%), 3.9% (0.9%, 6.9%) and 3.6% (0.2%, 6.9%) reduction in semen volume, total sperm number and total motile sperm count, respectively. Non-linear models including continuous BMI as a natural cubic spline function yielded similar results.Limitations, reasons for cautionOur study subjects were sperm donors who are typically young and healthy, and therefore not representative of the general male population. Caution should be paid in generalizing our results to other populations. Furthermore, we did not measure the donors' weight repeatedly along with each semen donation; instead, we only measured it once during the screening, which may cause bias due to the variations of weight across time.Wider implications of the findingsOur study provides evidence that underweight and overweight are associated with lower semen quality, and highlights the importance of maintaining a normal weight for men.Study funding/competing interest(s)This work was supported by the Health and Family Planning Commission of Hubei Province (Grant number WJ2015MA027), the Hubei Provincial Committee of the Communist Youth League of China, and Center for Global and Regional Environmental Research at the University of Iowa. The authors declare that there are no conflicts of interest.Trial registration numberN/A.

Dataset Information

High quality RNA in semen and sperm: isolation, analysis and potential application in clinical testing.

Publications

High quality RNA in semen and sperm: isolation, analysis and potential application in clinical testing.

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