Optimization of sperm RNA processing for developmental research

Abstract Recent studies have demonstrated the significance of sperm RNA function as a transporter of important information directing the course of life. To determine the message contained in sperm RNA, it is necessary to optimize transcriptomic research tools. The current study was performed to opti...

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Autores principales:	Won-Ki Pang, Saehan Kang, Do-Yeal Ryu, Md Saidur Rahman, Yoo-Jin Park, Myung-Geol Pang
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2cbdee5e3adf4d5bad7cc78597fdc6aa
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spelling	oai:doaj.org-article:2cbdee5e3adf4d5bad7cc78597fdc6aa2021-12-02T15:33:10ZOptimization of sperm RNA processing for developmental research10.1038/s41598-020-68486-12045-2322https://doaj.org/article/2cbdee5e3adf4d5bad7cc78597fdc6aa2020-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-68486-1https://doaj.org/toc/2045-2322Abstract Recent studies have demonstrated the significance of sperm RNA function as a transporter of important information directing the course of life. To determine the message contained in sperm RNA, it is necessary to optimize transcriptomic research tools. The current study was performed to optimize the processing of sperm RNA from sample storage to quantitative real-time PCR and assess the corresponding method with to evaluate male fertility and its representative markers, equatorin (EQTN) and peroxiredoxin (PRDX). Following successive steps of the Minimum Information for Publication of Quantitative Real-Time PCR Experiments guidelines, several options were compared using boar spermatozoa. To evaluate the optimized procedures, the relationship between mRNA expression of EQTN and PRDX in spermatozoa and the fertility (litter size) of 20 individual boars was assessed. Unexpectedly, DNase treatment during RNA isolation had the deleterious effect by decreasing the RNA concentration by 56% and eliminating the correlation between EQTN and PRDX4 mRNA expression and male fertility. Moreover, when sperm RNA was processed using the corresponding method, the results showed the highest exon sequence expression, male fertility prediction power, and consistency. This optimized protocol for predicting male fertility can be used to study the transport of messages directing the life course from spermatozoon to offspring.Won-Ki PangSaehan KangDo-Yeal RyuMd Saidur RahmanYoo-Jin ParkMyung-Geol PangNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Won-Ki Pang Saehan Kang Do-Yeal Ryu Md Saidur Rahman Yoo-Jin Park Myung-Geol Pang Optimization of sperm RNA processing for developmental research
description	Abstract Recent studies have demonstrated the significance of sperm RNA function as a transporter of important information directing the course of life. To determine the message contained in sperm RNA, it is necessary to optimize transcriptomic research tools. The current study was performed to optimize the processing of sperm RNA from sample storage to quantitative real-time PCR and assess the corresponding method with to evaluate male fertility and its representative markers, equatorin (EQTN) and peroxiredoxin (PRDX). Following successive steps of the Minimum Information for Publication of Quantitative Real-Time PCR Experiments guidelines, several options were compared using boar spermatozoa. To evaluate the optimized procedures, the relationship between mRNA expression of EQTN and PRDX in spermatozoa and the fertility (litter size) of 20 individual boars was assessed. Unexpectedly, DNase treatment during RNA isolation had the deleterious effect by decreasing the RNA concentration by 56% and eliminating the correlation between EQTN and PRDX4 mRNA expression and male fertility. Moreover, when sperm RNA was processed using the corresponding method, the results showed the highest exon sequence expression, male fertility prediction power, and consistency. This optimized protocol for predicting male fertility can be used to study the transport of messages directing the life course from spermatozoon to offspring.
format	article
author	Won-Ki Pang Saehan Kang Do-Yeal Ryu Md Saidur Rahman Yoo-Jin Park Myung-Geol Pang
author_facet	Won-Ki Pang Saehan Kang Do-Yeal Ryu Md Saidur Rahman Yoo-Jin Park Myung-Geol Pang
author_sort	Won-Ki Pang
title	Optimization of sperm RNA processing for developmental research
title_short	Optimization of sperm RNA processing for developmental research
title_full	Optimization of sperm RNA processing for developmental research
title_fullStr	Optimization of sperm RNA processing for developmental research
title_full_unstemmed	Optimization of sperm RNA processing for developmental research
title_sort	optimization of sperm rna processing for developmental research
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/2cbdee5e3adf4d5bad7cc78597fdc6aa
work_keys_str_mv	AT wonkipang optimizationofspermrnaprocessingfordevelopmentalresearch AT saehankang optimizationofspermrnaprocessingfordevelopmentalresearch AT doyealryu optimizationofspermrnaprocessingfordevelopmentalresearch AT mdsaidurrahman optimizationofspermrnaprocessingfordevelopmentalresearch AT yoojinpark optimizationofspermrnaprocessingfordevelopmentalresearch AT myunggeolpang optimizationofspermrnaprocessingfordevelopmentalresearch
_version_	1718387072943783936

Optimization of sperm RNA processing for developmental research

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