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Assessment of microRNA expression in mouse epididymal epithelial cells and spermatozoa by next generation sequencing.

Anderson AL, Stanger SJ, Mihalas BP, Tyagi S, Holt JE, McLaughlin EA, Nixon B - Genom Data (2015)

Bottom Line: Through the use of Next Generation Sequencing technology we have demonstrated that both epididymal epithelial cells and spermatozoa harbour a complex repertoire of miRNAs that have substantially different expression profiles along the length of the tract.Ultimately such information promises to inform our understanding of the aetiology of male infertility.Herein we provide a detailed description of the methodology used to generate these important data.

View Article: PubMed Central - PubMed

Affiliation: Reproductive Science Group, School of Environmental and Life Sciences, Faculty of Science and IT, University of Newcastle, Callaghan, New South Wales, Australia.

ABSTRACT
The mammalian epididymis is a highly specialized region of the male reproductive tract that is lined with a continuous layer of epithelial cells that display a remarkable level of regionalized secretory and absorptive activity. The luminal environment created by this combined secretory and absorptive activity is directly responsible for promoting the functional maturation of spermatozoa and their maintenance in a quiescent and viable state prior to ejaculation. This study was designed to identify the complement of microRNAs (miRNAs) that are expressed within the mouse epididymal epithelial cells and the maturing populations of spermatozoa. Through the use of Next Generation Sequencing technology we have demonstrated that both epididymal epithelial cells and spermatozoa harbour a complex repertoire of miRNAs that have substantially different expression profiles along the length of the tract. These data, deposited in the Gene Expression Omnibus (GEO) with the accession numbers GSE70197 and GSE70198, afford valuable insight into the post-transcriptional control of gene expression within the epididymis and provide the first evidence for the dynamic transformation of the miRNA content of maturing sperm cells. Ultimately such information promises to inform our understanding of the aetiology of male infertility. Herein we provide a detailed description of the methodology used to generate these important data.

No MeSH data available.


Related in: MedlinePlus

Experimental design. Adult mouse epididymal tissue was dissected into regions corresponding to the caput, corpus and cauda. From each region, the epididymal epithelial cells were isolated, spermatozoa purified, or the tissue left intact. Total RNA was extracted from each cell (or tissue) sample. Subsequently, the RNA was processed for Illumina HiSeq Next-Generation sequencing, or stored and used for validation of the expression profiles of selected miRNAs.
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f0005: Experimental design. Adult mouse epididymal tissue was dissected into regions corresponding to the caput, corpus and cauda. From each region, the epididymal epithelial cells were isolated, spermatozoa purified, or the tissue left intact. Total RNA was extracted from each cell (or tissue) sample. Subsequently, the RNA was processed for Illumina HiSeq Next-Generation sequencing, or stored and used for validation of the expression profiles of selected miRNAs.

Mentions: At autopsy, epididymal tissue was dissected from adult Swiss male mice and immediately separated into three major anatomical regions corresponding to the caput, corpus and cauda epididymis. The tissue was either left intact or alternatively, the epithelial cells and spermatozoa were isolated from each region as described below. Total RNA was extracted from each sample and processed for Illumina HiSeq Next-Generation Sequencing for small non-coding miRNAs. The expression profile of selected miRNAs was then validated using TaqMan Real-Time PCR (Fig. 1).


Assessment of microRNA expression in mouse epididymal epithelial cells and spermatozoa by next generation sequencing.

Anderson AL, Stanger SJ, Mihalas BP, Tyagi S, Holt JE, McLaughlin EA, Nixon B - Genom Data (2015)

Experimental design. Adult mouse epididymal tissue was dissected into regions corresponding to the caput, corpus and cauda. From each region, the epididymal epithelial cells were isolated, spermatozoa purified, or the tissue left intact. Total RNA was extracted from each cell (or tissue) sample. Subsequently, the RNA was processed for Illumina HiSeq Next-Generation sequencing, or stored and used for validation of the expression profiles of selected miRNAs.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664737&req=5

f0005: Experimental design. Adult mouse epididymal tissue was dissected into regions corresponding to the caput, corpus and cauda. From each region, the epididymal epithelial cells were isolated, spermatozoa purified, or the tissue left intact. Total RNA was extracted from each cell (or tissue) sample. Subsequently, the RNA was processed for Illumina HiSeq Next-Generation sequencing, or stored and used for validation of the expression profiles of selected miRNAs.
Mentions: At autopsy, epididymal tissue was dissected from adult Swiss male mice and immediately separated into three major anatomical regions corresponding to the caput, corpus and cauda epididymis. The tissue was either left intact or alternatively, the epithelial cells and spermatozoa were isolated from each region as described below. Total RNA was extracted from each sample and processed for Illumina HiSeq Next-Generation Sequencing for small non-coding miRNAs. The expression profile of selected miRNAs was then validated using TaqMan Real-Time PCR (Fig. 1).

Bottom Line: Through the use of Next Generation Sequencing technology we have demonstrated that both epididymal epithelial cells and spermatozoa harbour a complex repertoire of miRNAs that have substantially different expression profiles along the length of the tract.Ultimately such information promises to inform our understanding of the aetiology of male infertility.Herein we provide a detailed description of the methodology used to generate these important data.

View Article: PubMed Central - PubMed

Affiliation: Reproductive Science Group, School of Environmental and Life Sciences, Faculty of Science and IT, University of Newcastle, Callaghan, New South Wales, Australia.

ABSTRACT
The mammalian epididymis is a highly specialized region of the male reproductive tract that is lined with a continuous layer of epithelial cells that display a remarkable level of regionalized secretory and absorptive activity. The luminal environment created by this combined secretory and absorptive activity is directly responsible for promoting the functional maturation of spermatozoa and their maintenance in a quiescent and viable state prior to ejaculation. This study was designed to identify the complement of microRNAs (miRNAs) that are expressed within the mouse epididymal epithelial cells and the maturing populations of spermatozoa. Through the use of Next Generation Sequencing technology we have demonstrated that both epididymal epithelial cells and spermatozoa harbour a complex repertoire of miRNAs that have substantially different expression profiles along the length of the tract. These data, deposited in the Gene Expression Omnibus (GEO) with the accession numbers GSE70197 and GSE70198, afford valuable insight into the post-transcriptional control of gene expression within the epididymis and provide the first evidence for the dynamic transformation of the miRNA content of maturing sperm cells. Ultimately such information promises to inform our understanding of the aetiology of male infertility. Herein we provide a detailed description of the methodology used to generate these important data.

No MeSH data available.


Related in: MedlinePlus