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Evaluation of the effectiveness of semen storage and sperm purification methods for spermatozoa transcript profiling.

Authors
Mao-S; Goodrich-RJ; Hauser-R; Schrader-SM; Chen-Z; Krawetz-SA
Source
Syst Biol Reprod Med 2013 Oct; 59(5):287-295
NIOSHTIC No.
20043001
Abstract
Different semen storage and sperm purification methods may affect the integrity of isolated spermatozoal RNA. RNA-Seq was applied to determine whether semen storage methods (pelleted vs. liquefied) and somatic cell lysis buffer (SCLB) vs. PureSperm (PS) purification methods affect the quantity and quality of sperm RNA. The results indicate that the method of semen storage does not markedly impact RNA profiling whereas the choice of purification can yield significant differences. RNA-Seq showed that the majority of mitochondrial and mid-piece associated transcripts were lost after SCLB purification, which indicated that the mid-piece of spermatozoa may have been compromised. In addition, the number of stable transcript pairs from SCLB-samples was less than that from the PS samples. This study supports the view that PS purification better maintains the integrity of spermatozoal RNAs.
Keywords
Spermatozoa; Storage-facilities; Cell-function; Cell-biology; Cellular-function; Sampling; Analytical-processes; Author Keywords: mitochondrial RNA; preferentially isolated transcripts; stable transcript pairs
Contact
Stephen A. Krawetz, Department of Obstetrics and Gynecology, Wayne State University, 271 C.S. Mott Center, 275 E. Hancock Ave., Detroit, MI, 48201
CODEN
SBRMDP
Publication Date
20131001
Document Type
Journal Article
Email Address
steve@compbio.med.wayne.edu
Fiscal Year
2014
NTIS Accession No.
NTIS Price
Identifying No.
M082013
Issue of Publication
5
ISSN
1939-6368
NIOSH Division
DART
Priority Area
Manufacturing
Source Name
Systems Biology in Reproductive Medicine
State
MI; MA; MD
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