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United States Department of Agriculture

Agricultural Research Service

Research Project: DEVELOPMENT OF NEW TECHNOLOGIES AND METHODS TO ENHANCE THE UTILIZATION AND LONG-TERM STORAGE OF POULTRY, SWINE AND FISH GERMPLASM Title: Isolation of RNA from striped bass (Morone saxatilis) spermatozoa: implications for teleost male fertility and beyond?

Authors
item Woods, Iii, L -
item Guthrie, Howard
item Welch, Glenn
item Fuller, A -

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: August 31, 2012
Publication Date: N/A

Technical Abstract: Vertebrate sperm has been shown to provide more than paternal genomic material to the oocyte. For example, specific transcripts have been identified in bull sperm associated with fertility and with motility in boar sperm. Very little is currently known about functional, residual RNA in spermatozoa although residual RNA provides insight into the gene expression that took place during spermatogenesis. Recently, studies using human, mouse, boar, and bull sperm have confirmed functional mRNA in sperm and correlated identified transcripts to critical biological functions. Residual mRNA expression patterns have also been correlated to sperm morphology motility, and metabolism as well as to their potential role in embryonic development. While these patterns may be used to monitor or explain past events during spermatogenesis, it is the contribution of the spermatozoa to the zygote with functional, paternal mRNA and of the resulting individual’s phenotypic expressions that we are most interested in. To our knowledge, no one has successfully purified RNA from fish spermatozoa. The objective of our research was to evaluate and optimize the methodology to isolate RNA from freshly collected as well as frozen, shipped striped bass spermatozoa. We successfully purified and quantified total RNA, using an Agilent 2100 bioanalyzer, from sperm (Fig.1) and testis (not shown) of striped bass. Striped bass sperm RNA was most efficiently extracted using a trizol based procedure; resulting in repeatable quantities of 3-5 ng/uL of total RNA per 12 x 109 sperm. These results demonstrate that RNA can be isolated from small teleost spermatozoa and provide the foundation for functional genomics and epigenetic research with male teleosts of biological and economic importance.

Last Modified: 8/29/2014
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