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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: Effects of hypothermic storage of striped bass (Morone saxatilis) sperm on intracellular calcium, reactive oxygen species formation, mitochondrial function, motility, and viability

Authors
item Guthrie, Howard
item Woods, L -
item Welch, Glenn

Submitted to: Poultry Science Association Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: March 8, 2010
Publication Date: July 11, 2010
Citation: Guthrie, H.D., Woods, L.C., Welch, G.R. 2010. Effects of hypothermic storage of striped bass (Morone saxatilis) sperm on intracellular calcium, reactive oxygen species formation, mitochondrial function, motility, and viability. Poultry Science Association Meeting Abstract.

Technical Abstract: Experiments were conducted to determine the effect of hypothermic 24 h storage of striped bass sperm cells (Morone saxatilis) on viability, intracellular Ca2+ [Ca2+]i, mitochondrial membrane potential (''m), and reactive oxygen species (ROS) formation as determined by flow cytometry; motion activation and ATP concentration as determined by Luciferin-Luciferase bioluminescence assay. Semen was stored for 1 or 24 h at 4 oC in an O2 atmosphere undiluted (raw) or diluted 1:4 (one volume semen with 3 volumes) with T350 (20 mM TRIS base-NaCl, 350 mOsm/mL, pH = 8) or seminal plasma (SP) in the presence of various treatments. Viability (% cells excluding propidium iodide) approached 100% after 1 h storage raw or in T350 and SP. After 1 h of storage Fluo-3 fluorescence (marker for [Ca2+]) was detected in only 3% of sperm cells in raw and T350 or SP extended semen. However after 24 h, the incidence Fluo-3 fluorescence in viable sperm cells was > 50% in raw and T350 or SP extended semen and fluorescence intensity was greatly increased; the presence of 1 mM EGTA maintained viability and prevented increased Fluo-3 fluorescence. Activation of sperm motility was 82% after 1 h in T350 and decreased to 30% after 24 h. However, activation failed in the presence of EGTA at 1 or 24 h. During storage ''m and ATP did not change significantly between 1 and 24 h; however in the presence of EGTA ATP, but not ''m decreased between 1 and 24 h. While ROS formation was induced by menadione treatment, there was no evidence of storage-induced ROS formation (oxidation of hydroethidine dye to ethidium). The reduced expression of motility following activation after 24 h of storage or its absence in the presence of EGTA cannot be explained wholly in terms of ROS formation, or differences in Ca2+ flux, or ATP content.

Last Modified: 10/23/2014
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