. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. M. G. BENTLEY ET AL ]. o x |1 I 1 1 0 100 1000 10000 100000 BSA Concentration (ng/ml) Figure 3. Minimum concentrations of (a) 1,14-eicosatnenoic acid, and (b) prostomial homogcnatc required to bring about sperm activation in the presence of bovine serum albumin (BSA). The fatty acid concentration, or the concentration of prostomial extract required to bring about sperm activation //; wm>. increases with the concentra- tion of dissolved BSA. Data shown are the mean (±SE) minimum con- centrations required to bring a
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. M. G. BENTLEY ET AL ]. o x |1 I 1 1 0 100 1000 10000 100000 BSA Concentration (ng/ml) Figure 3. Minimum concentrations of (a) 1,14-eicosatnenoic acid, and (b) prostomial homogcnatc required to bring about sperm activation in the presence of bovine serum albumin (BSA). The fatty acid concentration, or the concentration of prostomial extract required to bring about sperm activation //; wm>. increases with the concentra- tion of dissolved BSA. Data shown are the mean (±SE) minimum con- centrations required to bring about sperm activation in three replicated experiments. Prostaglandins remain in the pH Tris buffer and would be recovered only in an ethyl acetate fraction from Tris buffer at pH This indicates that SMF activity is not recovered with prostaglandins but is recovered in the fatty acid fraction. A parallel approach to the purification of SMF has been carried out using Sep-Pak® cartridges and a succes- sion of aqueous and organic solvents. Table III shows that SMF activity is recovered in the same fractions as the 8,11,14-eicosatrienoic acid standard. Gas chromaiographic analysis n/'prnstumial lipids The results of separation of methyl esters of prostomial lipid extracts are shown in Figure 5. Three peaks with retention times corresponding to methyl esters of 5,8,1 l,14-eicosatetraenoicacid{ min), 8,11,14-eico- satrienoic acid ( min), ,17-eicosatrienoic acid ( min), can be identified. This clearly indicates the presence of 8,11,14-eicosatrienoic acid in prostomial lipid extracts obtained from prostomia showing SMF ac- tivity in vitro. Discussion The results obtained from thin layer chromatography of prostomial total lipid extracts described above showed that SMF activity co-migrated with 20-carbon fatty acid standards. In particular, it is associated with eicosatrie- noic acids (demonstrated by argentation TLC). The bio- assay of C20 fatty acids show tha
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
