Sperm from the Fruit Fly,Drosophila Melanogaster, Cannot Be Dispersed by Conventional Methods.

摘要

Several physical and chemical methods were used in attempts to disperse sperm from the fruit fly,Drosophila melanogaster into a single-cell suspension. We are developing flow cytometric techniques to determine DNA variability in mammalian sperm from animals exposed to physical and chemical mutagens,carcinogens,and teratogens. Because many genetic variants of fruit flies are available,the sperm from these variants could be a critical test object for our method of measuring relative DNA content in sperm. Dispersal procedures included coating the cells with bovine serum albumin (BSA) and sodium dodecyl sulfate (SDS) to reduce stickiness, sonicating sperm suspensions,applying dithiothreitol (DTT) and SDS or cetyl trimethyl ammonium bromide (CTAB) to dissolve the tails,and digesting with tryspin. All these methods were ineffective. When sperm were placed in a crude extract of spermatheca and seminal receptacles from the female,the cells became less sticky. Although many of the sperm masses were loosened,the yield of free sperm was low. (ERA citation 04:024612)