The effect of bovine serum albumin and cytohelicase on surface-spread synaptonemal complexes of rye (Secale cereale).
Synaptonemal complexes of rye meiocytes were spread on plastic coated slides for electron microscopic observation. Two proteins generally used in synaptonemal complex spreading techniques, bovine serum albumin and cytohelicase, were applied separately or in combination in an isotonic protoplast medium at concentrations of 0.1-5%. At high concentrations these proteins proved to enhance notably the ultimate number of cells with synaptonemal complexes in the preparations. Also under this condition, centromere structures became stainable with silver nitrate in both the synaptonemal complexes of pollen mother cells and in interphase nuclei of other cell types. Since the true action of cytohelicase under appropriate spreading conditions was uncertain, the putative enzymatic digestion of cell walls was determined in a series of experiments using the fluorochrome calcofluor white as a stain of callose walls. Obvious breakdown of the cell walls was not observed before 8 min of treatment under standard conditions. This made it plausible that the prime effect of cytohelicase is that of a nonspecific protein interacting with the chromatin and improving the adhesion of synaptonemal complexes to the hydrophobic plastic film. The differential staining of the centromere structures in the presence of bovine serum albumin and cytohelicase probably reflects a reduced spreading of these structures due to preferential binding between these proteins and centromeric proteins.
de Jong
J H
JH
Wageningen Agricultural University, Department of Genetics, The Netherlands.
van Eden
J
J
D000818:Animals; D002503:Centromere / Q000648:ultrastructure; D006372:Helix, Snails / Q000201:enzymology*; D008540:Meiosis; D010944:Plants / Q000648:ultrastructure*; D012434:Secale; D012710:Serum Albumin, Bovine; D013194:Staining and Labeling / Q000379:methods; D013573:Synaptonemal Complex
D012710:Serum Albumin, Bovine