Transmission and scanning electron microscopic studies of the human sperm chromatin decondensed by micrococcal nuclease and salt.
The human sperm chromatin was gently decondensed by treating the sperm head sequentially with micrococcal nuclease and 2 M NaCl. All histones, about 10% of DNA, and a small amount of degraded protamines were released into the soluble fraction, leaving mainly nucleoprotamines in the pellet fraction. Transmission and scanning electron microscopic studies revealed that the nucleoprotamine pellet consisted of chromatin cords of two dimensions, viz., 330- to 420-A and 650- to 1200-A thick cords laced together by very fine strands of 60- to 80-A fibers; both types of cord appeared knobby and had zig-zag patterns throughout their length. It appeared that these cords were derived from two types of sperm heads of approximately equal population; one type contained chiefly the thick cords and the other chiefly the thin cords. Further treatment of the pellet nuclease-NaCl with urea and mercaptoethanol resulted in the dissociation of the thick into the thin cords and unravelling of the thin cords into smaller sized fibers; whereas the treatment of the pellet nuclease-NaCl with DNAase I resulted in the disappearance of the 60- to 80-A fibers, and the remaining cords were chiefly of thick type together with the sperm head exoskeletons. From these results the packing order of the chromatin in human sperm was proposed.
D000328:Adult; D002843:Chromatin / Q000648:ultrastructure*; D003850:Deoxyribonuclease I; D003851:Deoxyribonucleases / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism; D006801:Humans; D008297:Male; D008623:Mercaptoethanol; D008836:Micrococcal Nuclease / Q000378:metabolism*; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D009994:Osmolar Concentration; D013077:Sperm Head / Q000648:ultrastructure*; D013094:Spermatozoa / Q000648:ultrastructure*; D014508:Urea
D002843:Chromatin; D008623:Mercaptoethanol; D014508:Urea; D003851:Deoxyribonucleases; D004720:Endonucleases; D003850:Deoxyribonuclease I; D008836:Micrococcal Nuclease