Isolation of the inner acrosomal-nuclear membrane complex from rabbit spermatozoa |
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Authors: | Hamid Rahi Gholamreza Sheikhnejade Prakash N Srivastava |
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Abstract: | Rabbit spermatozoa were passed through a Bio-Glas 2500 (BG) column to yield pure cytoplasmic droplets and consequently the droplet-free spermatozoa. Rapid decondensation of sperm nuclei was achieved by 1 mM dithiothreitol (DTT) and 1.5 M NaCl. This treatment caused removal of the plasma and the outer acrosomal membranes. The viscosity of decondensed nuclei was reduced by pancreatic DNase. Further elution of the suspension from BG column yielded a membrane complex. On the basis of electron microscopic observations and the enzyme analysis, these membranes appeared to be the inner acrosomal-nuclear membrane complex (IANC). The IANC was also prepared from isolated sperm head by DTT-NaCl and Dnasetreatment and low-speed centrifugation. |
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Keywords: | sperm cytoplasmic droplet membrane acrosome nucleus decondensation rabbit Bio-Glas 2500 |
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