[Biochemical and ultrastructural analyses of the synaptonemal complex in mammalian spermatocytes]

In order to study the molecular organization of synaptonemal complex (SC), a preparative method for isolation of relatively purified SC from rat, mouse and hamster testes was elaborated which involves isolation of SC-containing (pachytene) nuclei, their lysis, DNAase digestion of DNA and fractionation of nuclear elements by the discontinuous sucrose density gradient centrifugation. Electron microscopy revealed a rather good preservation of the SC structure after the isolation procedure. Effects of the dissociating agents on the SC structural integrity were studied. It has been demonstrated that the treatment with 2M NaCl, Triton X-100, sodium deoxycholate, 6M urea, and with a buffer containing 2% SDS and 5% mercaptoetthanol does not lead to a complete SC dissociation, though it results in some structural chanes. Possible reasons of the high resistance of SC to dissociating treatment are discussed.