Heyting C, Dietrich A J, Redeker E J, Vink A C
Eur J Cell Biol. 1985 Mar;36(2):307-14.
Synaptonemal complexes (SCs) (structures involved in chromosome pairing during meiosis) were isolated and purified from rat spermatocytes for the purpose of biochemical and morphological analysis. Spermatocytes were lysed in a medium, containing Triton X-100, EDTA and DTT; the resulting swollen nuclei were disrupted by DNAse II, and the suspension was centrifuged through 1.5 M sucrose. The resulting preparation consisted for at least 60% of free SCs, as judged from electron micrographs of agar filtrates. The purified SCs still possessed lateral and transversal elements and attachment plaques. A small fraction also contained a central element. Particularly in diplotene SCs, the lateral elements clearly consisted of two subelements, which are connected by thinner fibres. The lateral elements may fall apart into a network of thinner fibres, presumably as a result of degradation during isolation. On SDS-polyacrylamide gels, the major protein components of purified SCs had relative mobilities (Mrs) of 67 to 60 and 57 to 55 kDa; in addition, there were minor proteins with Mrs of 90, 35, 33, 28, and 26 kDa, and varying amounts of histones. The 67 to 60 kDa proteins comigrate with lamins of rat liver pore complexes and laminae. A possible relationship between SCs and pore complexes and laminae is discussed.
为了进行生化和形态学分析，从大鼠精母细胞中分离并纯化了联会复合体（SCs）（减数分裂过程中参与染色体配对的结构）。将精母细胞在含有 Triton X - 100、EDTA 和 DTT 的培养基中裂解；所得肿胀的细胞核用 DNA 酶 II 破坏，然后将悬浮液通过 1.5 M 蔗糖进行离心。从琼脂滤液的电子显微照片判断，所得制剂中至少 60% 是游离的 SCs。纯化后的 SCs 仍具有侧元件、横向元件和附着斑。一小部分还含有中央元件。特别是在双线期的 SCs 中，侧元件明显由两个亚元件组成，它们由更细的纤维连接。侧元件可能会分解成更细纤维的网络，这可能是由于分离过程中的降解所致。在 SDS - 聚丙烯酰胺凝胶上，纯化后的 SCs 的主要蛋白质成分的相对迁移率（Mr）为 67 至 60 kDa 和 57 至 55 kDa；此外，还有 Mr 为 90、35、33、28 和 26 kDa 的次要蛋白质，以及不同量的组蛋白。67 至 60 kDa 的蛋白质与大鼠肝孔复合体和核纤层的核纤层蛋白共迁移。本文讨论了 SCs 与孔复合体和核纤层之间可能的关系。
