Tandler C J, Solari A J
Instituto de Biología Celular, Facultad de Medicina, Buenos Aires, Argentina.
Genetica. 1991;84(1):39-49. doi: 10.1007/BF00123983.
This study reports the persistence of axis-like structures in the centromeric region of both homologues during the metaphase-I and anaphase-I stages of meiotic division of mouse spermatocytes. A novel type of silver 'argentaffin' technique (NH4-Ag) is employed. This technique includes the treatment of glutaraldehyde-fixed tissues with dilute ammonium hydroxide followed by a reduction of aldehyde groups with sodium borohydride. Staining is accomplished with ammoniacal silver nitrate in darkness followed by sulfite washing. The lateral elements of synaptonemal complexes and the single chromosomal axes of diplotene spermatocytes show a prominent reactivity with this technique. The pattern of very small grains over condensed chromatin is uniform and gives only a light opacity to the electron beam. The presence of an axis-like structure is seen in every centromeric end of meiotic chromosomes at metaphase I and anaphase I. The chromatin (heterochromatin) that surrounds the centromeric filament and some material distributed in irregular linear arrays along some of the homologues also showed a higher electron opacity than the bulk of deoxyribonucleoprotein. While the former is related to C+ heterochromatin, the latter could represent dispersed material of diplotene axes. It is suggested that the disposal of axial material is differentially delayed at the centromeric regions. The present evidence supports the hypothesis that axial fragments or lateral-element segments persisting at these regions contribute to the cohesiveness of centromeres of sister chromatids during normal disjunction.
本研究报告了在小鼠精母细胞减数分裂的中期I和后期I阶段,同源染色体的着丝粒区域中轴样结构的持续性。采用了一种新型的银“嗜银”技术(NH4-Ag)。该技术包括用稀氢氧化铵处理戊二醛固定的组织,然后用硼氢化钠还原醛基。在黑暗中用氨性硝酸银染色,随后进行亚硫酸盐洗涤。联会复合体的侧生元件和双线期精母细胞的单条染色体轴对此技术表现出显著的反应性。浓缩染色质上非常小的颗粒图案是均匀的,仅使电子束产生轻微的不透明度。在中期I和后期I的减数分裂染色体的每个着丝粒末端都可见轴样结构的存在。围绕着丝粒细丝的染色质(异染色质)以及沿着一些同源染色体以不规则线性阵列分布的一些物质,也显示出比大部分脱氧核糖核蛋白更高的电子不透明度。前者与C+异染色质有关,后者可能代表双线期轴的分散物质。有人提出,轴物质在着丝粒区域的处理存在差异延迟。目前的证据支持这样的假设,即在这些区域持续存在的轴片段或侧生元件片段有助于姐妹染色单体着丝粒在正常分离过程中的黏连。