Department of Molecular and Cell Biology and Biochemistry, Division of Biology and Medicine, Brown University;
J Vis Exp. 2024 Apr 19(206). doi: 10.3791/66751.
Laboratory stocks of the lower dipteran fly, Bradysia (Sciara) coprophila, have been maintained for over a century. Protocols for laboratory upkeep of B. coprophila are presented here. These protocols will be useful for the rapidly increasing number of laboratories studying B. coprophila to take advantage of its unique biological features, which include (1) a monopolar spindle in male meiosis I; (2) non-disjunction of the X dyad in male meiosis II; (3) chromosome imprinting to distinguish maternal from paternal homologs; (4) germ line-limited (L) chromosomes; (5) chromosome elimination (paternal chromosomes in male meiosis I; one to two X chromosomes in early embryos; L chromosomes from the soma in early embryos); (6) sex determination by the mother (there is no Y chromosome); and (7) developmentally regulated DNA amplification at the DNA puff loci in larval salivary gland polytene chromosomes. It is now possible to explore these many unique features of chromosome mechanics by using the recent advances in sequencing and assembly of the B. coprophila genome and the development of transformation methodology for genomic engineering. The growing scientific community that uses B. coprophila for research will benefit from the protocols described here for mating the flies (phenotypic markers for mothers that will have only sons or only daughters; details of mass mating for biochemical experiments), checking embryo hatch, feeding larvae, and other comments on its rearing.
实验室饲养的低级双翅目蝇,Bradysia(Sciara)coprophila,已经维持了一个多世纪。这里介绍了 B. coprophila 实验室维护的方案。这些方案将对越来越多的研究 B. coprophila 的实验室有用,以利用其独特的生物学特征,包括(1)雄性减数分裂 I 中的单极纺锤体;(2)雄性减数分裂 II 中 X 二联体的不分离;(3)染色体印迹以区分母本和父本同源物;(4)生殖系限制(L)染色体;(5)染色体消除(雄性减数分裂 I 中的父本染色体;早期胚胎中的一个到两个 X 染色体;早期胚胎中的体细胞中的 L 染色体);(6)由母亲决定性别(没有 Y 染色体);(7)在幼虫唾液腺多线染色体的 DNA 膨突位点发育调控的 DNA 扩增。现在,通过使用 B. coprophila 基因组测序和组装的最新进展以及基因组工程转化方法的发展,可以探索染色体力学的这些许多独特特征。使用 B. coprophila 进行研究的不断增长的科学社区将受益于这里描述的用于交配蝇(将只有儿子或只有女儿的母亲的表型标记;用于生化实验的大量交配的细节)、检查胚胎孵化、喂养幼虫的方案和其他饲养建议。