PERMEABILIZATION, STAINING AND CULTURE OF LIVING DROSOPHILA EMBRYOS
PERMEABILIZATION, STAINING AND CULTURE OF LIVING DROSOPHILA EMBRYOS
复制标题
DOI：
10.3109/10520299409106257
复制
发表时间：
1994-01-01
影响因子：
1.6
通讯作者：
HAM, D
中科院分区：
文献类型：
--
作者：
STRECKER, TR;MCGHEE, S;HAM, D
有机溶剂辛烷通常用于渗透果蝇胚胎周围的疏水性卵黄膜，从而使小分子进入卵中。我们目前的证据表明，己烷是一个更有效的透化剂比辛烷和比较这些溶剂的透化和胚胎活力的均匀性的影响。比较了每种溶剂使胚胎易于接触一系列生物染色剂的能力。在早期胚胎发生的七个不同阶段，辛烷与己烷渗透对随后的胚胎活力的影响进行了测定。我们发现，虽然己烷是一个上级溶剂渗透卵黄膜，它降低了生存能力的胚胎暴露之间的0和3小时的年龄。用己烷或辛烷处理的较老的胚胎通常是可行的。我们还表明，分子质量为984道尔顿或更高的分子没有扩散到胚胎后，无论是己烷或辛烷处理。这里提出的结果挑战相分区模型，已提出解释的分子基础的果蝇卵透化。另一种模型，以及透化和染色果蝇胚胎在早期胚胎发生过程中的任何阶段，同时保持后续培养的活力优化协议。
The organic solvent octane has been used routinely to permeabilize the hydrophobic vitelline membrane surrounding the Drosophila embryo, thereby allowing the movement of small molecules into the egg. We present evidence that hexane is a more effective permeabilizing agent than octane and compare the effects of these solvents on uniformity of permeabilization and embryonic viability. The ability of each solvent to make the embryo accessible to a range of biological stains was compared. The effect of octane versus hexane permeabilization on subsequent embryonic viability was measured at seven different stages during early embryogenesis. We found that although hexane is a superior solvent for permeabilizing the vitelline membrane, it decreases the viability of embryos exposed between 0 and 3 hr of age. Older embryos treated with either hexane or octane are usually viable. We also showed that molecules with a molecular mass of 984 Daltons or more did not diffuse into the embryo following treatment with either hexane or octane. Results presented here challenge a phase-partition model that has been proposed previously to explain the molecular basis of permeabilization of the Drosophila egg. An alternative model is described as well as an optimized protocol for permeabilizing and staining Drosophila embryos at any stage during early embryogenesis while maintaining viability for subsequent culture.