Urbán-Duarte David, Tomita Shuichiro, Sakai Hiroki, Sezutsu Hideki, De La Torre-Sánchez José Fernando, Kainoh Yooichi, Furukawa Seiichi, Uchino Keiro
Centro Nacional de Recursos Genéticos, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Tepatitlán de Morelos 47600, Jalisco, Mexico.
Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 1-2 Owashi, Tsukuba 305-8634, Ibaraki, Japan.
Bioengineering (Basel). 2023 May 8;10(5):563. doi: 10.3390/bioengineering10050563.
The lipid layer surrounding the vitelline membrane of insect eggs has a critical role in the waterproofing and desiccation resistance of embryos. However, this lipid layer also prevents the flux of chemicals into the embryos, such as cryoprotectants, which are required for successful cryopreservation. The permeabilization studies of silkworm embryos remain insufficient. Therefore, in this study, we developed a permeabilization method to remove the lipid layer in the silkworm, , and examined factors affecting the viability of dechorionated embryos, including the types and exposure times of chemicals and embryonic stages. Among the chemicals used, hexane and heptane were effective for permeabilization, whereas Triton X-100 and Tween-80 were less effective. Regarding the embryonic stages, there were significant differences between 160 and 166 h after egg laying (AEL) at 25 °C. Consequently, we found that the treatment of 160 AEL embryos with hexane for 30 s was the best condition for the permeability and viability of embryos, in which over 62% of the permeabilized embryos grew up to the second larval instar and their moths could lay fertilized eggs. Our method can be used for various purposes, including permeability investigations using other chemicals and embryonic cryopreservation.
昆虫卵黄膜周围的脂质层在胚胎的防水和抗干燥方面起着关键作用。然而，这层脂质层也会阻止化学物质进入胚胎，比如成功进行冷冻保存所需的冷冻保护剂。家蚕胚胎的透化研究仍然不足。因此，在本研究中，我们开发了一种去除家蚕脂质层的透化方法，并研究了影响去壳胚胎活力的因素，包括化学物质的类型和暴露时间以及胚胎发育阶段。在所使用的化学物质中，己烷和庚烷对透化有效，而吐温80和曲拉通X-100效果较差。关于胚胎发育阶段，在25℃下产卵后160至166小时之间存在显著差异。因此，我们发现用己烷处理160小时的胚胎30秒是胚胎通透性和活力的最佳条件，其中超过62%的透化胚胎发育到二龄幼虫，并且它们的蛾能够产下受精卵。我们的方法可用于各种目的，包括使用其他化学物质进行通透性研究以及胚胎冷冻保存。
