Ogura Toshihiko
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Takayama, Ikoma, Japan.
Differentiation. 2002 Jun;70(4-5):163-71. doi: 10.1046/j.1432-0436.2002.700406.x.
One of the key techniques in developmental biology is introducing transgenes into tissues and analyzing their subsequent effects on morphogenesis and organogenesis. In mammals, the transgenic approach is a way to misexpress foreign genes in various tissues and organs. However, targeting expression to certain tissues is totally dependent on the availability of specific promoters. Hence, it is not an easy task to control transgene expression temporally and spatially during embryogenesis. Further, if the transgene is toxic, embryonic development can be disrupted, resulting in premature death before the desired stages of development. As alternative systems, Xenopus and zebrafish are used frequently. In these vertebrate models, overexpression of genes can be carried out by injecting synthetic RNAs into eggs. However, genetic techniques in these systems are limited only to early development, prohibiting the precise analysis of gene effects on organogenesis in later stages. In contrast, the chick embryo has long served as a powerful and useful model system, holding a unique position in the field of developmental biology. Although trials of transgenic chicks have never been successful, easy accessibility to the developing embryo through a window opened in an eggshell enables performance of a variety of techniques, such as time-lapse cinephotomatography, microsurgical manipulations (including chick/quail chimeras), transplantation of cells and tissues, New's in vitro culture, etc. (Bortier et al., 1996; Douarin et al., 1996; Selleck, 1996). In addition to these experimental advantages, retrovirus-mediated gene delivery, and recently, adenovirus-mediated misexpression have been employed routinely in chick embryos (Leber et al., 1996; Morgan and Fekete, 1996).
发育生物学的关键技术之一是将转基因导入组织并分析其随后对形态发生和器官发生的影响。在哺乳动物中,转基因方法是在各种组织和器官中错误表达外源基因的一种方式。然而,将表达靶向特定组织完全依赖于特定启动子的可用性。因此,在胚胎发育过程中在时间和空间上控制转基因表达并非易事。此外,如果转基因有毒,胚胎发育可能会受到干扰,导致在期望的发育阶段之前过早死亡。作为替代系统,非洲爪蟾和斑马鱼被频繁使用。在这些脊椎动物模型中,可以通过将合成RNA注射到卵中来实现基因的过表达。然而,这些系统中的遗传技术仅限于早期发育,无法精确分析基因在后期对器官发生的影响。相比之下,鸡胚长期以来一直是一个强大且有用的模型系统,在发育生物学领域占据独特地位。尽管转基因鸡的试验从未成功,但通过在蛋壳上开一个窗口可以方便地接触到发育中的胚胎,从而能够进行各种技术操作,如延时电影摄影、显微手术操作(包括鸡/鹌鹑嵌合体)、细胞和组织移植、纽氏体外培养等(博蒂尔等人,1996年;杜瓦兰等人,1996年;塞莱克,1996年)。除了这些实验优势外,逆转录病毒介导的基因传递以及最近腺病毒介导的错误表达已在鸡胚中常规使用(勒伯等人,1996年;摩根和费凯特,1996年)。