Holash J A, Pasquale E B
La Jolla Cancer Research Foundation, California 92037, USA.
Dev Biol. 1995 Dec;172(2):683-93. doi: 10.1006/dbio.1995.8039.
Receptor protein tyrosine kinases of the Eph subfamily have been proposed to play roles in pattern formation based on their distribution during embryonic development. Cek5 (chicken embryo kinase 5) and Cek8 (chicken embryo kinase 8) are Eph-related kinases highly expressed in the chicken embryonic retina. To assess their potential roles in the development of the visual pathway, we examined their distribution by immunoperoxidase labeling. Cek8 is expressed throughout the pathway of the retinal ganglion cell axons, including the nerve fiber layer of the retina, optic nerve, optic chiasm, and stratum opticum of the tectum. Cek5 immunoreactivity is highly concentrated in only a portion of the optic nerve and optic chiasm, and in retinal cultures, Cek5 is detected in neurons. This prompted us to examine the regional distribution of Cek5 in the developing retina and led to the observation that Cek5 is most concentrated in the ventral aspect. RT-PCR established that the differential regulation of Cek5 expression in different portions of the retina occurs at the transcriptional level. Immunoblotting analysis revealed that this unusual expression pattern is distinctive for Cek5, as three other members of the Eph subfamily, Cek4, Cek8, and Cek9, are evenly expressed across the dorsal-ventral axis of the retina. Both Cek5 and Cek8 are distributed in manners which are consistent with their regulating the outgrowth of retinal ganglion cell axons to the tectum. Furthermore, Cek5 represents the first signal transduction molecule found to exhibit the polarized pattern of expression predicted for proteins that control the specificity of the retinotectal projections.
Eph亚家族的受体蛋白酪氨酸激酶,因其在胚胎发育过程中的分布情况,被认为在模式形成中发挥作用。Cek5(鸡胚激酶5)和Cek8(鸡胚激酶8)是在鸡胚视网膜中高表达的Eph相关激酶。为了评估它们在视觉通路发育中的潜在作用,我们通过免疫过氧化物酶标记来检测它们的分布。Cek8在视网膜神经节细胞轴突的整个通路上均有表达,包括视网膜的神经纤维层、视神经、视交叉以及顶盖的视层。Cek5免疫反应性高度集中于视神经和视交叉的一部分,并且在视网膜培养物中,在神经元中检测到Cek5。这促使我们研究Cek5在发育中的视网膜中的区域分布,并观察到Cek5最集中于腹侧。逆转录聚合酶链反应表明,视网膜不同部分Cek5表达的差异调节发生在转录水平。免疫印迹分析显示,这种不寻常的表达模式是Cek5所特有的,因为Eph亚家族的其他三个成员Cek4、Cek8和Cek9在视网膜的背腹轴上均匀表达。Cek5和Cek8的分布方式均与其调节视网膜神经节细胞轴突向顶盖的生长一致。此外,Cek5是第一个被发现呈现出预测用于控制视网膜顶盖投射特异性的蛋白质所具有的极化表达模式的信号转导分子。