Cann G M, Bradshaw A D, Gervin D B, Hunter A W, Clegg D O
Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara 93106, USA.
Dev Biol. 1996 Nov 25;180(1):82-96. doi: 10.1006/dbio.1996.0286.
During extension of axons, critical neuronal interactions with extracellular matrix (ECM) and other cells are thought to be mediated in part by heterodimeric beta1 integrin receptors. In this report, we examine the expression and function of beta1 integrins in the developing chick retina. Expression of the beta1 subunit, assayed by in situ hybridization and antibody staining of dissociated cells, was widespread in undifferentiated neuroepithelial cells, before the initiation of axons. Expression persisted in most retinal cell layers throughout embryonic development, during and after axon extension. The repertoire of beta1-associated alpha subunits was examined using reverse transcription-polymerase chain reaction. In addition to the alpha6 and alpha8 subunits previously reported, chick homologues of the alpha2 and alpha4 subunits were detected. Developmental Northern blots revealed varying patterns of integrin subunit expression and showed that expression of beta1 and the mRNAs of its associated alpha subunits are not always coregulated during retinal development. The timing and distribution of expression suggested that beta1 integrins may be involved in other developmental events in addition to axon extension. To address functions carried out by beta1 integrins in the early retina, explanted eye cups were incubated in the presence of function blocking anti-beta1 antibody and migration of newly born retinal ganglion cells (RGCs) was assessed. RGC migration from the ventricular zone to the vitreal border was significantly inhibited, suggesting that beta1 integrins play a role in neuroblast migration in the retina.
在轴突延伸过程中,关键的神经元与细胞外基质(ECM)及其他细胞的相互作用被认为部分是由异二聚体β1整合素受体介导的。在本报告中,我们研究了β1整合素在发育中的鸡视网膜中的表达和功能。通过原位杂交和对解离细胞进行抗体染色来检测β1亚基的表达,结果显示在轴突起始之前,其在未分化的神经上皮细胞中广泛表达。在整个胚胎发育过程中,在轴突延伸期间及之后,β1亚基的表达在大多数视网膜细胞层中持续存在。使用逆转录-聚合酶链反应检测了与β1相关的α亚基种类。除了先前报道的α6和α8亚基外,还检测到了α2和α4亚基的鸡同源物。发育Northern印迹显示整合素亚基的表达模式各异,并表明在视网膜发育过程中,β1及其相关α亚基的mRNA表达并不总是协同调节的。表达的时间和分布表明,β1整合素可能除了轴突延伸外还参与其他发育事件。为了研究β1整合素在早期视网膜中的功能,将分离的眼杯在功能阻断性抗β1抗体存在的情况下孵育,并评估新生视网膜神经节细胞(RGCs)的迁移。RGC从室管膜区向玻璃体边界的迁移受到显著抑制,这表明β1整合素在视网膜神经母细胞迁移中发挥作用。