Pax-6, Prox 1, and Chx10 homeobox gene expression correlates with phenotypic fate of retinal precursor cells.
Pax-6, Prox 1, and Chx10 homeobox gene expression correlates with phenotypic fate of retinal precursor cells.
复制标题
DOI：
--
复制
发表时间：
1997-06
影响因子：
4.4
通讯作者：
Ten Belecky-Adams;Stanislav I. Tomarev;Hua-Shun;Li;Lynda Ploder;Roderick R. Mdnnes;OlofSundin;Ruben Adler
中科院分区：
文献类型：
--
作者：
Ten Belecky-Adams;Stanislav I. Tomarev;Hua-Shun;Li;Lynda Ploder;Roderick R. Mdnnes;OlofSundin;Ruben Adler
目的 研究同源盒基因 Pax-6、Prox 1 和 Chx10 在体内和体外雏鸡视网膜发育过程中的表达模式。方法 在一系列发育阶段获得多聚甲醛固定、石蜡包埋的眼睛切片。使用地高辛标记的正义和反义 RNA 探针在组织切片上进行原位杂交，这些探针可识别鸡 Pax-6 和 Prox 1（其序列已可用）和鸡 Chx10（作为本研究的一部分进行克隆和测序）。还通过使用 Pax-6 和 Prox 1 抗体的免疫细胞化学以及使用 32P 标记探针的 Northern 印迹分析来研究选定的发育阶段。结果 直到胚胎 (ED) 5 天，原位杂交显示所有三个基因广泛、弥散分布。然而，在 ED 6 和 ED 8 之间，它们获得了不同的、地形特定的表达模式。 Prox 1 信号主要在神经上皮的预期水平细胞层中表达，玻璃体减弱，并且在神经节细胞和预期感光层中不存在。 Pax-6 仅在同一阶段的预期神经节细胞和无长突细胞区域中强烈表达，并且在预期光感受器中未检测到。 Chx10 表达集中在未来内核层的双极细胞区域。细胞分化发生后，相似的模式在 ED 15 至 ED 18 期间保持不变。 Pax-6 和 Prox 1 免疫反应材料显示出核定位和层状分布模式，相当于原位杂交所见。结论 这些结果表明视网膜前体细胞的分化命运可能受到 Pax-6、Prox 1 或 Chx10 的影响，目前正在使用离解的鸡胚视网膜细胞培养物来测试这一假设。
PURPOSE To study the expression patterns of the homeobox genes Pax-6, Prox 1, and Chx10 during chick retinal development in vivo and in vitro. METHODS Sections of paraformaldehyde-fixed, paraffin-embedded eyes were obtained at a range of developmental stages. In situ hybridization was carried out on tissue sections using digoxigenin-labeled sense and antisense RNA probes that recognize chicken Pax-6 and Prox 1 (whose sequences were already available), and chicken Chx10 (which was cloned and sequenced as part of this study). Selected developmental stages were also studied by immunocytochemistry with antibodies against Pax-6 and Prox 1, and by Northern blot analysis using 32P-labeled probes. RESULTS Until embryonic day (ED) 5, in situ hybridization shows widespread, diffuse distribution of all three genes. Between ED 6 and ED 8, however, they acquire distinct, topographically specific patterns of expression. The Prox 1 signal is predominantly expressed in the prospective horizontal cell layer of the neuroepithelium, decreases vitreally, and is absent from ganglion cells and the prospective photoreceptor layer. Pax-6 is strongly expressed only in the prospective ganglion-cell and amacrine-cell regions at the same stages, and is not detected in prospective photoreceptors. Chx10 expression becomes concentrated in the future bipolar-cell region of the inner nuclear layer. Similar patterns are maintained by ED 15 through ED 18, after cell differentiation has taken place. Pax-6 and Prox 1 immunoreactive materials showed nuclear localization and a pattern of laminar distribution equivalent to that seen by in situ hybridization. CONCLUSIONS These results suggest that the differentiated fate of retinal precursor cells may be influenced by Pax-6, Prox 1, or Chx10, this hypothesis is now being tested using dissociated chick embryo retinal cell cultures.