Goetz Jillian J, Laboissonniere Lauren A, Wester Andrea K, Lynch Madison R, Trimarchi Jeffrey M
Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa, United States of America.
Ames High School, Ames, Iowa, United States of America.
PLoS One. 2016 Mar 7;11(3):e0150878. doi: 10.1371/journal.pone.0150878. eCollection 2016.
During retinogenesis seven different cell types are generated in distinct yet overlapping timepoints from a population of retinal progenitor cells. Previously, we performed single cell transcriptome analyses of retinal progenitor cells to identify candidate genes that may play roles in the generation of early-born retinal neurons. Based on its expression pattern in subsets of early retinal cells, polo-like kinase 3 (Plk3) was identified as one such candidate gene. Further characterization of Plk3 expression by in situ hybridization revealed that this gene is expressed as cells exit the cell cycle. We obtained a Plk3 deficient mouse and investigated changes in the retina's morphology and transcriptome through immunohistochemistry, in situ hybridization and gene expression profiling. These experiments have been performed initially on adult mice and subsequently extended throughout retinal development. Although morphological studies revealed no consistent changes in retinogenesis upon Plk3 loss, microarray profiling revealed potential candidate genes altered in Plk3-KO mice. Further studies will be necessary to understand the connection between these changes in gene expression and the loss of a protein kinase such as Plk3.
在视网膜生成过程中,七种不同的细胞类型在不同但有重叠的时间点从一群视网膜祖细胞中产生。此前,我们对视网膜祖细胞进行了单细胞转录组分析,以鉴定可能在早期生成的视网膜神经元产生过程中发挥作用的候选基因。基于其在早期视网膜细胞亚群中的表达模式, polo样激酶3(Plk3)被鉴定为这样一个候选基因。通过原位杂交对Plk3表达的进一步表征显示,该基因在细胞退出细胞周期时表达。我们获得了一只Plk3缺陷小鼠,并通过免疫组织化学、原位杂交和基因表达谱分析研究了视网膜形态和转录组的变化。这些实验最初在成年小鼠上进行,随后扩展到整个视网膜发育过程。尽管形态学研究显示Plk3缺失后视网膜生成没有一致的变化,但微阵列分析揭示了Plk3基因敲除小鼠中潜在的候选基因发生了改变。需要进一步研究以了解这些基因表达变化与蛋白激酶如Plk3缺失之间的联系。