Université Pierre et Marie Curie-Paris 6, UMR 7622, Paris, France.
PLoS One. 2010 Feb 19;5(2):e9285. doi: 10.1371/journal.pone.0009285.
In Drosophila, each external sensory organ originates from the division of a unique precursor cell (the sensory organ precursor cell or SOP). Each SOP is specified from a cluster of equivalent cells, called a proneural cluster, all of them competent to become SOP. Although, it is well known how SOP cells are selected from proneural clusters, little is known about the downstream genes that are regulated during SOP fate specification.
METHODOLOGY/PRINCIPAL FINDINGS: In order to better understand the mechanism involved in the specification of these precursor cells, we combined laser microdissection, toisolate SOP cells, with transcriptome analysis, to study their RNA profile. Using this procedure, we found that genes that exhibit a 2-fold or greater expression in SOPs versus epithelial cells were mainly associated with Gene Ontology (GO) terms related with cell fate determination and sensory organ specification. Furthermore, we found that several genes such as pebbled/hindsight, scabrous, miranda, senseless, or cut, known to be expressed in SOP cells by independent procedures, are particularly detected in laser microdissected SOP cells rather than in epithelial cells.
CONCLUSIONS/SIGNIFICANCE: These results confirm the feasibility and the specificity of our laser microdissection based procedure. We anticipate that this analysis will give new insight into the selection and specification of neural precursor cells.
在果蝇中,每个外部感觉器官都起源于一个独特的前体细胞(感觉器官前体细胞或 SOP)的分裂。每个 SOP 都是由一个称为神经前簇的同源细胞簇指定的,所有这些细胞都有能力成为 SOP。尽管人们已经很清楚 SOP 细胞是如何从神经前簇中选择出来的,但对于在 SOP 命运指定过程中受到调控的下游基因知之甚少。
方法/主要发现:为了更好地理解这些前体细胞指定中涉及的机制,我们结合激光显微切割来分离 SOP 细胞,并进行转录组分析以研究它们的 RNA 谱。使用这种方法,我们发现 SOP 与上皮细胞相比表达倍数为 2 倍或更高的基因主要与细胞命运决定和感觉器官指定相关的基因本体论(GO)术语相关。此外,我们发现了几个基因,如 pebble/d 、 scabrous 、 miranda 、 senseless 或 cut ,它们通过独立的程序被证明在 SOP 细胞中表达,而在激光显微切割的 SOP 细胞中比在上皮细胞中更能被检测到。
结论/意义:这些结果证实了我们基于激光显微切割的程序的可行性和特异性。我们预计,这项分析将为神经前体细胞的选择和指定提供新的见解。