Microarray analysis of fiber cell maturation in the lens - PubMed (original) (raw)

Microarray analysis of fiber cell maturation in the lens

Dmitry Ivanov et al. FEBS Lett. 2005.

Abstract

The mammalian lens consists of an aged core of quiescent cells enveloped by layers of mature fully elongated cells and younger, continuously elongating transcriptionally active cells. The fiber cell maturation is initiated when fiber cells cease to elongate. The process of maturation represents a radical switch from active elongation to a life-long quiescence and has not been studied previously. It may also include critical stages of preparation for the organelle removal and denucleation. In the present study, we used laser capture microdisection (LCM) microdissection and RNA amplification to compare global gene expression profiles of young elongating and mature, non-elongating fiber cells. Analysis of microarray data from three independent dye-swap experiments identified 65 differentially expressed genes (FDR<0.1) with greater than 2-fold change in expression levels. Microarray array results for a group of randomly selected genes were confirmed by quantitative RT-PCR. These microarray results provide clues to understanding the molecular pathways underlying lens development. The identified changes in the profile of gene expression reflected a shift in cell physiology characterizing the lens fiber maturation.

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Figures

Fig. 1

Fig. 1

Maturing and young fibers were discriminated in the TgN(GFPU5)Nagy mouse lenses using contrasting GFP labeling patterns (A). Mature fibers (Mt) were localized within the region of uniform GFP labeling, whereas young (Yg) fibers localized exclusively to the variegated region at the lens periphery. Paraffin sections of P5 lenses were microdissected by LCM using measurements performed on the contralateral eye (red and blue arrows correspond to the inner borders of the maturing and young fibers) (B). Cells cut from each of the two regions were collected. Bar is 50 mkm.

Fig. 2

Fig. 2

The quality of the extracted RNA from elongating and maturing zones in the lens was determined by RT-PCR using primers for mouse Actb (β-actin), Gadd45b, App, Cd9, Srcasm.

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