Profiling of genes differentially expressed in a rat of early and later gestational ages with high-density oligonucleotide DNA array.
The early gestational fetus heals dermal wounds rapidly and scarlessly. This phenomenon appears to be intrinsic to fetal skin and is probably modulated by interplay of many genes. We ventured to study differences in gene expression between earlier gestational skin (EGS) and later gestational skin (LGS) with the aid of high-density oligonucleotide DNA array to explore the molecular mechanism underlying scarless healing. Total RNA was isolated from fetal Wistar rat skin of the scarless (E15) and scar-forming (E18) periods of gestation (term=21.5 days), and purified to mRNAs. Both the mRNAs from EGS and LGS were reversely transcribed to cDNAs, and were labeled with the incorporation of fluorescent dCTP for preparing the hybridization probes through single primer amplification reaction and Klenow labeling methods. The mixed probes were then hybridized to the oligonucleotide DNA arrays that contained 5,705 DNA fragments representing 5,705 rat genes. After highly stringent washing, the microarray was scanned for fluorescent signals to display the differentially expressed genes between two groups of tissues. Among 5,705 rat genes, there were 53 genes (0.93%) with differentially expressed levels between EGS and LGS; 27 genes, including fibroblast growth factor 8 and follistatin, were up-regulated (0.47%); and 26 genes, containing lymphoid enhancer binding factor-1 and beta-catenin, were down-regulated (0.46%) in fetal skin of scarless period vs. scar-forming period. Analyses of genes related to ion channels, growth factors, extracellular matrix and cellular skeleton, and movement confirmed that our molecular data obtained by oligonucleotide DNA array were consistent with the published biochemical and clinical findings of fetal scarless healing. Stronger expression of fibroblast growth factor 8, follistatin, and weaker expression of lymphoid enhancer binding factor-1 and beta-catenin in EGS vs. LGS were also testified with reverse transcription-polymerase chain reaction and Western blotting methods. Oligonucleotide DNA array was a powerful tool for investigating different gene expression between scarless and scar-forming periods of gestation in the rat fetal skin. Many genes were involved in the phenotypic transition from scarless to scar-forming wound repair during gestation. Further analysis of the obtained genes will help to understand the molecular mechanism of fetal scarless healing.
Chen
Wei
W
Wound Healing and Cell Biology Laboratory, Burns Institute, The First Affiliated Hospital (304 Hospital) of the General Hospital of PLA, Trauma Center of Postgraduate Medical College, Beijing, China.
Fu
Xiaobing
X
Ge
Shili
S
Sun
Tongzhu
T
Zhou
Gang
G
Han
Bing
B
Li
Haihong
H
Sheng
Zhiyong
Z
D000818:Animals; D002921:Cicatrix / Q000235:genetics*; D005333:Fetus; D020869:Gene Expression Profiling; D005865:Gestational Age; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D017208:Rats, Wistar; D012867:Skin / Q000196:embryology*; D012879:Skin Physiological Phenomena / Q000235:genetics*; D046509:Tissue Culture Techniques; D014945:Wound Healing / Q000235:genetics*