Xiaodong Long1,2, Zhong Yang3, Yi Zeng1,2, Hongli Li3, Yangyun Han2, Chao You1

1Department of Neurosurgery, West China Hospital, Sichuan University, Chengdu   610041, Sichuan Province, China
2Department of Neurosurgery, People’s Hospital of Deyang City, Deyang   618000, Sichuan Province, China
3Department of Neurobiology, Third Military Medical University of Chinese PLA, Chongqing   400038, China

Xiaodong Long★, Master, Attending physician, Department of Neurosurgery, West China Hospital, Sichuan University, Chengdu 610041, Sichuan Province, China; Department of Neurosurgery, People’s Hospital of Deyang City, Deyang   618000, Sichuan Province, China

Supported by: the Scientific and Technological Foundation of Sichuan Public Health Bureau in 2008, No. 080128*

Long XD, Yang Z, Zeng Y, Li HL, Han YY, You C. Gene expression in retinoic acid-induced neural tube defects: a cDNA microarray analysis. Neural Regen Res. 2009;4(3):165-170.

Abstract

BACKGROUND: Neural tube defects can be induced by abnormal factors in vivo or in vitro during development. However, the molecular mechanisms of neural tube defect induction, and the related gene expression and regulation are still unknown.

OBJECTIVE: To compare the differences in gene expression between normal embryos and those with neural tube defects.

DESIGN, TIME AND SETTING: A neural development study was performed at the Department of Neurobiology, Third Military Medical University of Chinese PLA between January 2006 and October 2007.

MATERIALS: Among 120 adult Kunming mice, 60 pregnant mice were randomly and evenly divided into a retinoic acid group (n = 30) and a normal control group (n =30). The retinoic acid was produced by Sigma, USA, the gene microarray by the Amersham Pharmacia Company, Hong Kong, and the gene sequence was provided by the Incyte database, USA.

METHODS: Retinoic acid was administered to prepare models of neural tube defects, and corn oil was similarly administered to the normal control group. Total RNA was extracted from embryonic tissue of the two groups using a Trizol kit, and a cDNA microarray containing 1 100 known genes was used to compare differences in gene expression between the normal control group and the retinoic acid group on embryonic (E) day 10.5 and 11.5. Several differentially expressed genes were randomly selected from the two groups for Northern blotting, to verify the results of the cDNA microarray.

MAIN OUTCOME MEASURES: Morphological changes and differential gene expression between the normal control group and the retinoic acid group.

RESULTS: Anatomical microscopy demonstrated that an intact closure of the brain was formed in the normal mouse embryos by days E10.5 and E11.5. The cerebral appearance was full and smooth, and the surface of the spine was intact. However, in the retinoic acid group on days E10.5 and E11.5, there were more dead embryos. Morphological malformations typically included non-closure at the top of the cranium and abnormal changes of the metencephalon and face. cDNA microarray analysis suggested that the changes in expression of seven different genes were similar on both days E10.5 and E11.5. These were downregulation of Nek7, Igfbp5, Zw10, Csf3r, Psmc6 and Rb1, and upregulation of Apoa-4. This study also indicated that Cdk5 expression was downregulated in the retinoic acid group on day E11.5. The results of the cDNA microarray analysis were partly confirmed by Northern blotting.

CONCLUSION: Cdk5, Nek7, Igfbp5, Zw10, Csf3r, Psmc6, Rb1 and Apoa-4 may be key factors in retinoic acid-induced neural tube defects.

Key Words: neural tube defects; neurulation; cDNA microarray; retinoic acid

[Full Text]Click