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Relative Expression of Type II MAGE Genes During Retinoic Acid-Induced Neural Differentiation of Mouse Embryonic Carcinoma P19 Cells: A Comparative Real-Time PCR Analysis

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Abstract

In mammals, the type II melanoma antigen (MAGE) protein family is constituted by at least ten closely related members, but our understanding of their function in the developing nervous system remains poor. To systematically study the expression pattern of type II MAGE genes during neurogenesis, we employed mouse embryonic carcinoma P19 cells as an in vitro model for neural differentiation by retinoic acid (RA) induction. The expression of type II MAGE genes was investigated under distinct steps of differentiation by a comparative ΔΔC T paradigm of real-time quantitative reverse-transcription PCR (qRT-PCR). The relative levels of each gene expression at various steps of differentiation were expressed as a fold change compared with that in RA-untreated P19 cells. The results revealed that: (1) the expression of MAGE-E1, E2, and Necdin transcripts was steadily increased, and the relative levels of MAGE-D1, D2, D3, F1, G1, and H1 mRNA were fluctuantly elevated after the RA-treatment at embryoid body and neural stages; (2) during RA-treatment and subsequent differentiation, the expression of MAGE-L2 mRNA was decreased. Therefore, our results suggested that MAGE-D1, D2, D3, E1, E2, F1, G1, H1, and Necdin might be involved in the early process of neurogenesis, and MAGE-L2 connected with maintenance of pluripotency of stem cells. These studies may present some clues for a better understanding of the fundamental aspects of type II MAGE genes during neurogenesis.

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Abbreviations

MAGE:

Melanoma antigen

RA:

Retinoic acid

qRT-PCR:

Quantitative reverse-transcription PCR

MHD:

MAGE homology domain

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GAP43:

Growth-associated protein 43

C T :

Threshold cycle

EBs:

Embryonic bodies

h:

Hour

d:

Day

RT:

Room temperature

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Acknowledgments

This study was supported by grants from Chinese National Basic Research Program (Grant No. 2009CB918301), and Beijing Municipal Natural Science Foundation (Grant No. 5112027).

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Authors and Affiliations

  1. State Key Laboratory of Proteomics, Department of Neurobiology, Institute of Basic Medical Sciences, 27 Taiping Road, Beijing, 100850, People’s Republic of China

    Yong Liu, Shuguang Yang, Jingwen Yang, Haiping Que & Shaojun Liu

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  1. Yong Liu
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  2. Shuguang Yang
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  3. Jingwen Yang
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  4. Haiping Que
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Correspondence to Yong Liu.

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Liu, Y., Yang, S., Yang, J. et al. Relative Expression of Type II MAGE Genes During Retinoic Acid-Induced Neural Differentiation of Mouse Embryonic Carcinoma P19 Cells: A Comparative Real-Time PCR Analysis. Cell Mol Neurobiol 32, 1059–1068 (2012). https://doi.org/10.1007/s10571-012-9826-2

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