Selection of Suitable Reference Genes for MRNA Quantification Studies Using Common Marmoset Tissues

Overview

Journal Mol Biol Rep

Specialty Molecular Biology

Date 2013 Sep 27

PMID 24068436

Citations 8

Authors

Yoshinori Shimamoto

Hiroshi Kitamura

Kimie Niimi

Yasunaga Yoshikawa

Fumio Hoshi

Mayumi Ishizuka

Eiki Takahashi

Affiliations

Soon will be listed here.

Abstract

The common marmoset (Callithrix jacchus) is increasingly being used as a non-human primate animal model in biomedical research. To perform accurate quantitative analysis of gene expression by quantitative reverse transcription polymerase chain reaction, reliable reference genes should be selected. In this study, we evaluated the expressions of 11 widely used reference genes: ACTB, ATP5F1, B2M, GAPDH, HPRT1, PGK1, PPIA, RN18S1, RPLP0, TBP and UBC in 12 tissues and five brain areas of healthy common marmosets. NormFinder and geNorm indicated that the most suitable reference genes for cross-sectional studies of the 17 tissues were RN18S1 and RPLP0. Conversely, ACTB and PPIA were the most suitable for analyzing brain samples; however, the expression of PGK1 fluctuated among brain areas. These results indicate that suitable reference genes differ between the tissues examined. This study provides fundamental information for gene expression studies of the common marmoset and highlights the importance of validating reference genes before quantification of target mRNAs.

Citing Articles

Highly Selective PPARα (Peroxisome Proliferator-Activated Receptor α) Agonist Pemafibrate Inhibits Stent Inflammation and Restenosis Assessed by Multimodality Molecular-Microstructural Imaging.

Iwata H, Osborn E, Ughi G, Murakami K, Goettsch C, Hutcheson J J Am Heart Assoc. 2021; 10(20):e020834.

PMID: 34632804 PMC: 8751880. DOI: 10.1161/JAHA.121.020834.

Interferon Regulatory Factors IRF1 and IRF7 Directly Regulate Gene Expression in Bats in Response to Viral Infection.

Irving A, Zhang Q, Kong P, Luko K, Rozario P, Wen M Cell Rep. 2020; 33(5):108345.

PMID: 33147460 PMC: 8755441. DOI: 10.1016/j.celrep.2020.108345.

Trait Anxiety Mediated by Amygdala Serotonin Transporter in the Common Marmoset.

Quah S, McIver L, Roberts A, Santangelo A J Neurosci. 2020; 40(24):4739-4749.

PMID: 32393533 PMC: 7294798. DOI: 10.1523/JNEUROSCI.2930-19.2020.

Insula serotonin 2A receptor binding and gene expression contribute to serotonin transporter polymorphism anxious phenotype in primates.

Santangelo A, Sawiak S, Fryer T, Hong Y, Shiba Y, Clarke H Proc Natl Acad Sci U S A. 2019; 116(29):14761-14768.

PMID: 31266890 PMC: 6642374. DOI: 10.1073/pnas.1902087116.

Generation of transgenic marmosets expressing genetically encoded calcium indicators.

Park J, Zhang X, Choi S, Okahara J, Sasaki E, Silva A Sci Rep. 2016; 6:34931.

PMID: 27725685 PMC: 5057151. DOI: 10.1038/srep34931.

References

Tong Z, Gao Z, Wang F, Zhou J, Zhang Z . Selection of reliable reference genes for gene expression studies in peach using real-time PCR. BMC Mol Biol. 2009; 10:71. PMC: 3224724. DOI: 10.1186/1471-2199-10-71. View

Aerts J, Gonzales M, Topalian S . Selection of appropriate control genes to assess expression of tumor antigens using real-time RT-PCR. Biotechniques. 2004; 36(1):84-6, 88, 90-1. DOI: 10.2144/04361ST04. View

Rozen S, Skaletsky H . Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol. 1999; 132:365-86. DOI: 10.1385/1-59259-192-2:365. View

Bustin S, Penning L . Improving the analysis of quantitative PCR data in veterinary research. Vet J. 2011; 191(3):279-81. DOI: 10.1016/j.tvjl.2011.06.044. View

Nygard A, Jorgensen C, Cirera S, Fredholm M . Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR. BMC Mol Biol. 2007; 8:67. PMC: 2000887. DOI: 10.1186/1471-2199-8-67. View

Schmid H, Cohen C, Henger A, Irrgang S, Schlondorff D, Kretzler M . Validation of endogenous controls for gene expression analysis in microdissected human renal biopsies. Kidney Int. 2003; 64(1):356-60. DOI: 10.1046/j.1523-1755.2003.00074.x. View

Kuchipudi S, Tellabati M, Nelli R, White G, Perez B, Sebastian S . 18S rRNA is a reliable normalisation gene for real time PCR based on influenza virus infected cells. Virol J. 2012; 9:230. PMC: 3499178. DOI: 10.1186/1743-422X-9-230. View

t Hart B, van Meurs M, Brok H, Massacesi L, Bauer J, Boon L . A new primate model for multiple sclerosis in the common marmoset. Immunol Today. 2000; 21(6):290-7. DOI: 10.1016/s0167-5699(00)01627-3. View

Baker H, Ridley R, DUCHEN L, Crow T, Bruton C . Experimental induction of beta-amyloid plaques and cerebral angiopathy in primates. Ann N Y Acad Sci. 1993; 695:228-31. DOI: 10.1111/j.1749-6632.1993.tb23057.x. View

10.

Thellin O, Zorzi W, Lakaye B, de Borman B, Coumans B, Hennen G . Housekeeping genes as internal standards: use and limits. J Biotechnol. 2000; 75(2-3):291-5. DOI: 10.1016/s0168-1656(99)00163-7. View

11.

Robinson T, Sutherland I, Sutherland J . Validation of candidate bovine reference genes for use with real-time PCR. Vet Immunol Immunopathol. 2006; 115(1-2):160-5. DOI: 10.1016/j.vetimm.2006.09.012. View

12.

Andersen C, Jensen J, Orntoft T . Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res. 2004; 64(15):5245-50. DOI: 10.1158/0008-5472.CAN-04-0496. View

13.

Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A . Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2002; 3(7):RESEARCH0034. PMC: 126239. DOI: 10.1186/gb-2002-3-7-research0034. View

14.

Mansfield K . Marmoset models commonly used in biomedical research. Comp Med. 2003; 53(4):383-92. View

15.

Dheda K, Huggett J, Chang J, Kim L, Bustin S, Johnson M . The implications of using an inappropriate reference gene for real-time reverse transcription PCR data normalization. Anal Biochem. 2005; 344(1):141-3. DOI: 10.1016/j.ab.2005.05.022. View

16.

Gnanalingham K, Smith L, Hunter A, Jenner P, Marsden C . Alterations in striatal and extrastriatal D-1 and D-2 dopamine receptors in the MPTP-treated common marmoset: an autoradiographic study. Synapse. 1993; 14(2):184-94. DOI: 10.1002/syn.890140212. View

17.

Penning L, Vrieling H, Brinkhof B, Riemers F, Rothuizen J, Rutteman G . A validation of 10 feline reference genes for gene expression measurements in snap-frozen tissues. Vet Immunol Immunopathol. 2007; 120(3-4):212-22. DOI: 10.1016/j.vetimm.2007.08.006. View

18.

Remington E, Osmanski M, Wang X . An operant conditioning method for studying auditory behaviors in marmoset monkeys. PLoS One. 2012; 7(10):e47895. PMC: 3480461. DOI: 10.1371/journal.pone.0047895. View

19.

Goidin D, Mamessier A, Staquet M, Schmitt D, Berthier-Vergnes O . Ribosomal 18S RNA prevails over glyceraldehyde-3-phosphate dehydrogenase and beta-actin genes as internal standard for quantitative comparison of mRNA levels in invasive and noninvasive human melanoma cell subpopulations. Anal Biochem. 2001; 295(1):17-21. DOI: 10.1006/abio.2001.5171. View

20.

Abbott D, Barnett D, Colman R, Yamamoto M, Schultz-Darken N . Aspects of common marmoset basic biology and life history important for biomedical research. Comp Med. 2003; 53(4):339-50. View