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Unanticipated Changes in Steady-state mRNA Levels for Glyceraldehyde-3-Phosphate Dehydrogenase in Rat Tibiae

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Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA levels are commonly used as an internal control to normalize gene expression data based on the belief that this gene is constitutively expressed. However, GAPDH mRNA levels increased by more than 2.5-fold in tibiae of hindlimb unloaded female rats compared to L32 mRNA levels. Similarly, GAPDH mRNA levels increased compared to 18S ribosomal RNA. Treatment with growth hormone and alcohol show no disparity in GAPDH mRNA levels whereas in some experiments, parathyroid hormone and 17β-estradiol increased GAPDH mRNA levels. Taken together, these findings indicate that it is essential to demonstrate that GAPDH expression is not altered prior to using the gene as an internal control.

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References

  1. CH Turner (1998) ArticleTitleThree rules for bone adaptation to mechanical stimuli. Bone 23 399–407 Occurrence Handle10.1016/S8756-3282(98)00118-5 Occurrence Handle1:STN:280:DyaK1M%2FktVOkuw%3D%3D Occurrence Handle9823445

    Article  CAS  PubMed  Google Scholar 

  2. RT Turner (2000) ArticleTitleInvited review: What do we know about the effects of spaceflight on bone?. J Appl Physiol 89 840–847

    Google Scholar 

  3. BP Halloran DD Bikle CM Cone E Morey-Holton (1988) ArticleTitleGlucocorticoids and inhibition of bone formation induced by skeletal unloading. Am J Physiol 255 E875–879 Occurrence Handle1:CAS:528:DyaL1MXltFGjsg%3D%3D Occurrence Handle3202163

    CAS  PubMed  Google Scholar 

  4. TM Skerry (1999) ArticleTitleIdentification of novel signaling pathways during functional adaptation of the skeleton to mechanical loading: the role of glutamate as a paracrine signaling agent in the skeleton. J Bone Miner Metab 17 66–70

    Google Scholar 

  5. TM Skerry MJ Pead LE Lanyon (1991) ArticleTitleModulation of bone loss during disuse by pulsed electromagnetic fields. J Orthop Res 9 600–608 Occurrence Handle1:STN:280:By6B2MbjsV0%3D Occurrence Handle2045987

    CAS  PubMed  Google Scholar 

  6. KC Westerlind TJ Wronski EL Ritman ZP Luo KN An NH Bell RT Turner (1997) ArticleTitleEstrogen regulates the rate of bone turnover but bone balance in ovariectomized rats is modulated by prevailing mechanical strain. Proc Natl Acad Sci USA 94 4199–4204 Occurrence Handle10.1073/pnas.94.8.4199 Occurrence Handle1:CAS:528:DyaK2sXis1eisrk%3D Occurrence Handle9108129

    Article  CAS  PubMed  Google Scholar 

  7. AD Leblanc VS Schneider HJ Evans DA Engelbretson JM Krebs (1990) ArticleTitleBone mineral loss and recovery after 17 weeks of bed rest. J Bone Miner Res 5 843–850 Occurrence Handle1:STN:280:By6D2M%2Fnt1U%3D Occurrence Handle2239368

    CAS  PubMed  Google Scholar 

  8. JM Cavolina GL Evans SA Harris M Zhang KC Westerlind RT Turner (1997) ArticleTitleThe effects of orbital spaceflight on bone histomorphometry and messenger ribonucleic acid levels for bone matrix proteins and skeletal signaling peptides in ovariectomized growing rats. Endocrinology 138 1567–1576 Occurrence Handle1:CAS:528:DyaK2sXitVGmur4%3D Occurrence Handle9075717

    CAS  PubMed  Google Scholar 

  9. TJ Wronski ER Morey (1983) ArticleTitleEffect of spaceflight on periosteal bone formation in rats. Am J Physiol 244 R305–309 Occurrence Handle1:STN:280:BiyC2M%2FisFM%3D Occurrence Handle6402940

    CAS  PubMed  Google Scholar 

  10. ER Morey-Holton RK Globus (2002) ArticleTitleHindlimb unloading rodent model: technical aspects. J Appl Physiol 92 1367–1377

    Google Scholar 

  11. P Chomczynski N Sacchi (1987) ArticleTitleSingle-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162 156–159 Occurrence Handle1:CAS:528:DyaL2sXitFSns7Y%3D Occurrence Handle2440339

    CAS  PubMed  Google Scholar 

  12. RT Turner TJ Wronski M Zhang LS Kidder SA Bloomfield JD Sibonga (1998) ArticleTitleEffects of ethanol on gene expression in rat bone: transient dose-dependent changes in mRNA levels for matrix proteins, skeletal growth factors, and cytokines are followed by reductions in bone formation. Alcohol Clin Exp Res 22 1591–1599 Occurrence Handle1:CAS:528:DyaK1cXnt1ensr0%3D Occurrence Handle9802546

    CAS  PubMed  Google Scholar 

  13. RT Turner GL Evans M Zhang A Maran JD Sibonga (1999) ArticleTitleIs resveratrol an estrogen agonist in growing rats?. Endocrinology 140 50–54 Occurrence Handle1:CAS:528:DyaK1MXhtFWgsQ%3D%3D Occurrence Handle9886806

    CAS  PubMed  Google Scholar 

  14. LS Kidder IU Schmidt GL Evans RT Turner (1997) ArticleTitleEffects of growth hormone and low dose estrogen on bone growth and turnover in long bones of hypophysectomized rats. Calcif Tissue Int 61 327–335 Occurrence Handle10.1007/s002239900343 Occurrence Handle1:CAS:528:DyaK2sXmsFCltbg%3D Occurrence Handle9312204

    Article  CAS  PubMed  Google Scholar 

  15. IA Rose JV Warms (1966) ArticleTitleControl of glycolysis in the human red blood cell. J Biol Chem 241 4848–4854 Occurrence Handle1:CAS:528:DyaF28XkvFehtrg%3D Occurrence Handle4288723

    CAS  PubMed  Google Scholar 

  16. K Kumar XL Wu (1995) ArticleTitleExpression of beta-actin and alpha-tubulin mRNA in gerbil brain following transient ischemia and reperfusion up to 1 month. Mol Brain Res 30 149–157 Occurrence Handle10.1016/0169-328X(94)00286-N Occurrence Handle1:CAS:528:DyaK2MXlsFGntL4%3D Occurrence Handle7609636

    Article  CAS  PubMed  Google Scholar 

  17. JE Tanner G Tosato (1992) ArticleTitleRegulation of B-cell growth and immunoglobulin gene transcription by interleukin-6. Blood 79 452–459 Occurrence Handle1:CAS:528:DyaK38XhtVCis7c%3D Occurrence Handle1370387

    CAS  PubMed  Google Scholar 

  18. SA Melki NA Abumrad (1993) ArticleTitleExpression of the adipocyte fatty acid-binding protein in streptozotocin-diabetes: effects of insulin deficiency and supplementation. J Lipid Res 34 1527–1534 Occurrence Handle1:STN:280:ByuD2crisFY%3D Occurrence Handle7693843

    CAS  PubMed  Google Scholar 

  19. P Fort L Marty M Piechaczyk S el Sabrouty C Dani P Jeanteur JM Blanchard (1985) ArticleTitleVarious rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. Nucleic Acids Res 13 1431–1442 Occurrence Handle1:CAS:528:DyaL2MXhvVSjtbc%3D Occurrence Handle2987824

    CAS  PubMed  Google Scholar 

  20. M Piechaczyk JM Blanchard L Marty C Dani F Panabieres S El Sabouty P Fort P Jeanteur (1984) ArticleTitlePost-transcriptional regulation of glyceraldehyde-3-phosphate-dehydrogenase gene expression in rat tissues. Nucleic Acids Res 12 6951–6963 Occurrence Handle1:CAS:528:DyaL2cXmt1ertb4%3D Occurrence Handle6548307

    CAS  PubMed  Google Scholar 

  21. T Fagan D Morse JW Hastings (1999) ArticleTitleCircadian synthesis of a nuclear-encoded chloroplast glyceraldehyde-3-phosphate dehydrogenase in the dinoflagellate Gonyaulax polyedra is translationally controlled. Biochemistry 38 7689–7695 Occurrence Handle10.1021/bi9826005 Occurrence Handle1:CAS:528:DyaK1MXjt1aqurs%3D Occurrence Handle10387008

    Article  CAS  PubMed  Google Scholar 

  22. ML Shinohara JJ Loros JC Dunlap (1998) ArticleTitleGlyceraldehyde-3-phosphate dehydrogenase is regulated on a daily basis by the circadian clock. J Biol Chem 273 446–452 Occurrence Handle1:CAS:528:DyaK1cXjvFeisQ%3D%3D Occurrence Handle9417102

    CAS  PubMed  Google Scholar 

  23. IU Schmidt H Dobnig RT Turner (1995) ArticleTitleIntermittent parathyroid hormone treatment increases osteoblast number, steady state messenger ribonucleic acid levels for osteocalcin, and bone formation in tibial metaphysis of hypophysectomized female rats. Endocrinology 136 5127–5134 Occurrence Handle1:CAS:528:DyaK2MXovFOltLg%3D Occurrence Handle7588250

    CAS  PubMed  Google Scholar 

  24. RT Turner LS Kidder M Zhang SA Harris KC Westerlind A Maran TJ Wronski (1999) ArticleTitleEstrogen has rapid tissue-specific effects on rat bone. J Appl Physiol 86 1950–1958

    Google Scholar 

  25. GL Evans E Morey-Holton RT Turner (1998) ArticleTitleSpaceflight has compartment- and gene-specific effects on mRNA levels for bone matrix proteins in rat femur. J Appl Physiol 84 2132–2137

    Google Scholar 

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Acknowledgements

This research is supported by grants from the National Aeronautics and Space Administration (NAG9-1150), National Institute of Health (AR 048833) and the Mayo Foundation.

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Authors and Affiliations

  1. Dept. of Orthopedics, Mayo Clinic, Rochester, MN 55905, USA

    A. Maran, T. E. Hefferan, M. Zhang & R. T. Turner

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  1. A. Maran
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  2. T. E. Hefferan
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  3. M. Zhang
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  4. R. T. Turner
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Correspondence to A. Maran.

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Maran, A., Hefferan, T., Zhang, M. et al. Unanticipated Changes in Steady-state mRNA Levels for Glyceraldehyde-3-Phosphate Dehydrogenase in Rat Tibiae . Calcif Tissue Int 74, 204–207 (2004). https://doi.org/10.1007/s00223-002-1098-2

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  • DOI: https://doi.org/10.1007/s00223-002-1098-2

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