Abstract
Green fluorescent protein (GFP) transgenic animals are widely used in biomedical research. We observed that the commonly used β-actin-GFP transgenic mouse has renal defects with proteinuria starting as early as 5 weeks of age. Histological analysis reveals a widespread increase in glomerular extracellular matrix, occasional mesangiolysis, and secondary tubulointerstitial injury. Electron microscopic (EM) analysis reveals dramatic thickening of the glomerular basement membrane (GBM). Several other transgenic strains with GFP on ubiquitous promoters including β-actin (with insertion in a different location) and ubiquitin C show no renal abnormalities. Western blot analysis on crude glomerular preparations from several GFP transgenic strains revealed that higher levels of GFP expression might be responsible for the observed pathogenesis. Mapping of the transgene insertion site by inverse PCR indicates that the β-actin GFP transgene does not cause insertional mutagenesis nor does it modify the transcription level of adjacent genes. Taken together, this strain of β-actin-GFP transgenic mouse may be used to study the mechanism of GBM expansion. Moreover, experiments using this strain of GFP mouse should be hereafter carefully planned because its renal pathology may interfere with data interpretation.
References
Baens M, Noels H, Broeckx V, Hagens S, Fevery S, Billiau AD, Vankelecom H, Marynen P (2006) The dark side of EGFP: defective polyubiquitination. PLoS ONE 1:e54
Devgan V, Rao MR, Seshagiri PB (2004) Impact of embryonic expression of enhanced green fluorescent protein on early mouse development. Biochem Biophys Res Commun 313:1030–1036
Giepmans BN, Adams SR, Ellisman MH, Tsien RY (2006) The fluorescent toolbox for assessing protein location and function. Science 312:217–224
Guo JK, Schedl A, Krause DS (2006) Bone marrow transplantation can attenuate the progression of mesangial sclerosis. Stem Cells 24:406–415
Haseloff J, Amos B (1995) GFP in plants. Trends Genet 11:328–329
Huang WY, Aramburu J, Douglas PS, Izumo S (2000) Transgenic expression of green fluorescence protein can cause dilated cardiomyopathy. Nat Med 6:482–483
Liu HS, Jan MS, Chou CK, Chen PH, Ke NJ (1999) Is green fluorescent protein toxic to the living cells? Biochem Biophys Res Commun 260:712–717
Misra RP (1972) Isolation of glomeruli from mammalian kidneys by graded sieving. Am J Clin Pathol 58:135–139
Novak A, Guo C, Yang W, Nagy A, Lobe CG (2000) Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon Cre-mediated excision. Genesis 28:147–155
Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y (1997) ‘Green mice’ as a source of ubiquitous green cells. FEBS Lett 407:313–319
Schaefer BC, Schaefer ML, Kappler JW, Marrack P, Kedl RM (2001) Observation of antigen-dependent CD8+ T-cell/ dendritic cell interactions in vivo. Cell Immunol 214:110–122
Stahl RA, Thaiss F, Kahf S, Schoeppe W, Helmchen UM (1990) Immune-mediated mesangial cell injury – biosynthesis and function of prostanoids. Kidney Int 38:273–281
Stewart CN Jr (2006) Go with the glow: fluorescent proteins to light transgenic organisms. Trends Biotechnol 24:155–162
Acknowledgements
We thank Stephanie Donaldson and Yale School of Medicine YARC facility for excellent mouse care. We thank Drs. Anil Karihaloo and Roland Schmitt for helpful discussions and reading through the manuscript. JKG was supported by grant # T32 DK007276 from the NIH/NIDDK.
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Guo, JK., Cheng, EC., Wang, L. et al. The commonly used β-actin-GFP transgenic mouse strain develops a distinct type of glomerulosclerosis. Transgenic Res 16, 829–834 (2007). https://doi.org/10.1007/s11248-007-9107-x
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DOI: https://doi.org/10.1007/s11248-007-9107-x