Sensitive Detection of GFP Utilizing Tyramide Signal Amplification to Overcome Gene Silencing

Overview

Journal Exp Cell Res

Specialty Cell Biology

Date 2007 Apr 13

PMID 17428464

Citations 9

Authors

Zsuzsanna E Toth

Tal Shahar

Ronen Leker

Ildiko Szalayova

Andras Bratincsak

Sharon Key

Anna Lonyai

Krisztian Nemeth

Eva Mezey

Affiliations

Soon will be listed here.

Abstract

The green fluorescent protein (GFP) is among the most commonly used expression markers in biology. GFP-tagged cells have played a particularly important role in studies of cell lineage. Sensitive detection of GFP is crucially important for such studies to be successful, and problems with detection may account for discrepancies in the literature regarding the possible fate choices of stem cells. Here we describe a very sensitive technique for visualization of GFP. Using it we can detect about 90% of cells of donor origin while we could only see about 50% of these cells when we employ the methods that are in general use in other laboratories. In addition, we provide evidence that some cells permanently silence GFP expression. In the case of the progeny of bone marrow stem cells, it appears that the more distantly related they are to their precursors, the more likely it is that they will turn off the lineage marker.

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References

Xia H, Mao Q, Paulson H, Davidson B . siRNA-mediated gene silencing in vitro and in vivo. Nat Biotechnol. 2002; 20(10):1006-10. DOI: 10.1038/nbt739. View

Shimomura O . The discovery of aequorin and green fluorescent protein. J Microsc. 2005; 217(Pt 1):1-15. DOI: 10.1111/j.0022-2720.2005.01441.x. View

Bobrow M, Harris T, Shaughnessy K, Litt G . Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays. J Immunol Methods. 1989; 125(1-2):279-85. DOI: 10.1016/0022-1759(89)90104-x. View

Novak A, Guo C, Yang W, Nagy A, Lobe C . Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon Cre-mediated excision. Genesis. 2000; 28(3-4):147-55. View

Tamura A, Graham D, McCulloch J, Teasdale G . Focal cerebral ischaemia in the rat: 2. Regional cerebral blood flow determined by [14C]iodoantipyrine autoradiography following middle cerebral artery occlusion. J Cereb Blood Flow Metab. 1981; 1(1):61-9. DOI: 10.1038/jcbfm.1981.7. View

Beilhack G, Scheffold Y, Weissman I, Taylor C, Jerabek L, Burge M . Purified allogeneic hematopoietic stem cell transplantation blocks diabetes pathogenesis in NOD mice. Diabetes. 2002; 52(1):59-68. DOI: 10.2337/diabetes.52.1.59. View

Amarzguioui M, Rossi J, Kim D . Approaches for chemically synthesized siRNA and vector-mediated RNAi. FEBS Lett. 2005; 579(26):5974-81. DOI: 10.1016/j.febslet.2005.08.070. View

Rinne A, Littwitz C, Kienitz M, Gmerek A, Bosche L, Pott L . Gene silencing in adult rat cardiac myocytes in vitro by adenovirus-mediated RNA interference. J Muscle Res Cell Motil. 2006; 27(5-7):413-21. DOI: 10.1007/s10974-006-9087-0. View

Hofstetter C, Schwarz E, Hess D, Widenfalk J, El Manira A, Prockop D . Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery. Proc Natl Acad Sci U S A. 2002; 99(4):2199-204. PMC: 122342. DOI: 10.1073/pnas.042678299. View

10.

Hadjantonakis A, Gertsenstein M, Ikawa M, Okabe M, Nagy A . Generating green fluorescent mice by germline transmission of green fluorescent ES cells. Mech Dev. 1998; 76(1-2):79-90. DOI: 10.1016/s0925-4773(98)00093-8. View

11.

Tiscornia G, Singer O, Ikawa M, Verma I . A general method for gene knockdown in mice by using lentiviral vectors expressing small interfering RNA. Proc Natl Acad Sci U S A. 2003; 100(4):1844-8. PMC: 149921. DOI: 10.1073/pnas.0437912100. View

12.

Ito T, Suzuki A, Okabe M, Imai E, Hori M . Application of bone marrow-derived stem cells in experimental nephrology. Exp Nephrol. 2001; 9(6):444-50. DOI: 10.1159/000052644. View

13.

Hunyady B, Krempels K, Harta G, Mezey E . Immunohistochemical signal amplification by catalyzed reporter deposition and its application in double immunostaining. J Histochem Cytochem. 1996; 44(12):1353-62. DOI: 10.1177/44.12.8985127. View

14.

Lambert J, Liu M, Colvin G, Dooner M, McAuliffe C, Becker P . Marrow stem cells shift gene expression and engraftment phenotype with cell cycle transit. J Exp Med. 2003; 197(11):1563-72. PMC: 2193900. DOI: 10.1084/jem.20030031. View

15.

Moore K, Lemischka I . Stem cells and their niches. Science. 2006; 311(5769):1880-5. DOI: 10.1126/science.1110542. View

16.

Adams J . Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains. J Histochem Cytochem. 1992; 40(10):1457-63. DOI: 10.1177/40.10.1527370. View

17.

Jackson K, Majka S, Wulf G, Goodell M . Stem cells: a minireview. J Cell Biochem Suppl. 2002; 38:1-6. DOI: 10.1002/jcb.10045. View

18.

Massengale M, Wagers A, Vogel H, Weissman I . Hematopoietic cells maintain hematopoietic fates upon entering the brain. J Exp Med. 2005; 201(10):1579-89. PMC: 2212913. DOI: 10.1084/jem.20050030. View

19.

Walter I, Fleischmann M, Klein D, Muller M, Salmons B, Gunzburg W . Rapid and sensitive detection of enhanced green fluorescent protein expression in paraffin sections by confocal laser scanning microscopy. Histochem J. 2000; 32(2):99-103. DOI: 10.1023/a:1004014211408. View

20.

Shichinohe H, Kuroda S, Lee J, Nishimura G, Yano S, Seki T . In vivo tracking of bone marrow stromal cells transplanted into mice cerebral infarct by fluorescence optical imaging. Brain Res Brain Res Protoc. 2004; 13(3):166-75. DOI: 10.1016/j.brainresprot.2004.04.004. View