Detection of Autophagy in Tissue by Standard Immunohistochemistry: Possibilities and Limitations

Transmission electron microscopy (TEM) is currently the standard method to monitor autophagy in tissue. Because TEM is labor intensive, we recently questioned whether marker proteins could be found for unambiguous detection of autophagy in tissue using standard immunohistochemical techniques. Our findings indicated that the identification of autophagy-specific biomarkers for tissue is highly compromised due to lack of differential gene expression. In this respect, TEM remains an indispensable technique for evaluation of autophagy in situ. Nevertheless, immunohistochemical staining of microtubule-associated protein 1 light chain 3 (LC3) appeared to be a valuable technique to detect autophagosome formation in tissue but only when this protein is overexpressed, e.g. in GFP-LC3 transgenic animals. Furthermore, demonstration of granular cytoplasmic ubiquitin inclusions by immunohistochemistry may be an attractive technique to measure autophagic cell degeneration in some human pathologies such as neurodegenerative diseases, heart failure and atherosclerosis. Addenda to: In Situ Detection of Starvation-Induced Autophagy W. Martinet, G.R.Y. De Meyer, L. Andries, A.G. Herman and M.M. Kockx J Histochem Cytochem 2005; In press

[1]  D. Schrijvers,et al.  7-Ketocholesterol Induces Protein Ubiquitination, Myelin Figure Formation, and Light Chain 3 Processing in Vascular Smooth Muscle Cells , 2004, Arteriosclerosis, thrombosis, and vascular biology.

[2]  C. Duve,et al.  Functions of lysosomes. , 1966, Annual review of physiology.

[3]  T. Ueno,et al.  Lysosomal Turnover, but Not a Cellular Level, of Endogenous LC3 is a Marker for Autophagy , 2005, Autophagy.

[4]  Rapamycin Induces the G0 Program of Transcriptional Repression in Yeast by Interfering with the TOR Signaling Pathway , 1998, Molecular and Cellular Biology.

[5]  Masaaki Komatsu,et al.  Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice , 2005, The Journal of cell biology.

[6]  J. Schaper,et al.  Myocytes Die by Multiple Mechanisms in Failing Human Hearts , 2003, Circulation research.

[7]  Y. Kitaura,et al.  Autophagic degeneration as a possible mechanism of myocardial cell death in dilated cardiomyopathy. , 2001, Japanese circulation journal.

[8]  H. P. Ostendorff,et al.  Orchestrating nuclear functions: ubiquitin sets the rhythm. , 2003, Trends in biochemical sciences.

[9]  Takeshi Tokuhisa,et al.  Dissection of Autophagosome Formation Using Apg5-Deficient Mouse Embryonic Stem Cells , 2001, The Journal of cell biology.

[10]  M. Figueiredo-Pereira,et al.  Ubiquitin, cellular inclusions and their role in neurodegeneration , 1998, Trends in Neurosciences.

[11]  M. Matsui,et al.  In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. , 2003, Molecular biology of the cell.

[12]  G. D. De Meyer,et al.  In Situ Detection of Starvation-induced Autophagy , 2006, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[13]  Takeshi Noda,et al.  LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing , 2000, The EMBO journal.

[14]  T. Powers,et al.  Regulation of ribosome biogenesis by the rapamycin-sensitive TOR-signaling pathway in Saccharomyces cerevisiae. , 1999, Molecular biology of the cell.

[15]  D. Klionsky,et al.  Dissection of Autophagosome Biogenesis into Distinct Nucleation and Expansion Steps , 2000, The Journal of cell biology.

[16]  M J Davies,et al.  Apoptotic versus autophagic cell death in heart failure. , 2001, Cardiovascular research.

[17]  B. Lardeux,et al.  Mechanism and regulation of protein degradation in liver. , 1989, Diabetes/metabolism reviews.

[18]  N. Mizushima.  Methods for monitoring autophagy. , 2004, The international journal of biochemistry & cell biology.