Original Papers
12 May 2010
11
0
4
0
Smart Citations
11
0
4
0
Citing PublicationsSupportingMentioningContrasting
View Citations

See how this article has been cited at scite.ai

scite shows how a scientific paper has been cited by providing the context of the citation, a classification describing whether it supports, mentions, or contrasts the cited claim, and a label indicating in which section the citation was made.

Histochemical detection of GM1 ganglioside using cholera toxin-B subunit. Evaluation of critical factors optimal for in situ detection with special emphasis to acetone pre-extraction

Authors

A comparison of histochemical detection of GM1 ganglioside in cryostat sections using cholera toxin B-subunit after fixation with 4% formaldehyde and dry acetone gave tissue-dependent results. In the liver no pre-treatment showed detectable differences related to GM1 reaction products, while studies in the brain showed the superiority of acetone pre-extraction (followed by formaldehyde), which yielded sharper images compared with the diffuse, blurred staining pattern associated with formaldehyde. Therefore, the aim of our study was to define the optimal conditions for the GM1 detection using cholera toxin B-subunit. Ganglioside extractability with acetone, the ever neglected topic, was tested comparing anhydrous acetone with acetone containing admixture of water. TLC analysis of acetone extractable GM1 ganglioside from liver sections did not exceed 2% of the total GM1 ganglioside content using anhydrous acetone at -20°C, and 4% at room temperature. The loss increased to 30.5% using 9:1 acetone/water. Similarly, photometric analysis of lipid sialic acid, extracted from dried liver homogenates with anhydrous acetone, showed the loss of gangliosides into acetone 3.0±0.3% only. The loss from dried brain homogenate was 9.5±1.1%. Thus, anhydrous conditions (dry tissue samples and anhydrous acetone) are crucial factors for optimal in situ ganglioside detection using acetone pre-treatment. This ensures effective physical fixation, especially in tissues rich in polar lipids (precipitation, prevention of in situ diffusion), and removal of cholesterol, which can act as a hydrophobic blocking barrier.

Altmetrics

Downloads

Citations

Crossref
8
Scopus
0
Crossref Logo
Moeko Kohyama, Akira Yabuki, Kenji Ochiai, Yuya Nakamoto, Kazuyuki Uchida, Daisuke Hasegawa, Kimimasa Takahashi, Hiroaki Kawaguchi, Masaya Tsuboi, Osamu Yamato (2016)
In situ detection of GM1 and GM2 gangliosides using immunohistochemical and immunofluorescent techniques for auxiliary diagnosis of canine and feline gangliosidoses. BMC Veterinary Research, 12(1).
Crossref Logo
Allisandra K. Rha, Shih-Hsin Kan, Perla Andrade-Heckman, Chloe L. Christensen, Jerry F. Harb, Raymond Y. Wang (2024)
Base editing of the GLB1 gene is therapeutic in GM1 gangliosidosis patient-derived cells. Molecular Genetics and Metabolism, 143(1-2), 108568.
Crossref Logo
Christopher J. Day, Adrienne W. Paton, Melanie A. Higgins, Lucy K. Shewell, Freda E.-C. Jen, Benjamin L. Schulz, Brock P. Herdman, James C. Paton, Michael P. Jennings (2017)
Structure aided design of a Neu5Gc specific lectin. Scientific Reports, 7(1).
Crossref Logo
Jiyue Chen, Neal K. Devaraj (2021)
Synthetic probes and chemical tools in sphingolipid research. Current Opinion in Chemical Biology, 65, 126.
Crossref Logo
Juan A. Estrada, Mathew A. Barlow, Darice Yoshishige, Arthur G. Williams, H. Fred Downey, Robert T. Mallet, James L. Caffrey (2016)
δ-Opioid receptors: Pivotal role in intermittent hypoxia-augmentation of cardiac parasympathetic control and plasticity. Autonomic Neuroscience, 198, 38.
Crossref Logo
Ángel Gaudioso, Teresa P. Silva, María Dolores Ledesma (2022)
Models to study basic and applied aspects of lysosomal storage disorders. Advanced Drug Delivery Reviews, 190, 114532.
Crossref Logo
Sooyeon Lee, Stephanie Amici, Hagai Tavori, Waylon M. Zeng, Steven Freeland, Sergio Fazio, Lucia Notterpek (2014)
PMP22 Is Critical for Actin-Mediated Cellular Functions and for Establishing Lipid Rafts. The Journal of Neuroscience, 34(48), 16140.
Crossref Logo
T. PETR, V. ŠMÍD, V. KUČEROVÁ, K. VÁŇOVÁ, M. LENÍČEK, L. VÍTEK, F. ŠMÍD, L. MUCHOVÁ (2014)
The Effect of Heme Oxygenase on Ganglioside Redistribution Within Hepatocytes in Experimental Estrogen-Induced Cholestasis. Physiological Research, 359.

How to Cite



Histochemical detection of GM1 ganglioside using cholera toxin-B subunit. Evaluation of critical factors optimal for in situ detection with special emphasis to acetone pre-extraction. (2010). European Journal of Histochemistry, 54(2), e23. https://doi.org/10.4081/ejh.2010.e23