Document Detail


Methanol-acetic anhydride: an efficient blocking agent for electron microscope cytochemistry. Its application to mouse testis and other tissues.
MedLine Citation:
PMID:  6186647     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A mixture of absolute methanol and acetic anhydride (MA) (5:1, v/v; 24 h at 25 degrees C) which is known to methylate and acetylate--respectively--free carboxyl and amino groups in proteins, was tested for its effectiveness as a blocking agent in glutaraldehyde-fixed mouse testis and skeletal muscle. In young spermatids the staining of the acrosome with either uranyl acetate (UA) or ethanolic phosphotungstic acid (PTA) was completely prevented by prior treatment with MA. Esterification of carboxyl groups with 0.1 N HCl in methanol (24 h at 30 degrees C) eliminated the UA staining without affecting that due to PTA. It is concluded that--COOH groups are responsible for UA binding and that acetylation (of amino groups) prevented PTA binding. MA also abolishes the strong affinity of PTA to the lateral elements of the synaptonemal complex in meiotic chromosomes, the axoneme and fibrous sheath of the spermatid tail and the Z band in skeletal muscle. Reactivity was diminished in nucleoli and remained unaffected in chromatin, the outer coarse fibers of the flagellum and collagen fibrils. Different functional groups may participate in PTA staining. The ultrastructure was well preserved in all cases.
Authors:
C J Tandler; A J Solari
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Histochemistry     Volume:  76     ISSN:  0301-5564     ISO Abbreviation:  Histochemistry     Publication Date:  1982  
Date Detail:
Created Date:  1983-04-21     Completed Date:  1983-04-21     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0411300     Medline TA:  Histochemistry     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  351-61     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Acetic Acids / diagnostic use*
Acetic Anhydrides / diagnostic use*
Acetylation
Animals
Crithidia / analysis,  ultrastructure
Histocytochemistry / methods*
Indicators and Reagents
Male
Methanol / diagnostic use*
Methylation
Mice
Microscopy, Electron
Staining and Labeling
Testis / analysis*,  ultrastructure
Trypanosoma cruzi / analysis,  ultrastructure
Chemical
Reg. No./Substance:
0/Acetic Acids; 0/Acetic Anhydrides; 0/Indicators and Reagents; 67-56-1/Methanol

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