| Conversions of formaldehyde-modified 2'-deoxyadenosine 5'-monophosphate in conditions modeling formalin-fixed tissue dehydration. | |
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MedLine Citation:
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PMID: 16116034 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Formalin-fixed, paraffin-embedded specimens typically provide molecular biologists with low yields of extractable nucleic acids that exhibit extensive strand cleavage and covalent modification of nucleic acid bases. This study supports the idea that these deleterious effects are promoted by the first step in formalin-fixed tissue processing--i.e., tissue dehydration with a graded series of alcohols. We analyzed the conversions of formaldehyde-modified 2'-deoxyadenosine 5'-monophosphate (dAMP) by reverse-phase ion-pair, high-performance liquid chromatography and found that dehydration does not stabilize N-methylol groups in the modified nucleotide. Furthermore, spontaneous demodification in a dry state or in anhydrous ethanol can be as fast as it is in aqueous solutions if the preparation is contaminated with salts of orthophosphoric acid. In ethanol, orthophosphates also catalyze formation of abundant N6-ethoxymethyl-dAMP, as well as cross-linking and depurination of nucleotides present in the mixture. Identification of the products was performed using ultraviolet absorbance spectroscopy and electrospray ionization Fourier-transform ion cyclotron resonance mass spectrometry. Alternatives to the traditional processing of formalin-fixed tissues are discussed. |
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Authors:
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Vladimir K Rait; Qingrong Zhang; Daniele Fabris; Jeffrey T Mason; Timothy J O'Leary |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't Date: 2005-08-22 |
Journal Detail:
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Title: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society Volume: 54 ISSN: 0022-1554 ISO Abbreviation: J. Histochem. Cytochem. Publication Date: 2006 Mar |
Date Detail:
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Created Date: 2006-02-20 Completed Date: 2006-04-06 Revised Date: 2012-01-24 |
Medline Journal Info:
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Nlm Unique ID: 9815334 Medline TA: J Histochem Cytochem Country: United States |
Other Details:
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Languages: eng Pagination: 301-10 Citation Subset: IM |
Affiliation:
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Department of Biophysics, Armed Forces Institute of Pathology, Rockville, Maryland, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Acetals
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chemistry Adenosine Monophosphate / analogs & derivatives, chemistry Catalysis Chromatography, High Pressure Liquid Cross-Linking Reagents / chemistry Deoxyadenine Nucleotides / chemistry* Ethanol Fixatives / chemistry* Formaldehyde / chemistry* Fourier Analysis Paraffin Embedding Phosphates Solutions Spectrometry, Mass, Electrospray Ionization Spectrophotometry, Ultraviolet Tissue Fixation / methods* Water |
| Grant Support | |
ID/Acronym/Agency:
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1R21CA91227/CA/NCI NIH HHS; 1R33 CA107844/CA/NCI NIH HHS; R33 CA107844-02/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Acetals; 0/Cross-Linking Reagents; 0/Deoxyadenine Nucleotides; 0/Fixatives; 0/N(6)-ethoxymethyl-2'-deoxyadenosine 5'-monophosphate; 0/Phosphates; 0/Solutions; 50-00-0/Formaldehyde; 61-19-8/Adenosine Monophosphate; 64-17-5/Ethanol; 653-63-4/2'-deoxy-5'-adenosine monophosphate; 7732-18-5/Water |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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