Document Detail


Technical report: laser microdissection and pressure catapulting is superior to conventional manual dissection for isolating pure spiral ganglion fractions from the cochlea.
MedLine Citation:
PMID:  17980526     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Isolating cells from the cochlea to perform molecular biology assessment presents a challenge, because it is not possible to dissect pure cell pools by conventional methods. Thus, we set out to demonstrate that laser microdissection and pressure catapulting (LMPC) is superior to conventional manual cochlea dissection for this purpose. Spiral ganglions (SG) were isolated from neonatal rat cochleae by manual dissection and LMPC. Also, modioli were manually dissected. Total RNA was isolated from all three cell pools. In order to demonstrate contamination of the dissected cell pool, we determined the expression of type II iodothyronine deiodinase (D2), claudin 11 (Cld-11), neurofilament light chain (NF-L) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) transcripts by RT-PCR. The results showed that LMPC is not only a suitable method for selectively dissecting cochlear tissues, but in addition the molecular markers confirmed pure spiral ganglion cell pools without indication for any contamination by other cells. This indicates that LMPC is capable of providing a pure SG cell pool in contrast to conventional manual dissection. Therefore, LMPC presents a new technique for cochlear tissue separation improving the validity of molecular biological studies of the inner ear.
Authors:
Attila Torkos; Kirsten Wissel; Athanasia Warnecke; Thomas Lenarz; Timo Stöver
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-09-29
Journal Detail:
Title:  Hearing research     Volume:  235     ISSN:  0378-5955     ISO Abbreviation:  Hear. Res.     Publication Date:  2008 Jan 
Date Detail:
Created Date:  2007-12-25     Completed Date:  2008-03-06     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7900445     Medline TA:  Hear Res     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  8-14     Citation Subset:  IM    
Affiliation:
Department of Otolaryngology, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
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MeSH Terms
Descriptor/Qualifier:
Animals
Animals, Newborn
Cell Separation / methods*
Cochlea / chemistry*,  cytology,  enzymology
Frozen Sections
Glyceraldehyde-3-Phosphate Dehydrogenases / analysis
Iodide Peroxidase / analysis
Lasers*
Microdissection*
Nerve Tissue Proteins / analysis
Neurofilament Proteins / analysis
Phenotype
Pressure
RNA, Messenger / analysis*
Rats
Rats, Sprague-Dawley
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Spiral Ganglion / chemistry*,  enzymology
Chemical
Reg. No./Substance:
0/Cldn11 protein, rat; 0/Nerve Tissue Proteins; 0/Neurofilament Proteins; 0/RNA, Messenger; 0/neurofilament protein L; EC 1.11.1.-/iodothyronine deiodinase type II; EC 1.11.1.8/Iodide Peroxidase; EC 1.2.1.-/Glyceraldehyde-3-Phosphate Dehydrogenases

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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