Expression and localization of proteins in tissue determines anatomy and physiology, and thus provides a clue to pathology. However, it is strenuous to identify the expression and localization of multiple targets using conventional immunostaining technique, especially when there is no specific antibody, nor explicit assumption, for target proteins. Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry is a proteomic technique to simultaneously map multiple targets in a frozen tissue section. Using this technique, we analyzed protein expression in heart, and in testis as a reference sample, in 129/sv mice. Spatial distribution of at least 57 individual proteins was simultaneously identified at 100 um resolution. Among them, cardiac alpha actin, cardiac troponin T, tropomyosin alpha-1 chain, titin, myosin light chain 1 (MYL1), myosin light chain 3 (MYL3), myosin-6 were exclusively expressed in the heart. MYL1 and MYL3, the altered expression of which is associated with heart failure, were localized in the atrium and in the ventricle, respectively, as expected. Interestingly, protein expression profile of cysteine-rich protein 2 and myomesin-1 is exactly as predicted from mRNA expression, namely more in the heart and less in the testis. Expression level of housekeeping genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta actin, was similar between the heart and the testis. These results suggest that MALDI imaging may provide a new platform for analysis of pathology and subsequent application to diagnosis of heart diseases.
Kanchipuram Escorts 🥰 8617370543 Call Girls Offer VIP Hot Girls
Simultaneous mapping of multiple proteins in heart using matrix-assisted laser desorption/ionization imaging mass spectrometry
1. Simultaneous mapping of multiple proteins
in heart using matrix-assisted laser
desorption/ionization imaging mass
spectrometry
Ken Takahashi
Department of Cardiovascular Physiology
Graduate School of Medicine, Dentistry and Pharmaceutical Sciences
Okayama University, Japan
2. Background
• Expression and localization of proteins in
tissue determines anatomy and physiology,
and thus provides a clue to pathology.
• However, it is strenuous to identify the
expression and localization of multiple targets
using conventional immunostaining technique.
Expression and localization of proteins in tissue determines anatomy and physiology, and thus provides a clue to pathology.
However, it is strenuous to identify the expression and localization of multiple targets using conventional immunostaining technique, especially when there is no specific antibody, nor explicit assumption, for target proteins.
Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry is a proteomic technique to simultaneously map multiple targets in a frozen tissue section.
Using this technique, we analyzed protein expression in heart, and in testis as a reference sample, in 129/sv mice.
Spatial distribution of at least 64 individual proteins was simultaneously identified at 100 um resolution.
Mass spectrometry (MS) has been successfully applied to study microbiological colonies (1-3), plants (4), insects (5), vertebrates including whole animals (6-8), human cells (9,10)
and tissues (11,12).
This method allows the analysis of proteins (13,14), peptides (15,16), lipids or phospholipids (6,17-19), carbohydrates or glycoconjugates (20) and exogenous or endogenous small molecules, especially molecules involved in drug metabolism (21-27).
Among the identified proteins, cardiac alpha actin, cardiac troponin T, tropomyosin alpha-1 chain, titin, myosin light chain 1 (MYL1), myosin light chain 3 (MYL3), myosin-6 were exclusively expressed in the heart.
MYL1 and MYL3, the altered expression of which is associated with heart failure, were localized in the atrium and in the ventricle, respectively, as expected.
Interestingly, protein expression profile of cysteine-rich protein 2 and myomesin-1 is exactly as predicted from mRNA expression, namely more in the heart and less in the testis.
Expression level of housekeeping genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta actin, was similar between the heart and the testis.
These results suggest that MALDI imaging may provide a new platform for analysis of pathology and subsequent application to diagnosis of heart diseases.
This work was supported by JSPS KAKENHI Grant Number JP16K01356.
**