Data di Pubblicazione:
2006
Citazione:
Comparative proteomic profile of sciatic nerve and muscle tissue in old and young rats / D. Capitanio, M. Vasso, M. Moriggi, A. Viganò, S. De Palma, M. Ripamonti, V. Magnaghi, C. Gelfi, D. Capitanio. ((Intervento presentato al 1. convegno ItPA congress tenutosi a Pisa nel 2006.
Abstract:
Using proteomics, we investigated the temporal expression profiles of proteins in rat sciatic nerve in
course of aging by 2D-DIGE and mass spectrometry. The differential proteomic expression was
compared with that of the surrounding muscle tissue.
The proteomic investigation was performed by DIGE and mass spectrometry on sciatic nerves and
muscles of 4 rats 8 and 24 month-old, respectively. Ground frozen nerves and muscles were
solubilized, centrifuged and quantitated. Protein labelling with cyanine dyes (Cy3 and Cy5) was
performed according to the manufacturer's instructions (GE Healthcare). These protocols suggest an
internal standard be run on all gels within an experiment. The internal standard is created by
pooling an aliquot of all biological samples in the experiment in order to warrant that every protein
from all samples will be represented. Each sample was labelled with Cy5 dye and internal standard
with Cy3 dye (400 pmol dye/50 ug sample). After labelling, protein samples were mixed with an
equal volume of 2X sample buffer (Urea 8M, DTT 130 mM, CHAPS 4% and IPG buffer pH 3-10
2%).Gels were scanned directly between low fluorescence plates using a Typhoon 9200 laser
scanner (GE Healthcare) according to the manufacturer's recommendations (532 and 633 nm laser;
emission filters of 520 and 670 nm).Determination of protein spot abundance was performed using
DeCyder software (version 5.0, GE Healthcare).A standard 2D map of sciatic nerve was defined
and compared to a standard map of rat skeletal muscle. Qualitative comparison shows a low degree
of similarity in proteome expression between nerve and muscle tissues. Nevertheless, we can
identify some common proteins. Proteins differentially expressed in sciatic nerve and in muscle
tissue in course of aging were characterized by MALDI TOF and ESI MS/MS.Quantitative
differential expression provides up and down regulated proteins in common between the two
tissues. Differences in protein expression identified during aging in sciatic nerve were retained also
in muscle samples indicating the muscle could be a possible target for neurodegenerative studies.
Tipologia IRIS:
14 - Intervento a convegno non pubblicato
Elenco autori:
D. Capitanio, M. Vasso, M. Moriggi, A. Viganò, S. De Palma, M. Ripamonti, V. Magnaghi, C. Gelfi, D. Capitanio
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