A Research Design for the Quantification of the Neuropeptides Substance P and Calcitonin Gene-Related Peptide in Rat Skin Using Western Blot Analysis

A Research Design for the Quantification of the Neuropeptides Substance P and Calcitonin Gene-Related Peptide in Rat Skin Using Western Blot Analysis

Author Lapin, Guilherme Abbud Franco Autor UNIFESP Google Scholar
Hochman, Bernardo Autor UNIFESP Google Scholar
Nishioka, Michele Akemi Autor UNIFESP Google Scholar
Maximino, Jessica Ruivo Autor UNIFESP Google Scholar
Chadi, Gerson Autor UNIFESP Google Scholar
Ferreira, Lydia Masako Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
IB CNPq Brazilian Natl Council Sci & Technol Dev
Abstract OBJECTIVE: To describe and standardize a protocol that overcomes the technical limitations of Western blot (WB) analysis in the quantification of the neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) following nociceptive stimuli in rat skin.DESIGN: Male Wistar rats (Rattus norvegicus albinus) weighing 250 to 350 g were used in this study. Elements of WB analysis were adapted by using specific manipulation of samples, repeated cycles of freezing and thawing, more thorough maceration, and a more potent homogenizer; increasing lytic reagents; promoting greater inhibition of protease activity; and using polyvinylidene fluoride membranes as transfer means for skin-specific protein. Other changes were also made to adapt the WB analysis to a rat model.SETTING: University research center.MAIN OUTCOME MEASURE: Western blot analysis adapted to a rat model.RESULTS: This research design has proven effective in collecting and preparing skin samples to quantify SP and CGRP using WB analysis in rat skin.CONCLUSION: This study described a research design that uses WB analysis as a reproducible, technically accessible, and cost-effective method for the quantification of SP and CGRP in rat skin that overcomes technical biases.
Keywords neuropeptides
substance P
calcitonin gene-related peptide
Western blot analysis
Language English
Date 2015-06-01
Published in Advances In Skin & Wound Care. Philadelphia: Lippincott Williams & Wilkins, v. 28, n. 6, p. 259-265, 2015.
ISSN 1527-7941 (Sherpa/Romeo, impact factor)
Publisher Lippincott Williams & Wilkins
Extent 259-265
Origin http://dx.doi.org/10.1097/01.ASW.0000465373.42350.c1
Access rights Closed access
Type Article
Web of Science ID WOS:000354885100004
URI http://repositorio.unifesp.br/11600/42125

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