Lapin GA, Hochman B, Nishioka MA, Maximino JR, Chadi G, Ferreira LM, et al.
Advances in skin & wound care. Date of publication 2015 Jun 1;volume 28(6):259-65.
1. Adv Skin Wound Care. 2015 Jun;28(6):259-65. doi:
10.1097/01.ASW.0000465373.42350.c1.
A research design for the quantification of the neuropeptides substance p and
calcitonin gene-related Peptide in rat skin using Western blot analysis.
Lapin GA(1), Hochman B, Nishioka MA, Maximino JR, Chadi G, Ferreira LM.
Author information:
(1)Guilherme Abbud Franco Lapin, MD, is a graduate student in the Division of
Plastic Surgery, Universidade Federal de São Paulo (UNIFESP), São Paulo, SP,
Brazil. Bernardo Hochman, MD, PhD, is an Affiliated Professor in the Division of
Plastic Surgery, UNIFESP, São Paulo, SP, Brazil. Michele Akemi Nishioka, FT, MS,
is a graduate student in the Division of Plastic Surgery, UNIFESP, São Paulo, SP,
Brazil. Jessica Ruivo Maximino, PhD, is a Neuroscience Researcher, UNIFESP, São
Paulo, SP, Brazil. Gerson Chadi, MD, PhD, is a Full Professor in the Experimental
Neurology Department of UNIFESP, head of the Laboratory of Pathophysiology-LIM45
in the Neurosurgery Department of UNIFESP, and Coordinator of the Study Group on
Neuroregeneration, also at UNIFESP, São Paulo, SP, Brazil. Lydia Masako Ferreira,
MD, PhD, is a Full Professor in the Plastic Surgery Division at UNIFESP, São
Paulo, SP, Brazil; Researcher IB CNPq (Brazilian National Council for Scientific
and Technological Development); and Assistant Coordinator of MED III CAPES
(Coordination for the Improvement of Higher Education).
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.
DOI: 10.1097/01.ASW.0000465373.42350.c1
PMID: 25988735 [Indexed for MEDLINE]
