• Title and Introduction
• Methods
• Statistical Analysis of Change in Gene Expression in Denervated Muscle (Table 1)
• Changes in expression of Denervated vs Innervated Muscle (Graph 1)
• Acetylcholine Receptors are Up-Regulated in Denervated Muscle (Graph 2)
• Fold Change of Denervated over Innervated Muscle (Table 2)
• Aquaporin-4 is Strongly Down-Regulated in Denervated Muscle (Graph 3)
• No Significant Changes are observed in the Dystroglycan Protein Complex (Graph 4)
• Conclusions
• Werle Lab Home

GENE ARRAY OF FOUR DAY DENERVATED MOUSE GASTROCNEMIUS MUSCLE

M.J. Werle¹, K. Brownback¹, M.N. VanSaun¹, M. Rodova¹, S.R. Svojanovsky<sup>2

1</sup>Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City , KS , USA
²Dept. of Molecular & Integrative Physiology, University of Kansas Medical Center, Kansas City , KS , USA

Introduction

In order to observe changes in the distribution of gene expression in mouse muscles following denervation we employed the following protocol: Three male C57/Bl6 mice (60 days old) were anesthetized and the right sciatic nerve was sectioned at mid thigh. After 4 days the mice were sacrificed, and total RNA was isolated from the gastrocnemius muscles from the denervated side; the contralateral innervated side served as a control. Probes from the RNA were generated and applied to Affymetrix gene chip arrays (Chips MG_U74Av2, MG_U74Bv2 and MG_U74Cv2). The Affymetrix gene chips covered approximately 36,000 mouse genes and ESTs. Affymetrix data were transferred from Affymetrix Microarray Suite (MAS) into Data Mining Tool (DMT) via MicroDB. Genes were first identified as being present in the samples. The genes that were present were then analyzed by comparing the signal in each denervated muscle to the contralateral control signal from the same mouse. Thus, we obtained the fold increase in signal from each mouse. The data was statistically analyzed by T-test with an alpha of 0.10 and a replicate of 3. This analysis resulted in the identification of 3,423transcripts which were increased and 3,559 which were decreased. Our results compared well with published results on the upregulation of the AChR alpha subunit. We found a 25 fold, 24 fold and 15 fold increase in AChR alpha subunit in our three mice (Average 20.82 fold; n=3). Other AChR subunits were also increased, and the variation from mouse to mouse was remarkably low. The entire data set for these experiments are available for download at http://www2.kumc.edu/mrrc/kbrin/KBC_Links.html . In conclusion, there are dramatic changes in gene expression four days following denervation in the mouse gastrocnemius muscle and these results are available for your analysis.

Methods >>

 

For more information about this study, contact Michael J. Werle, mwerle@kumc.edu, (913) 588-7491.