Effects of gene therapy on muscle 18S rRNA expression in mouse model of ALS.
BACKGROUND
The efficiency of gene therapy experiments is frequently evaluated by measuring the impact of the treatment on the expression of genes of interest by quantitative real time PCR (qRT-PCR) and by normalizing these values to those of housekeeping (HK) genes constitutively expressed throughout the experiment. The objective of this work was to study the effects of muscle gene therapy on the expression of 18 S ribosomal RNA (Rn18S), a commonly used HK gene.


RESULTS
Mouse model of motor neuron disease (SOD1-G93A) was injected intramuscularly with Brain-derived neurotrophic factor (BDNF-TTC) encoding or control naked DNA plasmids. qRT-PCR expression analysis was performed for BDNF and HK genes Rn18 S, glyceraldehyde-3-phosphate dehydrogenase (Gapdh) and β-actin (Actb). We report that elevated BDNF expression in the injected muscle was accompanied with increased Rn18 S expression, whereas Gapdh and Actb were not affected. Increased "ribosomal output" upon BDNF stimulation was supported by increased steady-state levels of ribosomal protein mRNAs.


CONCLUSIONS
Ribosomal RNA transcription may be directly stimulated by administration of trophic factors. Caution should be taken in using Rn18 S as a HK gene in experiments where muscle metabolism is likely to be altered by therapeutic intervention.
LAGENBIO-I3A, Veterinary Faculty, Aragon Institute of Health Sciences (IACS), Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain. osta@unizar.es.
María
M
Moreno-Igoa
Raquel
R
Manzano
Sara
S
Oliván
Ana C
AC
Calvo
Janne M
JM
Toivonen
Rosario
R
Osta