We have undertaken a systematic study of the progression of atrophy following denervation in mouse skeletal muscles: adult CD1 mice were denervated by unilaterally cutting the sciatic nerve; 1, 3, 7 and 14 days after denervation animals were sacrificed and Soleus, EDL and Tibialis anterior (TA) from denervated and controlateral leg were dissected out and utilized for physiological experiments or for gene expression studies. Target cDNAs from TA muscles were hybridized on a dedicated cDNA microarray of muscle genes. Seventy genes were found differentially expressed. Of those, many are constituents of the muscle fibers or are involved in energy production through the oxidative metabolism. Real-time PCR was carried out to confirm the microarray results and to monitor the expression of a few important genes that were absent on our array. Electrophoresis and Western blot showed changes in myofibrillar protein expression, partially confirming changes in gene expression. Functional studies showed an increase in twitch tension, a prolongation in twitch time parameters and reduction in fatigue in denervated muscles. Calcium sensitivity of skinned fibres increased. Denervation caused also an increase of resting membrane potential with a reduction of action potential amplitude.

Physiological changes and temporal expression profiling in mouse fast skeletal muscles following denervation

RAFFAELLO, ANNA;LAVEDER, PAOLO;ROMUALDI, CHIARA;TONIOLO, LUANA;GERMINARIO, ELENA;DANIELI, DANIELA;MEGIGHIAN, ARAM;REGGIANI, CARLO;LANFRANCHI, GEROLAMO
2004

Abstract

We have undertaken a systematic study of the progression of atrophy following denervation in mouse skeletal muscles: adult CD1 mice were denervated by unilaterally cutting the sciatic nerve; 1, 3, 7 and 14 days after denervation animals were sacrificed and Soleus, EDL and Tibialis anterior (TA) from denervated and controlateral leg were dissected out and utilized for physiological experiments or for gene expression studies. Target cDNAs from TA muscles were hybridized on a dedicated cDNA microarray of muscle genes. Seventy genes were found differentially expressed. Of those, many are constituents of the muscle fibers or are involved in energy production through the oxidative metabolism. Real-time PCR was carried out to confirm the microarray results and to monitor the expression of a few important genes that were absent on our array. Electrophoresis and Western blot showed changes in myofibrillar protein expression, partially confirming changes in gene expression. Functional studies showed an increase in twitch tension, a prolongation in twitch time parameters and reduction in fatigue in denervated muscles. Calcium sensitivity of skinned fibres increased. Denervation caused also an increase of resting membrane potential with a reduction of action potential amplitude.
JOURNAL OF MUSCLE RESEARCH AND CELL MOTILITY
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11577/1343749
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