unloading adaptation experimental models of decreased use – (immobilization) – (hindlimb...
TRANSCRIPT
Unloading Adaptation• Experimental models of decreased use
– (Immobilization)– (Hindlimb suspension)– Denervation– Spinal isolation
• Factors contributing to atrophy• Clinical consequences of immobilization
Denervation• Nerve transection
– Remove coordinated descending input– Potential mobility in surrounding muscles
• Repair processes– Nerve regrowth: Same fibers? Same junction?– Muscle-derived signals?
• Muscle remodeling– Inactivityatrophy– Neuromuscular junction remodeling
Degeneration-Regeneration• Initial insult
– Reduced protein synthesis/Elevated degradation– Fiber deconstruction/death
• Recovery– SC activation– Restored
protein syn
• Reinnervation– Fiber reorg– Relative
hypertrophy
Goldspink, 1976
Degradation/mg
Synthesis/mg
Degradation/muscle
Synthesis/muscle
Schwann CellAxon
Synaptic cleft: Primary Secondary
Control 1 Week
3 Weeks (reinnervation
Axon dies rapidly, Schwann cell & ECM remain.Secondary synaptic clefts shrink & separate
Saito & Zacks, 1969
Muscle wasting• Myofiber size
decrease• Connective tissue
hypertrophy• Adipocyte invasion
Soleus, denervated 7 months
Soleus, denervated 7 weeks
Adipocytes
Myofiber degeneration• Dramatic loss of myofibrils & myofibril order
Soleus structure after 21 days denervation (Tomanek & Lund, 1973)
Fiber-type specific• Fast Fibers, esp in fast muscle, degenerate• Mass & function preserved
by electrical stim
Niederle & Mayr, 1978 Dow & al., 2004
Mechanisms of degeneration• Increased proteolysis
– Increase MuRF/MAFbx & proteasome– Increase cathepsins– Decrease PGC-1a
• Reduced metabolic capacity– Decrease glycolysis (LDH, PK, triose isomerase)– Decrease ETC (NADH, malate dehydrogenase, ATP
synthase)
• Increase ECM– Collagen, fibronectin, fibrillin
Regeneration
Borisov & al., 2001
Laminin NCAMEmbMHCNew, small myofibers develop either as discrete structures outside the basal lamina (left), or as separate appendages inside the BL (right)
Regeneration
SlowMHC(mature fiber)
EmbMHC(regenerating fiber)
Laminin(fiber boundaries)
Borisov & al., 2001
Three relatively mature fibers with faint laminin boundaries within thicker laminin shell of (presumably) original fiber
Small, immature (EmbMHC+) fiber adjacent to (presumably) preserved original fiber
Reinnervation• Muscle-nerve match• Axon-fiber not matched• Loss of contractile specialization
– MU innervation ratio– Fiber size:phenotype
Motor Unit territories before & after reinnrvation (Bodine-Fowler & al 1993)
Twitch contraction records contralateral and reinnervated LG & Sol (Gillespie & al. 1986)
Electrical stim preserves morphology• Rat EDL, 2 mos; 200x 0.2 s @100 Hz/day
Kostrominova & al., 2005
Gene expression altered by ES• Degen/Regen
– AML1NCAM– Myogenin/MRF4/MyoD– Reduced by ES
• Myosin– Den: IIbIIa– Stim: IIaIIb
Kostrominova & al., 2005
Electrical stimulation of denervated muscle
• Neural cell adhesion molecule– Normal: only NMJ nuclei– Denervated: all nuclei
• Potential benefits– Increased ‘receptivity’ of muscle– Increase axonal branching/guidance
Normal Denervated Denervated+ES
NCAM influences nerve growth• Culture neurons on muscle
slices• Processes follow cell surface• Greater growth on denervated
(high NCAM)
Covault &al., 1987
NeuronNCAM
Axon growth stops on NCAM plaques
Electrical stimulation of damaged nerve
• Low intensity; no force• Retrograde transmission of AP• Improves reinnervation
Al-Majed & al., 2000
Denervation summary• Degeneration-Regeneration
– Increased protein degradation and synthesis– “Moderating” of phenotype (IIIa; IIbIIa)– Loss of mass and order– Loss of myonuclear specialization (NMJ)
• Reinnervation– Usually original MEP– Muscle-specific, not fiber-specific– Disrupts Size Principle– Loss of proprioception
Spinal Isolation• Transect spinal cord
– Proximal to muscle of interest: no descending input– Distal: no ascending reflex
• Transect dorsal roots– Sensory– Reduce reflex hyperactivity
• Muscle inactive, nerve intact• Spinal cord injury model
Hyatt & al., 2003
MU properties post-SI
FF-Pre
FF-Post
FR-Pre
FR-Post
Slower,Less sag,Less force,Larger Tw/Tet
Physiological Response to SI• Grossly similar to denervation
– Slow muscle fast– Fast muscle slow
• Moderating of metabolic processes– Lower SDH in slow muscles– Higher GPDH in slow muscles
• Inactive muscles revert to a ‘neutral’ phenotype
SI response is weaker than denveration• Rate and extent of mass/force decline lower• Upregulation of MRFs lower & shorter
Hyatt & al., 2003
Tibialis Anterior Medial Gastrocnemius
• Less SC activationin SI than DEN
DAPI (nucleus)M-Cadherin (SC)BrDU (DNA synthesis)
Spinal isolation summary• Limited Degeneration-Regeneration
– “Moderating” of phenotype (IIIa; IIbIIa)– Loss of mass, but structure is preserved
• Spinal neurons don’t repair
Training and spinal transection• Careful training, tapering weight support
– Spontaneous weight support(standing)
– Treadmill-assisted leg motion(stepping)
Belanger & al., 1996
Post-mortem spinal cord, showing complete lesion
Pre/post step postures
0
500
1000
1500
2000
2500
Po
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
Mass
Roy & al., 1998
Summary• Muscle wasting program: active degeneration
– FOXOMuRF/Atrogin-1– Proteasome proteins (ubiquitin, S26)– Autophagy proteins (cathepsin)
• Decreased metabolic capacity– Mitochondrial apoptosis– Reduced PGC-1a
• Loss of fiber type specialization• Atrophy is its own program, separate from
absence of hypertrophy