Muscle: molecular and cellular basis of contraction Flashcards
What do the Z lines define?
determine where the sarcomere is
How do actin and myosin generate force?
Move relative to one another
How to identify the A band?
It appears dark and light cannot penetrate through it easily, head of myosin sticking out there is no clear passageway for light
- anisotropic band
How to identify the I band?
Z line in the middle, just contains actin which are a thin filament arranged in a very regular spatial pattern
- isotropic as light can pass through it
What is the M line?
Middle of the myosin area
What is nebulin?
Essentially a molecular ruler that sets the length of the actin filaments, starts at Z disc
What is titin?
Starts as Z disc but goes as far as the M line, forms a measurement for the length of a sarcomere
How to define fibre type of muscle?
Particular isoform of myosin that it contains
What is the sliding filament theory of muscle contraction?
Z discs get closer to each other when the muscle contracts, when the muscle is stimulated and contracts
- achieved by myosin heads moving actin filaments towards the centre towards the M line
In relaxed state Z discs are fairly far apart
Why is force generation dependant on filament overlap?
In the optimal case, we have substantial overlap, but not too much of the actin in the myosin and that produces OPTIMAL RESTING LENGTH, can produce the most force
As the actin is pulled away from the myosin, the amount of force that can be generated falls until the point where the actin and myosin donβt overlap at all, zero ability to produce force
If the actin is driven too far into the myosin, we also get a drop off in force, particularly if the actin overlaps, force production is essentially nil because the structure cannot function as it should
What is the excitation-contraction coupling?
- Stimulus produces action potential moves along to motor end plate. Where the motor neurone branches and the branches connect with muscle fibre, specialised membrane formation forming the motor end plate
- Terminal of motor neurone contains lots of mitochondria and vesicles with neurotransmitter (Ach)
- When a stimulus reaches the end of the motor neurone causes an opening of the Ca2+ channels, allowing Ca2+ to fuse with the edge of the terminal releases Ach into synaptic cleft and binds to Ach receptors on muscle membrane
- Binding causes depolarisation stimulates action potential which travels along the sarcolemma through a system of tubules in the endoplasmic reticulum pockets
How does calcium released from the sarcoplasmic reticulum aid muscle contraction?
As action potential spreads across sarcolema it goes down T tubules and triggers the opening of voltage gated Ca2+ channel (dihydroperidine receptor DHP), allows Ca2+ to enter the sarcoplasm of muscle fibres
- triggers opening in hydnodinc receptors that are present within the temrinal cisternae of sarcoplasmic reticulum
Ca2+ ATPases in sacroplasmic reticulum that withdraw Ca2+ from sacroplasm and allow muscle to go back to its resting state
What proteins are involved in muscle contraction?
Actin filamet = π - helical arrangement of actin monomers
- Tropomyosin
- Tropnin complex:
- TnT - binds tropomyosin
- TnC - binds calcium
- TnI - inhibits tropomyosin
How are myosin binding sites exposed?
Ca2+ binds to troponin C, tropomyosin moves exposing myosin binding sites
What is the cross bridge cycle?
- Without ATP, crossbridge is bound strongly to actin
- ATP binds to myosin causing crossbridge to detach and moves the position of the head
- ATP is hydrolysed into ADP and Pi which remain bound
- In the presence of actin, ADP and Pi are displayed and the power stroke occurs
What are the muscle fibre types?
Type I
Type IIa
Type IIx(b)
What are features of Type I muscle fibres?
Slow oxidative (SO) High myoglobin Small diameter Many capillaries Used when tension is needed for a prolonged period Soleus muscle
What are features of Type IIa muscle fibres?
Fast oxidative glycolytic (FOG)
Intermediate
Quadriceps are a fast muscle
What are features of Type IIx (b) muscle fibres?
Fast Glycolytic (FG)
Low myoglobin
Large diameter
Few capillaries
What are four major cellular functional compartments?
- Membrane excitation
- Excitation - contraction coupling
- Contraction
- Energy supply
How to identify fibre type? (Classically)
Reveal ATPase of the myosin head, incubate with appropriate solutions @ different pH, could identify type I or IIa
How to identify fibre type? (Now)
Use antibodies that recognise specific myosin isoforms
- fast v slow myosin
What are advantages of myosin immuno-staining?
Co-expression
Labelling lasts
Post mortem
What is the triple labelling system for multiple monoclonal antibodies?
By labelling each antibody with a different flurophore/colour reagent, can identify variety of different myosin isoforms
- fast = red
- slow - green
- neonatal = pink
Some fibres express more than one type of myosin, seen in disease or where there has been a training effect that might have altered the balance of myosin isoforms
