What is the cross bridge stiffness for non-muscle myosin II (NMM II) ?

I'm standardizing the protocol for isolating muscle fiber, but am having trouble adjusting the time I leave the muscle in the solution of type I collagenase 0,2%, my muscle is being degraded. Another question is whether during the period of incubation in collagenase solution I should leave stirring or not?

In case of mTOR how many of those can be there in a muscle cell, and can there be opposite pathways in one muscle cell or compartiments?

Could a muscle cell have for instance activation of mTORs at certain places and inhibition at others?

What about compartimentalization?

For instance a part where there are mitochondria and catabolic organelles and parts were more anabolic work is done, for instance near myofibrils?

Can high carbohydrate ingestion lead to diffusional problems around the sarcolemma due to increased subsarcolemmal glycogen granules?

Can it lead to storage of glucose in the form of glycogen which would be less functional and cause diffusional problems around the sarcolemma, or lead to specific concentration gradient, which would blunt normal muscle cell functioning?

Can isolated glycogen granules occur in the sarcoplasm which are less functional, and while they lead to increase of muscle cell volume (form of hypertrophy) they are less functional?

Can there be an increase in amount of granules, and/or would you expect glycogen granules to be filled in more tiers, or not?

In length:

I assume a sarcomere gets stretched, this can be to a very long extend. This sarcomere is not functioning anymore very optimal. The muscle will, I guess form a cytoskeleton, with z-disk, and alfa-actin, and to this there will be added myosin and actin. The sarcomere can be added, I think, at the end or beginning, near the tendon. However, it can also happen inside the muscle fiber, near the middle (?) At least I reckon that there is stretch and lengthening there also.

In parallel:

I assume a sarcomere get microdamaged some myofilaments will have structural damage and this is restored and probably made stronger, so the myofilaments will enlarge in volume.

However, is this in parallel adding?

It is claimed the vastus lateralis will have a increased pennation angle, and more sarcomeres are added in parallel. However, how does this work? I assume there will be an adding of sarcomeres at some parts of the muscle fiber, so there would not be an parallel structure, as we see in books. It will be more like this : __|-|___|-|___

Is there any literature about this?

A lot of thanks.

Nitrogen oxide (NO) and creatine interact in skeletal muscle cells. So is supplement of creatine and NO helpfull?

When my biceps contracts, and imagine all my slow twitch motor units are involved. They are actively contracting. The muscle fibers of the MU type I's are divided through the muscle within fascicles.

What is happening at the same time with the MF connected to fast twitch MU's. Are they also overlapping at the sacromere level, and creating potentially cross bridges? Are they not changing [sacromeres stay at same length], but are structures like titin, z-disks, and connective tissues altered in length. Is both the case?

In most books or articles I found contraction on the sarcomere level, and the MF level, but not in relationship through the whole muscle.

Very much appreciated.

I'm comparing two muscle types using subtractive hybridization in order to evaluate differentially expressed genes and the protocols I'm looking at say to perform a second self-subtraction. Im not sure what they mean by self-subtraction and to me it seems counterintuitive to isolating differentially expressed sequences.

what is reliable and valid method of measuring hamstring muscle length?

objectively.

as tool used in research to measure the outcome of interventions used for increasing hamstring length or flexibility.