When we dissected our mice after treatment we found that the skeletal muscle in the treatment group were 'dry' and 'less fleshy' as compared to placebo.The mice were otherwise active and healthy when they were alive. Does anyone have any ideas on why this would be.?
I believe we need more details in order to attempt to answer your question. One, what was the treatment? How long was the treatment? What was the effect of the treatment on food and water intake? Two, how was the muscle tissue processed? Describe from euthansia on. Three, what muscles were excised? Four, what does "less fleshy" mean? Lower muscle mass?
Thanks,
Gordon Warren
The mice (n=8) were treated with intraperitoneal metformin for 16 days. They lost significant weight. They were killed by giving general anesthesia and decapitation. Their food and water intake was comparable to placebo group. their cholestrol was raised significantly. The muscle was gastrocnemius muscle. we havent processed it yet. its frozen at -80. but when we were dissecting it out, it had a fibrous feel to it. as compared to palcebo group (n=8). i am sorry i dont know how else to describe it. The muscles were just dry-ish. muscle mass was not visibly reduced as comapred to palcebo
If I were you, I would want to confirm your perceptions of dryness or "less fleshy". To do so, you could look at the ratios of dry weight to wet weight or protein content to wet weight. I don't know what your plans are for the muscle tissue but the latter ratio would be doable if you plan to do protein work.
i wanted to see genetic expression various genes by real time pcr. could you send me some information about the technique you are mentioning. May be you can guide me?
Measuring muscle dry weight or total protein content is not compatible with doing gene expression qPCR. About all you can do is to obtain wet weights immediately prior to isolating RNA. If the treated muscles are truly "dry" or "less fleshy", one would expect their wet weights to be less. If this turns out to be the case, in future experiments, you could plan to measure dry weight or total protein content in addition to wet weight. In a "dry" muscle, I would expect the ratio of dry to wet weight to be elevated. Same goes for the ratio of total protein content to wet weight.
Ok thanks. i will look these things up. (i have no idea yet what is dry weight.) also, i will be processing some of it for western blot. Do you think theres anything i can find out in that?
No Dr Mahdy there isnt. Hence i am interested in somehow quantifying my findings which were unexpected for me
we just took gastrocnemius mucle sample, we didn't really notice others. but i think they were all affected.
I am not familiar with muscle, but there are water-binding proteins f.ex. in cartilage. Perhaps you can measure the gene expression of water-binding proteins which are expressed in muscle tissue.
Melanie thats a great idea. could you send me some links of papers to familiarize myself with these proteins
https://www.ncbi.nlm.nih.gov/pubmed/27578148
This is one publication about perlecan, a protein involved in proteoglycan synthesis. It is related to cartilage, but perhaps you can also find some links to muscle in pubmed.
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