Can anyone please help me translate it in simple words ?
Parkin is recruited selectively to depolarized mitochondria and directs mitophagy. HeLa cells transfected with HA-Parkin were treated with CCCP for the indicated times( there was a figure in which times were indicated). Mitochondria were stained by anti-TOM20 and a mitochondrial membrane potential-dependent Mitotracker. Parkin was stained with anti-HA. Without treatment mitochondria are intact and stained by both mitochondrial markers
please help , i am not a biology student but I have been asked to read a Nature paper , these are lines from that paper
wow , you are good :) after that he says ..
After 2 hours of CCCP treatment, mitochondria are depolarized as shown by the loss of mitochondrial staining. After 24hr of CCCP treatment, massive loss of mitochondria is observed as shown by the disappearance of the mitochondrial marker. Only parkin positive cells show mitochondrial clustering and clearance, in contrast to transfected cells.
here what is
depolarization of mitochondria , parkin positive and mitochondrial clustering and clearance ??
you can also download this paper from http://www.nature.com/ncb/journal/v12/n2/abs/ncb20...
I think when you see the figures it would be much easy for you .. thank you thank you thank you so much for your help ::)
- RyanLv 48 years agoFavorite Answer
I don't know what parts specifically you don't understand but I'll try to help you out.
1.) HeLa cells are a widely used line of cells for cell culture
2.) HA-Parkin is probably a combination protein made of Parkin linked to hemagluttinin from influenza virus.
3.) CCCP- I had to look this one up but this is carbonyl cyanide m-chlorophenyl hydrazone. It seems to prevent oxidative phosphorylation.
4.) anti-TOM20 is some type of antibody directed toward the protein TOM20. I don't know what this protein is but it's probably a surface marker specific for mitochondria
5.) Mitotracker is a fluorescent probe to label live mitochondria
So basically it looks like the researchers are testing the effect of CCCP on mitochondrial health. Mitophagy literally translates to "eating of mitochondria". Cells cycle through their organelles to eliminate old and/or functionless ones.
Mitochondria were observed using TOM20 and antibodies directed toward it and by Mitotracker, which is a commercial fluorescent tracker that diffuses into live mitochondria. The researchers state that without the treatment, stated as CCCP, the mitochondria are intact and stained by both markers, TOM20 and mitotracker. So without CCCP the organelles are healthy.Source(s): B.S. in Microbiology