I want to acutely prevent budding of vesicles from the Golgi. Does anyone know of a good way to do this? I know that Brefeldin a works after 1 hour, but I want to know whether I can treat them for shorter time...
What concentration of BFA you are using and which cells you are talking about? I have seen the Golgi effect of BFA after 10mcg/ml, when Golgi gets disorganized. I am pretty ensure there no any exocytosis due to trans-Golgi collapse too.
Thanks. I use 1 microgram/ml as I read this prevented exocytosis in hepatocytes. I use MDCK cells and I want to do acute studies, so I want to treat them for as short time as possible. So I want to know how rapidly it prevents exocytosis.
Well, never worked with that concentration, but to the best of my knowledge, BFA was considered initially as a specific inhibitor of exocytosis. So, I would recommend use any exocytosis marker at the early time of treatment – 3 min, and later. I am confident, even at that dosage you should see some effect. Also, I would simultaneously monitor Golgi structure by Giantin or TGN46 proteins. Good luck.
Hello Emma,
We used a drug called EXO-1 (Tocris) to prevent release of glutamate from retinal bipolar cells. We added it at 50 microM to the solution in the recording pipette. It did the job within seconds, but of course we applied intracellularly (Tooker et al, 2013, JNeurosci).
Nonetheless, it is stated that it prevents vesicular traffic between Golgi and ER, somewhat similar to Brefeldin (Feng et al, 2003, PNAS).
You may want to look into it.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line