Transfecting neurons with GFP-tagged DNA - (Sep/15/2006 )

Hey guys,

I have transfected some neuronal cultures with GFP-tagged DNA yesterday but the expression after nearly 24h is too low for the experiment I was planning on doing today. Is it possible that the GFP expression would increase over the week-end?
Also, I treated those neurons with AP-5 at the end of my transfection, could that have anything to do with the low GFP expression? And will the AP-5 have a long-term effect or any bad effect on my neurons if I keep them for longer?

Thanks a lot for your replies!

We used to c an increase in GFP fluorescence from 24 to 48 hrs.

The GFP fluorescence could increase over the weekend. Also its possible that it doesnt increase as much as u expect it to. It depends on the promoter and since u have GFP tagged, this could also alter fluorescence.

how did u transfect ur neuronal cultures ? If u want high expression of ur protein within a day, u need to use SFV or AAV-1 (viruses).

What is AP-5 ?

I did a lipofection (it´s the technique used in my lab). Viral transfections sounds good but I hear that viruses for transfections are difficult to make or to get so I don´t think I am gonna be able to use it anytime soon.

AP-5 is a NMDA receptor blocker.

I tried lipofection once with neuronal cultures, the efficiency was bad so dropped the idea.

But i still think u will c an increase during the next 24 hrs. For some promoters, GFP fluorescence increases till 4 days after transfection and then stabilises.