Fluorescent L1 and L2 worms

Model Organism: Worms Scientific Discussion
1

As nematodes (strain NW1229) develop from the L1 to L2 stages, will all their neurons tagged with GFP be visible?

2

The neurons in NW1229 are visible even in the embryo, and continue to be visible throughout larval life. However, neurons that are born in larvae (or embryos) may not be visible with GFP until a couple of hours after birth, since it takes time for the GFP to be made and accumulate enough to see.

Hope this is useful,

Janet

3

Thank you

4

May i know if anyone has performed experiment for the GFP half-life in vivo (in c. elegans, for example?) – like how long time needed for it to express etc (although, yah, largely will be depending onto the nature of the target gene to be studied)? or how long time will it needed for the GFP to fade off (if the gene tagged with is being suppresses?)

just thought of that suddenly…

5

I did a bit of searching and found a reported half-life of 26 hours for GFP (in mammalian cells). If you want a shorter-lived GFP, there are several worms papers that use GFP-PEST (with a half-life of 1 hour).

Using a heat shock promoter, a review by Hobert & Loria reports a delay of 30 minutes between heat shock and detectable GFP.

Janet

6

Thanks Janet :smiley:

Yah, i think that should be taken into consideration when doing fluorescent assay, so that not to under/over-estimate the real scene? because what we are measuring (GFP intensity) might not reflect the actual expression at that time point (could be the fading GFP from previous time point etc…)