Crossing worms

Hello everyone,

We did some crossing with N2 males (stained with Mito tracker Red) and N2 hermaphrodite and then wait for 12 hrs for the worms to breed.
After 12hrs we dissect the hermaphrodite to take the eggs out and look under the microscopy.
I can see the millions of MT red sperm from males inside the hermaphrodite but they are not fertilizing the egg.
I crossed 3 young adults (even tried L4 staged worms) with 12 males and still we are not able to see the sperm fertilizing the egg.
I don’t know what is going on? Any suggestions would be really helpful


Deep in my brain, I have this feeling that maternal mitochondria win out, so you might not be seeing mito transfer because they aren’t. Are you getting male progeny out? Could you try another marker, like a histone GFP or some such in the males? Wait, maybe males don’t have histones, so that could be out. Tubulin GFP? Some other DNA binding protein? ama-1?


Hi Rakesh,
Check out this paper on the fate of paternal mitochondria:

Science. 2011 Nov 25;334(6059):1144-7. doi: 10.1126/science.1211878. Epub 2011 Oct 27.
Postfertilization autophagy of sperm organelles prevents paternal mitochondrial DNA transmission.
Al Rawi S, Louvet-Vallée S, Djeddi A, Sachse M, Culetto E, Hajjar C, Boyd L, Legouis R, Galy V


Can you use a centriole marker maybe? doi: 10.1016/j.cub.2013.06.043