Looks like our liquid culture protocol still needs some adjusting! First, the concentration of worms in the culture needs to be significantly increased to make plating them easier, so I’m hoping reducing the volume of the culture and increasing the amount of eggs added will solve this problem. Second, even with the small number of worms I was able to look at, there were hardly any dauer. Hannah thinks reducing the bacteria in the culture might help by preventing the response to the ascarosides from being reduced by the high availability of food. We really only need enough bacteria to keep the worms from arresting at L1. I’m going to give it a try and see if we get better results!
In the meantime I’ll be doing some trials for one of Renee’s experiments while she’s on vacation. Earlier on, did some mating assays for her which looked to see if male response to hermaphrodites or location of the vulva was impaired by any of the neuronal mutations she was looking at. This time around, I’ll be doing leaving assays which are similar in design. I’ll be working with tra-2 mutant males, which means they have pan-neural feminization. Any sex-specific roles of the neurons should be reversed, which may impact/reverse some sex-specific behaviors.When placed on a food source, males are known to be more likely to take excursions from the food or even leave it completely (presumably looking for a mate), where hermaphrodites are more likely to remain happily on the food. Tra-2 mutant males are known to have the leaving behavior of a hermaphrodite. I’ll be observing the leaving behavior of a few worms with additional mutations to see if any of these mutants will behave more like wild type males. This will help us take a look at which genes are involved in sex-specific leaving behavior.
Until next time! 🙂