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Real-time egg laying dynamics in Caenorhabditis elegans

Abstract

Caenorhabditis elegans (C. elegans) is a powerful model organism for aging studies due to its short generation time and the ease of performing genetic screens. Assaying egg-laying window and output in the worm complements prior lifespan studies because the effects of aging on dividing and non-diving tissues can be studied in tandem. However worms are routinely sterilized in multiday studies because separating a large number of adults from their progeny quickly becomes impractical.

We demonstrate a liquid culture system to deliver and remove food and pharmacological agents from individually housed worms. Worm reproduction is a sensitive indicator of health, so we show worms lay a comparable amount of eggs in our liquid culture system as on agar. Our fast environmental control allows us to withhold food for a short enough time to avoid matricidal hatching, but a long enough time to extend reproductive span. To solve biofilm issues within the system, we identified a biofilm-deficient strain of E. coli that the worms thrive in. This system counts progeny in real-time, measuring embryo size as well as allowing for embryo recovery from individual mothers for further developmental assays. Loaded worms can be examined for their whole lifespan, and they can be recovered to do fixation for morphological analysis or other molecular biology.

It is our hope that this system will allow reexamination of many lifespan extending factors

to see how their timed dosing affects reproduction.

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