AMPK Methods and Protocols

Chapter 36

Analysis of Transgenerational Phenotypes Following Acute

Starvation in AMPK-DeficientC.elegans

Emilie Demoinet and Richard Roy

Abstract

Environmental variation experienced early in life can result in long-term reproductive consequences. We
have recently identified an important role for AMPK in the prevention of transgenerational defects
following starvation of L1 stage larvae inC. elegans. Here we describe a means of analyzing these
transgenerational defects following a single exposure to energy stress during early larval development. We
also provide methods to quantify the histone modifications that are affected by this stress, along with the
resulting reproductive defects that arise in later generations.

Key wordsC. elegans, AMPK, L1 starvation, Transgenerational defects, Epigenetics, Chromatin,

Immunostaining

1 Introduction

Throughout evolution, the ability of organisms to adjust to envi-

ronmental challenges has been critical in the selection of adapta-

tions that enhance survival and/or fitness. Some of these

adaptations result in evolutionary trade-offs that nevertheless

improve survival in the generations of animals that were subjected

to a given challenge. Others fall into a class of short-term modifica-

tions of gene expression that affect survival of subsequent genera-

tions through epigenetic mechanisms.

These epigenetic changes have been observed downstream of

multiple developmental/physiological contexts and can have last-

ing multigenerational effects on gene expression affecting diverse

processes. In mammalian organisms these transgenerational epige-

netic phenomena can be correlated with changes that affect the

methylation state of the DNA and also in modifications of the

unstructured histone tails that comprise the overall chromatin

landscape [1–4]. Changes in DNA methylation have been identi-

fied between siblings born during periods of famine versus abun-

dance suggesting that these changes reflect epigenetic marks that

Dietbert Neumann and Benoit Viollet (eds.),AMPK:MethodsandProtocols, Methods in Molecular Biology, vol. 1732,
https://doi.org/10.1007/978-1-4939-7598-3_36,©Springer Science+Business Media, LLC 2018

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