Embryonic heat conditioning in chicks induces transgenerational heat/immunological resilience via methylation on regulatory elements

Tali Rosenberg, Asaf Marco, Tatiana Kisliouk, Amit Haron, Dmitry Shinder, Shelly Druyan, Noam Meiri

Research output: Contribution to journalArticlepeer-review

1 Scopus citations


The question of whether behavioral traits are heritable is under debate. An obstacle in demonstrating transgenerational inheritance in mammals originates from the maternal environment's effect on offspring phenotype. Here, we used in ovo embryonic heat conditioning (EHC) of first-generation chicks, demonstrating heredity of both heat and immunological resilience, confirmed by a reduced fibril response in their untreated offspring to either heat or LPS challenge. Concordantly, transcriptome analysis confirmed that EHC induces changes in gene expression in the anterior preoptic hypothalamus (APH) that contribute to these phenotypes in the offspring. To study the association between epigenetic mechanisms and trait heritability, DNA-methylation patterns in the APH of offspring of control versus EHC fathers were evaluated. Genome-wide analysis revealed thousands of differentially methylated sites (DMSs), which were highly enriched in enhancers and CCCTC-binding factor (CTCF) sites. Overlap analysis revealed 110 differentially expressed genes that were associated with altered methylation, predominantly on enhancers. Gene-ontology analysis shows pathways associated with immune response, chaperone-mediated protein folding, and stress response. For the proof of concept, we focused on HSP25 and SOCS3, modulators of heat and immune responses, respectively. Chromosome conformational capture (3C) assay identified interactions between their promoters and methylated enhancers, with the strongest frequency on CTCF binding sites. Furthermore, gene expression corresponded with the differential methylation patterns, and presented increased CTCF binding in both hyper- and hypomethylated DMSs. Collectively, we demonstrate that EHC induces transgenerational thermal and immunological resilience traits. We propose that one of the mechanisms underlying inheritance depends on three-dimensional (3D) chromatin reorganization.

Original languageEnglish
Article numbere22406
JournalFASEB Journal
Issue number7
StatePublished - Jul 2022
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2022 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.


We are grateful to the Volcani Center's chicken farm staff for their dedicated work. We would like to thank Rotem Raz for her guidance and support in using the Agricultural Research Organization's core facilities for bioinformatics. This research was supported by Grant No. 2020663 from the United States‐Israel Binational Science Foundation (BSF) and by Grant No. 2041923 from the United States National Science Foundation (NSF).

FundersFunder number
National Science Foundation
United States-Israel Binational Science Foundation2041923


    • cross-tolerance
    • embryonic heat conditioning
    • epigenetics
    • heredity
    • hypothalamus
    • stress


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