Genomic regulation of Krüppel-like-factor family members by corticosteroid receptors in the rat brain

Clare L M Kennedy; Emily M Price; Karen R Mifsud; Silvia Salatino; Eshita Sharma; Simon Engledow; John Broxholme; Hannah M Goss; Johannes M H M Reul

doi:10.1016/j.ynstr.2023.100532

Genomic regulation of Krüppel-like-factor family members by corticosteroid receptors in the rat brain

Clare L M Kennedy, Emily M Price, Karen R Mifsud, Silvia Salatino, Eshita Sharma, Simon Engledow, John Broxholme, Hannah M Goss, Johannes M H M Reul^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

6 Citations (Scopus)

Abstract

Hippocampal mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs) mediate glucocorticoid hormone (GC) action in the hippocampus. These receptors bind to glucocorticoid responsive elements (GREs) within target genes, eliciting transcriptional effects in response to stress and circadian variation. Until recently, little was known about the genome-wide targets of hippocampal MRs and GRs under physiological conditions. Following on from our genome-wide MR and GR ChIP-seq and Ribo-Zero RNA-seq studies on rat hippocampus, we investigated the Krüppel-like factors (KLFs) as targets of MRs and GRs throughout the brain under circadian variation and after acute stress. In particular, Klf2, Klf9 and Klf15 are known to be stress and/or GC responsive and play a role in neurobiological processes including synaptic plasticity and neuronal differentiation. We found increased binding of MR and GR to GREs within Klf2, Klf9 and Klf15 in the hippocampus, amygdala, prefrontal cortex, and neocortex after acute stress and resulting from circadian variation, which was accompanied by upregulation of corresponding hnRNA and mRNA levels. Adrenalectomy abolished transcriptional upregulation of specific Klf genes. These results show that MRs and GRs regulate Klf gene expression throughout the brain following exposure to acute stress or in response to circadian variation, likely alongside other transcription factors.

Original language	English
Article number	100532
Journal	Neurobiology of Stress
Volume	23
Early online date	7 Mar 2023
DOIs	https://doi.org/10.1016/j.ynstr.2023.100532
Publication status	Published - 7 Mar 2023

Bibliographical note

Funding Information:
Our study also provides evidence for a more widespread action of the MR in the brain than expected in view of early radioligand binding and in vitro autoradiography studies that detected hardly any MRs in the amygdala (except the central amygdala), PFC, and neocortex (Reul and de Kloet, 1985, 1986; Reul et al., 1987). While our findings of MR ChIP-qPCR at Klf GREs do support the evidence that levels of MRs are highest in hippocampal regions, we detected comparable levels of MR binding to Klf GREs in the amygdala and substantial MR binding to Klf GREs in the PFC and neocortex, indicating a less restricted expression than previously thought. Therefore, although our studies do not demonstrate absolute expression levels, the immunoprecipitation of both receptors at Klf GREs does confirm their presence in these tissues.In contrast to Klf9, the role of Klf15 has hardly been studied in the brain in relation to GC hormone activity. In this study, enrichment levels of MRs and GRs at Klf15 GREs were slightly lower compared with levels seen at Klf2 and Klf9 GREs and, in contrast to the other Klfs, the induction of Klf15 RNA did not exceed 2-fold in any of the brain regions investigated. Klf15 expression in the rat brain has been shown to be relatively low in comparison to other tissues (Yu et al., 2014), possibly making it more difficult to detect, hence the relatively lower MR and GR enrichment and RNA expression observed. Our findings of expression and GC regulation of the Klf15 gene in the brain, however, support a role of this KLF in the CNS.This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC; Grant Numbers BB/K007408/1 and BB/N015045/1), a Wellcome Trust Neural Dynamics PhD studentship (to Clare L.M. Kennedy), and a BBSRC SWBio DTP PhD studentship (to Emily M. Price). We thank the Oxford Genomics Centre at the Wellcome Centre for Human Genetics (supported by the Wellcome Trust Core Award Grant Number 203141/Z/16/Z) for the generation and initial processing of the sequencing data.

Funding Information:
This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC ; Grant Numbers BB/K007408/1 and BB/N015045/1 ), a Wellcome Trust Neural Dynamics PhD studentship (to Clare L.M. Kennedy), and a BBSRC SWBio DTP PhD studentship (to Emily M. Price). We thank the Oxford Genomics Centre at the Wellcome Centre for Human Genetics (supported by the Wellcome Trust Core Award Grant Number 203141/Z/16/Z ) for the generation and initial processing of the sequencing data.

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© 2023 The Authors

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Role of corticosteroid receptor DNA binding in stress-induced hippocampal gene transcription in relation to glucocorticoid and behavioural responses
Mifsud, K. R. (Co-Principal Investigator) & Reul, J. M. H. M. (Principal Investigator)
1/10/16 → 31/08/20
Project: Research
Epigenetic control of gene transcriptional and behavioural responses to stress in the dentate gyrus
Reul, J. M. H. M. (Principal Investigator)
1/10/13 → 1/10/16
Project: Research

Cite this

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title = "Genomic regulation of Kr{\"u}ppel-like-factor family members by corticosteroid receptors in the rat brain",

abstract = "Hippocampal mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs) mediate glucocorticoid hormone (GC) action in the hippocampus. These receptors bind to glucocorticoid responsive elements (GREs) within target genes, eliciting transcriptional effects in response to stress and circadian variation. Until recently, little was known about the genome-wide targets of hippocampal MRs and GRs under physiological conditions. Following on from our genome-wide MR and GR ChIP-seq and Ribo-Zero RNA-seq studies on rat hippocampus, we investigated the Kr{\"u}ppel-like factors (KLFs) as targets of MRs and GRs throughout the brain under circadian variation and after acute stress. In particular, Klf2, Klf9 and Klf15 are known to be stress and/or GC responsive and play a role in neurobiological processes including synaptic plasticity and neuronal differentiation. We found increased binding of MR and GR to GREs within Klf2, Klf9 and Klf15 in the hippocampus, amygdala, prefrontal cortex, and neocortex after acute stress and resulting from circadian variation, which was accompanied by upregulation of corresponding hnRNA and mRNA levels. Adrenalectomy abolished transcriptional upregulation of specific Klf genes. These results show that MRs and GRs regulate Klf gene expression throughout the brain following exposure to acute stress or in response to circadian variation, likely alongside other transcription factors. ",

author = "Kennedy, {Clare L M} and Price, {Emily M} and Mifsud, {Karen R} and Silvia Salatino and Eshita Sharma and Simon Engledow and John Broxholme and Goss, {Hannah M} and Reul, {Johannes M H M}",

note = "Funding Information: Our study also provides evidence for a more widespread action of the MR in the brain than expected in view of early radioligand binding and in vitro autoradiography studies that detected hardly any MRs in the amygdala (except the central amygdala), PFC, and neocortex (Reul and de Kloet, 1985, 1986; Reul et al., 1987). While our findings of MR ChIP-qPCR at Klf GREs do support the evidence that levels of MRs are highest in hippocampal regions, we detected comparable levels of MR binding to Klf GREs in the amygdala and substantial MR binding to Klf GREs in the PFC and neocortex, indicating a less restricted expression than previously thought. Therefore, although our studies do not demonstrate absolute expression levels, the immunoprecipitation of both receptors at Klf GREs does confirm their presence in these tissues.In contrast to Klf9, the role of Klf15 has hardly been studied in the brain in relation to GC hormone activity. In this study, enrichment levels of MRs and GRs at Klf15 GREs were slightly lower compared with levels seen at Klf2 and Klf9 GREs and, in contrast to the other Klfs, the induction of Klf15 RNA did not exceed 2-fold in any of the brain regions investigated. Klf15 expression in the rat brain has been shown to be relatively low in comparison to other tissues (Yu et al., 2014), possibly making it more difficult to detect, hence the relatively lower MR and GR enrichment and RNA expression observed. Our findings of expression and GC regulation of the Klf15 gene in the brain, however, support a role of this KLF in the CNS.This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC; Grant Numbers BB/K007408/1 and BB/N015045/1), a Wellcome Trust Neural Dynamics PhD studentship (to Clare L.M. Kennedy), and a BBSRC SWBio DTP PhD studentship (to Emily M. Price). We thank the Oxford Genomics Centre at the Wellcome Centre for Human Genetics (supported by the Wellcome Trust Core Award Grant Number 203141/Z/16/Z) for the generation and initial processing of the sequencing data. Funding Information: This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC ; Grant Numbers BB/K007408/1 and BB/N015045/1 ), a Wellcome Trust Neural Dynamics PhD studentship (to Clare L.M. Kennedy), and a BBSRC SWBio DTP PhD studentship (to Emily M. Price). We thank the Oxford Genomics Centre at the Wellcome Centre for Human Genetics (supported by the Wellcome Trust Core Award Grant Number 203141/Z/16/Z ) for the generation and initial processing of the sequencing data. Publisher Copyright: {\textcopyright} 2023 The Authors",

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AU - Kennedy, Clare L M

AU - Price, Emily M

AU - Mifsud, Karen R

AU - Salatino, Silvia

AU - Sharma, Eshita

AU - Engledow, Simon

AU - Broxholme, John

AU - Goss, Hannah M

AU - Reul, Johannes M H M

N1 - Funding Information: Our study also provides evidence for a more widespread action of the MR in the brain than expected in view of early radioligand binding and in vitro autoradiography studies that detected hardly any MRs in the amygdala (except the central amygdala), PFC, and neocortex (Reul and de Kloet, 1985, 1986; Reul et al., 1987). While our findings of MR ChIP-qPCR at Klf GREs do support the evidence that levels of MRs are highest in hippocampal regions, we detected comparable levels of MR binding to Klf GREs in the amygdala and substantial MR binding to Klf GREs in the PFC and neocortex, indicating a less restricted expression than previously thought. Therefore, although our studies do not demonstrate absolute expression levels, the immunoprecipitation of both receptors at Klf GREs does confirm their presence in these tissues.In contrast to Klf9, the role of Klf15 has hardly been studied in the brain in relation to GC hormone activity. In this study, enrichment levels of MRs and GRs at Klf15 GREs were slightly lower compared with levels seen at Klf2 and Klf9 GREs and, in contrast to the other Klfs, the induction of Klf15 RNA did not exceed 2-fold in any of the brain regions investigated. Klf15 expression in the rat brain has been shown to be relatively low in comparison to other tissues (Yu et al., 2014), possibly making it more difficult to detect, hence the relatively lower MR and GR enrichment and RNA expression observed. Our findings of expression and GC regulation of the Klf15 gene in the brain, however, support a role of this KLF in the CNS.This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC; Grant Numbers BB/K007408/1 and BB/N015045/1), a Wellcome Trust Neural Dynamics PhD studentship (to Clare L.M. Kennedy), and a BBSRC SWBio DTP PhD studentship (to Emily M. Price). We thank the Oxford Genomics Centre at the Wellcome Centre for Human Genetics (supported by the Wellcome Trust Core Award Grant Number 203141/Z/16/Z) for the generation and initial processing of the sequencing data. Funding Information: This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC ; Grant Numbers BB/K007408/1 and BB/N015045/1 ), a Wellcome Trust Neural Dynamics PhD studentship (to Clare L.M. Kennedy), and a BBSRC SWBio DTP PhD studentship (to Emily M. Price). We thank the Oxford Genomics Centre at the Wellcome Centre for Human Genetics (supported by the Wellcome Trust Core Award Grant Number 203141/Z/16/Z ) for the generation and initial processing of the sequencing data. Publisher Copyright: © 2023 The Authors

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N2 - Hippocampal mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs) mediate glucocorticoid hormone (GC) action in the hippocampus. These receptors bind to glucocorticoid responsive elements (GREs) within target genes, eliciting transcriptional effects in response to stress and circadian variation. Until recently, little was known about the genome-wide targets of hippocampal MRs and GRs under physiological conditions. Following on from our genome-wide MR and GR ChIP-seq and Ribo-Zero RNA-seq studies on rat hippocampus, we investigated the Krüppel-like factors (KLFs) as targets of MRs and GRs throughout the brain under circadian variation and after acute stress. In particular, Klf2, Klf9 and Klf15 are known to be stress and/or GC responsive and play a role in neurobiological processes including synaptic plasticity and neuronal differentiation. We found increased binding of MR and GR to GREs within Klf2, Klf9 and Klf15 in the hippocampus, amygdala, prefrontal cortex, and neocortex after acute stress and resulting from circadian variation, which was accompanied by upregulation of corresponding hnRNA and mRNA levels. Adrenalectomy abolished transcriptional upregulation of specific Klf genes. These results show that MRs and GRs regulate Klf gene expression throughout the brain following exposure to acute stress or in response to circadian variation, likely alongside other transcription factors.

AB - Hippocampal mineralocorticoid receptors (MRs) and glucocorticoid receptors (GRs) mediate glucocorticoid hormone (GC) action in the hippocampus. These receptors bind to glucocorticoid responsive elements (GREs) within target genes, eliciting transcriptional effects in response to stress and circadian variation. Until recently, little was known about the genome-wide targets of hippocampal MRs and GRs under physiological conditions. Following on from our genome-wide MR and GR ChIP-seq and Ribo-Zero RNA-seq studies on rat hippocampus, we investigated the Krüppel-like factors (KLFs) as targets of MRs and GRs throughout the brain under circadian variation and after acute stress. In particular, Klf2, Klf9 and Klf15 are known to be stress and/or GC responsive and play a role in neurobiological processes including synaptic plasticity and neuronal differentiation. We found increased binding of MR and GR to GREs within Klf2, Klf9 and Klf15 in the hippocampus, amygdala, prefrontal cortex, and neocortex after acute stress and resulting from circadian variation, which was accompanied by upregulation of corresponding hnRNA and mRNA levels. Adrenalectomy abolished transcriptional upregulation of specific Klf genes. These results show that MRs and GRs regulate Klf gene expression throughout the brain following exposure to acute stress or in response to circadian variation, likely alongside other transcription factors.

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DO - 10.1016/j.ynstr.2023.100532

M3 - Article (Academic Journal)

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JO - Neurobiology of Stress

JF - Neurobiology of Stress

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ER -

Genomic regulation of Krüppel-like-factor family members by corticosteroid receptors in the rat brain

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Role of corticosteroid receptor DNA binding in stress-induced hippocampal gene transcription in relation to glucocorticoid and behavioural responses

Epigenetic control of gene transcriptional and behavioural responses to stress in the dentate gyrus

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