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Oxytocin modulates markers of the unfolded protein response in Caco2BB gut cells

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Cell Stress and Chaperones Aims and scope

Abstract

We have shown that oxytocin receptor (OTR) expression in neonatal rat enterocytes is robust from birth to weaning, but OTR function during this period is unknown. We previously reported that oxytocin (OT) stimulation of Caco2BB cells (enterocytes in vitro) inhibits the mammalian target of rapamycin complex 1 (mTORC1) signaling. The unfolded protein response (UPR) is known to protectively reduce translation during endoplasmic reticulum (ER) stress. Because the mTORC1 pathway is linked to cellular stress, we investigated markers of UPR in OT-stimulated Caco2BB cells. We report that OT modulates several factors involved in sensing and translation of ER stress. High OT (62.5 nM) reduced translation initiation factor 4E-BP1 phosphorylation (Ser65), which is known to inhibit cap-dependent translation via its rate-limiting eukaryotic translation initiation factor 4E (eIF4E). Importantly, high OT increased phosphorylation of eukaryotic translation initiation factor 2a (eIF2a) phospho-Ser51, which inhibits eIF2a. High OT also increased protein kinase RNA-like endoplasmic reticulum kinase phosphorylation, a sensor of ER stress and a kinase of eIF2a. Both high and low OT activated inositol requiring enzyme1 (IRE1), which generates the transcription factor X-box binding protein 1 (XBP1) and induces the UPR. We also show that OT modulates XBP1 splicing and induces tribbles 3 (TRIB3; a negative regulator of Akt and protein involved in autophagy) and immunoglobulin binding protein (BiP; ER-chaperone). Taken together, these results indicate that OT modulates sensors of ER stress and autophagy. These findings support our hypothesis that transiently elevated OTR expression in neonatal gut may serve a protective function during a critical postnatal developmental period.

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Abbreviations

4E-BP1:

Translation initiation factor 4E-binding protein 1

ASD:

Autism spectrum disorder

ATF4:

Activated transcription factor 4

ATF6:

Activation transcription factor 6

BiP:

Immunoglobulin binding protein

CHOP:

C/EBP homology protein

eIF2a:

Eukaryotic translation initiation factor 2a

eIF4E:

Eukaryotic translation initiation factor 4E

FGM:

Fresh growth medium

IBD:

Irritable bowel disease

IRE1:

Inositol requiring enzyme 1

ER:

Endoplasmic reticulum

GADPH:

Glyceraldehyde-3-phosphate dehydrogenase

mTORC1:

Mammalian target of rapamycin complex 1

OT:

Oxytocin

OTR:

Oxytocin receptor

PDK1:

Phosphoinositide-dependent kinase-1

PERK:

Protein kinase RNA-like endoplasmic reticulum kinase

PI3K:

Phosphoinositol-3 kinase

PTEN:

Phosphatase and tensin homolog

TRIB3:

Tribbles 3

TSC2:

Tuberosclerosis protein

UPR:

Unfolded protein response

XBP1:

X-box binding protein 1

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Acknowledgments

We thank Chris Heger for Simon™ automated western blotting consultation and analysis, as well as with help in manuscript preparation. We thank Maurice Manning for supplying the oxytocin antagonist. The research was funded by the Einhorn Family Charitable Trust (UL1 RR024156).

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Authors and Affiliations

  1. Department of Psychiatry, Columbia University College of Physicians and Surgeons, New York, NY, 10032, USA

    Benjamin Y. Klein, Robert J. Ludwig & Martha G. Welch

  2. Department of Pathology & Cell Biology, Columbia University College of Physicians and Surgeons, New York, NY, 10032, USA

    Hadassah Tamir, David L. Hirschberg & Martha G. Welch

  3. Center for Infection and Immunology, Columbia University College of Physicians and Surgeons, New York, NY, 10032, USA

    David L. Hirschberg

  4. Division of Molecular Imaging and Neuropathology, New York State Psychiatric Institute, New York, NY, USA

    Hadassah Tamir

  5. EB Sciences, Oakland, CA, 94611, USA

    Sara B. Glickstein

  6. New York State Psychiatric Institute, 1051 Riverside Drive, Unit 40, New York, NY, 10032, USA

    Benjamin Y. Klein & Martha G. Welch

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  1. Benjamin Y. Klein
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  2. Hadassah Tamir
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Correspondence to Benjamin Y. Klein or Martha G. Welch.

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Klein, B.Y., Tamir, H., Hirschberg, D.L. et al. Oxytocin modulates markers of the unfolded protein response in Caco2BB gut cells. Cell Stress and Chaperones 19, 465–477 (2014). https://doi.org/10.1007/s12192-013-0473-4

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