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X, but not Y, Chromosomal Complement Contributes to Stroke Sensitivity in Aged Animals

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Abstract

Post-menopausal women become vulnerable to stroke and have poorer outcomes and higher mortality than age-matched men, and previous studies suggested that sex chromosomes play a vital role in mediating stroke sensitivity in the aged. It is unknown if this is due to effects of the X or Y chromosome. The present study used the XY* mouse model (with four genotypes: XX and XO gonadal females and XY and XXY gonadal males) to compare the effect of the X vs. Y chromosome compliment in stroke. Aged (18–20 months) and gonadectomized young (8–12 weeks) mice were subjected to a 60-min middle cerebral artery occlusion. Infarct volume and behavioral deficits were quantified 3 days after stroke. Microglial activation and infiltration of peripheral leukocytes in the aged ischemic brain were assessed by flow cytometry. Plasma inflammatory cytokine levels by ELISA, and brain expression of two X chromosome–linked genes, KDM6A and KDM5C by immunochemistry, were also examined. Both aged and young XX and XXY mice had worse stroke outcomes compared to XO and XY mice, respectively; however, the difference between XX vs. XXY and XO vs. XY aged mice was minimal. Mice with two copies of the X chromosome showed more robust microglial activation, higher brain-infiltrating leukocytes, elevated plasma cytokine levels, and enhanced co-localization of KDM6A and KDM5C with Iba1+ cells after stroke than mice with one X chromosome. The number of X chromosomes mediates stroke sensitivity in aged mice, which might be processed through the X chromosome–linked genes and the inflammatory responses.

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Abbreviations

ACA:

Anterior cerebral artery

CNS:

Central nervous system

CV:

Crystal violet

DiI:

1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate

ELISA:

Enzyme-linked immunosorbent assay

FC:

Flow cytometry

FCG:

Four core genotypes

FISH:

Fluorescence in situ hybridization

IHC:

Immunohistochemistry

IL-1β:

Interleukin-beta

iNOS:

Inducible NO synthase

KDM5C:

Lysine demethylase 5C

KDM6A:

Lysine-specific demethylase 6A

MCA:

Middle cerebral artery

MCAO:

Middle cerebral artery occlusion

NDS:

Neurological deficit scores

PCA:

Posterior cerebral artery

PFA:

Paraformaldehyde

TNFα:

Tumor necrosis factor alpha

XCI:

X chromosome inactivation

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Funding

This work was supported by funding from the National Institutes of Health: NS108779 (Fudong Liu).

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Author notes

  1. Shaohua Qi and Conelius Ngwa contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, McGovern Medical School, The University of Texas Health Science Center at Houston, McGovern Medical School, 6431 Fannin Street, Houston, TX, 77030, USA

    Shaohua Qi, Conelius Ngwa, Abdullah Al Mamun, Sharmeen Romana, Ting Wu, Sean P. Marrelli, Louise D. McCullough & Fudong Liu

  2. Department of Integrative Biology and Physiology, UCLA, 610 Charles Young Drive South, Los Angeles, CA, 90095, USA

    Arthur P. Arnold

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  1. Shaohua Qi
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Contributions

S.Q., L.D.M., A.P.A, and F.L. designed research; S.Q., A.A.M., C.N. and S.R. performed research; T.W. and S.P.M. contributed to new reagents and data analysis; S.Q. analyzed data; S.Q. and F.L. drafted the manuscript; L.D.M and A.P.A edited the manuscript.

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Correspondence to Fudong Liu.

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All animal experimental procedures were performed in accordance with NIH guidelines for the care and use of laboratory animals and were approved by the Institutional Animal care and use committee of the University of Texas Health Science Center.

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Qi, S., Ngwa, C., Al Mamun, A. et al. X, but not Y, Chromosomal Complement Contributes to Stroke Sensitivity in Aged Animals. Transl. Stroke Res. 14, 776–789 (2023). https://doi.org/10.1007/s12975-022-01070-z

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