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Hematogenous Macrophages Contribute to Fibrotic Scar Formation After Optic Nerve Crush

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Abstract

Although glial scar formation has been extensively studied after optic nerve injury, the existence and characteristics of traumatic optic nerve fibrotic scar formation have not been previously characterized. Recent evidence suggests infiltrating macrophages are involved in pathological processes after optic nerve crush (ONC), but their role in fibrotic scar formation is unknown. Using wild-type and transgenic mouse models with optic nerve crush injury, we show that macrophages infiltrate and associate with fibroblasts in the traumatic optic nerve lesion fibrotic scar. We dissected the role of hematogenous and resident macrophages, labeled with Dil liposomes intravenously administered, and observed that hematogenous macrophages (Dil+ cells) specifically accumulate in the center of traumatic fibrotic scar while Iba-1+ cells reside predominantly at the margins of optic nerve fibrotic scar. Depletion of hematogenous macrophages results in reduced fibroblast density and decreased extracellular matrix deposition within the fibrotic scar area following ONC. However, retinal ganglion cell degeneration and function loss after optic nerve crush remain unaffected after hematogenous macrophage depletion. We present new and previously not characterized evidence that hematogenous macrophages are selectively recruited into the fibrotic core of the optic nerve crush site and critical for this fibrotic scar formation.

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Abbreviations

CD3:

Cluster of differentiation 3

CD68:

Cluster of differentiation 68

CNS:

Central nervous system

GCL:

Ganglion cell layer

GFP:

Green fluorescent protein

GFAP:

Glial fibrillary acidic protein

Iba-1:

Ionized calcium binding adaptor molecule 1

IL-6:

Interleukin-6

IL-13:

Interleukin-13

ONC:

Optic nerve crush

PBS:

Phosphate buffered saline

PDGF:

Platelet derived growth factor

P-ERG:

Pattern electroretinogram

PFA:

Paraformaldehyde

RBPMS:

RNA-binding protein with multiple splicing

RGC:

Retinal ganglion cells

SCI:

Spinal cord injury

TGF-β1:

Transforming growth factor-beta1

Tmem119:

Transmembrane protein 119

TON:

Traumatic optic neuropathy

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Funding

This work was supported by the National Institutes of Health Center Core Grant (P30EY014801) and an unrestricted Research to Prevent Blindness Unrestricted Grant to the Bascom Palmer Eye Institute. R.K.L. is partially supported by the Walter G. Ross Foundation. This work was partly supported by the Gutierrez Family Research Fund and the Camiener Foundation Glaucoma Research Fund.

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Author notes

  1. Huiyi Jin and Yuan Liu contributed equally to this work.

Authors and Affiliations

  1. Department of Ophthalmology, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, 33136, USA

    Huiyi Jin, Yuan Liu, Xiangxiang Liu, Mohamed M. Khodeiry, Jae K. Lee & Richard K. Lee

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  1. Huiyi Jin
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  2. Yuan Liu
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  3. Xiangxiang Liu
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Contributions

RK conceived and designed the research project. HJ, YL, and XL performed the experimental protocols. HJ and YL did the data collection. HJ, MM, and XL analyzed the data. HJ, RK, and JK wrote and revised the paper. RK obtained the funding and supervised the study. All authors have read and approved the final manuscript for publication.

Corresponding author

Correspondence to Richard K. Lee.

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All animals were treated in compliance with the guidelines from the Association for Research in Vision and Ophthalmology (ARVO) Statement for the Use of Animals in Ophthalmic and Vision Research. All animal procedures were performed in accordance with the ethical standards of the institution or practice of the University of Miami Institutional Animal Care and Use Committee and NIH guidelines. No human participant was involved in this study.

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The authors declare no competing interests.

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Jin, H., Liu, Y., Liu, X. et al. Hematogenous Macrophages Contribute to Fibrotic Scar Formation After Optic Nerve Crush. Mol Neurobiol 59, 7393–7403 (2022). https://doi.org/10.1007/s12035-022-03052-6

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