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Reactive Fibroblasts in Response to Optic Nerve Crush Injury

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Abstract

Traumatic optic neuropathy leads to bidirectional degeneration of retinal ganglion cells and axons and results in optic nerve scaring, which inhibits the regeneration of damaged axons. Compared with its glial counterpart, the fibrotic response causing nerve scar tissue is poorly permissive to axonal regeneration. Using collagen1α1-GFP reporter mice, we characterize the development of fibrotic scar formation following optic nerve crush injury. We observe that perivascular collagen1α1 cells constitute a major cellular component of the fibrotic scar. We demonstrate that extracellular molecules and monocytes are key factors contributing to the pathogenesis of optic nerve fibrotic scar formation, with a previously unrecognized encapsulation of this scar. We also characterize the distribution of collagen1α1 cells in the retina after optic nerve crush injury based on in vivo and whole-mount retinal imaging. Our results identify collagen1α1 cells as a major component of fibrotic scarring following ONC and are a potential molecular target for promoting axonal regeneration after optic nerve injury.

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Funding

The Bascom Palmer Eye Institute is supported by the NIH Center Core Grant P30EY014801 and a Research to Prevent Blindness Unrestricted Grant. R.K. Lee is supported by the Walter G. Ross Foundation. This work was partly supported by the Gutierrez Family Research Fund.

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Author notes

  1. Xiangxiang Liu and Yuan Liu contributed equally to this work.

Authors and Affiliations

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

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

  2. Beijing Tongren Eye Center, Beijing Tongren Hospital, Beijing Eye Institute, Capital Medical University, Beijing, China

    Xiangxiang Liu & Ningli Wang

  3. Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China

    Huiyi Jin

  4. Department of Ophthalmology, Research Institute of Ophthalmology, Giza, 12557, Egypt

    Mohamed M. Khodeiry

  5. School of Life Science, Nanjing University, Nanjing, 210023, Jiangsu, China

    Weizheng Kong

  6. Miami Project to Cure Paralysis, Department of Neurological Surgery, University of Miami Miller School of Medicine, Miami, FL, 33136, USA

    Jae K. Lee

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  1. Xiangxiang Liu
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Supplemental Fig. 1

Time course of GS distribution throughout the ON following ONC injury. GS negative area was marked with blue dotted line. The fibrotic scar was marked with yellow dotted line. d-i At 3 dpi and 7 dpi, a large area with absence of GS expression was observed with a faint border (marked with blue dotted lines), while accumulation of Col1α1+-GFP expressing cells were present in the ONC crush site. j-o By 10 dpi and 14 dpi, a clearly demarcated GS deficient area was observed at the ONC lesion site with fibrotic components expressed within the area. Scale bar, 200 μm. (PNG 2941 kb)

High Resolution Image (TIF 12110 kb)

Supplemental Fig. 2

Time course of vimentin distribution throughout the ON following ONC injury. Lesion area stained by vimentin was marked with blue dotted line, the fibrotic scar was marked with white dotted line. a-c In the uninjured optic nerve, an even distribution of fibrillary patterned vimentin was present throughout the ON. d-f At 3 dpi, a faint border of vimentin deficient area was observed (marked with blue dotted line) with residual vimentin expression within the lesion area, while infiltrating Col1α1+-GFP positive cells began to appear in the scar center. g-i At 7 dpi, a number of vimentin expressing cells together with Col1α1+-GFP cells were observed at the center of the lesion site, but vimentin and Col1α1+-GFP expressing cells are entirely not co-localized. j-o By 10 dpi and 14 dpi, as the Col1α1+-GFP cells form an encapsulated fibrotic scar (k, n), a cluster of high density vimentin expressing cells are present at the fibrotic scar area. Scale bar, 200 μm. (PNG 2454 kb)

High Resolution Image (TIF 11297 kb)

Supplemental Fig. 3

Distribution of F4/80 in the mature fibrotic scar. F4/80+ cells accumulate at the ONC site acutely and reach a peak within the ONC fibrotic scar region by 10 dpi, some F4/80+ cells are also expressed around the fibrotic scar, Col1α1+ cells do not co-express F4/80. (PNG 2066 kb)

High Resolution Image (TIF 5981 kb)

Supplemental Fig. 4

Time course of Col1α1+-GFP (green), CD13 (red) and PDGFR-ß (magenta) expression in the whole-mount retina images. a-e Col1α1+-GFP expressing cells decreased significantly between 10dpi to 14dpi. CD13 (f-j) and PDGFR-ß expressing cells (k-o) did not show significant changes following ONC injury. (PNG 2119 kb)

High Resolution Image (TIF 8977 kb)

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Liu, X., Liu, Y., Jin, H. et al. Reactive Fibroblasts in Response to Optic Nerve Crush Injury. Mol Neurobiol 58, 1392–1403 (2021). https://doi.org/10.1007/s12035-020-02199-4

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