Assessment of necroptosis in the retina in a repeated primary ocular blast injury mouse model

Research output: Contribution to journalArticle


  • Ella Courtie
  • Alexandra Bernardo-Colón
  • Gareth Essex
  • Tonia S Rex
  • Richard J Blanch

Colleges, School and Institutes

External organisations

  • Neuroscience and Ophthalmology, Institute of Inflammation and Ageing, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
  • Institute for Inflammation and Ageing
  • School of Biomedical Sciences, Monash University
  • Institute of Clinical Sciences
  • Department of Ophthalmology
  • Departments of Cardiology, Sandwell and West Birmingham Hospitals NHS Trust and University Hospitals Birmingham NHS Foundation Trust, Birmingham, UK.
  • Vanderbilt University Medical Center
  • University of Birmingham Microbiome Treatment Centre, University of Birmingham, Birmingham, United Kingdom; Department of Gastroenterology, University Hospital Birmingham, Birmingham, United Kingdom. Electronic address:
  • Academic Department of Military Surgery and Trauma (ADMST)


Primary blast injury (caused by the initial rapid increase in pressure following an explosive blast) to the retina and optic nerve (ON) causes progressive visual loss and neurodegeneration. Military personnel are exposed to multiple low-overpressure blast waves, which may be in quick succession, such as during breacher training or in combat. We investigated the necroptotic cell death pathway in the retina in a mouse repeated primary ocular blast injury (rPBI) model using immunohistochemistry. We further evaluated whether intravitreal injections of a potent necroptosis inhibitor, Necrostatin-1s (Nec-1s), protects the retina and ON axons by retinal ganglion cells (RGC) counts, ON axonal counting and optical coherence tomography (OCT) analysis of vitreous haze. Receptor interacting protein kinase (RIPK) 3, increased in the inner plexiform layer 2 days post injury (dpi) and persisted until 14 dpi, whilst RIPK1 protein expression did not change after injury. The number of degenerating ON axons was increased at 28 dpi but there was no evidence of a reduction in the number of intact ON axons or RNA-binding protein with multiple splicing (RBPMS)+ RGC in the retina by 28 dpi in animals not receiving any intravitreal injections. But, when intravitreal injections (vehicle or Nec-1s) were given there was a significant reduction in RBPMS+ RGC numbers, suggesting that rPBI with intraocular injections is damaging to RGC. There were fewer RGC lost after Nec-1s than vehicle injection, but there was no effect of Nec-1s or vehicle treatment on the number of degenerating axons. OCT analysis demonstrated no effect of rPBI on vitreous haze, but intravitreal injection combined with rPBI increased vitreous haze (P = 0.004). Whilst necroptosis may be an active cell death signalling pathway after rPBI, its inhibition did not prevent cell death, and intravitreal injections in combination with rPBI increased vitreous inflammation and reduced RBPMS+ RGC numbers, implying intravitreal injection is not an ideal method for drug delivery after rPBI.

Bibliographic note

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Original languageEnglish
Pages (from-to)108102
JournalExperimental Eye Research
Publication statusE-pub ahead of print - 6 Jun 2020