June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
RNA sequencing of transcriptome analyses of juvenile rabbit cornea and iris/ciliary body following lensectomy with intraocular lens insertion
Author Affiliations & Notes
  • Jonathon Young
    Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Ross F Collery
    Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
    Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Christine Skumatz
    Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Iris S Kassem
    Ophthalmology & Visual Sciences, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
    Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, Wisconsin, United States
  • Footnotes
    Commercial Relationships   Jonathon Young, None; Ross Collery, None; Christine Skumatz, None; Iris Kassem, None
  • Footnotes
    Support  NIH K08EY024645
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 5002. doi:
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      Jonathon Young, Ross F Collery, Christine Skumatz, Iris S Kassem; RNA sequencing of transcriptome analyses of juvenile rabbit cornea and iris/ciliary body following lensectomy with intraocular lens insertion. Invest. Ophthalmol. Vis. Sci. 2020;61(7):5002.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Juvenile rabbits have robust postoperative inflammation and fibrin formation after lensectomy with intraocular lens (IOL) insertion, similar to children. The underlying cause of this response remains unknown. Previous work using proteomics revealed increased abundance of coagulation and complement proteins in the aqueous humor following surgery. This project evaluated changes in the transcriptome for inflammatory and coagulation-related genes in the cornea and iris/ciliary body (CB) in response to lensectomy with primary IOL.

Methods : All experiments were approved with the Institutional Animal Care Committee of MCW. Under anesthesia with isoflurane, 3 juvenile New Zealand White rabbits had unilateral clear-cornea lens extraction surgery with insertion of an acrylic IOL (Alcon). Following clinical examination on day 3, the cornea and iris/CB were dissected, homogenized with bead beater disruption, RNA extracted using the RNeasy kit (Qiagen), and transcriptomes profiled. Equivalent tissues from 3 eyes from surgically naïve rabbits served as controls. Gene expression values were classified by biological process using Gene Set Enrichment Analysis (GSEA) public database.

Results : The highest enrichment scores for biological processes comparing operated versus unoperated eyes revealed significant changes in genes associated with vasculature development and the inflammatory response (Figure 1). Individual genes related to coagulation and inflammation were also evaluated. Fragments per kilobase million were significantly increased for Complement C3 and complement factor H following surgery in the cornea (6.51 and 3.69 fold increase respectively, p < 0.05), but not the iris/CB. Transforming Growth Factor Beta-2 expression decreased in the iris/CB (2.16 fold decrease, p<0.05), but not in the cornea.

Conclusions : The majority of genes related to the processes of inflammation, cytokine production, and the response to wounding are significantly enriched in the cornea and iris/CB tissues following surgery, and correlate with proteomic analysis of aqueous humor. A variety of gene expression level changes differ in the cornea versus the iris/CB and may provide insight into the source of aqueous humor proteomic differences in response to juvenile lensectomy.

This is a 2020 ARVO Annual Meeting abstract.

 

Ranked normalized enrichment scores from GSEA analyses, all p-values and false discovery rates <0.001.

Ranked normalized enrichment scores from GSEA analyses, all p-values and false discovery rates <0.001.

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