Lipid-derived and other oxidative modifications of retinal proteins in a rat model of Smith-Lemli-Opitz syndrome

Becky Kapphahn, Michael J. Richards, Deborah A Ferrington, Steven J. Fliesler

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Oxidative modification of proteins can perturb their structure and function, often compromising cellular viability. Such modifications include lipid-derived adducts (e.g., 4-hydroxynonenal (HNE) and carboxyethylpyrrole (CEP)) as well as nitrotyrosine (NTyr). We compared the retinal proteome and levels of such modifications in the AY9944-treated rat model of Smith-Lemli-Opitz syndrome (SLOS), in comparison to age-matched controls. Retinas harvested at 3 months of age were either subjected to proteomic analysis or to immuno-slot blot analysis, the latter probing blots with antibodies raised against HNE, CEP, and NTyr, followed by quantitative densitometry. HNE modification of retinal proteins was markedly (>9-fold) higher in AY9944-treated rats compared to controls, whereas CEP modification was only modestly (≤2-fold) greater, and NTyr modification was minimal and exhibited no difference as a function of AY9944 treatment. Anti-HNE immunoreactivity was greatest in the plexiform and ganglion cell layers, but also present in the RPE, choroid, and photoreceptor outer segment layer in AY9944-treated rats; control retinas showed minimal HNE labeling. 1D-PAGE/Western blot analysis of rod outer segment (ROS) membranes revealed HNE modification of both opsin and β-transducin. Proteomic analysis revealed the differential expression of several retinal proteins as a consequence of AY9944 treatment. Upregulated proteins included those involved in chaperone/protein folding, oxidative and cellular stress responses, transcriptional regulation, and energy production. βA3/A1 Crystallin, which has a role in regulation of lysosomal acidification, was down-regulated. Hence, oxidative modification of retinal proteins occurs in the SLOS rat model, in addition to the previously described oxidation of lipids. The results are discussed in the context of the histological and physiological changes that occur in the retina in the SLOS rat model.

Original languageEnglish (US)
Pages (from-to)247-254
Number of pages8
JournalExperimental Eye Research
Volume178
DOIs
StatePublished - Jan 1 2019

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trans-1,4-Bis(2-chlorobenzaminomethyl)cyclohexane Dihydrochloride
Smith-Lemli-Opitz Syndrome
Lipids
Retina
Proteins
Proteomics
Transducin
Opsins
Rod Cell Outer Segment
Crystallins
Choroid
Densitometry
Protein Folding
Proteome
Ganglia
Oxidative Stress
Western Blotting
Membranes
Antibodies

Keywords

  • Cholesterol
  • Oxidation
  • Oxidative stress
  • Protein modification
  • Retina
  • Smith-Lemli-Opitz

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

Cite this

Lipid-derived and other oxidative modifications of retinal proteins in a rat model of Smith-Lemli-Opitz syndrome. / Kapphahn, Becky; Richards, Michael J.; Ferrington, Deborah A; Fliesler, Steven J.

In: Experimental Eye Research, Vol. 178, 01.01.2019, p. 247-254.

Research output: Contribution to journalArticle

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