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Related Experiment Video

Updated: Jun 8, 2025

Efficient Dissection and Culture of Primary Mouse Retinal Pigment Epithelial Cells
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Prominin-1 Knockdown Causes RPE Degeneration in a Mouse Model.

Sujoy Bhattacharya1, Tzushan Sharon Yang2, Bretton P Nabit2

  • 1Department of Ophthalmology and Visual Sciences, Vanderbilt University Medical Center, Nashville, TN 37232, USA.

Cells
|November 8, 2024
PubMed
Summary

Prominin-1 (Prom1) is crucial for retinal pigment epithelial (RPE) cell health. Disrupting Prom1 in mice caused RPE defects and vision loss, suggesting new therapeutic targets for atrophic AMD.

Keywords:
adeno-associated virus (AAV2/1)atrophic age-related macular degenerationgeographic atrophylysosomal pathwaysmicrogliamitochondria

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Area of Science:

  • Ophthalmology
  • Cell Biology
  • Genetics

Background:

  • Atrophic age-related macular degeneration (aAMD) involves retinal pigment epithelial (RPE) cell loss with no current effective treatments.
  • Prominin-1 (Prom1), a photoreceptor structural protein, has an emerging, yet unclear, role in RPE physiology and disease.
  • Pathogenic Prom1 mutations are linked to macular diseases, but its broader function in RPE homeostasis requires in vivo investigation.

Purpose of the Study:

  • To investigate the in vivo expression and function of Prominin-1 (Prom1) in mouse RPE (mRPE) cells.
  • To elucidate the role of Prom1 in maintaining RPE homeostasis and its potential contribution to retinal degeneration.
  • To explore Prom1 as a potential therapeutic target for atrophic AMD (aAMD).

Main Methods:

  • Utilized RNAscope assays and immunogold electron microscopy (EM) to characterize Prom1 mRNA and protein expression in mouse retinal sections.
  • Analyzed human RPE single-cell RNA-sequencing data to confirm Prom1 expression in human RPE.
  • Performed RPE-specific Prom1 knockdown (KD) in mice using adeno-associated virus (AAV)-mediated CRISPR/Cas9 gene editing.

Main Results:

  • Confirmed Prom1 mRNA and protein expression in mouse RPE cells, localized to the cytoplasm and mitochondria.
  • Demonstrated that RPE-specific Prom1 KD in vivo led to abnormal RPE morphology, subretinal fluid, and photoreceptor loss.
  • Observed RPE cell death and reduced a-wave amplitude in Prom1-KD mice, indicative of retinal degeneration and aAMD-like pathology.

Conclusions:

  • Prominin-1 plays a critical cell-autonomous role in maintaining mouse RPE homeostasis.
  • RPE defects and retinal degeneration caused by Prom1 knockdown in mice highlight its significance in aAMD pathogenesis.
  • Targeting Prom1 may offer a novel therapeutic strategy for treating atrophic AMD.