# Cilium-associated structures in rod cells

> **NIH NIH R01** · BAYLOR COLLEGE OF MEDICINE · 2021 · $159,948

## Abstract

The goal of this Supplement request is to acquire a Typhoon 5 to enable the experiments planned for project
R01-EY026545, Cilium-Associated Structures in Photoreceptors, as well as those for R01-EY031949 and other
NEI-funded projects at Baylor College of Medicine. An instrument of this kind is essential for rigorous
quantification of proteins, nucleic acids, lipids and other small molecules separated by gel electrophoresis or
thin-layer chromatography. It will eliminate the need for outmoded x-ray film-based detection and allow routine
rapid quantification of immunoblots, a workhorse technique for our NEI projects. A condensed summary of the
parent grant follows.
The goal of this project is to develop a thorough understanding of the structural and molecular basis of p
function of the rod sensory cilium, and to understand the molecular mechanisms of rod cell death in
ciliopathies. There are three Specific Aims: 1. Use cryo-electron tomography (cryo-ET) and recent
developments in sub-tomogram averaging to determine the three-dimensional structure to nanometer
resolution of repeating structures of the rod cell connecting cilium and basal body, including microtubule
doublets and triplets, microtubule inner proteins, “Y-shaped links”, transition fibers and appendages. Our goal
is to apply recent developments in hardware and software to rod cells in both wild type retinas and in animal
models of retinal degeneration. 2. Use superresolution fluorescence to test hypotheses about trafficking of
specific proteins and about the roles of IFT (intraflagellar transport) particles and the BBSome (a coat-forming
protein complex implicated in the blinding ciliopathy, Bardet-Biedl syndrome) in ciliary trafficking in rods. Two-
color superresolution fluorescence and quantitative interaction analysis will be used to assess putative
interactions between IFT proteins or BBS proteins and outer segment membrane proteins, as well as well as
proteins normally excluded from the outer segment which mis-accumulate there in BBS-deficient mice. These
experiments will test the hypothesis that specific membrane proteins are actively trafficked through the
connecting cilium membrane through their association with IFT particles, whereas others are transported via
alternative routes and excluded proteins are actively removed by the BBSome. 3. Use mouse models to test
the hypotheses that CEP290 is a major component of the “Y-shaped links” extending from the ciliary axoneme
to the membrane, using superresolution fluorescence, conventional TEM, and cryo-electron tomography with
timed gene disruption or gene restoration at different developmental stages to distinguish initiating as opposed
to secondary events in the development of the pathophysiology of ciliopathies associated with this protein

## Key facts

- **NIH application ID:** 10303692
- **Project number:** 3R01EY026545-06S1
- **Recipient organization:** BAYLOR COLLEGE OF MEDICINE
- **Principal Investigator:** THEODORE G WENSEL
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $159,948
- **Award type:** 3
- **Project period:** 2016-04-01 → 2025-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10303692

## Citation

> US National Institutes of Health, RePORTER application 10303692, Cilium-associated structures in rod cells (3R01EY026545-06S1). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10303692. Licensed CC0.

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