研究目的
To investigate the influence of cone signaling in ocular refractive development and myopia susceptibility in mice using a genetic mouse model of cone dysfunction.
研究成果
The absence of cone function does not affect normal refractive development in mice under normal visual conditions, but increases susceptibility to form-deprivation myopia. This suggests that cone pathways are not critical for emmetropization in mice, and both rods and cones contribute to visual signaling for responding to form-deprivation. Retinal dopamine levels were not altered by the mutation or form-deprivation, indicating that increased myopia susceptibility may be independent of dopamine changes.
研究不足
The use of Gnat2cplf3/cplf3 mice may not have total loss of cone function at P28, as some remaining cone function could provide a minimally required threshold for normal refractive development under laboratory conditions, while increasing susceptibility to form-deprivation. Additionally, limited resolution of OCT to detect RPE surface may affect axial length measurements.
1：Experimental Design and Method Selection:
Used Gnat2 cplf3/cplf3 (Gnat2-/-) mice as a genetic model of cone dysfunction to study refractive development and myopia susceptibility under normal and form-deprivation (FD) conditions. Methods included electroretinography to confirm cone dysfunction, photorefraction for refractive error, keratometry for corneal curvature, optical coherence tomography for axial length, and high-performance liquid chromatography for dopamine and DOPAC analysis.
2：Sample Selection and Data Sources:
Mice were from an in-house breeding colony with Gnat2 cplf3/cplf3 mice purchased from Jackson Laboratories (Stock number: 006795). Age-matched male and female wild-type (Gnat2+/+) and Gnat2-/- mice on C57BL/6J background were used. Sample sizes: normal visual conditions (Gnat2+/+: n=7, Gnat2-/-: n=9), FD conditions (Gnat2+/+ goggled n=5-7, na?ve n=5-6; Gnat2-/- goggled n=7-9, na?ve n=10-11).
3：5). Age-matched male and female wild-type (Gnat2+/+) and Gnat2-/- mice on C57BL/6J background were used. Sample sizes:
3. List of Experimental Equipment and Materials: Automated infrared photorefractor, photokeratometer, spectral-domain optical coherence tomography system (SD-OCT; Bioptigen Inc., Durham, NC), high-performance liquid chromatography (HPLC) system, head-mounted diffuser goggles, electroretinography equipment.
4：1). List of Experimental Equipment and Materials:
4. Experimental Procedures and Operational Workflow: Mice were kept in 12:12 hour light-dark cycles. For normal development, ocular parameters measured every 2 weeks from 4 to 14 weeks. For FD, monocular diffusers attached at 4 weeks, weekly measurements for 3 weeks. Retinas harvested 48h after final measurement, between 4 to 6h after light onset, frozen and analyzed by HPLC.
5：Experimental Procedures and Operational Workflow:
5. Data Analysis Methods: Data analyzed using two-way repeated-measures ANOVA and Holm-Sidak post-hoc tests for ocular parameters, two-way ANOVA for DA and DOPAC levels, using SigmaStat 3.5 software.
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