Optimization of the Retinal Vein Occlusion Mouse Model to Limit Variability

Crystal Col&#243;n Ortiz; Anna Potenski; Jaqueline M. Lawson; Jade Smart; Carol M. Troy

doi:10.3791/62980

Optymalizacja mysiego modelu niedrożności żyły siatkówki w celu ograniczenia zmienności

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Optimization of the Retinal Vein Occlusion Mouse Model to Limit Variability

Optymalizacja mysiego modelu niedrożności żyły siatkówki w celu ograniczenia zmienności

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07:23 min

August 6, 2021

DOI: 10.3791/62980-v

Crystal Colón Ortiz*¹, Anna Potenski*², Jaqueline M. Lawson¹, Jade Smart¹, Carol M. Troy^1,3,4

¹Department of Pathology & Cell Biology; Vagelos College of Physicians and Surgeons,Columbia University, ²Department of Molecular Pharmacology and Therapeutics; Vagelos College of Physicians and Surgeons,Columbia University, ³Department of Neurology; Vagelos College of Physicians and Surgeons,Columbia University, ⁴The Taub Institute for Research on Alzheimer’s Disease and the Aging Brain; Vagelos College of Physicians and Surgeons,Columbia University

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Overview

This study outlines an optimized protocol for retinal vein occlusion using rose bengal and a laser-guided retinal imaging microscope system. The model facilitates the investigation of vascular injury mechanisms and edema development in real-time without surgical intervention, allowing for more efficient treatment testing.

Key Study Components

Area of Science

Neuroscience
Ophthalmology
Vascular biology

Background

Retinal vein occlusion models are important for studying vascular injuries.
Rose bengal serves as a vital agent for enhancing imaging and occlusion efficacy.
The method allows for live observation of retinal damage and recovery.
An improved ability to quantify retinal responses using techniques like optical coherence tomography (OCT) is emphasized.

Purpose of Study

To develop a reproducible model for investigating retinal vascular injury.
To measure and observe changes in retinal blood vessel states after laser-induced occlusion.
To enhance knowledge on the effects of neurovascular injury on retinal function.

Methods Used

The main platform used is a laser-guided retinal imaging microscope.
The biological model consists of genetically modified mice used to assess vascular occlusions.
Critical steps involve image acquisition, laser control settings, and proper handling of rose bengal.
Measurements of laser power and appropriate timing after rose bengal administration were noted.
Protocols for anesthesia and recovery monitoring were specified to ensure animal welfare.

Main Results

The timing of laser application post-rose bengal dosage influenced the number of sustained occlusions.
Different occlusion states were observed, revealing important insights into hemorrhagic responses post-injury.
Key molecular changes were quantified, demonstrating the model's efficacy for studying vascular repair mechanisms.
OCT imaging was used to assess retinal edema and inner layer disorganization.

Conclusions

This protocol allows for detailed investigation of neurovascular disease mechanisms.
The ability to monitor retinal changes over time will aid in translating findings to human conditions.
Overall, the study improves understanding of vascular injuries and the potential for treatment interventions in retinal diseases.

Frequently Asked Questions

What are the advantages of this experimental model?

The model allows for live monitoring of retinal vascular changes and facilitates non-invasive treatment testing without surgical interventions.

How is the retinal vein occlusion implemented in the model?

Retinal vein occlusion is achieved by administering rose bengal followed by laser irradiation of targeted retinal vessels.

What types of data are collected in this study?

Data includes imaging of retinal vasculature, quantification of occlusions, and assessments of retinal state using optical coherence tomography.

How can this method be adapted for other studies?

The protocol could be modified to investigate different treatment regimens or to extend its application to other forms of vascular injury in various models.

Are there any limitations to this method?

Challenges may arise in accurately performing the tail vein technique and ensuring proper animal positioning, which requires practice and patience.

What types of ocular pathologies can be studied with this technique?

The technique offers insights into several ocular conditions, including retinal edema and hemorrhagic responses due to vascular occlusions.

What critical factors affect the success of occlusions?

Baseline laser power settings and timing post-rose bengal administration were critical in achieving successful vessel occlusions.

Tutaj opisujemy zoptymalizowany protokół do zamykania żył siatkówki za pomocą róży bengalskiej i sterowanego laserowo systemu obrazowania siatkówki z zaleceniami maksymalizacji jego odtwarzalności w genetycznie modyfikowanych szczepach.

Protokół stanowi model do badania mechanizmów uszkodzenia naczyń krwionośnych i późniejszego rozwoju obrzęku w sposób nieinwazyjny. Główną zaletą tej techniki jest to, że można śledzić uraz na żywo w czasie bez interwencji chirurgicznej. Daje to możliwość skuteczniejszego testowania zabiegów.

Nauczenie się, jak prawidłowo żyłkować ogon i prawidłowo umieścić zwierzę na platformie, może być trudne. W przypadku obu technik zalecam uzbroić się w cierpliwość i działać powoli. Aby rozpocząć, delikatnie obejmij się z światłowodowym i podłącz go do skrzynki sterowniczej lasera i adaptera laserowego mikroskopu do obrazowania siatkówki, a następnie włącz lampę mikroskopu do obrazowania siatkówki.

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