Developing a Translaminar Pressure Model Using a Translaminar Autonomous System
JoVE Encyclopedia of Experiments
Neuroscience
0 views • 3:18 min • April 28th, 2025
Begin with a human eye. Remove the sheath to expose the optic nerve.
Bisect the eye, then remove the vitreous humor from the posterior segment.
Trim the sclera, the outer layer, and spread the retina containing the retinal ganglion cells connected to the optic nerve via the lamina cribrosa, a mesh-like structure.
Place this segment over the dome of the lower chamber of the translaminar autonomous system, with the optic nerve facing upwards then seal it.
Connect the tubing and flow a perfusion medium.
The liquid's movement builds pressure inside the eye cup, resembling intraocular pressure.
Next, position the upper chamber covering the optic nerve and seal it.
Connect the tubing to flow the medium around the nerve, resembling intracranial pressure.
The pressure difference between these chambers establishes a translaminar pressure across the lamina cribrosa, enabling the study of retinal ganglion cell signal transmission to the optic nerve under this pressure condition.
To prepare a human whole eye globe sample, remove the optic nerve sheath and remove the vitreous humor from the posterior segment. To ensure a good fit on the round dome of the IOP chamber, trim additional sclera from the posterior segment as necessary and use forceps to ensure that the retina is spread evenly over the human posterior of the segment. Then place the segment into the IOP chamber of the TAS over the round dome, with the optic nerve facing up, and use an epoxy resin O-ring and 4 screws to seal the posterior segment.
To set up the IOP chamber, insert the tubing into in and out ports of the chamber and insert the IOP inflow syringe into the in port. Insert the empty IOP outflow syringe setup into the out port and use the push-pull method to slowly infuse the perfusion medium into the inflow port to fill the posterior eye cup while simultaneously slowly pulling the perfusion medium out through the outflow syringe to remove any air bubbles from the lines. Once both in and out tubes are void of air bubbles, stop the infusion and lock the stopcocks in the off position.
Remove the syringe from the IOP import filter assembly and refill the syringe with 30 additional milliliters of medium. Then reinsert the syringe setup into the filter assembly. To set up the ICP chamber, place the chamber over the back of the posterior segment, taking care that the optic nerve is within the top chamber, and seal the top chamber with 4 screws.
Then insert the tubing into in and out ports of the ICP chamber-- the ICP input syringe into the in port and the empty ICP outflow syringe into the out port to flush the system with medium, as just demonstrated.
Overview
This article demonstrates the preparation of human donor eye tissue for studying the effects of translaminar pressure gradients on retinal ganglion cell signaling. Using a translaminar autonomous system (TAS), the protocol simulates intraocular and intracranial pressures to investigate how pressure differences across the lamina cribrosa influence neural signal transmission from the retina to the optic nerve.
Key Study Components
Area of Science
- Ophthalmology
- Neuroscience
- Experimental Physiology
Background
- Translaminar pressure difference (TLPD) is the gradient between intraocular pressure (IOP) and intracranial pressure (ICP) across the lamina cribrosa.
- Altered TLPD is implicated in optic nerve diseases such as glaucoma.
- Retinal ganglion cells transmit visual signals through the optic nerve, passing through the lamina cribrosa.
- Studying TLPD effects requires controlled simulation of IOP and ICP in ex vivo human tissue.
Purpose of Study
- To develop and demonstrate a method for simulating IOP and ICP in human posterior eye segments.
- To enable controlled study of translaminar pressure effects on retinal ganglion cell signaling.
- To provide a reproducible protocol for future research on pressure-induced optic nerve pathophysiology.
Methods Used
- Dissection of human donor eyes to expose the optic nerve and posterior segment.
- Removal of the vitreous humor and trimming of the sclera for optimal chamber fit.
- Mounting the posterior segment onto the dome of the TAS lower chamber (IOP chamber) with the optic nerve facing up.
- Sealing the tissue using an epoxy resin O-ring and screws.
- Perfusion of the IOP chamber with medium to simulate intraocular pressure.
- Placement and sealing of the upper chamber (ICP chamber) over the optic nerve.
- Perfusion of the ICP chamber to simulate intracranial pressure.
- Establishment of a controlled translaminar pressure gradient across the lamina cribrosa.
Main Results
- The protocol enables precise simulation of IOP and ICP in human posterior eye segments.
- A stable translaminar pressure gradient can be established and maintained across the lamina cribrosa.
- The system allows for the study of retinal ganglion cell signal transmission under controlled pressure conditions.
- The method is reproducible and suitable for investigating pressure-related optic nerve pathologies.
Conclusions
- The TAS protocol provides a robust platform for ex vivo modeling of translaminar pressure effects in human eye tissue.
- This approach facilitates mechanistic studies of pressure-induced changes in retinal ganglion cell function.
- The method supports research into the pathophysiology of diseases such as glaucoma.
Frequently Asked Questions
Executive Industry Relevance
This method enables mechanistic de-risking of retinal ganglion cell targets by simulating physiologically relevant translaminar pressure conditions. It supports target validation in glaucoma research by providing a disease-relevant system for assessing functional responses under controlled pressure differentials. The approach enhances predictive confidence in preclinical models by isolating pressure as an independent variable in visual pathway signaling studies.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Scientific Value: Enables interrogation of retinal ganglion cell hypothesis under physiologic pressure conditions.
- Operational Value: Provides a reproducible system for functional target validation in posterior segment tissues.
- Strategic Value: Supports mechanistic de-risking of glaucoma targets by modeling translaminar pressure effects.
Screening & Assay Development
- Scientific Value: Generates quantitative readouts of retinal ganglion cell signaling under defined pressure gradients.
- Operational Value: Standardizes pressure-controlled conditions for assay reproducibility across experiments.
- Strategic Value: Enables scalable compound screening in a human-relevant posterior segment model.
Translational & Preclinical Research
- Scientific Value: Maintains disease relevance by using human donor posterior segments to study pressure-dependent signaling.
- Operational Value: Ensures translational continuity from discovery to preclinical validation via consistent pressure simulation.
- Strategic Value: Informs risk-adjusted advancement decisions by modeling a key pathophysiological variable in glaucoma.
Pipeline & Workflow Integration
The method integrates into the discovery continuum by enabling pressure-modulated functional assays following target identification and preceding lead optimization in neuroscience and ophthalmology programs.
- Discovery Biology: Supports hypothesis testing of pressure-sensitive pathways in retinal ganglion cells.
- Screening: Delivers quantitative, reproducible measurements of cellular responses to translaminar pressure.
- Analytics: Provides pressure-dependent readouts that allow comparison of experimental conditions.
- Translational Research: Connects to preclinical continuity through use of human tissue and clinically relevant pressure ranges.
- Enterprise Reuse: Establishes a reusable platform for pressure-modulated studies across ocular disease programs.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence by reducing mechanistic ambiguity in pressure-dependent signaling.
- Operational Value: Ensures standardization and reproducibility of pressure simulation across laboratories.
- Strategic Value: Improves go/no-go decisions by validating targets in a disease-relevant mechanical context.
- Portfolio Impact: Enables risk-adjusted prioritization of glaucoma targets based on functional validation under translaminar stress.
Implementation Considerations
- Requires expertise in ocular tissue handling and pressure system calibration.
- Depends on perfusion medium control and air bubble elimination for stable pressure maintenance.
- Necessitates standardization of sealing and tubing connections to prevent leaks.
- Involves adaptation considerations when applying to non-human or diseased tissue models.
- Limited by tissue viability duration and the need for precise pressure monitoring equipment.
Biopharma FAQs
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Last updated: 22 August 2026