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1. Consensus Recommendations for Studies of Outflow Facility and Intraocular Pressure Regulation Using Ex Vivo Perfusion Approaches.

Author

Acott, Ted, Fautsch, Michael, Mao, Weiming, Ethier, C, Huang, Alex, Kelley, Mary, Aga, Mini, Bhattacharya, Sanjoy, Borras, Terete, Bovenkamp, Diane, Chowdhury, Uttio, Clark, Abbot, Dibas, Mohammed, Du, Yiqin, Elliott, Michael, Faralli, Jennifer, Gong, Haiyan, Herberg, Samuel, Johnstone, Murray, Kaufman, Paul, Keller, Kate, Kelly, Ruth, Krizaj, David, Kuehn, Markus, Li, Hoi, Lieberman, Raquel, Lin, Shan, Liu, Yutao, McDonnell, Fiona, McDowell, Colleen, McLellan, Gillian, Mzyk, Philip, Nair, Kayarat, Overby, Darryl, Peters, Donna, Raghunathan, VijayKrishna, Rao, Ponugoti, Roddy, Gavin, Sharif, Najam, Shim, Myoung, Sun, Yang, Thomson, Benjamin, Toris, Carol, Willoughby, Colin, Zhang, Hao, Freddo, Thomas, Fuchshofer, Rudolf, Hill, Kamisha, Karimi, Alireza, Kizhatil, Krishnakumar, Kopcyznski, Casey, Liton, Paloma, Patel, Gaurang, Peng, Michael, Pattabiraman, Padmanabhan, Prasanna, Ganesh, Reina-Torres, Ester, Samples, E, Samples, John, Steel, Cynthia, Strohmaier, Clemens, Subramanian, Preeti, Sugali, Chenna, van Batenburg-Sherwood, Joseph, Wong, Cydney, Youngblood, Hannah, Zode, Gulab, White, Elizabeth, and Stamer, W

Subjects
Animals, Humans, Anterior Eye Segment, Aqueous Humor, Consensus, Glaucoma, Intraocular Pressure, Organ Culture Techniques, Perfusion, Trabecular Meshwork
Abstract

Intraocular pressure (IOP) elevation is the primary risk factor and currently the main treatable factor for progression of glaucomatous optic neuropathy. In addition to direct clinical and living animal in vivo studies, ex vivo perfusion of anterior segments and whole eyes is a key technique for studying conventional outflow function as it is responsible for IOP regulation. We present well-tested experimental details, protocols, considerations, advantages, and limitations of several ex vivo model systems for studying IOP regulation. These include: (1) perfused whole globes, (2) stationary anterior segment organ culture, (3) perfused human anterior segment organ culture, (4) perfused animal anterior segment organ culture, (5) perfused human corneal rims, and (6) perfused human anterior segment wedges. These methods, with due consideration paid to their strengths and limitations, comprise a set of very strong tools for extending our understanding of IOP regulation.

Published
2024

3. Consensus Recommendation for Mouse Models of Ocular Hypertension to Study Aqueous Humor Outflow and Its Mechanisms.

Author

McDowell, Colleen M, Kizhatil, Krishnakumar, Elliott, Michael H, Overby, Darryl R, van Batenburg-Sherwood, Joseph, Millar, J Cameron, Kuehn, Markus H, Zode, Gulab, Acott, Ted S, Anderson, Michael G, Bhattacharya, Sanjoy K, Bertrand, Jacques A, Borras, Terete, Bovenkamp, Diane E, Cheng, Lin, Danias, John, De Ieso, Michael Lucio, Du, Yiqin, Faralli, Jennifer A, Fuchshofer, Rudolf, Ganapathy, Preethi S, Gong, Haiyan, Herberg, Samuel, Hernandez, Humberto, Humphries, Peter, John, Simon WM, Kaufman, Paul L, Keller, Kate E, Kelley, Mary J, Kelly, Ruth A, Krizaj, David, Kumar, Ajay, Leonard, Brian C, Lieberman, Raquel L, Liton, Paloma, Liu, Yutao, Liu, Katy C, Lopez, Navita N, Mao, Weiming, Mavlyutov, Timur, McDonnell, Fiona, McLellan, Gillian J, Mzyk, Philip, Nartey, Andrews, Pasquale, Louis R, Patel, Gaurang C, Pattabiraman, Padmanabhan P, Peters, Donna M, Raghunathan, Vijaykrishna, Rao, Ponugoti Vasantha, Rayana, Naga, Raychaudhuri, Urmimala, Reina-Torres, Ester, Ren, Ruiyi, Rhee, Douglas, Chowdhury, Uttio Roy, Samples, John R, Samples, E Griffen, Sharif, Najam, Schuman, Joel S, Sheffield, Val C, Stevenson, Cooper H, Soundararajan, Avinash, Subramanian, Preeti, Sugali, Chenna Kesavulu, Sun, Yang, Toris, Carol B, Torrejon, Karen Y, Vahabikashi, Amir, Vranka, Janice A, Wang, Ting, Willoughby, Colin E, Xin, Chen, Yun, Hongmin, Zhang, Hao F, Fautsch, Michael P, Tamm, Ernst R, Clark, Abbot F, Ethier, C Ross, and Stamer, W Daniel

Subjects
Biomedical and Clinical Sciences, Ophthalmology and Optometry, Eye Disease and Disorders of Vision, Cardiovascular, Aging, Hypertension, Neurosciences, 2.1 Biological and endogenous factors, Aetiology, Animals, Aqueous Humor, Consensus, Disease Models, Animal, Glaucoma, Intraocular Pressure, Mice, Ocular Hypertension, Tonometry, Ocular, Trabecular Meshwork, mouse model, conventional outflow, glaucoma, intraocular pressure, ocular hypertension, outflow facility, Biological Sciences, Medical and Health Sciences, Ophthalmology & Optometry, Ophthalmology and optometry
Abstract

Due to their similarities in anatomy, physiology, and pharmacology to humans, mice are a valuable model system to study the generation and mechanisms modulating conventional outflow resistance and thus intraocular pressure. In addition, mouse models are critical for understanding the complex nature of conventional outflow homeostasis and dysfunction that results in ocular hypertension. In this review, we describe a set of minimum acceptable standards for developing, characterizing, and utilizing mouse models of open-angle ocular hypertension. We expect that this set of standard practices will increase scientific rigor when using mouse models and will better enable researchers to replicate and build upon previous findings.

Published
2022

5. AxoNet: an AI-based tool to count retinal ganglion cell axons

Author

Ritch, Matthew D., Hannon, Bailey G., Read, A. Thomas, Feola, Andrew J., Cull, Grant A., Reynaud, Juan, Morrison, John C., Burgoyne, Claude F., Pardue, Machelle T., and Ethier, C. Ross

Subjects
Quantitative Biology - Quantitative Methods, Electrical Engineering and Systems Science - Image and Video Processing
Abstract

Goal: In this work, we develop a robust, extensible tool to automatically and accurately count retinal ganglion cell axons in images of optic nerve tissue from various animal models of glaucoma. Methods: The U-Net convolutional neural network architecture was adapted to learn pixelwise axon count density estimates, which were then integrated over the image area to determine axon counts. The tool, termed AxoNet, was trained and evaluated using a dataset containing images of optic nerve regions randomly selected from complete cross sections of intact rat optic nerves and manually annotated for axon count and location. Both control and damaged optic nerves were used. This rat-trained network was then applied to a separate dataset of non-human primate (NHP) optic nerve images. AxoNet was then compared to two existing automated axon counting tools, AxonMaster and AxonJ, using both datasets. Results: AxoNet outperformed the existing tools on both the rat and NHP optic nerve datasets as judged by mean absolute error, R2 values when regressing automated vs. manual counts, and Bland-Altman analysis. Conclusion: The proposed tool allows for accurate quantification of axon numbers as a measure of glaucomatous damage. AxoNet is robust to variations in optic nerve tissue damage extent, image quality, and species of mammal. Significance: The deep learning method does not rely on hand-crafted image features for axon recognition. Therefore, this approach is not species-specific and can be extended to quantify additional optic nerve features. It will aid evaluation of optic nerve changes in glaucoma and potentially other neurodegenerative diseases., Comment: 11 pages, 11 figures

Published
2019
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6. The relationship between wall shear stress distributions and intimal thickening in the human abdominal aorta

Author

Butany Jagdish, Leask Richard L, Bonert Michael, Ethier C, Myers Jerry G, Johnston K, and Ojha Matadial

Subjects
Medical technology, R855-855.5
Abstract

Abstract Purpose The goal of this work was to determine wall shear stress (WSS) patterns in the human abdominal aorta and to compare these patterns to measurements of intimal thickness (IT) from autopsy samples. Methods The WSS was experimentally measured using the laser photochromic dye tracer technique in an anatomically faithful in vitro model based on CT scans of the abdominal aorta in a healthy 35-year-old subject. IT was quantified as a function of circumferential and axial position using light microscopy in ten human autopsy specimens. Results The histomorphometric analysis suggests that IT increases with age and that the distribution of intimal thickening changes with age. The lowest WSS in the flow model was found on the posterior wall inferior to the inferior mesenteric artery, and coincided with the region of most prominent IT in the autopsy samples. Local geometrical features in the flow model, such as the expansion at the inferior mesenteric artery (common in younger individuals), strongly influenced WSS patterns. The WSS was found to correlate negatively with IT (r2 = 0.3099; P = 0.0047). Conclusion Low WSS in the abdominal aorta is co-localized with IT and may be related to atherogenesis. Also, rates of IT in the abdominal aorta are possibly influenced by age-related geometrical changes.

Published
2003
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9. Consensus recommendations for trabecular meshwork cell isolation, characterization and culture

Author

Keller, Kate E, Bhattacharya, Sanjoy K, Borrás, Theresa, Brunner, Thomas M, Chansangpetch, Sunee, Clark, Abbott F, Dismuke, W Michael, Du, Yiqin, Elliott, Michael H, Ethier, C Ross, Faralli, Jennifer A, Freddo, Thomas F, Fuchshofer, Rudolf, Giovingo, Michael, Gong, Haiyan, Gonzalez, Pedro, Huang, Alex, Johnstone, Murray A, Kaufman, Paul L, Kelley, Mary J, Knepper, Paul A, Kopczynski, Casey C, Kuchtey, John G, Kuchtey, Rachel W, Kuehn, Markus H, Lieberman, Raquel L, Lin, Shan C, Liton, Paloma, Liu, Yutao, Lütjen-Drecoll, Elke, Mao, Weiming, Masis-Solano, Marisse, McDonnell, Fiona, McDowell, Colleen M, Overby, Darryl R, Pattabiraman, Padmanabhan P, Raghunathan, Vijay K, Rao, P Vasanth, Rhee, Douglas J, Chowdhury, Uttio Roy, Russell, Paul, Samples, John R, Schwartz, Donald, Stubbs, Evan B, Tamm, Ernst R, Tan, James C, Toris, Carol B, Torrejon, Karen Y, Vranka, Janice A, Wirtz, Mary K, Yorio, Thomas, Zhang, Jie, Zode, Gulab S, Fautsch, Michael P, Peters, Donna M, Acott, Ted S, and Stamer, W Daniel

Subjects
Biomedical and Clinical Sciences, Ophthalmology and Optometry, Eye Disease and Disorders of Vision, Neurosciences, Age Factors, Animals, Biomarkers, Cell Culture Techniques, Cell Separation, Consensus, Fetus, Guidelines as Topic, Humans, Tissue Donors, Tissue Preservation, Tissue and Organ Harvesting, Trabecular Meshwork, Aqueous humor dynamics, Conventional outflow, Glaucoma, Intraocular pressure, Ocular hypertension, Medical Biochemistry and Metabolomics, Opthalmology and Optometry, Ophthalmology & Optometry, Ophthalmology and optometry
Abstract

Cultured trabecular meshwork (TM) cells are a valuable model system to study the cellular mechanisms involved in the regulation of conventional outflow resistance and thus intraocular pressure; and their dysfunction resulting in ocular hypertension. In this review, we describe the standard procedures used for the isolation of TM cells from several animal species including humans, and the methods used to validate their identity. Having a set of standard practices for TM cells will increase the scientific rigor when used as a model, and enable other researchers to replicate and build upon previous findings.

Published
2018

14. Biomechanical aspects of axonal damage in glaucoma: A brief review.

Author

Stowell, Cheri, Burgoyne, Claude, Tamm, Ernst, and Ethier, C

Subjects
Animals, Axons, Biomechanical Phenomena, Cerebrospinal Fluid Pressure, Connective Tissue, Glaucoma, Humans, Intraocular Pressure, Optic Disk, Optic Nerve Diseases, Retinal Ganglion Cells
Abstract

The biomechanical environment within the optic nerve head (ONH) is complex and is likely directly involved in the loss of retinal ganglion cells (RGCs) in glaucoma. Unfortunately, our understanding of this process is poor. Here we describe factors that influence ONH biomechanics, including ONH connective tissue microarchitecture and anatomy; intraocular pressure (IOP); and cerebrospinal fluid pressure (CSFp). We note that connective tissue factors can vary significantly from one individual to the next, as well as regionally within an eye, and that the understanding of ONH biomechanics is hindered by anatomical differences between small-animal models of glaucoma (rats and mice) and humans. Other challenges of using animal models of glaucoma to study the role of biomechanics include the complexity of assessing the degree of glaucomatous progression; and inadequate tools for monitoring and consistently elevating IOP in animal models. We conclude with a consideration of important open research questions/challenges in this area, including: (i) Creating a systems biology description of the ONH; (ii) addressing the role of astrocyte connective tissue remodeling and reactivity in glaucoma; (iii) providing a better characterization of ONH astrocytes and non-astrocytic constituent cells; (iv) better understanding the role of ONH astrocyte phagocytosis, proliferation and death; (v) collecting gene expression and phenotype data on a larger, more coordinated scale; and (vi) developing an implantable IOP sensor.

Published
2017

15. Biological aspects of axonal damage in glaucoma: A brief review

Author

Tamm, Ernst R, Ethier, C Ross, Participants, The Lasker IRRF Initiative on Astrocytes and Glaucomatous Neurodegeneration, Dowling, John E, Downs, Crawford, Ellisman, Mark H, Fisher, Steven, Fortune, Brad, Fruttiger, Marcus, Jakobs, Tatjana, Lewis, Geoffrey, Masland, Richard H, Mitchell, Claire H, Morrison, John, Sharma, Sansar C, Sigal, Ian, Sofroniew, Michael, Wang, Lin, Wiggs, Janey, and Wu, Samuel

Subjects
Biomedical and Clinical Sciences, Ophthalmology and Optometry, Neurodegenerative, Neurosciences, Eye Disease and Disorders of Vision, Aging, Aetiology, 2.1 Biological and endogenous factors, Eye, Neurological, Animals, Astrocytes, Axons, Glaucoma, Gliosis, Humans, Intracranial Pressure, Intraocular Pressure, Nerve Degeneration, Optic Disk, Optic Nerve Diseases, Lasker/IRRF Initiative on Astrocytes and Glaucomatous Neurodegeneration Participants, Medical Biochemistry and Metabolomics, Opthalmology and Optometry, Ophthalmology & Optometry, Ophthalmology and optometry
Abstract

Intraocular pressure (IOP) is a critical risk factor in glaucoma, and the available evidence derived from experimental studies in primates and rodents strongly indicates that the site of IOP-induced axonal damage in glaucoma is at the optic nerve head (ONH). However, the mechanisms that cause IOP-induced damage at the ONH are far from understood. A possible sequence of events could originate with IOP-induced stress in the ONH connective tissue elements (peripapillary sclera, scleral canal and lamina cribrosa) that leads to an increase in biomechanical strain. In consequence, molecular signaling cascades might be activated that result in extracellular matrix turnover of the peripapillary sclera, changing its biomechanical properties. Peripapillary sclera strain might induce reactive changes in ONH astrocytes and cause astrogliosis. The biological changes that are associated with ONH astrocyte reactivity could lead to withdrawal of trophic or metabolic support for optic nerve axons and cause their degeneration. Alternatively, the expression of neurotoxic molecules might be induced. Unfortunately, direct experimental in vivo evidence for these or other scenarios is currently lacking. The pathogenic processes that cause axonal degeneration at the ONH in glaucoma need to be identified before any regenerative therapy is likely to succeed. Several topics and emerging techniques should be pursued to enhance our understanding of the mechanisms that are behind axonal degeneration. Among them are: Advanced imaging techniques, the development of in vivo markers to identify axonal injury, the generation of molecular approaches for in vivo detection of mechanosensitivity and for molecular manipulation of the ONH, a more complete characterization of retinal ganglion cells, the use of organ cultures, 3D-bioprinting, and the engineering of microdevices that can measure pressure. Questions that need to be answered relate to the specific roles of astrogliosis, neuroinflammation, blood flow and intracranial pressure in axonal degeneration at the ONH.

Published
2017

25. Aging and intraocular pressure homeostasis in mice

Author

Li, Guorong, primary, van Batenburg‐Sherwood, Joseph, additional, Safa, Babak N., additional, Fraticelli Guzmán, Nina Sara, additional, Wilson, Andrea, additional, Bahrani Fard, Mohammad Reza, additional, Choy, Kevin, additional, de Ieso, Michael L., additional, Cui, J. Serena, additional, Feola, Andrew J., additional, Weisz, Tara, additional, Kuhn, Megan, additional, Rickman, Catherine Bowes, additional, Farsiu, Sina, additional, Ethier, C. Ross, additional, and Stamer, W. Daniel, additional

Published
2024
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37. Consensus recommendations for trabecular meshwork cell isolation, characterization and culture

Author

Keller, Kate E., Bhattacharya, Sanjoy K., Borrás, Theresa, Brunner, Thomas M., Chansangpetch, Sunee, Clark, Abbott F., Dismuke, W. Michael, Du, Yiqin, Elliott, Michael H., Ethier, C. Ross, Faralli, Jennifer A., Freddo, Thomas F., Fuchshofer, Rudolf, Giovingo, Michael, Gong, Haiyan, Gonzalez, Pedro, Huang, Alex, Johnstone, Murray A., Kaufman, Paul L., Kelley, Mary J., Knepper, Paul A., Kopczynski, Casey C., Kuchtey, John G., Kuchtey, Rachel W., Kuehn, Markus H., Lieberman, Raquel L., Lin, Shan C., Liton, Paloma, Liu, Yutao, Lütjen-Drecoll, Elke, Mao, Weiming, Masis-Solano, Marisse, McDonnell, Fiona, McDowell, Colleen M., Overby, Darryl R., Pattabiraman, Padmanabhan P., Raghunathan, Vijay K., Rao, P. Vasanth, Rhee, Douglas J., Chowdhury, Uttio Roy, Russell, Paul, Samples, John R., Schwartz, Donald, Stubbs, Evan B., Tamm, Ernst R., Tan, James C., Toris, Carol B., Torrejon, Karen Y., Vranka, Janice A., Wirtz, Mary K., Yorio, Thomas, Zhang, Jie, Zode, Gulab S., Fautsch, Michael P., Peters, Donna M., Acott, Ted S., and Stamer, W. Daniel

Published
2018
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39. Aging and intraocular pressure homeostasis in mice

Author

Li, Guorong, primary, van Batenburg-Sherwood, Joseph, additional, Safa, Babak N., additional, Fraticelli Guzmán, Nina Sara, additional, Wilson, Andrea, additional, Bahrani Fard, Mohammad Reza, additional, Choy, Kevin, additional, De Ieso, Michael L., additional, Cui, J. Serena, additional, Feola, Andrew J, additional, Weisz, Tara, additional, Kuhn, Megan, additional, Rickman, Cathy Bowes, additional, Farsiu, Sina, additional, Ethier, C. Ross, additional, and Stamer, W. Daniel, additional

Published
2023
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43. Biomechanical aspects of axonal damage in glaucoma: A brief review

Author

Dowling, John E., Downs, Crawford, Ellisman, Mark H., Fisher, Steven, Fortune, Brad, Fruttiger, Marcus, Jakobs, Tatjana, Lewis, Geoffrey, Masland, Richard H., Mitchell, Claire H., Morrison, John, Sharma, Sansar C., Sigal, Ian, Sofroniew, Michael, Wang, Lin, Wiggs, Janey, Wu, Samuel, Stowell, Cheri, Burgoyne, Claude F., Tamm, Ernst R., and Ethier, C. Ross

Published
2017
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