Editors: Najam A. Sharif, Sunny E. Ohia

Research Protocols for Ophthalmic Disease Mechanisms and Therapeutics: Glaucoma - Ocular Hypertension

eBook: US $129 Special Offer (PDF + Printed Copy): US $207
Printed Copy: US $142
Library License: US $516
ISBN: 978-981-5324-34-1 (Print)
ISBN: 978-981-5324-33-4 (Online)
Year of Publication: 2025
DOI: 10.2174/97898153243341250101

Introduction

Research Protocols for Ophthalmic Disease Mechanisms and Therapeutics: Glaucoma – Ocular Hypertension offers a detailed, step-by-step protocols for isolating, characterizing, and analyzing cells and tissues of the eye’s anterior segment—where OHT originates—using techniques from molecular biology, pharmacology, and histology.

Across eight thematic sections, the book covers ocular anatomy, drug screening, receptor localization, cell-based assays, in vitro and in vivo models, drug formulation, and cutting-edge technologies such as gene editing, single-cell RNA sequencing, and stem cell therapies. Each protocol is designed to support reproducibility and translational relevance.

Key Features:

  • - Details validated protocols for drug discovery and disease modeling
  • - Demonstrates methods for studying receptor function and pharmacokinetics
  • - Provides animal models and ex vivo systems to evaluate therapeutic strategies
  • - Incorporates emerging tools such as CRISPR, metabolomics, and exosomes
  • - Bridges experimental design with clinical relevance for glaucoma research


Readership:

Researchers, clinicians, and industry experts working in ophthalmic drug development, vision science, and translational eye disease research.

Dedication

- Pp. i
Najam A. Sharif, Sunny E. Ohia
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Preface

- Pp. ii-iv (3)
Najam A. Sharif, Sunny E. Ohia
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List of Contributors

- Pp. v-ix (5)

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Basic Anatomy and Physiology of the Human Eye

- Pp. 1-8 (8)
Fatima Muili, Anthonia Okolie, Catherine A. Opere, Ya Fatou Njie-Mbye, Sunny E. Ohia*

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Introduction to Ocular Hypertension and Glaucoma and an Overview of Drug Screening Funnels for Discovering and Characterizing New Intraocular Pressure-Reducing Compounds

- Pp. 9-19 (11)
Najam A. Sharif*, Saima Chaudhry

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Protocols for Access to and Dose Selection of Selected IOP-Lowering Drugs

- Pp. 20-32 (13)
Suchismita Acharya*, Peter G. Klimko

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Protocols for Localizing Receptor mRNAs and Proteins by RT-PCR, In Situ Hybridization, and Autoradiography

- Pp. 33-48 (16)
Najam A. Sharif*, Saima Chaudhry

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Protocol for Generating and Utilizing Ocular Tissue Homogenates for Receptor Binding Studies

- Pp. 49-56 (8)
Nozhat Choudry*

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Protocols for Visualizing and Quantifying Receptors in Ocular Tissues Involved in Aqueous Humor Inflow and Outflow

- Pp. 57-66 (10)
Najam A. Sharif*, Saima Chaudhry

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Protocols for Prostaglandin Receptor Localization in Eye Tissues Using Immunohistochemistry

- Pp. 67-78 (12)
Ursula Schlötzer-Schrehardt*

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Isolation and Use of Trabecular Meshwork Cells

- Pp. 79-85 (7)
Najam A. Sharif*, Saima Chaudhry

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Protocols for TM Cell Replacement Using Differentiated TM-Like Induced Pluripotent Stem Cells (iPSC)

- Pp. 86-97 (12)
Mary J. Kelley*, Ted S. Acott

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Protocols to Isolate and Characterize Ciliary Muscle Cells and Schlemm’s Canal Cells and Detect Matrix Metalloproteases

- Pp. 98-110 (13)
Shahid Husain*

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A Protocol for Primary Culture of Non-Pigmented Ciliary Epithelium Cells

- Pp. 111-125 (15)
Mohammad Shahidullah*, Nicholas Delamere

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Anterior Segment Scleral Fibroblasts and Protocols for Ocular Cell Isolation and Characterization for Glaucoma Studies

- Pp. 126-151 (26)
Veluchamy A. Barathi*

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Protocols for Isolation and Characterization of Human Corneal Epithelial and Conjunctival Epithelial Cells

- Pp. 152-163 (12)
Hanna Rao*, Najam A. Sharif

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Overview and Examples of Drug Screening in Cell-and Tissue-Based Assay Systems: Screening Funnels for In Vitro and In Vivo Studies

- Pp. 164-176 (13)
Najam A. Sharif*, Saima Chaudhry

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Protocol for Studying Ocular Cell Relaxation / Contraction by Cell Impedance

- Pp. 177-194 (18)
Thomas R. Vollmer, Amy Chen, Ganesh Prasanna, Christopher W. Wilson*

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Protocols for Studying Ocular Cell Volume Changes in Aqueous Humor Outflow Regulating Cells In Vitro

- Pp. 195-207 (13)
Dorette Z. Ellis*

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Protocols for Studying Aquaporin Function Related to Ocular Hypertension In Vitro

- Pp. 208-222 (15)
Raj Patil*

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Measuring Elastic Moduli of Human Trabecular Meshwork Tissues, Cells, or Extracellular Matrices

- Pp. 223-243 (21)
VijayKrishna Raghunathan*

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Derivation of Primary Human Trabecular Meshwork Cell-Derived Matrices

- Pp. 244-252 (9)
VijayKrishna Raghunathan*

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Protocol for Studying TM Cell Function in a 3-D Matrix

- Pp. 253-265 (13)
Karen Y. Torrejon*, Andrea M. Unser, Pujhitha Ramesh

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Protocols for Studying Neurotransmitter Release from Isolated Ocular Tissues

- Pp. 266-273 (8)
Sunny E. Ohia*, Catherine A. Opere, Anthonia Okolie, Fatima Muili, Pratik Bhankele, Ya Fatou Njie-Mbye

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Protocols for Studying Contraction/Relaxation of Ocular Tissues In Vitro

- Pp. 274-280 (7)
Kalu K. Ngele, Sunny E. Ohia, Catherine A. Opere, Anthonia Okolie, Fatima Muili, Pratik Bhankele, Ya Fatou Njie-Mbye*

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Protocol for Studying Aqueous Outflow Facility from Anterior Eye Segments In Vitro as an Organ Culture Procedure

- Pp. 281-286 (6)
Najam A. Sharif*, Saima Chaudhry

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Human TM Cells as Models for Pseudo-Exfoliation Glaucoma (PEXG)

- Pp. 287-296 (10)
Aparna Rao*, Munmun Chakraborthy

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Protocol for Inducing IOP-Elevation (OHT) Using Hypertonic Saline Injection into Episcleral Veins of Rats

- Pp. 297-302 (6)
Shahid Husain*

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Protocol for Inducing Ocular Hypertension in Mice Using Magnetic Microbeads

- Pp. 303-310 (8)
Najam A. Sharif*, Saima Chaudhry

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Protocol for the Rat Episcleral Vein Cauterization Model of OHT/Glaucoma

- Pp. 311-314 (4)
Anthonia Okolie, Ya Fatou Njie Mbye*, Fatima Muili, Catherine A. Opere, Sunny E. Ohia

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Protocol for Silicone Oil-Induced Ocular Hypertension

- Pp. 315-319 (5)
Alyssa Hostert, Sunny E. Ohia, Ya Fatou Njie Mbye, Catherine A. Opere*

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Protocol for Inducing Ocular Hypertension in Rats and Mice Using Circumlimbal Sutures

- Pp. 320-326 (7)
Da Zhao*, Zheng He, Pei Ying Lee, Brianna Caroline Afiat, Anna Van Koeverden, Vickie Hoi Ying Wong, Christine Tram Oanh Nguyen, Bang Viet Bui*

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Protocol for Corticosteroid-Induced Ocular Hypertension and Glaucomatous Optic Neuropathy in Rodents

- Pp. 327-336 (10)
Najam A. Sharif*, Saima Chaudhry

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Protocol for Measuring Intraocular Pressure in Guinea Pigs

- Pp. 337-340 (4)
Alyssa Hostert, Catherine A. Opere*

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Protocol for Measuring Intraocular Pressure in Dogs

- Pp. 341-344 (4)
Pratik Bankhele, Catherine A. Opere*, Alyssa Hostert

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Laser Model of Ocular Hypertension in Nonhuman Primates

- Pp. 345-353 (9)
Carol B. Toris*, Shan Fan

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Protocol for Assessment of Aqueous Humor Dynamics in the Living Mouse

- Pp. 354-375 (22)
J. Cameron Millar*, Wei Zhang

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Aqueous Humor Dynamic Measurements in Nonhuman Primates

- Pp. 376-388 (13)
Carol B. Toris*, Shan Fan

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Protocols for Pre-Clinical Testing Microshunts to Drain AQH for Glaucoma

- Pp. 389-405 (17)
Yasushi Kato*, Len Pinchuk

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Protocols for the Use of DBA/2J Mice for Pigmentary Glaucoma Research

- Pp. 406-417 (12)
Abdelrahman Anter, Betsy Benitez, Sanjoy K. Bhattacharya*

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Protocols for Creating Genetically-Transduced Animal Models of Ocular Hypertension and Glaucoma

- Pp. 418-422 (5)
Najam A. Sharif*, Saima Chaudhry

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Protocol for Conditional Knockout Mouse Model of OHT and Glaucoma

- Pp. 423-439 (17)
Judith West-Mays*, Fatima Shirazee

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Protocols to Study the Drug Formulation and Stability Aspects of Anti-Ocular Hypertension and Anti-Glaucoma Drugs

- Pp. 440-451 (12)
Onkar N. Singh*

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Protocol for an Ocular Pharmacokinetic Study in Rabbits for an Ocular Anti-hypertensive Drug

- Pp. 452-461 (10)
James E. Chastain*

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Considerations for Ocular Toxicologic Pathology in Nonclinical Studies for the Development of Intraocular Biologics to Treat Glaucoma and Secondary Neurodegeneration

- Pp. 462-472 (11)
Vladimir Bantseev*, Rachel N. Andrews

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Gene Therapy Approaches and Protocols to Deliver Genes for IOP-Reduction in Animals

- Pp. 473-494 (22)
Adnan Dibas*, Samarendra Mohanty

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Protocol for the Delivery of Stem Cells as Trabecular Meshwork

- Pp. 495-500 (6)
Anthonia Okolie, Ya Fatou Njie Mbye, Fatima Muili, Catherine A. Opere, Sunny E. Ohia*

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Isolation and Characterization of Trabecular Meshwork Stem Cell Exosomes from the Conditioned Medium

- Pp. 501-521 (21)
Iswarya Radhakrishnan, Krishnadas Subbaiah, Muthukkaruppan Veerappan, Dharmalingam Kuppamuthu, Gowri Priya Chidambaranathan*

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Protocol for Single-Cell RNA-Sequence Analysis of Genetic Models of Glaucoma

- Pp. 522-531 (10)
Jack Hopkins, Alex Leung, Youichi Shinozaki, Schuichi Koizumi, Shin-ichi Ohnuma*

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Protocols for Studying Nanotubes and their Role in Human Ocular Cell Communication in the Anterior Chamber of the Eye

- Pp. 532-540 (9)
Saima Chaudhry*

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Protocol for Aqueous Humor Metabolite Measurements for Glaucoma Research

- Pp. 541-549 (9)
Anna Mueller, Isabella Moceri, Sanjoy K. Bhattacharya*

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Protocols for Performing CRISPR Gene Editing in Anterior Chamber Eye Cells

- Pp. 550-564 (15)
Sam Yacoub*, Charles E. Amankwa, Gulab Zode

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Subject Index

- Pp. 565-576 (12)
Najam A. Sharif, Sunny E. Ohia
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