TY - CHAP
T1 - Methods for analyzing endoplasmic reticulum stress in the trabecular meshwork of glaucoma models
AU - Maddineni, Prabhavathi
AU - Kasetti, Ramesh B.
AU - Zode, Gulab S.
N1 - Funding Information:
This work was supported by funding from the following grants and organizations: National Eye Institute, EY022077 (R00) and EY026177 (R01) to G. S. Zode and funding from the North Texas Eye Research Institute.
Publisher Copyright:
© 2018, Springer Science+Business Media LLC.
PY - 2018
Y1 - 2018
N2 - The pathological mechanisms underlying increased outflow resistance at the trabecular meshwork (TM) that is responsible for elevating intraocular pressure (IOP) have not been fully delineated. Recent studies have shown that progressive accumulation of misfolded proteins and induction of endoplasmic reticulum (ER) stress is associated with the pathophysiology of glaucomatous TM damage and IOP elevation. We have shown that known causes of human glaucoma, including expression of mutant myocilin or dexamethasone treatment induce abnormal protein accumulation and ER stress in the TM in vitro and in vivo models. To cope up with abnormal protein accumulation, TM cells activate a cytoprotective pathway of unfolded protein response (UPR). However, chronic ER stress can lead to TM dysfunction and IOP elevation. Using cell culture, mouse models, and human postmortem tissues as well as genetic and pharmacological manipulations, we have analyzed ER stress and UPR mediators in the glaucomatous TM damage and IOP elevation. In this chapter, we have described a detailed protocol for the analysis of protein misfolding and ER stress in TM cells and tissues and its association with glaucomatous TM damage and IOP elevation.
AB - The pathological mechanisms underlying increased outflow resistance at the trabecular meshwork (TM) that is responsible for elevating intraocular pressure (IOP) have not been fully delineated. Recent studies have shown that progressive accumulation of misfolded proteins and induction of endoplasmic reticulum (ER) stress is associated with the pathophysiology of glaucomatous TM damage and IOP elevation. We have shown that known causes of human glaucoma, including expression of mutant myocilin or dexamethasone treatment induce abnormal protein accumulation and ER stress in the TM in vitro and in vivo models. To cope up with abnormal protein accumulation, TM cells activate a cytoprotective pathway of unfolded protein response (UPR). However, chronic ER stress can lead to TM dysfunction and IOP elevation. Using cell culture, mouse models, and human postmortem tissues as well as genetic and pharmacological manipulations, we have analyzed ER stress and UPR mediators in the glaucomatous TM damage and IOP elevation. In this chapter, we have described a detailed protocol for the analysis of protein misfolding and ER stress in TM cells and tissues and its association with glaucomatous TM damage and IOP elevation.
KW - Chemical induction of ER stress in TM
KW - ER stress in glaucomatous TM
KW - Genetic manipulation of ER stress
KW - Glucocorticoid-induced ocular hypertension
KW - Myocilin induced ER stress
KW - Unfolded protein response (UPR) target genes
KW - XBP-1 splicing
UR - http://www.scopus.com/inward/record.url?scp=85037046278&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-7407-8_12
DO - 10.1007/978-1-4939-7407-8_12
M3 - Chapter
C2 - 29190024
AN - SCOPUS:85037046278
T3 - Methods in Molecular Biology
SP - 121
EP - 134
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -