Publication

Common steps in analysis of microRNA expression levels between different tissues, developmental stages, or disease states is to study microRNA expression levels by several methods as: NGS, microarray analysis, real-time PCR, Northern blots, in situ hybridization, and solution hybridization. Of these techniques, quantitative reverse transcription Polymerase Chain Reaction (qRT-PCR) is one the most sensitive and accurate method.

Purpose : FOXC1 is a transcription factor involved in heart, craniofacial and ocular development in vertebrates. Mutations in FOXC1, along with mutations in PITX2, cause Axenfeld-Rieger syndrome (ARS) and explain approximately half of the ARS cases. However, there is still a significant number of patients with an unknown genetic cause. Expression and activity of transcription factors involved in development are finely controlled by their regulatory elements that are often evolutionarily conserved. It has been shown that mutations in those regulatory elements could be also pathogenic.

Purpose : Mutations in the chromatin remodeling factor CHD7 are the predominant cause of CHARGE syndrome, a congenital disorder that frequently includes ocular coloboma. Although CHD7 is known to be required for proper ocular morphogenesis, its role in retinal development has not been thoroughly investigated. In this study, we characterize the expression pattern of chd7 in the developing zebrafish retina and begin to study its function using two chd7 mutant lines.

Purpose : The formation of retinal ganglion cells (RGCs) is a stepwise process subject to tight genetic control. Our purpose in this study is to use single cell ATAC-seq (scATAC-seq) to investigate how the epigenetic landscape changes along the RGC developmental trajectory. Methods : We used two knock-in mouse lines, Atoh7-zaGreen and Pou4f2-tdTomato, to enrich different cell populations by FACS. scATAC-seq and scRNA-seq libraries were then generated using the 10X Chromium platform and sequenced on an Illumina sequencer.

Purpose : A subset of mice in our Cfh knockout (Cfh-/-) colony exhibited rapid retinal degeneration, suggesting a spontaneous mutation occurred on mouse chromosome 1 (Chr 1). The retinal phenotype was similar to that in AdipoR1 knockout (AdipoR1-/-) mice, whose gene is located near the Cfh locus on Chr 1. We attempted to determine if a mutation in AdipoR1 occurred on the Cfh-/- background. Methods : We performed an allele complementation test with a cross between a Cfh-/- mouse with retinal degeneration and an AdipoR1-/- mouse with retinal degeneration.

Purpose : Soluble epoxide hydrolase (sEH) metabolizes pro-resolving epoxy fatty acids into diols. sEH is a potential therapeutic target for choroidal neovascularization (CNV) in wet age-related macular degeneration (AMD) and other eye diseases. Localization of sEH in the retina is contentious and cross-interpretation among different studies is complicated due to antibody limitations.

Purpose : The human cornea has been defined as our “external window” to the visual world that serves as a barrier against the outside environment and as the main refractive lens to focus light into the retina. The histological structure is defined by three layers of cellular elements (epithelium, stroma, endothelium) and two layers of extracellular membranes.

Purpose : Elevated intraocular pressure (IOP) is a major risk factor for the development and progression of primary open angle glaucoma and is due to trabecular meshwork (TM) damage. We investigated the role of an endogenous Toll-like receptor 4 (TLR4) ligand, FN-EDA, in the development of glaucoma utilizing a transgenic mouse strain (B6.EDA+/+) that constitutively expresses only FN containing the EDA isoform.

Purpose : Mechanisms of axonal insult within the ONH in glaucoma are not fully understood. This study aimed to delineate ONH molecular alterations in chronic stages of glaucoma, in an inherited feline model with ONH structure comparable to humans. Methods : ONH tissues from 10 LTBP2mut/mut cats with glaucoma and 5 wt control cats (age 1-3 yrs) were used to generate cDNA libraries for RNAseq. Weekly intraocular pressure (IOP) data and optic nerve axon counts were available for all subjects.

Purpose : Human corneal epithelial stem cells or limbal stem cells (LSCs), have been recognized to locate in corneal limbus for three decades. However, the molecular identity and definitive markers of LSCs are still elusive. This study aimed to uncover novel cell types in heterogenous basal limbus of human cornea for identifying LSC population at single cell resolution. Methods : Single cells of human limbal basal epithelium were isolated from young donor corneas.