Quantification of Zonula Occludens-1 mRNA expression in cultu-red bovine retinal endothelial cells, using SYBR green real-time PCR

• Main Authors: Ismail, I.S., Muniandy, S., Koon, C.Y., Bee, J.C., Rayappan, A.V., Kum, T.W., Qvist, R.
• Format: Article
• Language: English
• Published: Biomedical Research Press 2008
• Subjects: R Medicine (General)
• Online Access: http://eprints.um.edu.my/10273/1/Quantification_of_Zonula_Occludens-1_mRNA_expression_in_cultu-red_bovine_retinal_endothelial_cells%2C_using_SYBR_green_real-time_PCR.pdf; http://eprints.um.edu.my/10273/

A simple and rapid real-time reverse transcription-PCR (RT-PCR) technique using SYBR green fluorescence was developed to quantify Zonula Occludens-1 (ZO-1) mRNA expression in histamine-treated cultured bovine retinal microvessel endothelial cells. There was no difference between the quantity of ZO-1 mRNA expression in histamine-treated (10-7M to 10-9 M) and untreated bovine retinal endothelial cells at high magnesium levels. Immunohisto-chemistry staining and western blot assay unequivocally demonstrated the approximately similar levels of ZO-1 protein in both histamine-treated and non-treated endothelial cells at high magnesium concentrations.