Angiotensin II Receptor Types 1A, 1B, and 2 in Murine Neuroblastoma Neuro‐2a Cells

Abstract

In this study, the mouse neuroblastoma cell line Neuro‐2a was analyzed for expression of angiotensin II receptors. Reverse‐transcriptase polymerase chain reaction (RT–PCR) showed that Neuro‐2a cells express mRNA of angiotensin II (AngII) receptor subtypes AT_1A, AT_1B, and AT₂. Analysis of Neuro‐2a cells by Western blotting revealed AT₁ and AT₂ receptor protein expression. The predominant molecular weights were determined to be 50.4 kDa for the AT₁ receptor and 62.4 kDa for the AT₂ receptor. Observation of AT₁ and AT₂ receptor localization within Neuro‐2a cells using immunocytochemistry showed distribution similar to other G‐protein coupled receptors with diffuse distribution in the cytosol, perinuclear enrichment and accumulation of receptors on the outer cellular periphery with extension into the neurites. Furthermore, we observed InsP₃ formation following AngII induction that could be abolished in presence of the AT_1A receptor antagonist losartan. The results clearly show expression of the AngII receptor types AT_1A and AT₂ in the Neuro‐2a cell line. We conclude that Neuro‐2a cells represent an interesting model cell line for study of mechanisms that control the interplay between these receptors.

Key Words: :

• Angiotensin receptors
• Losartan
• Neuro‐2a
• RT‐PCR
• Western Blot