Characterization of the Specificity of Imidazoline I-1 Receptor Antibody for Subtype of Imidazoline Receptors In Vitro

L.-Y. Wang; P.-M. Ku; S.-H. Chen; L.-J. Chen; Y.-M. Yu; J.-T. Cheng

doi:10.1055/s-0033-1333765

Hormone and Metabolic Research, Table of Contents

Horm Metab Res 2013; 45(07): 485-489
DOI: 10.1055/s-0033-1333765

Original Basic

Characterization of the Specificity of Imidazoline I-1 Receptor Antibody for Subtype of Imidazoline Receptors In Vitro

L.-Y. Wang

¹Department of Pediatrics, Department of Medical Research, Chi-Mei Medical Center, Yong Kang, Tainan City, Taiwan

²The Center of General Education, Chia Nan University of Pharmacy & Science, Rende Dist, Tainan City, Taiwan

,

P.-M. Ku

³Department of Cardiology, Department of Medical Research, Chi-Mei Medical Center, Yong Kang, Tainan City, Taiwan

,

S.-H. Chen

⁴Department of Obstetrics and Gynecology, Department of Medical Research, Chi-Mei Medical Center, Yong Kang, Tainan City, Taiwan

⁵Department of Medical Research, Chi-Mei Medical Center, Yong Kang, Tainan City, Taiwan

,

L.-J. Chen

⁶Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan City, Taiwan

,

Y.-M. Yu

⁷Department of Nutrition and Institute of Medical Science, College of Health Science, Chang Jung Christian University, Guei-Ren, Tainan City, Taiwan

,

J.-T. Cheng

⁵Department of Medical Research, Chi-Mei Medical Center, Yong Kang, Tainan City, Taiwan

⁶Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan City, Taiwan

› Author Affiliations

Abstract

Specific antibodies are essential in the study of receptor protein. Gene matching shows that Nischarin (NISCH) is a mouse homologue of human imidazoline receptor antisera-selective (IRAS) protein, a viable candidate for imidazoline I-1 receptor. However, selectivity of this antibody against imidazoline I-2 or imidazoline I-3 receptors remained obscure. At first, an intracerebroventricular (ICV) injection of anti-NISCH antibody blocked the blood pressure lowering action of rilmenidine (I-1 receptor agonist) in spontaneous hypertensive rat (SHR). However, the same injection of anti-NISCH antibody showed no effect in SHR treated with clonidine (α₂ agonist). In order to clarify the selectivity of anti-NISCH antibody for each subtype of imidazoline receptors, this anti-NISCH antibody was subjected to the lysate of organs isolated from Wistar rats including cortex, hippocampus, cerebellum, and brain stem as central nervous tissues, and heart, liver, pancreas, skeletal muscle, kidney, prostate, and bladder as peripheral tissues. The results show that anti-NISCH antibody positively reacted with all tissues including heart, pancreas, skeletal muscle, kidney and bladder by Western blot analysis. Also, the blotting spots for anti-NISCH antibody show a concentration-dependent manner. Moreover, anti-NISCH antibody blocked the action of glucose uptake induced by 2-BFI (I-2 receptor agonist) in L6 cells. Taken together, the obtained data suggest that anti-NISCH antibody can be used not only for imidazoline I-1 receptor but also for I-2 and I-3 subtypes in immunoassays.

Key words

imidazoline I-1 receptor - imidazoline I-2 receptor - imidazoline I-3 receptor - Nischarin (HISCH) - imidazoline receptor antisera-selective (IRAS) protein

Full Text

References

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