期刊
JOURNAL OF UROLOGY
卷 183, 期 2, 页码 812-819出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.juro.2009.09.078
关键词
prostate; receptors, adrenergic, alpha-1; KMD 3213; fluorescent dyes; prostatic hyperplasia
资金
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- Smoking Research Foundation of Japan
- Organization for Life Science Advancement Programs, University of Fukui
Purpose: Although alpha(1L)-adrenoceptor is recognized as a target of alpha(1) antagonist therapy for benign prostatic hyperplasia, the most common techniques, such as immunohistochemistry and in situ hybridization, are not applicable to examine alpha(1L)-AR vs alpha(1A)-AR tissue distribution because alpha(1L)-AR is now considered another phenotype sharing the alpha(1A)-AR gene and protein molecule. We labeled the alpha(1A) and alpha(1L)-adrenoceptor selective antagonist silodosin (Kissei Pharmaceutical, Matsumoto, Japan) with the fluorophore Alexa Fluor (R) 488 (Alexa-488-silodosin) to visualize alpha(1L)-AR expression. Materials and Methods: Radioligand binding and functional bioassay experiments were done to assess alpha(1)-AR expression in Chinese hamster ovary cells and human prostate tissues. Confocal imaging was subsequently performed. Results: Although Alexa-488-silodosin had about 10 times lower affinity for all alpha(1)-AR subtypes than silodosin in binding and functional studies, it had high selectivity to alpha(1A) and alpha(1L)-ARs. Confocal imaging revealed clear localization of fluorescence on the membrane of Chinese hamster ovary cells expressing alpha(1A)-AR but not a(1B)-and alpha(1D)-ARs, and in the muscle layer of the human prostate. The fluorescent signal in Chinese hamster ovary cells disappeared in the presence of 3 nM prazosin but fluorescence was observed in the human prostate even in the presence of 100 nM prazosin. Conclusions: Alexa-488-silodosin is a powerful fluorescent probe with high selectivity to a1A and alpha(1L)-ARs. Thus, Alexa-488-silodosin successfully visualizes the site of alpha(1L)-ARs in the muscle layer of the human prostate without losing its distinct pharmacological profile.
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