کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6263475 | 1613897 | 2014 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Biophysical and pharmacological characterization of α6-containing nicotinic acetylcholine receptors expressed in HEK293 cells
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
علم عصب شناسی
علوم اعصاب (عمومی)
پیش نمایش صفحه اول مقاله

چکیده انگلیسی
Nicotinic acetylcholine receptors (nAChR's) containing the α6 subunit (α6â) are putative drug targets of relevance to Parkinson's disease and nicotine addiction. However, heterologous expression of α6â receptors has proven challenging which has stifled drug discovery efforts. Here, we investigate potential new avenues for achieving functional α6â receptor expression. Combinations of chimeric and mutated α6, β2 and β3 subunits were co-expressed in the human HEK293 cell line and receptor expression was assessed using Ca2+-imaging (FLIPRâ¢) and whole-cell patch-clamp electrophysiology. Transient transfections of a chimeric α6/α3 subunit construct in combination with β2 and β3V9'S gave rise to significant acetylcholine-evoked whole-cell currents. Increasing the β3V9'S:β2:α6/α3 cDNA ratio, resulted in a significantly higher fraction of cells with robust current levels. Using an excess of wild-type β3, significant functional expression of α6/α3β2β3 was also demonstrated. Comparing the acetylcholine concentration-response relationship of α6/α3β2β3V9'S to that of α6/α3β2β3 revealed the β3 point mutation to result in decreased current decay rate and increased ACh agonist potency. Ca2+-imaging experiments showed preservation of basic α6â receptor pharmacology. Our results establish that α6/α3β2β3V9'S replicate several basic features of native α6â receptors but also highlight several caveats associated with using this construct and may therefore provide guidance for future drug hunting efforts.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Brain Research - Volume 1542, 13 January 2014, Pages 1-11
Journal: Brain Research - Volume 1542, 13 January 2014, Pages 1-11
نویسندگان
Andreas H. Rasmussen, Dorte Strøbæk, Tino Dyhring, Marianne L. Jensen, Dan Peters, Morten Grunnet, Daniel B. Timmermann, Philip K. Ahring,