FUNCTIONAL EXPRESSION OF NICOTINIC ACETYLCHOLINE-RECEPTORS CONTAINING RAT ALPHA-7 SUBUNITS IN HUMAN SH-SY5Y NEUROBLASTOMA-CELLS

被引:93
|
作者
PUCHACZ, E [1 ]
BUISSON, B [1 ]
BERTRAND, D [1 ]
LUKAS, RJ [1 ]
机构
[1] UNIV GENEVA, CTR MED, DEPT PHYSIOL, CH-1211 GENEVA 4, SWITZERLAND
来源
FEBS LETTERS | 1994年 / 354卷 / 02期
关键词
ACETYLCHOLINE; NICOTINE; NICOTINIC RECEPTOR; ALPHA-BUNGAROTOXIN; SH-SY5Y CELL LINE;
D O I
10.1016/0014-5793(94)01108-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Neuronal nicotinic acetylcholine receptors (nAChR) are made from different combinations of subunits encoded by a diverse family of genes. However, the recently cloned alpha 7 gene codes for subunits that can form homooligomeric nAChR complexes when expressed in Xenopus oocytes. Electrophysiological studies reveal that these alpha 7-nAChR function as alpha-bungarotoxin (Bgt)-sensitive, quickly activating/inactivating ion channels with a unique pharmacological profile and an unusually high permeability to calcium ions. Although similar observations have been made in studies of Bgt-sensitive, functional nAChR subtypes that are naturally expressed in neuronal cells, all attempts until now to reconstitute functional alpha 7-nAChR in cell lines have failed. Here we report the successful use of SH-SY5Y human neuroblastoma cells, which naturally express low levels of endogenous alpha 7 transcripts, to stably overexpress heterologous rat nAChR alpha 7 transgenes. These transgenes are expressed as the appropriately-sized alpha 7 messages and protein, and stably transfected SH-SY5Y cells have over 30-times higher levels of specific Bgt binding sites than do wild-type cells. Whole cell current recordings confirm that transfected cells express functional nAChR that are sensitive to blockade by Bgt and display the typical physiological and pharmacological profiles of alpha 7-nAChR. We conclude that stable, functional expression of alpha 7 transgenes in a mammalian cell line has been achieved for the first time.
引用
收藏
页码:155 / 159
页数:5
相关论文
共 50 条
  • [31] Duplicated α7 nicotinic receptor gene product directly interacts with a 7 receptor in human SH-SY5Y neuroblastoma cells
    Takagi, K
    Tsuneki, H
    Kobayashi, S
    Ogawa, M
    Wada, T
    Sasaoka, T
    JOURNAL OF PHARMACOLOGICAL SCIENCES, 2006, 100 : 291P - 291P
  • [32] Modulation of α7-containing nicotinic receptor expression in SH-SY5Y cells and primary hippocampal cultures
    Ridley, DL
    Wonnacott, S
    BRITISH JOURNAL OF PHARMACOLOGY, 1998, 125 : U40 - U40
  • [33] NATIVE NICOTINIC ACETYLCHOLINE-RECEPTORS IN HUMAN IMR32 NEUROBLASTOMA-CELLS - FUNCTIONAL, IMMUNOLOGICAL AND PHARMACOLOGICAL PROPERTIES
    GOTTI, C
    BRISCINI, L
    VERDERIO, C
    OORTGIESEN, M
    BALESTRA, B
    CLEMENTI, F
    EUROPEAN JOURNAL OF NEUROSCIENCE, 1995, 7 (10) : 2083 - 2092
  • [34] Functional α2C-adrenoceptors in human neuroblastoma SH-SY5Y cells
    Parsley, S
    Gazi, L
    Bobirnac, I
    Loetscher, E
    Schoeffter, P
    EUROPEAN JOURNAL OF PHARMACOLOGY, 1999, 372 (01) : 109 - 115
  • [35] GLIAL CONDITIONED MEDIUM ALTERS THE EXPRESSION OF AMYLOID PRECURSOR PROTEIN IN SH-SY5Y NEUROBLASTOMA-CELLS
    HIROSE, Y
    IMAI, Y
    NAKAJIMA, K
    TAKEMOTO, N
    TOYA, S
    KOHSAKA, S
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1994, 198 (02) : 504 - 509
  • [36] Effect of HFE Variants on Sphingolipid Expression by SH-SY5Y Human Neuroblastoma Cells
    F. Ali-Rahmani
    J. A. Hengst
    J. R. Connor
    C.-L. Schengrund
    Neurochemical Research, 2011, 36 : 1687 - 1696
  • [37] Modulation of tyrosine hydroxylase expression by melatonin in human SH-SY5Y neuroblastoma cells
    McMillan, Catherine R.
    Sharma, Rohita
    Ottenhof, Tom
    Niles, Lennard P.
    NEUROSCIENCE LETTERS, 2007, 419 (03) : 202 - 206
  • [38] Expression and Pharmacology of Endogenous Cav Channels in SH-SY5Y Human Neuroblastoma Cells
    Sousa, Silmara R.
    Vetter, Irina
    Ragnarsson, Lotten
    Lewis, Richard J.
    PLOS ONE, 2013, 8 (03):
  • [39] Propofol increases μ-opioid receptor expression in SH-SY5Y human neuroblastoma cells
    Li, Zuojun
    Pei, Qi
    Cao, Lijun
    Xu, Linyong
    Zhang, Bikui
    Liu, Shikun
    MOLECULAR MEDICINE REPORTS, 2012, 6 (06) : 1333 - 1336
  • [40] COUPLING OF MUSCARINIC RECEPTORS TO THE MOBILIZATION OF INTRACELLULAR CA-2+ STORES IN PERMEABILIZED SH-SY5Y HUMAN NEUROBLASTOMA-CELLS
    WOJCIKIEWICZ, RJH
    SAFRANY, ST
    CHALLISS, RAJ
    STRUPISH, J
    NAHORSKI, SR
    BIOCHEMICAL JOURNAL, 1990, 272 (01) : 269 - 272