Functional acetylcholine receptor in PC12 cells reacts with a monoclonal antibody to brain nicotinic receptors

Nature ◽  
1987 ◽  
Vol 327 (6122) ◽  
pp. 515-518 ◽  
Author(s):  
P. J. Whiting ◽  
R. Schoepfer ◽  
L. W. Swanson ◽  
D. M. Simmons ◽  
J. M. Lindstrom
Keyword(s):  
Monoclonal Antibody ◽  
Pc12 Cells ◽  
Acetylcholine Receptor ◽  
Nicotinic Receptors ◽  
Brain Nicotinic Receptors

Effects of nerve growth factor and nicotine on the expression of nicotinic acetylcholine receptor subunits in PC12 cells

1999 ◽  
Vol 35 (3) ◽  
pp. 175-181 ◽  
Author(s):  
Tetsuya Takahashi ◽  
Hiroshi Yamashita ◽  
Sigenobu Nakamura ◽  
Hiroshi Ishiguro ◽  
Toshiharu Nagatsu ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Pc12 Cells ◽  
Acetylcholine Receptor ◽  
Nicotinic Acetylcholine Receptor ◽  
Nicotinic Acetylcholine ◽  
Nerve Growth

Monoclonal antibody to acetylcholine receptor: cell line established from thymus of patient with Myasthenia gravis

Science ◽  
1982 ◽  
Vol 215 (4535) ◽  
pp. 995-997 ◽  
Author(s):  
I Kamo ◽  
S Furukawa ◽  
A Tada ◽  
Y Mano ◽  
Y Iwasaki ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Myasthenia Gravis ◽  
Cell Line ◽  
Acetylcholine Receptor ◽  
Receptor Cell

Local Anesthetics Inhibit Muscarinic Receptor-mediated Activation of Extracellular Signal-regulated Kinases in Rat Pheochromocytoma PC12 Cells 

1999 ◽  
Vol 91 (4) ◽  
pp. 1014-1014 ◽  
Author(s):  
Zhiming Tan ◽  
Shuji Dohi ◽  
Kenji Ohguchi ◽  
Shigeru Nakashima ◽  
Yoshinori Nozawa
Keyword(s):  
Tyrosine Phosphorylation ◽  
Pc12 Cells ◽  
Acetylcholine Receptor ◽  
Local Anesthetics ◽  
Muscarinic Acetylcholine Receptor ◽  
Erk Activation ◽  
Muscarinic Acetylcholine ◽  
M3 Muscarinic Acetylcholine Receptor ◽  
Extracellular Signal ◽  
Pheochromocytoma Pc12

Background Because protein phosphorylation is a key mechanism for controlling cellular functions and extracellular signal-regulated kinase (ERK) plays a role in cellular signal transduction, the authors wanted to determine whether local anesthetics interfere with biochemical signaling molecules. Methods Protein tyrosine phosphorylation and ERK activation induced by carbachol, an agonist for muscarinic acetylcholine receptors, were examined in rat pheochromocytoma PC12 cells, a model for investigating signal transduction. Carbachol-induced tyrosine-phosphorylated proteins of 44 and 42 kd were determined by Western blot analysis and identified as activated ERK1 and ERK2 using anti-ERK antibody. The ERK activation was blocked by preincubation with atropine or an M3 muscarinic acetylcholine receptor antagonist 4-diphenyacetooxy-1, 1-dimethylpiperidinium, indicating that is was mediated by M3 muscarinic acetylcholine receptor activation. Then, in the presence of local anesthetic, the carbachol-induced tyrosine phosphorylation and ERK activation were evaluated. The effects of three Na+ current-modifying reagents on carbachol-induced ERK activation were also evaluated. Results Procaine (10(-4) to 10(-3) M) inhibited carbachol-induced tyrosine phosphorylation and ERK activation in a concentration-dependent manner. Although tetracaine, lidocaine, and bupivacaine similarly suppressed carbachol-induced tyrosine phosphorylation and ERK activation, neither tetrodotoxin, veratridine, nor ouabain affected the carbachol-induced ERKs activation. Both ERKs were also activated by 4beta-phorbol 12-myristate 13-acetate, an activator of protein kinase C, and fluoroaluminate (AlF4-), respectively, but procaine did not affect ERK activation induced by these two substances. The inhibition of carbachol-induced ERK activation by procaine was not modified by a phosphatase inhibitor, calyculin A. Conclusions The current results indicate that local anesthetics inhibit the activity of the signal-transducing molecule(s) leading to M3 muscarinic acetylcholine receptor-mediated ERK activation in PC12 cells. Such action is unlikely to be a result of the drug's action on Na+ channels or on the electrochemical gradients of the neuronal cell membrane.

The membrane skeleton of acetylcholine receptor domains in rat myotubes contains antiparallel homodimers of beta-spectrin in filaments quantitatively resembling those of erythrocytes

1995 ◽  
Vol 108 (9) ◽  
pp. 3145-3154 ◽  
Author(s):  
D.W. Pumplin
Keyword(s):  
Monoclonal Antibody ◽  
Acetylcholine Receptor ◽  
Acetylcholine Receptors ◽  
Cytoplasmic Membrane ◽  
Membrane Surface ◽  
Membrane Skeleton ◽  
Immunogold Labeling ◽  
Platinum Coating ◽  
High Concentrations ◽  
Receptor Clusters

I used immunogold labeling and quick-freeze, deep-etch, rotary replication to characterize the membrane skeleton at regions with high concentrations of acetylcholine receptor domains in receptor clusters of cultured rat muscle cells. This membrane skeleton consists of a network of filaments closely applied to the cytoplasmic membrane surface. The filaments are specifically decorated by immunogold labeling with a monoclonal antibody, VIIF7, that recognizes an isoform of beta-spectrin colocalizing with acetylcholine receptors. The filaments are 32 +/- 11 nm in length and three to four filaments (average 3.1-3.3) join at each intersection to form the network. These parameters are nearly identical to those reported previously for the membrane skeleton of erythrocytes. Depending on the amount of platinum coating, filament diameters range from 9 to 11 nm in diameter, and are 1.4 nm larger on average than spectrin filaments of erythrocytes replicated at the same time. Filaments are decorated with gold particles close to one end, consistent with the location of the epitope recognized by the monoclonal antibody. Computer modeling shows that all filament intersections in the membrane skeletal network are equally capable of being labeled by the monoclonal antibody. This pattern of labeling is consistent with a network containing antiparallel homodimers of beta-spectrin.

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