The properties of 5‐HT 3 receptors in clonal cell lines studied by patch‐clamp techniques

The properties of 5‐HT 3 receptors in clonal cell lines studied by patch‐clamp techniques 1 The characteristics of transmembrane currents evoked by 5‐hydroxytryptamine (5‐HT) in the neuroblastoma x Chinese hamster brain cell line NCB‐20 and neuroblastoma clonal cell line N1E‐115 have been studied under voltage‐clamp conditions by the whole‐cell recording and outside‐out membrane patch modes of the patch‐clamp technique. 2 In 73% of NCB‐20 cells examined (n = 221), and all N1E‐115 cells studied (n = 80), 5‐HT (10 μm) elicited a transient inward current at negative holding potentials, this being associated with an increase in membrane conductance. In both cell lines responses to 5‐HT reversed in sign at a potential of approximately −2mV and demonstrated inward rectification. 3 The reversal potential of 5‐HT‐induced currents (E5‐HT) recorded from either NCB‐20 or N1E‐115 cells was unaffected by total replacement of internal K+ by Cs+. In N1E‐115 cells, reducing internal K+ concentration from 140 to 20 mm produced a positive shift in E5‐HT of approximately 28 mV, whereas reducing external Na+ from 143 to 20 mm was associated with a negative shift in E5‐HT of about 37 mV. A large reduction in internal Cl− concentration (from 144 to 6 mm) had little effect on E5‐HT. 4 5‐HT‐induced currents of NCB‐20 cells were unaffected by methysergide (1 μm) or ketanserin (1 μm), but were reversibly antagonized by GR38032F (0.1–1.0 nm) with an IC50 of 0.25 nm. GR 38032F (0.3 nm) reduced 5‐HT‐induced currents in N1E‐115 cells to approximately 26% of their control value. 5 On outside‐out membrane patches excised from both NCB‐20 and N1E‐115 cells, 5‐HT induced small inward currents which could not be clearly resolved into discrete single channel events. Such responses were: (i) reversibly antagonized by GR 38032F (1 nm) (ii) reversed in sign at 0 mV, and (iii) subject to desensitization. 6 Fluctuation analysis of inward currents evoked by 5‐HT (1 μm) in N1E‐115 cells suggests that 5‐HT gates a channel with a conductance of approximately 310fS. Such a relatively small conductance could readily explain why the response of outside‐out membrane patches to 5‐HT cannot at present be resolved into clear single channel events. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png British Journal of Pharmacology Wiley

The properties of 5‐HT 3 receptors in clonal cell lines studied by patch‐clamp techniques

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Publisher
Wiley
Copyright
1989 British Pharmacological Society
ISSN
0007-1188
eISSN
1476-5381
DOI
10.1111/j.1476-5381.1989.tb11920.x
Publisher site
See Article on Publisher Site

Abstract

1 The characteristics of transmembrane currents evoked by 5‐hydroxytryptamine (5‐HT) in the neuroblastoma x Chinese hamster brain cell line NCB‐20 and neuroblastoma clonal cell line N1E‐115 have been studied under voltage‐clamp conditions by the whole‐cell recording and outside‐out membrane patch modes of the patch‐clamp technique. 2 In 73% of NCB‐20 cells examined (n = 221), and all N1E‐115 cells studied (n = 80), 5‐HT (10 μm) elicited a transient inward current at negative holding potentials, this being associated with an increase in membrane conductance. In both cell lines responses to 5‐HT reversed in sign at a potential of approximately −2mV and demonstrated inward rectification. 3 The reversal potential of 5‐HT‐induced currents (E5‐HT) recorded from either NCB‐20 or N1E‐115 cells was unaffected by total replacement of internal K+ by Cs+. In N1E‐115 cells, reducing internal K+ concentration from 140 to 20 mm produced a positive shift in E5‐HT of approximately 28 mV, whereas reducing external Na+ from 143 to 20 mm was associated with a negative shift in E5‐HT of about 37 mV. A large reduction in internal Cl− concentration (from 144 to 6 mm) had little effect on E5‐HT. 4 5‐HT‐induced currents of NCB‐20 cells were unaffected by methysergide (1 μm) or ketanserin (1 μm), but were reversibly antagonized by GR38032F (0.1–1.0 nm) with an IC50 of 0.25 nm. GR 38032F (0.3 nm) reduced 5‐HT‐induced currents in N1E‐115 cells to approximately 26% of their control value. 5 On outside‐out membrane patches excised from both NCB‐20 and N1E‐115 cells, 5‐HT induced small inward currents which could not be clearly resolved into discrete single channel events. Such responses were: (i) reversibly antagonized by GR 38032F (1 nm) (ii) reversed in sign at 0 mV, and (iii) subject to desensitization. 6 Fluctuation analysis of inward currents evoked by 5‐HT (1 μm) in N1E‐115 cells suggests that 5‐HT gates a channel with a conductance of approximately 310fS. Such a relatively small conductance could readily explain why the response of outside‐out membrane patches to 5‐HT cannot at present be resolved into clear single channel events.

Journal

British Journal of PharmacologyWiley

Published: May 1, 1989

References

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