Recombinant human serotonin 5A receptors stably expressed in C6 glioma cells couple to multiple signal transduction pathways

Mami Noda, Satsuki Yasuda, Mitsuko Okada, Haruhiro Higashida, Aki Shimada, Nakao Iwata, Norio Ozaki, Kaori Nishikawa, Sakiko Shirasawa, Mayumi Uchida, Shunsuke Aoki, Keiji Wada

Research output: Contribution to journalArticle

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Abstract

Human serotonin 5A (5-HT5A) receptors were stably expressed in undifferentiated C6 glioma. In 5-HT5A receptors-expressing cells, accumulation of cAMP by forskolin was inhibited by 5-HT as reported previously. Pertussis toxin-sensitive inhibition of ADP-ribosyl cyclase was also observed, indicating a decrease of cyclic ADP ribose, a potential intracellular second messenger mediating ryanodine-sensitive Ca2+ mobilization. On the other hand, 5-HT-induced outward currents were observed using the patch-clamp technique in whole-cell configuration. The 5-HT-induced outward current was observed in 84% of the patched 5-HT5A receptor-expressing cells and was concentration-dependent. The 5-HT-induced current was inhibited when intracellular K+ was replaced with Cs+ but was not significantly inhibited by typical K+ channel blockers. The 5-HT-induced current was significantly attenuated by 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) in the patch pipette. Depleting intracellular Ca2+ stores by application of caffeine or thapsigargin also blocked the 5-HT-induced current. Blocking G protein, the inositol triphosphate (IP3) receptor, or pretreatment with pertussis toxin, all inhibited the 5-HT-induced current. IP3 showed a transient increase after application of 5-HT in 5-HT5A receptor-expressing cells. It was concluded that in addition to the inhibition of cAMP accumulation and ADP-ribosyl cyclase activity, 5-HT5A receptors regulate intracellular Ca2+ mobilization which is probably a result of the IP3-sensitive Ca2+ store. These multiple signal transduction systems may induce complex changes in the serotonergic system in brain function.

Original languageEnglish
Pages (from-to)222-232
Number of pages11
JournalJournal of Neurochemistry
Volume84
Issue number2
DOIs
Publication statusPublished - 01-01-2003

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Signal transduction
Glioma
Signal Transduction
Serotonin
Induced currents
ADP-ribosyl Cyclase
Inositol 1,4,5-Trisphosphate Receptors
Pertussis Toxin
Cyclic ADP-Ribose
serotonin 5 receptor
Ryanodine
Ethane
Thapsigargin
Clamping devices
Second Messenger Systems
Patch-Clamp Techniques
Colforsin
Inositol
Caffeine
GTP-Binding Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Noda, Mami ; Yasuda, Satsuki ; Okada, Mitsuko ; Higashida, Haruhiro ; Shimada, Aki ; Iwata, Nakao ; Ozaki, Norio ; Nishikawa, Kaori ; Shirasawa, Sakiko ; Uchida, Mayumi ; Aoki, Shunsuke ; Wada, Keiji. / Recombinant human serotonin 5A receptors stably expressed in C6 glioma cells couple to multiple signal transduction pathways. In: Journal of Neurochemistry. 2003 ; Vol. 84, No. 2. pp. 222-232.
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abstract = "Human serotonin 5A (5-HT5A) receptors were stably expressed in undifferentiated C6 glioma. In 5-HT5A receptors-expressing cells, accumulation of cAMP by forskolin was inhibited by 5-HT as reported previously. Pertussis toxin-sensitive inhibition of ADP-ribosyl cyclase was also observed, indicating a decrease of cyclic ADP ribose, a potential intracellular second messenger mediating ryanodine-sensitive Ca2+ mobilization. On the other hand, 5-HT-induced outward currents were observed using the patch-clamp technique in whole-cell configuration. The 5-HT-induced outward current was observed in 84{\%} of the patched 5-HT5A receptor-expressing cells and was concentration-dependent. The 5-HT-induced current was inhibited when intracellular K+ was replaced with Cs+ but was not significantly inhibited by typical K+ channel blockers. The 5-HT-induced current was significantly attenuated by 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) in the patch pipette. Depleting intracellular Ca2+ stores by application of caffeine or thapsigargin also blocked the 5-HT-induced current. Blocking G protein, the inositol triphosphate (IP3) receptor, or pretreatment with pertussis toxin, all inhibited the 5-HT-induced current. IP3 showed a transient increase after application of 5-HT in 5-HT5A receptor-expressing cells. It was concluded that in addition to the inhibition of cAMP accumulation and ADP-ribosyl cyclase activity, 5-HT5A receptors regulate intracellular Ca2+ mobilization which is probably a result of the IP3-sensitive Ca2+ store. These multiple signal transduction systems may induce complex changes in the serotonergic system in brain function.",
author = "Mami Noda and Satsuki Yasuda and Mitsuko Okada and Haruhiro Higashida and Aki Shimada and Nakao Iwata and Norio Ozaki and Kaori Nishikawa and Sakiko Shirasawa and Mayumi Uchida and Shunsuke Aoki and Keiji Wada",
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Noda, M, Yasuda, S, Okada, M, Higashida, H, Shimada, A, Iwata, N, Ozaki, N, Nishikawa, K, Shirasawa, S, Uchida, M, Aoki, S & Wada, K 2003, 'Recombinant human serotonin 5A receptors stably expressed in C6 glioma cells couple to multiple signal transduction pathways', Journal of Neurochemistry, vol. 84, no. 2, pp. 222-232. https://doi.org/10.1046/j.1471-4159.2003.01518.x

Recombinant human serotonin 5A receptors stably expressed in C6 glioma cells couple to multiple signal transduction pathways. / Noda, Mami; Yasuda, Satsuki; Okada, Mitsuko; Higashida, Haruhiro; Shimada, Aki; Iwata, Nakao; Ozaki, Norio; Nishikawa, Kaori; Shirasawa, Sakiko; Uchida, Mayumi; Aoki, Shunsuke; Wada, Keiji.

In: Journal of Neurochemistry, Vol. 84, No. 2, 01.01.2003, p. 222-232.

Research output: Contribution to journalArticle

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T1 - Recombinant human serotonin 5A receptors stably expressed in C6 glioma cells couple to multiple signal transduction pathways

AU - Noda, Mami

AU - Yasuda, Satsuki

AU - Okada, Mitsuko

AU - Higashida, Haruhiro

AU - Shimada, Aki

AU - Iwata, Nakao

AU - Ozaki, Norio

AU - Nishikawa, Kaori

AU - Shirasawa, Sakiko

AU - Uchida, Mayumi

AU - Aoki, Shunsuke

AU - Wada, Keiji

PY - 2003/1/1

Y1 - 2003/1/1

N2 - Human serotonin 5A (5-HT5A) receptors were stably expressed in undifferentiated C6 glioma. In 5-HT5A receptors-expressing cells, accumulation of cAMP by forskolin was inhibited by 5-HT as reported previously. Pertussis toxin-sensitive inhibition of ADP-ribosyl cyclase was also observed, indicating a decrease of cyclic ADP ribose, a potential intracellular second messenger mediating ryanodine-sensitive Ca2+ mobilization. On the other hand, 5-HT-induced outward currents were observed using the patch-clamp technique in whole-cell configuration. The 5-HT-induced outward current was observed in 84% of the patched 5-HT5A receptor-expressing cells and was concentration-dependent. The 5-HT-induced current was inhibited when intracellular K+ was replaced with Cs+ but was not significantly inhibited by typical K+ channel blockers. The 5-HT-induced current was significantly attenuated by 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) in the patch pipette. Depleting intracellular Ca2+ stores by application of caffeine or thapsigargin also blocked the 5-HT-induced current. Blocking G protein, the inositol triphosphate (IP3) receptor, or pretreatment with pertussis toxin, all inhibited the 5-HT-induced current. IP3 showed a transient increase after application of 5-HT in 5-HT5A receptor-expressing cells. It was concluded that in addition to the inhibition of cAMP accumulation and ADP-ribosyl cyclase activity, 5-HT5A receptors regulate intracellular Ca2+ mobilization which is probably a result of the IP3-sensitive Ca2+ store. These multiple signal transduction systems may induce complex changes in the serotonergic system in brain function.

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