The mRNA Expression of Serotonin 2C Subtype Receptors Uncoupled With Inositol Hydrolysis in NG108-15 Cells

Tohda, Michihisa; Sukma, Monrudee; Nomura, Yasuyuki; Watanabe, Hiroshi

doi:10.1254/jjp.90.138

Cited by 9 publications

(10 citation statements)

References 29 publications

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“…Figure 1 shows that two major PCR fragments were made from the RT product of the high-temperaturestable enzyme. Sequence analysis revealed that the low molecular weight band corresponded to the same short variant 5-HT2CR mRNA as we previously reported (6). The high molecular band was shown to have the sequence of intact 5-HT2CR mRNA.…”

Section: Determination Of the Bases In The Position Of Rna Editingsupporting

confidence: 73%

“…Since the RNA editing site is included in the 95-nt deleted sequence of the short variant, it should be of interest to determine the relationship between editing and this deleted sequence. We previously reported that NG108-15 cells abundantly express the short variant mRNA, but show almost no expression of 5-HT2CR-type mRNA in the normal condition (6). To study the relationship and the cellular function of short variant, drug induced neuronal differentiated NG108-15 cells were used for detection of 5-HT2C mRNA and the base sequences at the editing site compared with nondifferentiated cells.…”

Section: Introductionmentioning

confidence: 99%

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RNA Editing and Short Variant of Serotonin 2C Receptor mRNA in Neuronally Differentiated NG108-15 Cells

2004

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Abstract. Two types of serotonin 2C subtype receptor mRNA, receptor-type and short variant, has been reported. The expression of the receptor-type mRNA could be detected as well as the short variant in NG108-15 cells by using a high temperature stable reversetranscriptase and the expression of the receptor-type mRNA was enhanced in drug-induced neuronal differentiated cells. The deleted sequence of the short variant include the RNA editing site by adenosine deaminase. Analysis of the sequence at the editing site revealed that the mRNA of undifferentiated cells was highly edited at sites A and B and that cytosine deaminase activity may also be involved in neuronal differentiation.

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Section: Determination Of the Bases In The Position Of Rna Editingsupporting

confidence: 73%

Section: Introductionmentioning

confidence: 99%

RNA Editing and Short Variant of Serotonin 2C Receptor mRNA in Neuronally Differentiated NG108-15 Cells

2004

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“…The RNA editing on 5-HT2CR is a rare case that generates on the coding site (23). The 95-bases fragment of the RNA including these editing sites is stripped off and the short variant without function as the receptor is produced (24). Moreover, brain-specific small nuclear RNA, HBII-52, is reported as small RNA combined with the editing area (25).…”

Section: Discussionmentioning

confidence: 99%

Serotonin 2C Receptor (5-HT2CR) mRNA Editing–Induced Down-Regulation of 5-HT2CR Function in Xenopus Oocytes: the Significance of Site C Editing

et al. 2010

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Abstract. Serotonin 2C receptor (5-HT2CR) mRNA receives editing at 5 nucleotide positions (sites A -E) located in the sequence encoding the second intracellular loop of 5-HT2CR. 5-HT2CR mRNA without editing and with editing at sites AB, ABD, ABC, ABCD, and C are translated to 6 isoforms of 5-HT2CR: INI(non-edited), VNI(AB), VNV(ABD), VSI(ABC), VSV(ABCD), and ISI(C), respectively. In this study, we investigated electrophysiologically the ability of these isoforms to couple with the G protein / phospholipase C (PLC) system using Xenopus oocytes injected with edited 5-HT2CR RNAs and muscarinic M 1 receptor (M1R) RNA. The efficacy with which 5-HT stimulated each isoform was calculated by comparing 5-HT-induced current with 100 μ M acetylcholine-induced M1R current. Stimulation with 5-HT of INI(non-edited), VNI(AB), VNV(ABD), VSI(ABC), VSV(ABCD), and ISI(C) expressed in Xenopus oocytes showed concentration-dependent responses with EC 50 values of 8.6, 17.2, 76,5, 22.0, 91.2, and 20.3 nM, respectively. No significant difference in the ability of 5-HT to induce currents among the oocytes expressing these isoforms was detected, but in the oocytes expressing VSI(ABC) or VSV(ABCD), 5-HT had a significantly reduced ability to induce currents. These results suggest that editing at site C together with sites A and B and/or D markedly reduces 5-HT2CR function by generating isoforms with reduced ability to activate PLC.

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“…The present findings about high sensitivity of EHNA against 5-HT2CR mRNA editing at sites C and D suggest that EHNA is useful to investigate the functional roles of these sites' mRNA editing such as the generation of short variant mRNA. 16,17) …”

Section: Discussionmentioning

confidence: 99%

Influence of an Adenosine Deaminase Inhibitor, Erythro-9-(2-hydroxy-3-nonyl) Adenine Hydrochloride, on 5-HT2CR mRNA Editing in Primary Cultured Cortical Cells

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Tohda

Tezuka

et al. 2010

Biological & Pharmaceutical Bulletin

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The adenosine to inosine RNA editing process mediated by adenosine deaminase acting on RNA (ADARs) has been reported to play critical roles in the functional development of the central nervous system by modulating the functions of neurotransmitter receptors. Serotonin 2C receptor (5-HT2CR) mRNA undergoes editing by these ADARs at 5 nucleotide positions (sites A-E) located in the sequence encoding the second intracellular loop of 5-HT2CR. This editing allows for the generation of 32 mRNA variants and 24 protein isoforms of the receptor differing in G-protein coupling efficiency.1) The involvement of 5-HT2CR mRNA editing in psychiatric disorders has been hypothesized based on etiological and pharmacological studies. Altered editing of 5-HT2CR mRNA was detected in the postmortem brains of patients with major depression, schizophrenia and bipolar disorder.2-4) Yang et al. 5) found that 5-HT2CR mRNA editing frequency in human glioblastoma cell lines was altered by treatment with interferon which causes depression as an adverse effect. These findings also raised the possibility that compounds with the ability to inhibit/stimulate the editing of 5-HT2CR mRNA are useful for clarifying the role of 5-HT2CR mRNA editing in psychiatric disorders.Our previous studies have shown that the editing efficacy of 5-HT2CR mRNA and ADAR-2 pre-mRNA and the level of ADAR-2 pre-mRNA are altered during the development of the brain in rats.6,7) These results suggested that 5-HT2CR mRNA editing and these enzymes play a role in the brain's maturation. To find a pharmacological tool with which to investigate the roles of 5-HT2CR mRNA editing, we examined the effect of erythro-9-(2-hydroxy-3-nonyl) adenine hydrochloride (EHNA), a well-known inhibitor of adenosine deaminase (ADA) which breaks extracellular adenosine, 8) on the editing of 5-HT2CR mRNA at sites A-D in cultured cortical cells. MATERIALS AND METHODSCultured cortical cells prepared from E20 embryos as described previously 6,7) were used for the experiments. The cells were exposed to vehicle (control) or EHNA included in culture media at concentrations of 30 and 60 mM for 6 d.After a 6-d cultivation period, total RNA was extracted, and RT-PCR was carried out as described previously.6,7) The specific primers for target genes to identify the bases at editing sites were: 5-HT2CR mRNA (NM_008312); 5Ј-atg tcc cta gcc att gct gat atg ctg gtg-3Ј (sense), 5Ј-atg cca cga agg acc cga tga gaa cga agt-3Ј (antisense), and Texas Red-labeled: 5Ј-ata ttt gtg ccc cgt cgt ga; ADAR-2 pre-mRNA (NW_047598), 5Ј-atg tgc tgg tga act cac ag-3Ј (sense), 5Ј-gtg gtg ccc aga aag agt gg-3Ј (antisense) and Texas Red-labeled: 5Ј-taa gta gga gag gca ata ggt ccg-3Ј; and GluR2 (NM_017261), 5Ј-tat atg agg agt gca gag cc-3Ј (sense), 5Ј-ata ctc cag caa cgt tgc tc-3Ј (antisense) and Texas Red-labeled: 5Ј-ctg tgt ttg tga gga cta cc-3Ј. Based on pilot studies where we ran specific primers for target genes, the proper cycle number for the amplification of each target gene was chosen (5-HT2CR mRNA including the editing sit...

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The mRNA Expression of Serotonin 2C Subtype Receptors Uncoupled With Inositol Hydrolysis in NG108-15 Cells

Cited by 9 publications

References 29 publications

RNA Editing and Short Variant of Serotonin 2C Receptor mRNA in Neuronally Differentiated NG108-15 Cells

RNA Editing and Short Variant of Serotonin 2C Receptor mRNA in Neuronally Differentiated NG108-15 Cells

Serotonin 2C Receptor (5-HT2CR) mRNA Editing–Induced Down-Regulation of 5-HT2CR Function in Xenopus Oocytes: the Significance of Site C Editing

Influence of an Adenosine Deaminase Inhibitor, Erythro-9-(2-hydroxy-3-nonyl) Adenine Hydrochloride, on 5-HT2CR mRNA Editing in Primary Cultured Cortical Cells

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