Cloning and expression of ligand-gated ion-channel receptor L2 in central nervous system

Houtani, Takeshi; Munemoto, Yumi; Kase, Masahiko; Sakuma, Satoru; Tsutsumi, Toshiyuki; Sugimoto, Tetsuo

doi:10.1016/j.bbrc.2005.07.079

Cited by 38 publications

(29 citation statements)

References 18 publications

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“…These classical members of this superfamily play important roles in physiology and pathology and thus constitute interesting drug targets in a wide range of disorders [2–5]. ZAC is the least understood member of this family, partly because the gene encoding for ZAC is a pseudogene in mouse and rat genomes, which has complicated explorations into the physiological function of the receptor [1,6]. …”

Section: Introductionmentioning

confidence: 99%

“…ZAC displays very little amino acid sequence similarity with other members in the superfamily, and thus it is classified within its own subgroup, the closest matches to ZAC being the human 5-HT3A, 5-HT3B and nACh α7 subunits that exhibit approximately 15% amino acid sequence identity to ZAC [1,6]. Expression studies have detected ZAC mRNA in human fetal whole brain, spinal cord, pancreas, placenta, prostate, thyroid, trachea and stomach [1,6].…”

Section: Introductionmentioning

confidence: 99%

“…Expression studies have detected ZAC mRNA in human fetal whole brain, spinal cord, pancreas, placenta, prostate, thyroid, trachea and stomach [1,6]. Furthermore, the presence of ZAC mRNA has been demonstrated by RT-PCR in adult human hippocampus, striatum, amygdala and thalamus [6].…”

Section: Introductionmentioning

confidence: 99%

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Copper and protons directly activate the zinc-activated channel

Trattnig

Gasiorek

Deeb

et al. 2016

Biochemical Pharmacology

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The zinc-activated channel (ZAC) is a cationic ion channel belonging to the superfamily of Cys-loop receptors, which consists of pentameric ligand-gated ion channels. ZAC is the least understood member of this family so in the present study we sought to characterize the properties of this channel further. We demonstrate that not only zinc (Zn2+) but also copper (Cu2+) and protons (H+) are agonists of ZAC, displaying potencies and efficacies in the rank orders of H+ > Cu2+ > Zn2+ and H+ > Zn2+ > Cu2+, respectively. The responses elicited by Zn2+, Cu2+ and H+ through ZAC are all characterized by low degrees of desensitization. In contrast, currents evoked by high concentrations of the three agonists comprise distinctly different activation and decay components, with transitions to and from an open state being significantly faster for H+ than for the two metal ions. The permeabilities of ZAC for Na+ and K+ relative to Cs+ are indistinguishable, whereas replacing all of extracellular Na+ and K+ with the divalent cations Ca2+ or Mg2+ results in complete elimination of Zn2+-activated currents at both negative and positive holding potentials. This indicates that ZAC is non-selectively permeable to monovalent cations, whereas Ca2+ and Mg2+ inhibit the channel. In conclusion, this is the first report of a Cys-loop receptor being gated by Zn2+, Cu2+ and H+. ZAC could be an important mediator of some of the wide range of physiological functions regulated by or involving Zn2+, Cu2+ and H+.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Copper and protons directly activate the zinc-activated channel

Trattnig

Gasiorek

Deeb

et al. 2016

Biochemical Pharmacology

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“…On the other hand, non-neuronal fibroblast cell lines (Vero, HaK) are also able to respond to pesticides with a considerable change of the cell membrane potential, as previously shown for Vero cells treated with either organophosphates or carbamates [14], an effect that has been partially attributed to pesticide interactions with the zinc receptors on the kidney cells [15].…”

Section: Methodsmentioning

confidence: 62%

Cellular bioelectric profiling: a novel method to differentially assess the in vitro toxicity of pesticides

2017

JCEBC

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We present a novel approach for assessing pesticide toxicity on mammalian cells, by measuring the response patterns of four cell lines (two neuroblastoma and two fibroblast) to three different pesticide groups (carbamates, organophosphates, pyrethroids) at a broad range of concentrations. Two different aspects of the cellular bioelectric response are measured, namely the potential and the combined resistance + capacitance of the cell suspension. Both the cell line and the mode of measurement affected the observed in vitro cellular responses, with each cell type providing a unique pattern. The measured of resistance + capacitance through amperometry provided more reproducible results than potentiometry. The results of the study demonstrate the potential of bioelectric profiling as a tool for developing novel toxicity assays and associated biosensors with superior analytical capacity, speed of assay and the ability to respond to a broad spectrum of toxicants, at the same time satisfying the demand for increased cost-efficiency and ease of operation.

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“…Parvalbumin cDNA and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) cDNA were ampliWed by PCR as described previously (Houtani et al 2005). BrieXy, PCR was performed using a 20 l volume with Eppendorf Mastercycler Gradient (parvalbumin: 95°C 10 s, 68°C 20 s, 72°C 30 s, 40 cycles; GAPDH: 95°C 10 s, 60°C 20 s, 72°C 30 s, 45 cycles) using Extaq HS (Takara, Ohtsu, Japan) and primers (parvalbumin: 5Ј-AAA CAA AGA CGC TGA TGG CTG CTG G-3Ј and 5Ј-GGT GTC ATT CGA GGG CCA TAA AGG A-3Ј; GAPDH: 5Ј-CCC TCA AGA TTG TCA GCA ATG C-3Ј and 5Ј-GTC CTC AGT GTA GCC CAG GAT-3Ј).…”

Section: Methodsmentioning

confidence: 99%

Laser microdissection combined with immunohistochemistry on serial thin tissue sections: a method allowing efficient mRNA analysis

Kase

Houtani

Sakuma

et al. 2006

Histochem Cell Biol

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Laser microdissection (LMD) with subsequent reverse transcription-PCR analysis is a powerful histochemical technique subserving the molecular characterization of specific cell types. We developed an efficient method for selective sampling of specific cell populations using immunohistochemistry coupled with LMD. The cerebral cortex of adult rats was cut into serial thin sections. Some sections were immunostained for parvalbumin. The adjacent sections were mounted on Cell Support Film for LMD and stained with neutral red. By comparison of the two adjacent sections, neuronal profiles representing parts of parvalbumin-immunopositive somata were identified in the neutral red-stained sections. These neuronal profiles were safely captured with LMD and analyzed on reverse transcription-PCR using extracted RNA. The method presented here can be applied to cell-type-specific characterizations using fixed cells under RNase-free conditions.

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Cloning and expression of ligand-gated ion-channel receptor L2 in central nervous system

Cited by 38 publications

References 18 publications

Copper and protons directly activate the zinc-activated channel

Copper and protons directly activate the zinc-activated channel

Cellular bioelectric profiling: a novel method to differentially assess the in vitro toxicity of pesticides

Laser microdissection combined with immunohistochemistry on serial thin tissue sections: a method allowing efficient mRNA analysis

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