R&amp;D of C-Band Accelerating Structure at SINAP

Gu, Qiang; Fang, Wencheng; Zhao, Zhentang; Tong, Dechun

doi:10.1142/9789814602105_0009

Cited by 2 publications

(8 citation statements)

References 6 publications

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“…Next, we examined zinc's effect on hippocampal neurons. Some neurons, presumably GABAergic interneurons in mature cultured hippocampal neurons, generating relatively high firing frequency due to the presence of Kv3 channels (Gu et al 2012), were also reversibly inhibited by extracellular zinc at 100 μM (Fig. 1B).…”

Section: Zinc Reversibly Reduces the Firing Frequency Of Cultured Neumentioning

confidence: 94%

“…Glass pipettes with diameter around 50 μm, made by adjusting the pulling parameters and filled with Hank's buffer containing different concentrations of ZnCl 2 , were used for perfusion of whole neurons. The procedure has been described previously (Gu et al 2012). The perfusion lasts for approximately 10 s. The bath volume is about 2 ml, which is replaced with new Hank's buffer at the rate of 4 ml min −1 .…”

Section: Current-clamp Recording From Cultured Neurons and Whole-neurmentioning

confidence: 99%

“…The internal solution and Hank's buffer have been previously described for the recording of primary cultured neurons (Gu et al 2012). Hank's buffer: 150 mM NaCl, 4 mM KCl, 1.2 mM MgCl 2 , 10 mg ml −1 glucose, 1 mM CaCl 2 , 20 mM Hepes (pH 7.4).…”

Section: Current-clamp Recording From Cultured Neurons and Whole-neurmentioning

confidence: 99%

“…The internal solution of electrical pipettes: (in mM) 122 KMeSO 4 , 20 NaCl, 5 Mg-ATP, 0.3 GTP and 10 Hepes (pH 7.2). The current-clamp procedure has been previous described (Gu et al 2012).…”

Section: Current-clamp Recording From Cultured Neurons and Whole-neurmentioning

confidence: 99%

“…Biophysical properties of various Kv3 channel constructs were determined with voltage-clamp recording on transfected cells from the human embryonic kidney cell line HEK293 with the same internal solution and Hank's buffer, as previously described (Gu et al 2012). The patch pipette resistance in the bath was about 2-2.5 M .…”

Section: Voltage-clamp Recording From Transfected Hek293 Cellsmentioning

confidence: 99%

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Kv3 channel assembly, trafficking and activity are regulated by zinc through different binding sites

Gu¹,

Barry

2013

The Journal of Physiology

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Key points• Kv3 channels play a critical role in neuronal fast spiking and their regulation can profoundly influence the functions of various neural circuits.• Zinc reversibly inhibits fast spiking in cultured neurons by inhibiting Kv3 channels. Surprisingly, we found that the zinc inhibition is not through its known site in the N-terminal T1 domain of Kv3.1.• We have identified a new zinc-binding site in the Kv3.1 C-terminus, involved in regulating channel activity and targeting, but not the zinc inhibition.• We have also identified the junction between the first transmembrane segment and the first extracellular loop as the major zinc-sensing site in regulating Kv3 activity. This result allows us to reconstitute zinc-resistant fast spiking.• Kv3 channel assembly, localization and activity are regulated by zinc through intracellular and extracellular binding sites.Abstract Zinc, a divalent heavy metal ion and an essential mineral for life, regulates synaptic transmission and neuronal excitability via ion channels. However, its binding sites and regulatory mechanisms are poorly understood. Here, we report that Kv3 channel assembly, localization and activity are regulated by zinc through different binding sites. Local perfusion of zinc reversibly reduced spiking frequency of cultured neurons most likely by suppressing Kv3 channels. Indeed, zinc inhibited Kv3.1 channel activity and slowed activation kinetics, independent of its site in the N-terminal T1 domain. Biochemical assays surprisingly identified a novel zinc-binding site in the Kv3.1 C-terminus, critical for channel activity and axonal targeting, but not for the zinc inhibition. Finally, mutagenesis revealed an important role of the junction between the first transmembrane (TM) segment and the first extracellular loop in sensing zinc. Its mutant enabled fast spiking with relative resistance to the zinc inhibition. Therefore, our studies provide novel mechanistic insights into the multifaceted regulation of Kv3 channel activity and localization by divalent heavy metal ions.

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Section: Zinc Reversibly Reduces the Firing Frequency Of Cultured Neumentioning

confidence: 94%

Section: Current-clamp Recording From Cultured Neurons and Whole-neurmentioning

confidence: 99%

Section: Current-clamp Recording From Cultured Neurons and Whole-neurmentioning

confidence: 99%

Section: Current-clamp Recording From Cultured Neurons and Whole-neurmentioning

confidence: 99%

Section: Voltage-clamp Recording From Transfected Hek293 Cellsmentioning

confidence: 99%

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Kv3 channel assembly, trafficking and activity are regulated by zinc through different binding sites

Gu¹,

Barry

2013

The Journal of Physiology

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Numerical investigation of tuning method for nonuniform disk-loaded waveguides

Ayzatsky¹,

Mytrochenko²

2016

Preprint

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R&D of C-Band Accelerating Structure at SINAP

Cited by 2 publications

References 6 publications

Kv3 channel assembly, trafficking and activity are regulated by zinc through different binding sites

Kv3 channel assembly, trafficking and activity are regulated by zinc through different binding sites

Numerical investigation of tuning method for nonuniform disk-loaded waveguides

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