Trafficking of Kv2.1 Channels to the Axon Initial Segment by a Novel Nonconventional Secretory Pathway

Camilla Stampe Jensen; Shoji Watanabe; Jeroen Ingrid Stas; Jessica Klaphaak; Ayaka Yamane; Nicole Schmitt; Søren-Peter Olesen; James S Trimmer; Hanne Borger Rasmussen; Hiroaki Misonou

doi:10.1523/jneurosci.3510-16.2017

Trafficking of Kv2.1 Channels to the Axon Initial Segment by a Novel Nonconventional Secretory Pathway

Camilla Stampe Jensen, Shoji Watanabe, Jeroen Ingrid Stas, Jessica Klaphaak, Ayaka Yamane, Nicole Schmitt, Søren-Peter Olesen, James S Trimmer, Hanne Borger Rasmussen, Hiroaki Misonou

27 Citations (Scopus)

Abstract

Kv2.1 is a major delayed-rectifier voltage-gated potassium channel widely expressed in neurons of the CNS. Kv2.1 localizes in high-density cell-surface clusters in the soma and proximal dendrites as well as in the axon initial segment (AIS). Given the crucial roles of both of these compartments in integrating signal input and then generating output, this localization of Kv2.1 is ideal for regulating the overall excitability of neurons. Here we used fluorescence recovery after photobleaching imaging, mutagenesis, and pharmacological interventions to investigate the molecular mechanisms that control the localization of Kv2.1 in these two different membrane compartments in cultured rat hippocampal neurons of mixed sex. Our data uncover a unique ability of Kv2.1 channels to use two molecularly distinct trafficking pathways to accomplish this. Somatodendritic Kv2.1 channels are targeted by the conventional secretory pathway, whereas axonal Kv2.1 channels are targeted by a nonconventional trafficking pathway independent of the Golgi apparatus. We further identified a new AIS trafficking motif in the C-terminus of Kv2.1, and show that putative phosphorylation sites in this region are critical for the restricted and clustered localization in the AIS. These results indicate that neurons can regulate the expression and clustering of Kv2.1 in different membrane domains independently by using two distinct localization mechanisms, which would allow neurons to precisely control local membrane excitability.

Original language	English
Journal	The Journal of neuroscience : the official journal of the Society for Neuroscience
Volume	37
Issue number	48
Pages (from-to)	11523-11536
Number of pages	14
ISSN	0270-6474
DOIs	https://doi.org/10.1523/jneurosci.3510-16.2017
Publication status	Published - 29 Nov 2017

Keywords

Animals
Axon Initial Segment
Cell Membrane
Cells, Cultured
Female
HEK293 Cells
Hippocampus
Humans
Male
Neurons
Protein Transport
Rats
Secretory Pathway
Shab Potassium Channels
Journal Article

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Jensen, C. S., Watanabe, S., Stas, J. I., Klaphaak, J., Yamane, A., Schmitt, N., Olesen, S-P., Trimmer, J. S., Rasmussen, H. B., & Misonou, H. (2017). Trafficking of Kv2.1 Channels to the Axon Initial Segment by a Novel Nonconventional Secretory Pathway. The Journal of neuroscience : the official journal of the Society for Neuroscience, 37(48), 11523-11536. https://doi.org/10.1523/jneurosci.3510-16.2017

Trafficking of Kv2.1 Channels to the Axon Initial Segment by a Novel Nonconventional Secretory Pathway. / Jensen, Camilla Stampe; Watanabe, Shoji; Stas, Jeroen Ingrid et al.

In: The Journal of neuroscience : the official journal of the Society for Neuroscience, Vol. 37, No. 48, 29.11.2017, p. 11523-11536.

Research output: Contribution to journal › Journal article › Research › peer-review

Jensen, CS, Watanabe, S, Stas, JI, Klaphaak, J, Yamane, A, Schmitt, N, Olesen, S-P, Trimmer, JS, Rasmussen, HB & Misonou, H 2017, 'Trafficking of Kv2.1 Channels to the Axon Initial Segment by a Novel Nonconventional Secretory Pathway', The Journal of neuroscience : the official journal of the Society for Neuroscience, vol. 37, no. 48, pp. 11523-11536. https://doi.org/10.1523/jneurosci.3510-16.2017

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abstract = "Kv2.1 is a major delayed-rectifier voltage-gated potassium channel widely expressed in neurons of the CNS. Kv2.1 localizes in high-density cell-surface clusters in the soma and proximal dendrites as well as in the axon initial segment (AIS). Given the crucial roles of both of these compartments in integrating signal input and then generating output, this localization of Kv2.1 is ideal for regulating the overall excitability of neurons. Here we used fluorescence recovery after photobleaching imaging, mutagenesis, and pharmacological interventions to investigate the molecular mechanisms that control the localization of Kv2.1 in these two different membrane compartments in cultured rat hippocampal neurons of mixed sex. Our data uncover a unique ability of Kv2.1 channels to use two molecularly distinct trafficking pathways to accomplish this. Somatodendritic Kv2.1 channels are targeted by the conventional secretory pathway, whereas axonal Kv2.1 channels are targeted by a nonconventional trafficking pathway independent of the Golgi apparatus. We further identified a new AIS trafficking motif in the C-terminus of Kv2.1, and show that putative phosphorylation sites in this region are critical for the restricted and clustered localization in the AIS. These results indicate that neurons can regulate the expression and clustering of Kv2.1 in different membrane domains independently by using two distinct localization mechanisms, which would allow neurons to precisely control local membrane excitability.",

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T1 - Trafficking of Kv2.1 Channels to the Axon Initial Segment by a Novel Nonconventional Secretory Pathway

AU - Jensen, Camilla Stampe

AU - Watanabe, Shoji

AU - Stas, Jeroen Ingrid

AU - Klaphaak, Jessica

AU - Yamane, Ayaka

AU - Schmitt, Nicole

AU - Olesen, Søren-Peter

AU - Trimmer, James S

AU - Rasmussen, Hanne Borger

AU - Misonou, Hiroaki

PY - 2017/11/29

Y1 - 2017/11/29

N2 - Kv2.1 is a major delayed-rectifier voltage-gated potassium channel widely expressed in neurons of the CNS. Kv2.1 localizes in high-density cell-surface clusters in the soma and proximal dendrites as well as in the axon initial segment (AIS). Given the crucial roles of both of these compartments in integrating signal input and then generating output, this localization of Kv2.1 is ideal for regulating the overall excitability of neurons. Here we used fluorescence recovery after photobleaching imaging, mutagenesis, and pharmacological interventions to investigate the molecular mechanisms that control the localization of Kv2.1 in these two different membrane compartments in cultured rat hippocampal neurons of mixed sex. Our data uncover a unique ability of Kv2.1 channels to use two molecularly distinct trafficking pathways to accomplish this. Somatodendritic Kv2.1 channels are targeted by the conventional secretory pathway, whereas axonal Kv2.1 channels are targeted by a nonconventional trafficking pathway independent of the Golgi apparatus. We further identified a new AIS trafficking motif in the C-terminus of Kv2.1, and show that putative phosphorylation sites in this region are critical for the restricted and clustered localization in the AIS. These results indicate that neurons can regulate the expression and clustering of Kv2.1 in different membrane domains independently by using two distinct localization mechanisms, which would allow neurons to precisely control local membrane excitability.

AB - Kv2.1 is a major delayed-rectifier voltage-gated potassium channel widely expressed in neurons of the CNS. Kv2.1 localizes in high-density cell-surface clusters in the soma and proximal dendrites as well as in the axon initial segment (AIS). Given the crucial roles of both of these compartments in integrating signal input and then generating output, this localization of Kv2.1 is ideal for regulating the overall excitability of neurons. Here we used fluorescence recovery after photobleaching imaging, mutagenesis, and pharmacological interventions to investigate the molecular mechanisms that control the localization of Kv2.1 in these two different membrane compartments in cultured rat hippocampal neurons of mixed sex. Our data uncover a unique ability of Kv2.1 channels to use two molecularly distinct trafficking pathways to accomplish this. Somatodendritic Kv2.1 channels are targeted by the conventional secretory pathway, whereas axonal Kv2.1 channels are targeted by a nonconventional trafficking pathway independent of the Golgi apparatus. We further identified a new AIS trafficking motif in the C-terminus of Kv2.1, and show that putative phosphorylation sites in this region are critical for the restricted and clustered localization in the AIS. These results indicate that neurons can regulate the expression and clustering of Kv2.1 in different membrane domains independently by using two distinct localization mechanisms, which would allow neurons to precisely control local membrane excitability.

KW - Animals

KW - Axon Initial Segment

KW - Cell Membrane

KW - Cells, Cultured

KW - Female

KW - HEK293 Cells

KW - Hippocampus

KW - Humans

KW - Male

KW - Neurons

KW - Protein Transport

KW - Rats

KW - Secretory Pathway

KW - Shab Potassium Channels

KW - Journal Article

U2 - 10.1523/jneurosci.3510-16.2017

DO - 10.1523/jneurosci.3510-16.2017

M3 - Journal article

C2 - 29042434

SN - 0270-6474

VL - 37

SP - 11523

EP - 11536

JO - The Journal of neuroscience : the official journal of the Society for Neuroscience

JF - The Journal of neuroscience : the official journal of the Society for Neuroscience

IS - 48

ER -

Trafficking of Kv2.1 Channels to the Axon Initial Segment by a Novel Nonconventional Secretory Pathway

Abstract

Keywords

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