βγ subunits mediate the NPY enhancement of ATP-stimulated inositol phosphate formation

Xinying Li; Tsuneya Ikezu; Terry D. Hexum

doi:10.1016/j.peptides.2003.12.019

βγ subunits mediate the NPY enhancement of ATP-stimulated inositol phosphate formation

Xinying Li
, Tsuneya Ikezu
, Terry D. Hexum

Neuroscience

Research output: Contribution to journal › Article › peer-review

Abstract

Neuropeptide Y (NPY) enhances ATP-stimulated inositol phosphate (InsP) formation in bovine chromaffin cells through an unknown mechanism. Chromaffin cells were transduced with the carboxyl terminus of β-adrenergic receptor kinase 1 (βARK1_CT), a Gβγ subunits scavenger, using a recombinant adenovirus system. The adenovirus also expresses a green fluorescent protein (GFP) which serves as an index of transduction. Flow cytometry showed that up to 80% of chromaffin cells were transduced by the virus. There was a direct correlation between the βARK1_CT inhibition of the NPY enhancement of ATP-stimulated InsP formation and the percent of cells expressing GFP (r²=0.9993). These results demonstrate that Gβγ subunits are required for the NPY enhancement of ATP-stimulated InsP formation in bovine chromaffin cells.

Original language	English (US)
Pages (from-to)	267-274
Number of pages	8
Journal	Peptides
Volume	25
Issue number	2
DOIs	https://doi.org/10.1016/j.peptides.2003.12.019
State	Published - Feb 2004

Keywords

ATP
CAMP
Chromaffin cell
Cyclic AMP
DMEM
Dulbecco's modified Eagle medium
GFP
Green fluorescent protein
Inositol phosphate
InsP
MOI
Multiplicity of infection
NPY
NPY receptor
Neuropeptide Y
P2Y
Purinergic
Purinergic receptor
Y1

ASJC Scopus subject areas

Biochemistry
Physiology
Endocrinology
Cellular and Molecular Neuroscience

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10.1016/j.peptides.2003.12.019

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title = "βγ subunits mediate the NPY enhancement of ATP-stimulated inositol phosphate formation",

abstract = "Neuropeptide Y (NPY) enhances ATP-stimulated inositol phosphate (InsP) formation in bovine chromaffin cells through an unknown mechanism. Chromaffin cells were transduced with the carboxyl terminus of β-adrenergic receptor kinase 1 (βARK1CT), a Gβγ subunits scavenger, using a recombinant adenovirus system. The adenovirus also expresses a green fluorescent protein (GFP) which serves as an index of transduction. Flow cytometry showed that up to 80\% of chromaffin cells were transduced by the virus. There was a direct correlation between the βARK1CT inhibition of the NPY enhancement of ATP-stimulated InsP formation and the percent of cells expressing GFP (r2=0.9993). These results demonstrate that Gβγ subunits are required for the NPY enhancement of ATP-stimulated InsP formation in bovine chromaffin cells.",

keywords = "ATP, CAMP, Chromaffin cell, Cyclic AMP, DMEM, Dulbecco's modified Eagle medium, GFP, Green fluorescent protein, Inositol phosphate, InsP, MOI, Multiplicity of infection, NPY, NPY receptor, Neuropeptide Y, P2Y, Purinergic, Purinergic receptor, Y1",

author = "Xinying Li and Tsuneya Ikezu and Hexum, \{Terry D.\}",

note = "Funding Information: This work was supported by U.S. Public Health Service grant HL65135 (T.D.H.) and a Vada Oldfield Alzheimer{\textquoteright}s Research Award (T.I.). We thank Dr. R. Lefkowitz, Duke University, Durham, NC, for providing the minigene encoding the carboxyl terminus of the bovine beta adrenergic receptor kinase 1 and Dr. Bert Vogelstein, Johns Hopkins University, Baltimore, MD, for providing the pAdEasy system. The authors also thank Dr. Keshore Bidasee for helpful suggestions in the preparation of this manuscript and April Walls and Desinee Drakulich for their excellent technical assistance.",

year = "2004",

month = feb,

doi = "10.1016/j.peptides.2003.12.019",

language = "English (US)",

volume = "25",

pages = "267--274",

journal = "Peptides",

issn = "0196-9781",

publisher = "Elsevier Inc.",

number = "2",

}

TY - JOUR

T1 - βγ subunits mediate the NPY enhancement of ATP-stimulated inositol phosphate formation

AU - Li, Xinying

AU - Ikezu, Tsuneya

AU - Hexum, Terry D.

N1 - Funding Information: This work was supported by U.S. Public Health Service grant HL65135 (T.D.H.) and a Vada Oldfield Alzheimer’s Research Award (T.I.). We thank Dr. R. Lefkowitz, Duke University, Durham, NC, for providing the minigene encoding the carboxyl terminus of the bovine beta adrenergic receptor kinase 1 and Dr. Bert Vogelstein, Johns Hopkins University, Baltimore, MD, for providing the pAdEasy system. The authors also thank Dr. Keshore Bidasee for helpful suggestions in the preparation of this manuscript and April Walls and Desinee Drakulich for their excellent technical assistance.

PY - 2004/2

Y1 - 2004/2

N2 - Neuropeptide Y (NPY) enhances ATP-stimulated inositol phosphate (InsP) formation in bovine chromaffin cells through an unknown mechanism. Chromaffin cells were transduced with the carboxyl terminus of β-adrenergic receptor kinase 1 (βARK1CT), a Gβγ subunits scavenger, using a recombinant adenovirus system. The adenovirus also expresses a green fluorescent protein (GFP) which serves as an index of transduction. Flow cytometry showed that up to 80% of chromaffin cells were transduced by the virus. There was a direct correlation between the βARK1CT inhibition of the NPY enhancement of ATP-stimulated InsP formation and the percent of cells expressing GFP (r2=0.9993). These results demonstrate that Gβγ subunits are required for the NPY enhancement of ATP-stimulated InsP formation in bovine chromaffin cells.

AB - Neuropeptide Y (NPY) enhances ATP-stimulated inositol phosphate (InsP) formation in bovine chromaffin cells through an unknown mechanism. Chromaffin cells were transduced with the carboxyl terminus of β-adrenergic receptor kinase 1 (βARK1CT), a Gβγ subunits scavenger, using a recombinant adenovirus system. The adenovirus also expresses a green fluorescent protein (GFP) which serves as an index of transduction. Flow cytometry showed that up to 80% of chromaffin cells were transduced by the virus. There was a direct correlation between the βARK1CT inhibition of the NPY enhancement of ATP-stimulated InsP formation and the percent of cells expressing GFP (r2=0.9993). These results demonstrate that Gβγ subunits are required for the NPY enhancement of ATP-stimulated InsP formation in bovine chromaffin cells.

KW - ATP

KW - CAMP

KW - Chromaffin cell

KW - Cyclic AMP

KW - DMEM

KW - Dulbecco's modified Eagle medium

KW - GFP

KW - Green fluorescent protein

KW - Inositol phosphate

KW - InsP

KW - MOI

KW - Multiplicity of infection

KW - NPY

KW - NPY receptor

KW - Neuropeptide Y

KW - P2Y

KW - Purinergic

KW - Purinergic receptor

KW - Y1

UR - https://www.scopus.com/pages/publications/1942420376

UR - https://www.scopus.com/pages/publications/1942420376#tab=citedBy

U2 - 10.1016/j.peptides.2003.12.019

DO - 10.1016/j.peptides.2003.12.019

M3 - Article

C2 - 15063008

AN - SCOPUS:1942420376

SN - 0196-9781

VL - 25

SP - 267

EP - 274

JO - Peptides

JF - Peptides

IS - 2

ER -

βγ subunits mediate the NPY enhancement of ATP-stimulated inositol phosphate formation

Abstract

Keywords

ASJC Scopus subject areas

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