Ryanodine and inositol trisphosphate receptors are differentially distributed and expressed in rat parotid gland

Xuejun Zhang, Jiayu Wen, Keshore R. Bidasee, Henry R. Besch, Richard J.H. Wojcikiewicz, Bumsup Lee, Ronald P. Rubin

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

The present study examines the cellular distribution of the ryanodine receptor/channel (RyR) and inositol 1,4,5-trisphosphate receptor (InsP3R) subtypes in parotid acini. Using fluorescently labelled 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene-3-propionic acid glycyl-ryanodine (BODIPY®-ryanodine) and confocal microscopy, RyRs were localized primarily to the perinuclear region (basal pole) of the acinar cell. Ryanodine, Ruthenium Red, cAMP and cADP ribose (cADPR) competed with BODIPY-ryanodine, resulting in a reduction in the fluorescence signal. However, inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] did not alter the binding of BODIPY-ryanodine. Using receptor-subtype-specific antisera, InsP3Rs (types I, II and III) were located predominantly in the apical pole of the parotid cell. The presence of these three subtypes was confirmed using reverse transcriptase PCR with RNA-specific oligonucleotide probes. Binding studies using a parotid cell-membrane fraction identified and characterized RyRs and InsP3Rs in terms of binding affinity (K(d)) and maximum binding capacity (B(max)) and confirmed that cADPR displaces ryanodine from its binding sites. Ruthenium Red and 8-Br-cADP-ribose blocked Ca2+ release in permeabilized acinar cells in response to cADPR and cAMP or forskolin, whereas Ins(1,4,5)P3-induced Ca2+ release was unaffected. The localization of the RyRs and InsP3Rs in discrete regions endow broad areas of the parotid cell with ligand-activated Ca2+ channels. The consequences of the dual activation of the RyRs and InsP3Rs by physiologically relevant stimuli such as noradrenaline (norepinephrine) are considered in relation to Ca2+ signalling in the parotid gland.

Original languageEnglish (US)
Pages (from-to)519-527
Number of pages9
JournalBiochemical Journal
Volume340
Issue number2
DOIs
StatePublished - Jun 1 1999

Fingerprint

Ryanodine
Cyclic ADP-Ribose
Parotid Gland
Inositol
Rats
Ruthenium Red
Acinar Cells
Poles
Norepinephrine
Inositol 1,4,5-Trisphosphate Receptors
Ryanodine Receptor Calcium Release Channel
Inositol 1,4,5-Trisphosphate
Oligonucleotide Probes
Confocal microscopy
RNA-Directed DNA Polymerase
Colforsin
Cell membranes
Reverse Transcriptase Polymerase Chain Reaction
Confocal Microscopy
Immune Sera

Keywords

  • Cyclic ADP-ribose
  • Cytochemical localization
  • InsP receptor
  • Parotid cell

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Ryanodine and inositol trisphosphate receptors are differentially distributed and expressed in rat parotid gland. / Zhang, Xuejun; Wen, Jiayu; Bidasee, Keshore R.; Besch, Henry R.; Wojcikiewicz, Richard J.H.; Lee, Bumsup; Rubin, Ronald P.

In: Biochemical Journal, Vol. 340, No. 2, 01.06.1999, p. 519-527.

Research output: Contribution to journalArticle

Zhang, Xuejun ; Wen, Jiayu ; Bidasee, Keshore R. ; Besch, Henry R. ; Wojcikiewicz, Richard J.H. ; Lee, Bumsup ; Rubin, Ronald P. / Ryanodine and inositol trisphosphate receptors are differentially distributed and expressed in rat parotid gland. In: Biochemical Journal. 1999 ; Vol. 340, No. 2. pp. 519-527.
@article{a50f5c1b329b4936aa3df106b3cf7cfa,
title = "Ryanodine and inositol trisphosphate receptors are differentially distributed and expressed in rat parotid gland",
abstract = "The present study examines the cellular distribution of the ryanodine receptor/channel (RyR) and inositol 1,4,5-trisphosphate receptor (InsP3R) subtypes in parotid acini. Using fluorescently labelled 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene-3-propionic acid glycyl-ryanodine (BODIPY{\circledR}-ryanodine) and confocal microscopy, RyRs were localized primarily to the perinuclear region (basal pole) of the acinar cell. Ryanodine, Ruthenium Red, cAMP and cADP ribose (cADPR) competed with BODIPY-ryanodine, resulting in a reduction in the fluorescence signal. However, inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] did not alter the binding of BODIPY-ryanodine. Using receptor-subtype-specific antisera, InsP3Rs (types I, II and III) were located predominantly in the apical pole of the parotid cell. The presence of these three subtypes was confirmed using reverse transcriptase PCR with RNA-specific oligonucleotide probes. Binding studies using a parotid cell-membrane fraction identified and characterized RyRs and InsP3Rs in terms of binding affinity (K(d)) and maximum binding capacity (B(max)) and confirmed that cADPR displaces ryanodine from its binding sites. Ruthenium Red and 8-Br-cADP-ribose blocked Ca2+ release in permeabilized acinar cells in response to cADPR and cAMP or forskolin, whereas Ins(1,4,5)P3-induced Ca2+ release was unaffected. The localization of the RyRs and InsP3Rs in discrete regions endow broad areas of the parotid cell with ligand-activated Ca2+ channels. The consequences of the dual activation of the RyRs and InsP3Rs by physiologically relevant stimuli such as noradrenaline (norepinephrine) are considered in relation to Ca2+ signalling in the parotid gland.",
keywords = "Cyclic ADP-ribose, Cytochemical localization, InsP receptor, Parotid cell",
author = "Xuejun Zhang and Jiayu Wen and Bidasee, {Keshore R.} and Besch, {Henry R.} and Wojcikiewicz, {Richard J.H.} and Bumsup Lee and Rubin, {Ronald P.}",
year = "1999",
month = "6",
day = "1",
doi = "10.1042/0264-6021:3400519",
language = "English (US)",
volume = "340",
pages = "519--527",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "2",

}

TY - JOUR

T1 - Ryanodine and inositol trisphosphate receptors are differentially distributed and expressed in rat parotid gland

AU - Zhang, Xuejun

AU - Wen, Jiayu

AU - Bidasee, Keshore R.

AU - Besch, Henry R.

AU - Wojcikiewicz, Richard J.H.

AU - Lee, Bumsup

AU - Rubin, Ronald P.

PY - 1999/6/1

Y1 - 1999/6/1

N2 - The present study examines the cellular distribution of the ryanodine receptor/channel (RyR) and inositol 1,4,5-trisphosphate receptor (InsP3R) subtypes in parotid acini. Using fluorescently labelled 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene-3-propionic acid glycyl-ryanodine (BODIPY®-ryanodine) and confocal microscopy, RyRs were localized primarily to the perinuclear region (basal pole) of the acinar cell. Ryanodine, Ruthenium Red, cAMP and cADP ribose (cADPR) competed with BODIPY-ryanodine, resulting in a reduction in the fluorescence signal. However, inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] did not alter the binding of BODIPY-ryanodine. Using receptor-subtype-specific antisera, InsP3Rs (types I, II and III) were located predominantly in the apical pole of the parotid cell. The presence of these three subtypes was confirmed using reverse transcriptase PCR with RNA-specific oligonucleotide probes. Binding studies using a parotid cell-membrane fraction identified and characterized RyRs and InsP3Rs in terms of binding affinity (K(d)) and maximum binding capacity (B(max)) and confirmed that cADPR displaces ryanodine from its binding sites. Ruthenium Red and 8-Br-cADP-ribose blocked Ca2+ release in permeabilized acinar cells in response to cADPR and cAMP or forskolin, whereas Ins(1,4,5)P3-induced Ca2+ release was unaffected. The localization of the RyRs and InsP3Rs in discrete regions endow broad areas of the parotid cell with ligand-activated Ca2+ channels. The consequences of the dual activation of the RyRs and InsP3Rs by physiologically relevant stimuli such as noradrenaline (norepinephrine) are considered in relation to Ca2+ signalling in the parotid gland.

AB - The present study examines the cellular distribution of the ryanodine receptor/channel (RyR) and inositol 1,4,5-trisphosphate receptor (InsP3R) subtypes in parotid acini. Using fluorescently labelled 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene-3-propionic acid glycyl-ryanodine (BODIPY®-ryanodine) and confocal microscopy, RyRs were localized primarily to the perinuclear region (basal pole) of the acinar cell. Ryanodine, Ruthenium Red, cAMP and cADP ribose (cADPR) competed with BODIPY-ryanodine, resulting in a reduction in the fluorescence signal. However, inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] did not alter the binding of BODIPY-ryanodine. Using receptor-subtype-specific antisera, InsP3Rs (types I, II and III) were located predominantly in the apical pole of the parotid cell. The presence of these three subtypes was confirmed using reverse transcriptase PCR with RNA-specific oligonucleotide probes. Binding studies using a parotid cell-membrane fraction identified and characterized RyRs and InsP3Rs in terms of binding affinity (K(d)) and maximum binding capacity (B(max)) and confirmed that cADPR displaces ryanodine from its binding sites. Ruthenium Red and 8-Br-cADP-ribose blocked Ca2+ release in permeabilized acinar cells in response to cADPR and cAMP or forskolin, whereas Ins(1,4,5)P3-induced Ca2+ release was unaffected. The localization of the RyRs and InsP3Rs in discrete regions endow broad areas of the parotid cell with ligand-activated Ca2+ channels. The consequences of the dual activation of the RyRs and InsP3Rs by physiologically relevant stimuli such as noradrenaline (norepinephrine) are considered in relation to Ca2+ signalling in the parotid gland.

KW - Cyclic ADP-ribose

KW - Cytochemical localization

KW - InsP receptor

KW - Parotid cell

UR - http://www.scopus.com/inward/record.url?scp=0033151932&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033151932&partnerID=8YFLogxK

U2 - 10.1042/0264-6021:3400519

DO - 10.1042/0264-6021:3400519

M3 - Article

C2 - 10333498

AN - SCOPUS:0033151932

VL - 340

SP - 519

EP - 527

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -