Differential coupling of the P2Y1 receptor to Gα14 and Gαq/11 proteins during the development of the rat salivary gland

Olga J. Baker, Jean M. Camden, Ann M. Ratchford, Cheikh Seye, Laurie Erb, Gary A. Weisman

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

In rat submandibular gland (SMG), the P2Y1 receptor (P2Y1R) mediates increases in the intracellular calcium concentration, [Ca2+]i that diminish as the animal ages from 1 to 4-6 weeks. However, P2Y1R mRNA levels do not change with age, suggesting that the age-dependent decrease in the [Ca2+]i response to P2Y1R agonists may be due to alterations in the activity of a component of the P2Y1R signalling pathway. Objectives: To assess whether the decrease in P2Y1R-mediated intracellular calcium signalling in SMG cells as rats age is due to a decrease in P2Y1R coupling to G proteins or to a decrease in the expression of a cognate G protein. Design: SMG cells were isolated from Sprague-Dawley rats. P2Y1R function was assessed by measuring 2-MeSADP-induced increases in [Ca2+]i and ERK1/2 activation. P2Y1R-mediated activation of G proteins was determined by the [35S]GTPγS binding assay. Gq protein expression was determined by RT-PCR, Northern, and Western analysis. Results: In SMG cells from 1-week-old rats, two bands (52 and 42 kDa) were detected using anti-Gα14 antibody, whereas in SMG cells from 4- to 6-week-old rats only the 42 kDa band was detected. Furthermore, 2-MeSADP-induced GTPγ35S binding to Gα14 and Gαq/11 decreases in SMG cells from 4- to 6-week-old rats as compared to 1-week-old rats. Conclusions: These findings suggest that the age-dependent decrease in P2Y1R-mediated intracellular calcium signalling in rat SMG cells is due to a loss of 52 kDa Gα14 and indicate the differential coupling of the P2Y1R to Gα14 and Gαq/11 as the gland develops.

Original languageEnglish (US)
Pages (from-to)359-370
Number of pages12
JournalArchives of Oral Biology
Volume51
Issue number5
DOIs
StatePublished - May 2006
Externally publishedYes

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Gq-G11 GTP-Binding Protein alpha Subunits
Purinergic P2Y1 Receptors
Salivary Glands
Submandibular Gland
GTP-Binding Proteins
Calcium Signaling
Sprague Dawley Rats
Anti-Idiotypic Antibodies

Keywords

  • ADP
  • Development
  • Gα protein
  • Gα protein
  • P2Y nucleotide receptors
  • Submandibular glands

ASJC Scopus subject areas

  • Dentistry(all)

Cite this

Differential coupling of the P2Y1 receptor to Gα14 and Gαq/11 proteins during the development of the rat salivary gland. / Baker, Olga J.; Camden, Jean M.; Ratchford, Ann M.; Seye, Cheikh; Erb, Laurie; Weisman, Gary A.

In: Archives of Oral Biology, Vol. 51, No. 5, 05.2006, p. 359-370.

Research output: Contribution to journalArticle

Baker, Olga J. ; Camden, Jean M. ; Ratchford, Ann M. ; Seye, Cheikh ; Erb, Laurie ; Weisman, Gary A. / Differential coupling of the P2Y1 receptor to Gα14 and Gαq/11 proteins during the development of the rat salivary gland. In: Archives of Oral Biology. 2006 ; Vol. 51, No. 5. pp. 359-370.
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abstract = "In rat submandibular gland (SMG), the P2Y1 receptor (P2Y1R) mediates increases in the intracellular calcium concentration, [Ca2+]i that diminish as the animal ages from 1 to 4-6 weeks. However, P2Y1R mRNA levels do not change with age, suggesting that the age-dependent decrease in the [Ca2+]i response to P2Y1R agonists may be due to alterations in the activity of a component of the P2Y1R signalling pathway. Objectives: To assess whether the decrease in P2Y1R-mediated intracellular calcium signalling in SMG cells as rats age is due to a decrease in P2Y1R coupling to G proteins or to a decrease in the expression of a cognate G protein. Design: SMG cells were isolated from Sprague-Dawley rats. P2Y1R function was assessed by measuring 2-MeSADP-induced increases in [Ca2+]i and ERK1/2 activation. P2Y1R-mediated activation of G proteins was determined by the [35S]GTPγS binding assay. Gq protein expression was determined by RT-PCR, Northern, and Western analysis. Results: In SMG cells from 1-week-old rats, two bands (52 and 42 kDa) were detected using anti-Gα14 antibody, whereas in SMG cells from 4- to 6-week-old rats only the 42 kDa band was detected. Furthermore, 2-MeSADP-induced GTPγ35S binding to Gα14 and Gαq/11 decreases in SMG cells from 4- to 6-week-old rats as compared to 1-week-old rats. Conclusions: These findings suggest that the age-dependent decrease in P2Y1R-mediated intracellular calcium signalling in rat SMG cells is due to a loss of 52 kDa Gα14 and indicate the differential coupling of the P2Y1R to Gα14 and Gαq/11 as the gland develops.",
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T1 - Differential coupling of the P2Y1 receptor to Gα14 and Gαq/11 proteins during the development of the rat salivary gland

AU - Baker, Olga J.

AU - Camden, Jean M.

AU - Ratchford, Ann M.

AU - Seye, Cheikh

AU - Erb, Laurie

AU - Weisman, Gary A.

PY - 2006/5

Y1 - 2006/5

N2 - In rat submandibular gland (SMG), the P2Y1 receptor (P2Y1R) mediates increases in the intracellular calcium concentration, [Ca2+]i that diminish as the animal ages from 1 to 4-6 weeks. However, P2Y1R mRNA levels do not change with age, suggesting that the age-dependent decrease in the [Ca2+]i response to P2Y1R agonists may be due to alterations in the activity of a component of the P2Y1R signalling pathway. Objectives: To assess whether the decrease in P2Y1R-mediated intracellular calcium signalling in SMG cells as rats age is due to a decrease in P2Y1R coupling to G proteins or to a decrease in the expression of a cognate G protein. Design: SMG cells were isolated from Sprague-Dawley rats. P2Y1R function was assessed by measuring 2-MeSADP-induced increases in [Ca2+]i and ERK1/2 activation. P2Y1R-mediated activation of G proteins was determined by the [35S]GTPγS binding assay. Gq protein expression was determined by RT-PCR, Northern, and Western analysis. Results: In SMG cells from 1-week-old rats, two bands (52 and 42 kDa) were detected using anti-Gα14 antibody, whereas in SMG cells from 4- to 6-week-old rats only the 42 kDa band was detected. Furthermore, 2-MeSADP-induced GTPγ35S binding to Gα14 and Gαq/11 decreases in SMG cells from 4- to 6-week-old rats as compared to 1-week-old rats. Conclusions: These findings suggest that the age-dependent decrease in P2Y1R-mediated intracellular calcium signalling in rat SMG cells is due to a loss of 52 kDa Gα14 and indicate the differential coupling of the P2Y1R to Gα14 and Gαq/11 as the gland develops.

AB - In rat submandibular gland (SMG), the P2Y1 receptor (P2Y1R) mediates increases in the intracellular calcium concentration, [Ca2+]i that diminish as the animal ages from 1 to 4-6 weeks. However, P2Y1R mRNA levels do not change with age, suggesting that the age-dependent decrease in the [Ca2+]i response to P2Y1R agonists may be due to alterations in the activity of a component of the P2Y1R signalling pathway. Objectives: To assess whether the decrease in P2Y1R-mediated intracellular calcium signalling in SMG cells as rats age is due to a decrease in P2Y1R coupling to G proteins or to a decrease in the expression of a cognate G protein. Design: SMG cells were isolated from Sprague-Dawley rats. P2Y1R function was assessed by measuring 2-MeSADP-induced increases in [Ca2+]i and ERK1/2 activation. P2Y1R-mediated activation of G proteins was determined by the [35S]GTPγS binding assay. Gq protein expression was determined by RT-PCR, Northern, and Western analysis. Results: In SMG cells from 1-week-old rats, two bands (52 and 42 kDa) were detected using anti-Gα14 antibody, whereas in SMG cells from 4- to 6-week-old rats only the 42 kDa band was detected. Furthermore, 2-MeSADP-induced GTPγ35S binding to Gα14 and Gαq/11 decreases in SMG cells from 4- to 6-week-old rats as compared to 1-week-old rats. Conclusions: These findings suggest that the age-dependent decrease in P2Y1R-mediated intracellular calcium signalling in rat SMG cells is due to a loss of 52 kDa Gα14 and indicate the differential coupling of the P2Y1R to Gα14 and Gαq/11 as the gland develops.

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KW - Development

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KW - P2Y nucleotide receptors

KW - Submandibular glands

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