Expression and regulation of P2U-purinergic receptor in human granulosa-luteal cells

Chen Jei Tai, Sung Keun Kang, Kwai Wa Cheng, Kyung Chul Choi, Parimal S. Nathwani, Peter C K Leung

研究成果: 雜誌貢獻文章

29 引文 (Scopus)

摘要

The P2U purinoceptor (P2UR) has been identified pharmacologically in the ovary. However, the expression and regulation of the P2UR messenger RNA (mRNA) in human ovarian cells are still poorly characterized. The present study was designed to examine the expression and regulation of the P2UR in human granulosa-luteal cells (hGLCs) by RT-PCR and Northern blot analysis. A PCR product corresponding to the expected 599-bp P2UR complementary DNA was obtained from hGLCs. Molecular cloning and sequencing of the PCR product revealed an identical sequence to the reported P2UR complementary DNA. Two mRNA transcripts of 2.0 kb and 4.6 kb were identified in hGLCs using Northern blot analysis. The expression of the P2UR mRNA was down-regulated by human CG in a dose- and time-dependent manner. Treatment with 8-bromo-cAMP and forskolin also attenuated P2UR mRNA levels. Calcium signaling following the activation of the P2UR in single hGLCs was studied using microspectrofluorimetry. It revealed that, like ATP, uridine triphosphate (UTP) also induced cytosolic calcium mobilization in a dose-dependent manner. These results demonstrate for the first time that the P2UR mRNA is expressed in hGLCs and that P2UR mRNA is regulated by human CG, cAMP, and forskolin. The P2UR expressed in hGLCs functional because activation of the P2UR by ATP or UTP resulted in rapid and transient mobilization of cytosolic calcium at the single cell level. These findings further support a potential role of this neurotransmitter receptor in the human ovary.

原文英語
頁(從 - 到)1591-1597
頁數7
期刊Journal of Clinical Endocrinology and Metabolism
85
發行號4
DOIs
出版狀態已發佈 - 2000
對外發佈Yes

指紋

Purinergic P2Y2 Receptors
Luteal Cells
Purinergic Receptors
Messenger RNA
Uridine Triphosphate
Colforsin
Calcium
Northern Blotting
Polymerase Chain Reaction
Ovary
Complementary DNA
Adenosine Triphosphate
Chemical activation
8-Bromo Cyclic Adenosine Monophosphate
Neurotransmitter Receptor
Calcium Signaling
Cloning
Molecular Cloning

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology, Diabetes and Metabolism

引用此文

Expression and regulation of P2U-purinergic receptor in human granulosa-luteal cells. / Tai, Chen Jei; Kang, Sung Keun; Cheng, Kwai Wa; Choi, Kyung Chul; Nathwani, Parimal S.; Leung, Peter C K.

於: Journal of Clinical Endocrinology and Metabolism, 卷 85, 編號 4, 2000, p. 1591-1597.

研究成果: 雜誌貢獻文章

Tai, Chen Jei ; Kang, Sung Keun ; Cheng, Kwai Wa ; Choi, Kyung Chul ; Nathwani, Parimal S. ; Leung, Peter C K. / Expression and regulation of P2U-purinergic receptor in human granulosa-luteal cells. 於: Journal of Clinical Endocrinology and Metabolism. 2000 ; 卷 85, 編號 4. 頁 1591-1597.
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abstract = "The P2U purinoceptor (P2UR) has been identified pharmacologically in the ovary. However, the expression and regulation of the P2UR messenger RNA (mRNA) in human ovarian cells are still poorly characterized. The present study was designed to examine the expression and regulation of the P2UR in human granulosa-luteal cells (hGLCs) by RT-PCR and Northern blot analysis. A PCR product corresponding to the expected 599-bp P2UR complementary DNA was obtained from hGLCs. Molecular cloning and sequencing of the PCR product revealed an identical sequence to the reported P2UR complementary DNA. Two mRNA transcripts of 2.0 kb and 4.6 kb were identified in hGLCs using Northern blot analysis. The expression of the P2UR mRNA was down-regulated by human CG in a dose- and time-dependent manner. Treatment with 8-bromo-cAMP and forskolin also attenuated P2UR mRNA levels. Calcium signaling following the activation of the P2UR in single hGLCs was studied using microspectrofluorimetry. It revealed that, like ATP, uridine triphosphate (UTP) also induced cytosolic calcium mobilization in a dose-dependent manner. These results demonstrate for the first time that the P2UR mRNA is expressed in hGLCs and that P2UR mRNA is regulated by human CG, cAMP, and forskolin. The P2UR expressed in hGLCs functional because activation of the P2UR by ATP or UTP resulted in rapid and transient mobilization of cytosolic calcium at the single cell level. These findings further support a potential role of this neurotransmitter receptor in the human ovary.",
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AU - Nathwani, Parimal S.

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