Prostaglandin-D synthetase induces transcription of the LH beta subunit in the primary culture of chicken anterior pituitary cells via the PPAR signaling pathway

Chen, L.-R.; Lee, S.-C.; Lin, Y.-P.; Hsieh, Y.-L.; Chen, Y.-L.; Yang, J.-R.; Liou, J.-F.; Chen, C.-F.; Lee, Y.-P.; Shiue, Y.-L.

doi:10.1016/j.theriogenology.2009.09.020

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Volume 73, Issue 3 , Pages 367-382, February 2010

Prostaglandin-D synthetase induces transcription of the LH beta subunit in the primary culture of chicken anterior pituitary cells via the PPAR signaling pathway

L.-R. Chen
- Division of Physiology, Taiwan Livestock Research Institute, Council of Agriculture, Tainan, Taiwan
- Institute of Biotechnology, National Cheng Kung University, Tainan, Taiwan
- Institute of Biotechnology, Southern Taiwan University of Technology, Tainan, Taiwan
- These authors contributed equally.
S.-C. Lee
- Department of Surgical Sciences, Zuoying Armed Forces General Hospital, Kaohsiung, Taiwan
- These authors contributed equally.
Y.-P. Lin
- Institute of Biomedical Science, National Sun Yat-sen University, Kaohsiung, Taiwan
- These authors contributed equally.
Y.-L. Hsieh
- Institute of Biomedical Science, National Sun Yat-sen University, Kaohsiung, Taiwan
Y.-L. Chen
- Institute of Biomedical Science, National Sun Yat-sen University, Kaohsiung, Taiwan
J.-R. Yang
- Division of Physiology, Taiwan Livestock Research Institute, Council of Agriculture, Tainan, Taiwan
J.-F. Liou
- Division of Physiology, Taiwan Livestock Research Institute, Council of Agriculture, Tainan, Taiwan
C.-F. Chen
- Department of Animal Science, National Chung-Hsing University, Taichung, Taiwan
Y.-P. Lee
- Department of Animal Science, National Chung-Hsing University, Taichung, Taiwan
Y.-L. Shiue
- Institute of Biomedical Science, National Sun Yat-sen University, Kaohsiung, Taiwan
- Corresponding author. Tel.: +886 7 5255818; fax: +886 7 5250197.

Received 14 February 2009; received in revised form 16 September 2009; accepted 24 September 2009. published online 02 December 2009.

Abstract

Downstream effects of prostaglandin-D synthetase (PGDS) in a primary culture of chicken (Gallus gallus) anterior pituitary cells were investigated to study how PGDS regulated laying in hens. Either PGDS downstream metabolite, PGD₂ or PGJ₂, elevated LHB mRNA and LHB protein levels in dose- and time-dependent manners, and treatment with arachidonic acid (1μM) alone upregulated 15-deoxy-Δ^12,14-PGJ₂ (15-d-PGJ₂; derived from PGJ₂)/PGJ₂, LHB mRNA, and LHB protein levels (P<0.05) in the primary culture of chicken pituitary anterior cells. Transfection of the plasmid Enhanced Green Fluorescent Protein-PGDS plasmid into these cells in medium containing 1μM arachidonic acid additionally increased 15-d-PGJ₂/PGJ₂, LHB mRNA, and LHB protein levels (P<0.05). In the hypothalamus/pituitary gland of laying hens, there was a high correlation (r=0.64; P<0.05) between PGDS and the peroxisome proliferator-activated receptor A (PPARA) mRNA level in a high egg production strain, the L2 Taiwan country chickens. In commercial Single-Comb White Leghorn layers, there were high correlation coefficients between PGDS and PPARA (r=0.65; P<0.05) and between PGDS and PPARG (r=0.67; P<0.01) mRNA levels. A broad-range PPARs agonist, GW9578 (5 to 500nM), enhanced LHB mRNA and LHB protein levels (P<0.05). The PPARA-specific (GW6471) and PPARG-specific (T0070907) antagonists suppressed endogenous LHB mRNA and LHB protein levels (P<0.05); in addition, both antagonists attenuated arachidonic acid–induced LHB mRNA levels (P<0.05) and PGDS-induced (in the presence of 1μM arachidonic acid) LHB mRNA and LHB protein (P<0.05) levels in the primary culture of chicken anterior pituitary cells. Higher LHB mRNA/LHB protein ratios in PGD₂-, PGJ₂-, arachidonic acid-, PGDS-, and GW9578-induced as well as GW6471- and T0070907-suppressed anterior pituitary cells suggested that LHB transcription occurred before translation. In conclusion, PGDS induced LHB transcription and subsequent translation via the PPAR signaling pathway.