The cPGDS induces LHB expression in primary culture of hen anterior pituitary cells via the PPAR signaling pathway

• Main Authors: Yi-lin Hsieh, 謝易霖
• Other Authors: Yow-Ling Shiue
• Format: Others
• Language: en_US
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/u956xd

碩士 === 國立中山大學 === 生物醫學研究所 === 97 === Our previous study identified one candidate transcript, chicken prostaglandin D2 synthase (cPGDS), significantly higher expressed in hens with high-egg production across several chicken strains including the Single-Comb White Leghorn layers. To further elucidate its underlying molecular mechanisms, the downstream effects of cPGDS in primary culture of chicken anterior pituitary cells were investigated. We successfully generated a highly specific rabbit anti-cPGDS polyclonal antibody. Of 13 examined tissues/cells, quantitative reverse transcription polymerase chain reaction and Western blotting analysis demonstrated that cPGDS mRNA and protein were highly expressed in intestine, kidney, liver and pituitary gland. In addition, significantly higher cPGDS mRNA and protein expression levels of the pituitary gland and ovary in hens than of the pituitary gland and testis in roosters with the same age were found. Transfection of pEGFP-cPGDS plasmid into primary culture of pituitary anterior cells in medium that contained 1 mM arachidonic acids induced luteinizing hormone beta subunit (LHB) transcription and subsequent translation. Treatments of two metabolites in the cyclooxygenase pathway, PGD2 and PGJ2 in the primary culture of chicken anterior pituitary cells induced LHB transcription and translation in both dose- and time-dependent manners. Treatment of peroxisome proliferator activated receptors (PPARs) agonist increased LHB transcription and translation in chicken anterior pituitary cells. On the other hand, specific inhibitions of PPARA and PPARG using antagonists dramatically suppressed PGJ2-induced LHB transcription level. Taken together; these data suggested PGDS upregulated LHB transcription in primary culture of chicken anterior pituitary cells via the PPAR signaling pathway.