Yang Wensheng, Diehl John R, Yerle Martine, Ford J Joe, Christenson Ronald K, Roudebush William E, Plummer William E
Poole Agricultural Center, Department of Animal & Veterinary Sciences, Clemson University, Clemson, South Carolina 29634, USA.
Mol Reprod Dev. 2003 Jan;64(1):4-12. doi: 10.1002/mrd.10217.
Although platelet-activating factor receptor (PAFr) gene was well characterized in the human, little was known about it in domestic animals. Porcine PAFr gene was mapped using fluorescence in situ hybridization (FISH). The structure of this gene was investigated using a 5' rapid amplification of cDNA ends (RACE) technique. Temporal expression of PAFr and estrogen receptor alpha genes (ER), and distribution of the PAFr transcripts in porcine endometrial and embryonic tissues on days 0, 10, 12, 14, 16, and 18 were analyzed using DNA competitors and reverse transcription and polymerase chain reaction (RT-PCR). The porcine PAFr gene was mapped to SSC6q26-27. Alternative splicing of primary transcripts of the PAFr gene produced two different transcripts. Transcript 1 was expressed in all tissues and cells, and transcript 2 was detected in all tissues but white blood cells. The temporal expression of the PAFr gene in endometrial (P > 0.05) and embryonic (P < 0.05) tissues of pregnant sows increased from day 10 to 16. The temporal expression of ER genes in endometrial tissues of pregnant sows decreased from day 10 to 18 (P < 0.05). In addition, ER expression was detectable in 20-60% of embryonic tissue samples, which generally decreased. In combination with previously obtained data on PAF and estradiol-17beta (E(2)) concentrations in pregnant uterine luminal fluids (pULF), endometrial and embryonic tissues, the present results indicated that the increasing PAFr transcripts were positively associated with increasing levels of PAF. Both ER transcripts and E(2) found in pULF decreased correspondingly from day 13 to 16. These results indicate that via PAFr, PAF could play a dominant role in peri-implantation development in pigs as compared to E(2).
尽管血小板活化因子受体（PAFr）基因在人类中已得到充分表征，但在家畜中对此了解甚少。利用荧光原位杂交（FISH）技术对猪PAFr基因进行了定位。使用5' cDNA末端快速扩增（RACE）技术研究了该基因的结构。利用DNA竞争物以及逆转录和聚合酶链反应（RT-PCR）分析了PAFr和雌激素受体α基因（ER）的时序表达，以及在第0、10、12、14、16和18天猪子宫内膜和胚胎组织中PAFr转录本的分布。猪PAFr基因定位于猪6号染色体q26 - 27区域。PAFr基因初级转录本的可变剪接产生了两种不同的转录本。转录本1在所有组织和细胞中均有表达，转录本2在除白细胞外的所有组织中均能检测到。妊娠母猪子宫内膜（P>0.05）和胚胎（P<0.05）组织中PAFr基因的时序表达从第10天到第16天增加。妊娠母猪子宫内膜组织中ER基因的时序表达从第10天到第18天下降（P<0.05）。此外，在20% - 60%的胚胎组织样本中可检测到ER表达，且其表达通常下降。结合先前获得的关于妊娠子宫腔液（pULF）、子宫内膜和胚胎组织中PAF和雌二醇-17β（E₂）浓度的数据，目前的结果表明，PAFr转录本的增加与PAF水平的升高呈正相关。在pULF中发现的ER转录本和E₂从第13天到第16天相应下降。这些结果表明，与E₂相比，PAF通过PAFr可能在猪着床前后的发育中起主导作用。
