Bowden Marissa A, Li Ying, Liu Yi-Xun, Findlay Jock K, Salamonsen Lois A, Nie Guiying
Prince Henry's Institute of Medical Research, PO Box 5152, Monash University, Clayton, Victoria 3168, Australia.
Reprod Biol Endocrinol. 2008 Jun 18;6:22. doi: 10.1186/1477-7827-6-22.
HTRA3 is a recently identified member of the mammalian serine protease family HTRA (high temperature requirement factor A). In both the rodent and the human HTRA3 is transcribed into two mRNA species (long and short) through alternative splicing. We have previously shown that HTRA3 is expressed in the mature rat ovary and may be involved in folliculogenesis and luteinisation. HTRA3 is also upregulated during mouse and human placental development. The current study investigated whether HTRA3 is also localised in the primate ovary (rhesus monkey n = 7). In addition, we examined the non-pregnant rhesus monkey endometrium (n = 4) and maternal-fetal interface during early pregnancy (n = 5) to further investigate expression of HTRA3 in primate endometrium and placentation.
HTRA3 mRNA levels in several rhesus monkey tissues was determined by semiquantitative RT-PCR. Protein expression and localisation of HTRA3 was determined by immunohistochemistry.
Long and short forms of HTRA3 mRNA were detected in the ovary, aorta, bladder, small intestine, skeletal muscle, heart and uterus but not the liver nor the kidney. HTRA3 protein was immunolocalised to the oocyte of all follicular stages in the rhesus monkey ovary. Protein expression in mural and cumulus granulosa cells of late secondary follicles increased significantly compared to granulosa cells of primordial, primary and secondary follicles. Mural and cumulus granulosa cells of antral follicles also showed a significant increase in expression. Staining intensity was higher in the granulosa-lutein cells compared to the theca-lutein cells of corpora lutea (n = 3). In the non-pregnant monkey endometrium, HTRA3 was detected in the glandular epithelium. The basalis endometrial glands showed higher staining intensity than functionalis endometrial glands. During early pregnancy, strong staining for HTRA3 protein was seen in both maternal decidual cells and glands.
We propose that HTRA3 may be involved in folliculogenesis and luteinisation in the primate ovary. Furthermore, similar to previous findings in the human, HTRA3 is possibly a factor involved in and potentially important for primate placentation.
HTRA3是最近鉴定出的哺乳动物丝氨酸蛋白酶家族HTRA(高温需求因子A)的成员。在啮齿动物和人类中,HTRA3通过可变剪接转录为两种mRNA亚型(长型和短型)。我们之前已经表明,HTRA3在成熟大鼠卵巢中表达,可能参与卵泡发生和黄体化过程。在小鼠和人类胎盘发育过程中,HTRA3也上调表达。本研究调查了HTRA3是否也定位于灵长类动物卵巢(恒河猴,n = 7)。此外,我们检查了未孕恒河猴子宫内膜(n = 4)以及妊娠早期的母胎界面(n = 5),以进一步研究HTRA3在灵长类动物子宫内膜和胎盘形成中的表达情况。
通过半定量RT-PCR测定几只恒河猴组织中HTRA3 mRNA水平。通过免疫组织化学确定HTRA3的蛋白表达和定位。
在卵巢、主动脉、膀胱、小肠、骨骼肌、心脏和子宫中检测到HTRA3 mRNA的长型和短型,但在肝脏和肾脏中未检测到。HTRA3蛋白在恒河猴卵巢所有卵泡阶段的卵母细胞中免疫定位。与原始卵泡、初级卵泡和次级卵泡的颗粒细胞相比,次级晚期卵泡的壁层和卵丘颗粒细胞中的蛋白表达显著增加。窦状卵泡的壁层和卵丘颗粒细胞中的表达也显著增加。黄体的颗粒黄体细胞中的染色强度高于膜黄体细胞(n =