Zhou Cindy, Wu Jean, Borillo Jason, Torres Lisa, McMahon John, Bao Yongde, Lou Ya-Huan
Department of Diagnostic Sciences, Dental Branch, School of Medicine, University of Texas Health Science Center at Houston, Houston, T 77030, USA.
Am J Reprod Immunol. 2005 May;53(5):238-48. doi: 10.1111/j.1600-0897.2005.00265.x.
Chemokine thymus-expressed chemokine (TECK), which is expressed exclusively in the thymus and small intestine, plays a critical role in T-cell development. Our previous study revealed its expression in the ovary also. This study investigated its ovarian expression during ovulatory process.
Super-ovulation was induced in young female CD1 mice by equine chorionic gonadotropin (eCG) and human chorionic gonadotropic (hCG). Ovarian TECK expression during ovulation was determined by: (1) reverse transcriptase-polymerase chain reaction (RT-PCR) at mRNA level, (2) Western blot and immunohistology at the protein level, and (3) leukocyte infiltration assay at the bioactive level.
A transient, high-level expression of TECK in murine ovaries at the mRNA level during hCG-induced ovulation was detected. Sequencing of directly cloned PCR product confirmed the ovarian expression of TECK. The peak expression of TECK was observed at 10-12 hr post-hCG injection; real-time PCR revealed an 800-fold increase during its expression peak over 0 hr. The expressed ovarian TECK protein was readily detectable by Western blot. Immunohistochemistry localized TECK expression to the ovarian interstitial tissue surrounding, or in the theca layer of the mature follicles undergoing ovulatory process. Expression of TECK receptor, the CC chemokine receptor (CCR9) was also detected in the ovulating ovaries. Using in vitro leukocyte infiltration assay, we first demonstrated that ovaries undergoing the ovulatory process were able to selectively chemoattract mononuclear cells. Importantly, neutralization of TECK by the antibody resulted in a 85% reduction in the chemotactic activities of the ovaries.
This study suggested that ovarian expression of TECK is under a tight hormonal regulation, and expressed TECK may be responsible for recruitment of mononuclear cells into the ovary to participate in the ovulatory process.
趋化因子胸腺表达趋化因子（TECK）仅在胸腺和小肠中表达，在T细胞发育中起关键作用。我们之前的研究也揭示了它在卵巢中的表达。本研究调查了其在排卵过程中的卵巢表达情况。
用马绒毛膜促性腺激素（eCG）和人绒毛膜促性腺激素（hCG）诱导年轻雌性CD1小鼠超排卵。通过以下方法确定排卵期间卵巢TECK的表达：（1）在mRNA水平进行逆转录聚合酶链反应（RT-PCR）；（2）在蛋白质水平进行蛋白质印迹和免疫组织学检测；（3）在生物活性水平进行白细胞浸润试验。
在hCG诱导排卵期间，检测到小鼠卵巢中TECK在mRNA水平有短暂的高水平表达。直接克隆的PCR产物测序证实了TECK在卵巢中的表达。TECK的表达峰值出现在hCG注射后10 - 12小时；实时PCR显示其表达峰值时比0小时增加了800倍。通过蛋白质印迹很容易检测到卵巢中表达的TECK蛋白。免疫组织化学将TECK的表达定位在排卵过程中成熟卵泡周围的卵巢间质组织或卵泡膜层。在排卵的卵巢中也检测到了TECK受体CC趋化因子受体（CCR9）的表达。使用体外白细胞浸润试验，我们首次证明正在经历排卵过程的卵巢能够选择性地趋化单核细胞。重要的是，用抗体中和TECK导致卵巢趋化活性降低85%。
本研究表明，卵巢中TECK的表达受到严格的激素调节，表达的TECK可能负责募集单核细胞进入卵巢参与排卵过程。
