Wang D, Ishimura R, Walia D S, Müller H, Dai G, Hunt J S, Lee N A, Lee J J, Soares M J
Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA.
J Endocrinol. 2000 Oct;167(1):15-28. doi: 10.1677/joe.0.1670015.
The uterus and placenta of the mouse and rat produce a member of the prolactin (PRL) family referred to as decidual/trophoblast PRL-related protein (d/tPRP). This cytokine/hormone has been hypothesized to regulate decidual cell activities needed for the establishment and maintenance of gestation. An alkaline phosphatase (AP)-tagging strategy was used to identify d/tPRP target cells. AP-d/tPRP bound to virtually all cells and tissues to which it was exposed, consistent with our earlier evidence that d/tPRP binds to heparin-containing molecules. Moreover, we found that co-incubation with heparin or pretreatment with heparitinase greatly decreased the binding of AP-d/tPRP to tissue sections. In addition, we observed that the AP-d/tPRP probe bound to the surface of Chinese hamster ovary (CHO) cells but not to heparan sulfate-deficient CHO-pgsD-677 cells. Potential unique non-heparin d/tPRP binding sites within mouse and rat uteroplacental tissues were identified by consecutively incubating sections with AP-d/tPRP followed by heparin. This strategy led to the identification of d/tPRP target cells associated with the uterus and the labyrinth zone of the chorioallantoic placenta. Within the uterus, d/tPRP specifically bound to eosinophils. d/tPRP-binding and eosinophil peroxidase activity were co-localized and showed similar patterns of distribution during the estrous cycle, pregnancy, and following hormonal manipulation. d/tPRP interactions with eosinophils were further demonstrated in the lung and intestine, with eosinophils isolated from the peritoneum, and in mice with generalized tissue eosinophilia. Collectively, these findings suggest that intercellular d/tPRP targeting is mediated through associations with heparin-containing molecules which help direct d/tPRP to specific interactions with eosinophils within the uterus and with the labyrinthine compartment of the chorioallantoic placenta.
小鼠和大鼠的子宫及胎盘会产生一种催乳素（PRL）家族成员，称为蜕膜/滋养层PRL相关蛋白（d/tPRP）。据推测，这种细胞因子/激素可调节妊娠建立和维持所需的蜕膜细胞活动。采用碱性磷酸酶（AP）标记策略来鉴定d/tPRP的靶细胞。AP-d/tPRP几乎能与所有接触到的细胞和组织结合，这与我们之前关于d/tPRP能与含肝素分子结合的证据一致。此外，我们发现与肝素共同孵育或用肝素酶预处理会大大降低AP-d/tPRP与组织切片的结合。另外，我们观察到AP-d/tPRP探针能结合到中国仓鼠卵巢（CHO）细胞表面，但不能结合到缺乏硫酸乙酰肝素的CHO-pgsD-677细胞表面。通过用AP-d/tPRP连续孵育切片，随后用肝素处理，确定了小鼠和大鼠子宫胎盘组织内潜在的独特非肝素d/tPRP结合位点。该策略鉴定出了与子宫及绒毛尿囊胎盘迷路区相关的d/tPRP靶细胞。在子宫内，d/tPRP特异性结合嗜酸性粒细胞。d/tPRP结合与嗜酸性粒细胞过氧化物酶活性共定位，且在发情周期、妊娠期间以及激素处理后呈现相似的分布模式。在肺和肠道、从腹膜分离出的嗜酸性粒细胞以及患有全身性组织嗜酸性粒细胞增多症的小鼠中，进一步证实了d/tPRP与嗜酸性粒细胞的相互作用。总体而言，这些发现表明细胞间d/tPRP靶向作用是通过与含肝素分子的结合介导的，这些分子有助于引导d/tPRP与子宫内的嗜酸性粒细胞以及绒毛尿囊胎盘迷路区进行特异性相互作用。
