Bowen J A, Bazer F W, Burghardt R C
Department of Veterinary Anatomy & Public Health, Texas A&M University, College Station 77843, USA.
Biol Reprod. 1996 Nov;55(5):1098-106. doi: 10.1095/biolreprod55.5.1098.
The spatial and temporal expressions of Muc-1, selected integrin subunits, and extracellular matrix components in porcine uterine epithelium from estrous (Days 0, 4, 8, 10-15) and early pregnant (Days 10-15 of pregnancy) gilts and from steroid-treated ovariectomized gilts were analyzed using indirect immunofluorescence analyses on cryosectioned tissues to identify potential components of uterine receptivity. Integrin subunit and extracellular matrix expressions were also examined in Day 11-15 conceptuses. Intense Muc-1 staining was detected on apical uterine epithelium on Day 0 but was absent by Day 10 in both cyclic and pregnant gilts. The result of estrogen treatment (E2; 100 micrograms/day for 10 days) was similar to that of the corn oil vehicle control, while treatment with progesterone (P4; 200 mg/day for 10 days) or E2 + P4 decreased Muc-1 staining in ovariectomized gilts. Immunostaining performed with antibodies directed against integrin subunits (alpha 1, alpha 3, alpha 4, alpha 5, alpha v, beta 1, and beta 3) in uterine epithelium revealed low (integrin subunits alpha 1 alpha 3), high (integrin subunits alpha v and beta 3), or modulated (integrin subunits alpha 4, alpha 5, and beta 1) expression, with the lowest expression on Day 0 and maximum expression by Days 10-15. Additionally, no differences due to pregnancy status were detected in staining of uterine epithelium on Days 10-15. Uterine epithelium from steroid-treated ovariectomized gilts had low expression of alpha 4, alpha 5, and beta 1 subunits in the presence of E2 that increased in response to P4 and E2 + P4 treatments. The expression of integrin subunits alpha 3, alpha v and beta 3 was not affected by sex steroids. Trophectoderm also expressed alpha 1, alpha 4, alpha 5, alpha v, beta 1, and beta 3, integrin subunits. Extracellular matrix constituents (fibronectin, vitronectin, laminin, and collagen type IV) were also examined. Fibronectin and vitronectin were present on trophectoderm, but only vitronectin was detected on uterine epithelium. The alpha 4, alpha 5, alpha v, beta 1, and beta 3 integrin subunits, vitronectin, and fibronectin were detected at sites of attachment between uterine epithelial cells and trophectoderm on Days 12-15 of pregnancy. These studies indicate that down-regulation of Muc-1 coincides with the transition of the prereceptive uterus to the receptive uterus. Additionally, the expression of alpha 4, alpha 5, alpha v, beta 1, and beta 3 integrin subunits along with the extracellular matrix components of fibronectin and vitronectin correlates with the time of implantation in swine.
利用间接免疫荧光分析法,对发情期(第0、4、8、10 - 15天)和妊娠早期(妊娠第10 - 15天)的母猪以及经类固醇处理的去卵巢母猪的冷冻切片组织进行分析,以确定猪子宫上皮中黏蛋白-1(Muc-1)、选定的整合素亚基和细胞外基质成分的时空表达,从而识别子宫接受性的潜在成分。还对妊娠第11 - 15天的胚胎进行了整合素亚基和细胞外基质表达的检测。在发情周期第0天,子宫顶端上皮检测到强烈的Muc-1染色,但在发情周期和妊娠母猪中,到第10天时染色消失。雌激素处理(E2;100微克/天,持续10天)的结果与玉米油载体对照相似,而孕酮处理(P4;200毫克/天,持续10天)或E2 + P4处理可降低去卵巢母猪的Muc-1染色。用针对子宫上皮中整合素亚基(α1、α3、α4、α5、αv、β1和β3)的抗体进行免疫染色,结果显示表达水平有低(整合素亚基α1、α3)、高(整合素亚基αv和β3)或有调节变化(整合素亚基α4、α5和β1),在第0天表达最低,在第10 - 15天表达最高。此外,在第10 - 15天,未检测到妊娠状态对子宫上皮染色的影响。经类固醇处理的去卵巢母猪的子宫上皮在存在E2时,α4、α5和β1亚基表达较低,而P4和E2 + P4处理后表达增加。整合素亚基α3、αv和β3的表达不受性类固醇的影响。滋养外胚层也表达α1、α4、α5、αv、β1和β3整合素亚基。还检测了细胞外基质成分(纤连蛋白、玻连蛋白、层粘连蛋白和IV型胶原)。纤连蛋白和玻连蛋白存在于滋养外胚层,但仅在子宫上皮中检测到玻连蛋白。在妊娠第12 - 15天,在子宫上皮细胞与滋养外胚层的附着部位检测到α4、α5、αvβ1和β3整合素亚基、玻连蛋白和纤连蛋白。这些研究表明,Muc-1的下调与子宫从非接受状态向接受状态的转变相一致。此外,α4、α5、αv、β1和β3整合素亚基以及纤连蛋白和玻连蛋白的细胞外基质成分的表达与猪的着床时间相关。