小鼠子宫内膜中鸡冠珊瑚树凝集素和双花藕豆凝集素受体在胚泡植入过程中的表达

目的 探讨小鼠子宫内膜鸡冠珊瑚树凝集素(ECL)受体和双花藕豆凝集素(DBA)受体与胚泡植入的关系.方法 以生物素标记的ECL和DBA来检测怀孕侧和输卵管结扎未孕侧小鼠孕早期子宫内膜中两种凝集 素受体的分布状况和变化规律.结果怀孕侧,ECL主要表达于胚胎滋养层细胞及其基膜,蜕膜细胞及其周围的细胞外基质(ECM);未孕侧,主要表达于子宫内膜上皮和腺上皮.在怀孕侧,DBA受体主要表达于胚胎滋养层细胞和子宫内膜血管基膜;在未孕侧,主要表达于子宫内膜血管的基膜.结论 ECL和DBA受体与小鼠胚泡植入过程密切相关.     

小鼠动情周期、妊娠和哺乳过程中凝集素PHA-E与UEA的受体在子宫内膜的分布

目的 探讨子宫内膜菜豆凝集素(PHA-E)和荆豆凝集素(UEA)的受体与胚泡植入的关系。方法 应用亲合细胞化学和图像分析方法检测PHA-E与UEA的受体在昆明小鼠动情周期、妊娠和哺乳过程中子宫内膜的分布状况和变化规律。结果 以上2种凝集素受体均存在于不同阶段的小鼠子宫内膜,但2种凝集素受体的数量和分布存在差异。PHA-E受体在孕早期,尤其是围植入期的水平显著高于动情周期组和哺乳期组;其广泛分布于围植入期子宫内膜腔上皮和腺上皮游离缘、胚胎组织、蜕膜细胞表面及其周围的细胞外基质(ECM)。UEA受体则呈现动情期水平最高,孕期逐渐下降的趋势,其主要分布于子宫内膜腺上皮游离缘。结论凝集素PHA-E受体与小鼠胚泡植入过程密切相关,对UEA受体与胚胞植入的关系尚难做出满意地解释。     

凝集素PHA-E、UEA受体在真孕小鼠和假孕模型小鼠孕早期子宫内膜中的变化

目的：探讨子宫内膜凝集素PHA-E和UEA的受体与胚泡植入的关系。方法：应用生物素标记的凝集素PHA-E和UEA,来检测真孕组、单纯假孕组和蜕膜化假孕组小鼠孕早期子宫内膜中两种凝集素受体的分布状况和变化规律。结果：PHA-E受体广泛分布于各组小鼠子宫内膜腔上皮和腺上皮、蜕膜细胞表面及其周围的ECM;其在真孕组的水平均显著高于假孕组,且在孕9d时,蜕膜化假孕组的水平又显著高于单纯假孕组。UEA受体则主要分布于子宫内膜腺上皮游离缘;其在真孕组的水平高于单纯假孕组,在孕6、9d时,真孕组的水平又明显低于蜕膜化假孕组。结论：凝集素PHA-E受体与小鼠胚泡植入过程密切相关,对UEA受体与胚泡植入的关系尚难做出满意的解释。     

妊娠早期小鼠子宫内膜层粘连蛋白定位、定量及对胚泡植入的作用

目的　研究妊娠早期小鼠子宫内膜层粘连蛋白（ｌａｍ ｉｎｉｎ,ＬＮ）定位和定量变化及其对胚泡着床的作用。方法　应用间接免疫荧光法、免疫组织化学ＡＢＣ技术及图像分析法进行ＬＮ定位和定量测定;利用子宫内注射ＬＮ抗体的方法探讨ＬＮ对小鼠胚泡植入的作用。　结果　动情期及妊娠早期小鼠子宫内膜内源性ＬＮ 在上皮和腺基膜及血管内皮基膜均有表达,并随着植入的发生基膜的ＬＮ 表达减弱,而内膜上皮中ＬＮ免疫染色逐渐增强;外源性ＬＮ 对小鼠胚泡粘附与植入子宫内膜不产生明显影响,层粘连蛋白抗体明显抑制小鼠胚泡着床。　结论　动情期及妊娠早期小鼠子宫内膜的层粘连蛋白主要存在于基膜,胚泡植入期间内膜上皮细胞内ＬＮ 的含量增加;ＬＮ在促进小鼠胚泡粘附与植入子宫内膜过程中起重要作用。     

RhoA在小鼠胚胎围着床期子宫内膜的表达及其意义

目的探讨RhoA在小鼠胚胎植入过程中的表达及其生物学作用。方法取胚胎围种植期（D0至D6天）昆明小鼠子宫作切片，用免疫组织化学方法和图像分析方法对RhoA在子宫内膜中的表达进行定位和半定量分析。结果RhoA主要分布于小鼠子宫内膜腺上皮、腔上皮和基质细胞。受精后，RhoA在子宫内膜的含量高于未孕组（P＜0．05）；受精后第4d（植入期），小鼠子宫内膜腔上皮中RhoA的含量明显增多，明显高于未孕及植入前各时间组（P＜0．05），并逐渐向基质细胞延伸，第5d广泛分布在蜕膜细胞和基质细胞，到第6天大部分子宫内膜基质区域及腺上皮的细胞中有大量的RhoA分布。RhoA在小鼠子宫内膜组织中的表达强度在受精后呈逐渐上升趋势。结论①RhoA可能参与了胚胎的种植和子宫内膜容受性的建立；②还可能间接参与植入后胚胎的滋养层扩展、分化及子宫内膜蜕膜化反应。     

T淋巴细胞侵袭转移诱导蛋白1在小鼠子宫内膜基质细胞-胚泡共培养体系中对胚泡黏附的影响

目的:检测不同浓度T淋巴细胞侵袭转移诱导蛋白1 (Tiaml)抗体下,小鼠着床窗子宫内膜基质细胞内的Tiaml蛋白表达及其对基质细胞-胚泡共培养体系中胚泡黏附的影响,探讨Tiaml对小鼠胚胎着床的影响.方法:提取着床窗小鼠子宫内膜基质细胞并构建基质细胞-胚泡共培养体系;分别用免疫细胞化学和免疫印迹法检测不同浓度抗Tiaml抗体时着床窗基质细胞Tiaml蛋白的表达及其对胚泡黏附的影响.结果:抗体干预后着床窗基质细胞内的Tiaml蛋白表达水平降低;在不同抗体浓度下胚泡黏附率有差异.结论:随Tiaml抗体浓度增加,着床窗小鼠子宫内膜基质细胞Tiaml蛋白表达量及胚泡黏附率降低.表明Tiarnl可能在胚泡植入过程起重要的作用.     

小鼠胚胎腭板发育过程中麦胚凝集素、蓖麻凝集素和鸡冠珊瑚树凝集素受体的表达

方法:以生物素标记的凝集素WGA、 RCA和ECL为探针,应用亲和组织化学显色方法检测上述3种凝集素受体在各组小鼠胚胎腭板上皮及其间充质的分布状况和变化规律.结果:在小鼠胚胎腭发育过程中3种凝集素受体在腭板上皮中的表达水平均高于在间充质中的表达水平.WGA受体在腭板上皮中的表达峰值出现于d1322-d148,即腭板上抬和水平生长期;而RCA和ECL受体的表达峰值出现于d158-d1522,即腭板融合期.结论:3种凝集素受体均可能不同程度地参与腭板正常发育的调节,WGA受体可能主要作用于腭板上抬和水平增长期,而RCA和ECL受体则可能主要作用于腭板融合期.     

细胞因子与着床过程中小鼠子宫内膜细胞凋亡的相关性研究

目的 探讨着床过程中小鼠子宫内膜细胞凋亡的调控机制，以及细胞因子在子宫内膜细胞凋亡发生中的生物学作用。方法 用TUNEL法原位检测着床过程中子宫内膜细胞凋亡状况，用原位杂交和免疫组织化学的方法，检测凋亡相关基因(bcl-2、bax)和细胞因子(EGF、bFGF和TGFβ1)在着床期小鼠子宫内膜中的表达，分析凋亡相关基因、细胞因子与子宫内膜细胞凋亡之间的关系，并在体外培养的子宫内膜细胞中，直接检测了上述因子对bcl-2和bax转录的影响。结果 孕4～5d，凋亡细胞主要分布于子宫内膜表面上皮和腺上皮，与此相对应，上皮细胞TGFβ1和bax表达增加，而bFGF和bCl-2表达减少；孕7～8d，凋亡细胞主要分布于胚泡着床部位周围的蜕膜中，此时胚泡周围的蜕膜中，EGF、bFGF和bcl-2表达明显降低，TGEβ1和bax表达明显增加。体外实验显示，培养基中加入抗EGF或bFGF抗体后，bcl-2／bax比率下降，而加入抗TGFβ-1抗体后，bcl-2／bax比率增加。结论 细胞因子、凋亡相关基因与着床期子宫内膜细胞凋亡密切相关，细胞因子可通过调节bcl-2和bax的表达参与细胞凋亡的调控。     

小鼠胚泡植入前、中、后子宫内膜nm23-M2动态表达

目的研究抑癌基因nm23-M2在小鼠胚泡植入前、中、后子宫内膜的动态表达。方法采用RT-PCR及免疫组织化学方法分别检测小鼠妊娠第2天（植入前）、第5天（植入中）、第7走（植入后）子宫内膜nm23-M2表达。结果RT-PER测得胚泡植入前、中、后子宫内膜中均存在nm23-M2的表达，但在妊娠第5天植入窗口期（植入中）nm23-M2 mRNA/β-actin mRNA的比值明显升高，与植入前和植入后相比较，具有显著差异（P〈0．01）。免疫组织化学分析显示妊娠第5天子宫内膜nm23-M2表达为强阳性，而植入前和植入后子宫内膜nm23-M2表达多为弱阳性。表明胚泡植入窗口期子宫内膜nm23-M2表达上调。结论nm23-M2可能参与了小鼠胚泡植入。     

妊娠早期小鼠卵巢、输卵管及子宫内诱导型一氧化氮合酶的分布

目的　了解胚泡着床前后妊娠昆明系小鼠卵巢、输卵管及子宫内诱导型一氧化氮合酶 (inducible ni-tric oxide synthase,i NOS)的分布。　方法　免疫细胞化学 L SAB法。　结果　妊娠 2～ 5 d小鼠的卵巢内 ,黄体细胞上有 i NOS的阳性表达 ;输卵管粘膜上有 i NOS的分布 ,肌层则为阴性 ;妊娠 2、3d的小鼠子宫内 ,阳性标记主要出现在子宫内膜上皮以及子宫内膜中的子宫腺上皮 ,内膜基质细胞为阴性 ;妊娠 4d的子宫内 ,子宫腺及蜕膜部分均有 i-NOS的分布 ,妊娠 5 d时 ,在小鼠胚泡的表面也检测到了 i NOS的存在。　结论　小鼠胚泡着床前后 ,在其卵巢、输卵管及子宫内均有 i NOS的存在 ,提示 i NOS在小鼠胚胎早期发育及着床过程中起作用。     

Ultrastructural studies of glycan changes in the apical surface of the uterine epithelium during pre-ovulatory and and pre-implantation stages in the marmoset monkey.

It has been postulated that carbohydrates are involved in a variety of cell-cell interactions including blastocyst implantation. In primates, there are only limited investigations on the ultrastructural localisation of the cyclic changes in uterine epithelial surface carbohydrates. Our aim was to investigate such changes during the pre-ovulatory and pre-implantation stages of the reproductive cycle in the marmoset monkey. After fixation of endometrial tissues, avidin-ferritin lectin cytochemistry was employed for apical surface glycan detection at the ultrastructural level. Five lectins were used including Canavalia ensiformis (Con A), Lotus tetragonolobus (LTA), Glycine max (SBA), Phytolacca americana (PWM) and Triticum vulgaris (WGA). Morphometry was used to quantitate changes in the intensity of lectin staining by determining the total number of ferritin particles per unit length of membrane. Surface and intra-cytoplasmic vesicles, stained by the lectins, were also examined. Quantitative ferritin assessment showed that 1 day before presumed implantation (days 11 to 12 after ovulation in the marmoset monkey) there was a significant increase in Con A, LTA and SBA staining on the apical uterine epithelial plasma membrane compared to the pre-ovulatory phase and earlier stages of pregnancy (days 4-8 after ovulation). A significant increase in PWM was also detected from early pregnancy to pre-implantation stages. All lectins except WGA produced reproducible staining within reproductive cycle groups. The greatest variation and intensity of epithelial surface staining was observed with WGA and the weakest with LTA. The patchy staining with LTA compared with thick coverage by WGA indicated the complexity of the carbohydrate arrangement in the glycocalyx of the uterine surface plasma membrane. Reduction of WGA reactivity after neuraminidase treatment suggested that the lectin binding might be related to the presence of heavily sialylated apical uterine membrane glycoconjugates. This is the first high-resolution study in primates to report quantitative cyclic changes in fucosyl, galactosyl, glucosyl, and mannosyl sugar residues of the apical uterine epithelial glycocalyx. The findings support the concept that uterine epithelial glycocalyx surface carbohydrates play a role in preparing a receptive uterine surface.     

Ultrastructural studies of glycan changes in the apical surface of the uterine epithelium during pre‐ovulatory and pre‐implantation stages in the marmoset monkey

It has been postulated that carbohydrates are involved in a variety of cell‐cell interactions including blastocyst implantation. In primates, there are only limited investigations on the ultrastructural localisation of the cyclic changes in uterine epithelial surface carbohydrates. Our aim was to investigate such changes during the pre‐ovulatory and pre‐implantation stages of the reproductive cycle in the marmoset monkey. After fixation of endometrial tissues, avidin‐ferritin lectin cytochemistry was employed for apical surface glycan detection at the ultrastructural level. Five lectins were used including Canavalia ensiformis (Con A), Lotus tetragonolobus (LTA), Glycine max (SBA), Phytolacca americana (PWM) and Triticum vulgaris (WGA). Morphometry was used to quantitate changes in the intensity of lectin staining by determining the total number of ferritin particles per unit length of membrane. Surface and intra‐cytoplasmic vesicles, stained by the lectins, were also examined. Quantitative ferritin assessment showed that 1 day before presumed implantation (days 11 to 12 after ovulation in the marmoset monkey) there was a significant increase in Con A, LTA and SBA staining on the apical uterine epithelial plasma membrane compared to the pre‐ovulatory phase and earlier stages of pregnancy (days 4–8 after ovulation). A significant increase in PWM was also detected from early pregnancy to pre‐implantation stages. All lectins except WGA produced reproducible staining within reproductive cycle groups. The greatest variation and intensity of epithelial surface staining was observed with WGA and the weakest with LTA. The patchy staining with LTA compared with thick coverage by WGA indicated the complexity of the carbohydrate arrangement in the glycocalyx of the uterine surface plasma membrane. Reduction of WGA reactivity after neuraminidase treatment suggested that the lectin binding might be related to the presence of heavily sialylated apical uterine membrane glycoconjugates. This is the first high‐resolution study in primates to report quantitative cyclic changes in fucosyl, galactosyl, glucosyl, and mannosyl sugar residues of the apical uterine epithelial glycocalyx. The findings support the concept that uterine epithelial glycocalyx surface carbohydrates play a role in preparing a receptive uterine surface. Anat Rec 255:241–251, 1999. © 1999 Wiley‐Liss, Inc.     

分泌期反复流产子宫内膜凝集素结合特性探讨

本文通过生物素分别标记的双花藕豆凝集素（ＤＢＡ）、大豆凝集素（ＳＢＡ）、麦胚凝集素（ＷＧＡ）及荆豆凝集素（ＵＥＡ－１）与子宫内膜的反应，应用Ｓ－Ｐ法研究２８例无明显原因反复流产的分泌期子宫内膜糖复合物的变化及与其组织形态学分期的相关性。结果：①宫内膜迟延发生率为３９．３％（１１／２８）；②四种凝集素中只有ＤＢＡ与子宫内膜的结合表现出阶段特异性。着床期，ＤＢＡ宫内膜结合率增强，且与宫内膜组织形态学分期呈有意义的直线正相关（ｒ＝０．４０４，Ｐ＜０．０５）。提示与ＤＢＡ特异结合的子宫内膜上的糖复合物与孕卵着床有关，这种特异结合可以作为评价分泌期子宫内膜分化成熟的一项指标     

Advanced oviductal development, transport to the preferred implantation site, and attachment of the blastocyst in captive-bred, short-tailed fruit bats, Carollia perspicillata

The final stages of embryonic development in the oviduct, transport of the embryo to the uterus, and the initial stages of implantation have been examined in captive-bred Carollia perspicillata at the light and electron microscopic levels. Development progressed to the expanded, zona pellucida-free, blastocyst stage in the oviduct. The abundance of microvilli on the exterior of the trophoblast varied with the degree of blastocyst expansion and cell shape, and may function in part as a membrane reservoir. Cells of the blastocyst also typically contained many lipid droplets and prominent areas of cytoplasm occupied by finely granular material (probably glycogen) instead of organelles. In most females, closure of the uterine lumen occurred prior to, or around the time of, transport of the blastocyst to the usual implantation site and appeared to play a role in preventing transport of the blastocyst too far distally in the uterus. This was associated with increased endometrial edema, particularly in the fundic region of the simplex uterus, and the extravasation of many erythrocytes into the endometrial stroma. Both of these changes began while the blastocyst was still being held in the oviduct and became pronounced during implantation. Engulfment of these erythrocytes by processes of the endometrial stromal cells and their phagocytosis by macrophages was also observed. Implantation was usually initiated within narrow tubular segments, lined by endometrium, that were located between the end of each oviduct and the main cavity of the uterus, or from immediately adjacent areas of the main cavity. During the early stages of implantation, the blastocyst was clasped by the endometrium at the implantation site, and this was associated with extensive interdigitation of the microvilli of the trophoblast and adjacent uterine epithelial cells. Initial adhesion of the trophoblast, which was still cellular rather than synctial, occurred over the apical intercellular junctions of the uterine epithelial cells.     

人及哺乳动物消化管中凝集素结合部位的形态观察

用三种辣根过氧化物酶标记的凝集素—WGA、SBA和PNA观察人及大小鼠的消化管各段中凝集素结合部位的分布。在各段消化管的粘膜上皮均有染色反应,但胃粘膜上皮细胞的核上区不被PNA着色。在胃底腺中,人的壁细胞只对WGA反应,主细胞对WGA和PNA都有反应,而大小鼠二种细胞对三种凝集素均有反应。杯状细胞对PNA不着色。肠腺反应与粘膜上皮相似。固有膜、平滑肌被WGA着色。神经元对WGA和PNA反应。由此提示糖蛋白分布的差异存在于各段消化管和同段消化管的不同区域并有种间差异。     

Immunohistochemical localization of receptors for progesterone and oestradiol-17 beta in the implantation site of the rhesus monkey

The aim of the present study was to examine the cellular basis of the involvement of oestradiol and progesterone in blastocyst implantation in the primate. To this end, the cellular distribution of receptors for oestradiol (ER) and progesterone (PR) in fetal trophoblast cells and in endometrial compartments of timed lacunar (pre-villous) and villous stages of placentation in primary implantation sites collected on days 13-22 of gestation were investigated in rhesus monkeys. Both in pre-villous stage tissue and in villous stage tissue, cytotrophoblast cells and syncytiotrophoblast cells and other trophoblast derived cells were PR positive, while they were generally ER negative. Maternal endometrial cells were ER negative, while epithelial cells, stromal cells and vascular endothelial cells in maternal endometrium showed heterogeneous staining patterns for PR depending on their relative location; these patterns, however, correlated well with glandular hyperplasia and differentiation, stromal-decidual transformation and vascular response seen during blastocyst implantation.