Assessment of Maternal Vascular Remodeling During Pregnancy in the Mouse Uterus

The placenta mediates the exchange of factors such as gases and nutrients between mother and fetus and has specific demands for supply of blood from the maternal circulation. The maternal uterine vasculature needs to adapt to this temporary demand and the success of this arterial remodeling process has implications for fetal growth. Cells of the maternal immune system, especially natural killer (NK) cells, play a critical role in this process. Here we describe a method to assess the degree of remodeling of maternal spiral arteries during mouse pregnancy. Hematoxylin and eosin-stained tissue sections are scanned and the size of the vessels analysed. As a complementary validation method, we also present a qualitative assessment for the success of the remodeling process by immunohistochemical detection of smooth muscle actin (SMA), which normally disappears from within the arterial vascular media at mid-gestation. Together, these methods enable determination of an important parameter of the pregnancy phenotype. These results can be combined with other endpoints of mouse pregnancy to provide insight into the mechanisms underlying pregnancy-related complications.     

表皮生长因子对鼠胚成纤维细胞细胞周期的调节作用

本文观察了表皮生长因子（ＥＧＦ）对小鼠胚胎成纤维细胞Ｃ_３Ｈ／１０Ｔ_１／２ＣＬ_８（简称Ｃ_３Ｈ／１０）细胞周期的影响。结果表明：ＥＧＦ使Ｓ期提前,细胞周期缩短。进一步探讨了ＥＧＦ对细胞周期影响的机制,发现ＥＧＦ可活化在细胞周期调节中起重要作用的Ｐ３４~（ｃｄｃ２）激酶（简称ＣＤ２Ｋ）,使ＣＤ２Ｋ在周期中活性高峰出现的时间提前,提示ＥＧＦ对细胞周期的影响可能通过作用于ＣＤ２Ｋ实现的。     

Epidermal Growth Factor in Synaptosomal Fractions of Mouse Cerebral Cortex

Using a specific and sensitive epidermal growth factor radioimmunoassay (EGF-RIA) we measured EGF concentrations in whole brain, cerebral cortex, and cerebral cortical synaptosomal (pinched-off presynaptic nerve terminals) fractions of 26-day-old mouse brain. The relative EGF concentration in synaptosomal fractions was significantly greater than the growth factor concentrations in whole brain or cerebral cortex. Intracerebral injection, in an amount of EGF, several-fold greater than whole brain EGF content, did not appreciably increase synaptosomal EGF concentration, suggesting that no artifact was involved. The high synaptosomal EGF content suggests a neurotransmitter or a neuromodulator role for EGF in the CNS.     

Thrombospondin is Sequentially Expressed and then De-expressed During Early Pregnancy in the Rat Uterus

The expression of thrombospondin on Day 1, Day 3 and Day 6 of pregnancy has been examined in the rat, using light microscopic immunoperoxidase and electron immunogold techniques. The glycoprotein was expressed in the apical, lateral and basal uterine epithelium on Days 1 and 3 but was then de-expressed at the time of implantation on Day 6. We propose that these data suggest a role for thrombospondin in remodelling the uterine epithelium during the plasma membrane transformation, but that it does not play a part in attachment and implantation.     

IK细胞因子在小鼠早孕期子宫内膜的表达规律及其在胚胎着床中的作用

通过Real-time PCR、Western blot及免疫组织化学方法分析了IK细胞因子(IK cytokine)在早孕小鼠(妊娠D1~D7)子宫内膜中的表达规律及宫角注射IK细胞因子反义寡聚脱氧核苷酸后对胚胎着床的影响。结果显示,IK细胞因子mRNA表达在D1~D4逐渐升高,于D4达到高峰(P<0.05);Western blot和免疫组织化学结果与Real-time PCR结果基本一致,其蛋白表达在D1~D5逐渐升高,于D5达到高峰(P<0.05);IK细胞因子在D5胚胎着床点的表达显著高于着床旁组织;假孕小鼠子宫内膜IK细胞因子蛋白表达明显低于正常妊娠,且整个假孕过程中没有表达高峰;宫角注射IK细胞因子反义寡聚脱氧核苷酸后24 h和48 h(即D4和D5)子宫内膜IK细胞因子表达明显受到抑制,MHCⅡ抗原表达增强,且胚胎着床数量明显减少(P<0.05),提示IK细胞因子在胚胎着床中发挥着重要作用。     

表皮生长因子及受体在甲状腺肿瘤中的表达

目的:研究表皮生长因子(Epidermal Growth Factor,EGF)及受体(Epidermal Growth Factor Receptor,EGFR)及在甲状腺肿瘤中的表达。方法:应用免疫组织化学法检测91例甲状腺病变组织中EGFR和EGF的表达情况。结果:结节性甲状腺肿、甲状腺腺瘤、分化型甲状腺癌标本中EGFR表达的阳性率分别为15%、25%、68.62%,EGF表达的阳性率分别为10%、15%、68.62%,其中EGFR、EGF在分化型甲状腺癌与其余两组间差异均有统计学意义(P<0.05)。EGFR和EGF在甲状腺乳头状癌中的表达与性别、年龄、肿瘤大小、淋巴结转移、临床分期等临床因素无明显相关。结论:EGF和EGFR的表达可作为鉴别甲状腺肿瘤良恶性的一个指标。     

Urinary Oestrogens in Heifers Treated with Prednisolone During the Oestrous Cycle

Heiferis treated with a glucocorticoid during the oestrous cycle, displayed increased follicular activity and small corpora lutea compared to untreated controls (Tomasgard 1976a), but showed no signs of oestrogen production during the treatment.     

Epidermal Growth Factor Signaling and Mitogenesis in Plcg1 Null Mouse Embryonic Fibroblasts

Gene targeting techniques and early mouse embryos have been used to produce immortalized fibroblasts genetically deficient in phospholipase C (PLC)-γ1, a ubiquitous tyrosine kinase substrate. Plcg1^−/− embryos die at embryonic day 9; however, cells derived from these embryos proliferate as well as cells from Plcg1^+/+ embryos. The null cells do grow to a higher saturation density in serum-containing media, as their capacity to spread out is decreased compared with that of wild-type cells. In terms of epidermal growth factor receptor activation and internalization, or growth factor induction of mitogen-activated protein kinase, c-fos, or DNA synthesis in quiescent cells, PLcg1^−/− cells respond equivalently to PLcg1^+/+ cells. Also, null cells are able to migrate effectively in a wounded monolayer. Therefore, immortalized fibroblasts do not require PLC-γ1 for many responses to growth factors.     

Osteopontin Is Expressed in the Mouse Uterus during Early Pregnancy and Promotes Mouse Blastocyst Attachment and Invasion In Vitro

Embryo implantation into the maternal uterus is a decisive step for successful mammalian pregnancy. Osteopontin (OPN) is a member of the small integrin-binding ligand N-linked glycoprotein family and participates in cell adhesion and invasion. In this study, we showed that Opn mRNA levels are up-regulated in the mouse uterus on day 4 and at the implantation sites on days 5 and 8 of pregnancy. Immunohistochemistry localized the OPN protein to the glandular epithelium on day 4 and to the decidual zone on day 8 of pregnancy. OPN mRNA and proteins are induced by in vivo and in vitro decidualization. OPN expression in the endometrial stromal cells is regulated by progesterone, a key regulator during decidualization. As a secreted protein, the protein level of OPN in the uterine cavity is enriched on day 4, and in vitro embryo culturing has indicated that OPN can facilitate blastocyst hatching and adhesion. Knockdown of OPN attenuates the adhesion and invasion of blastocysts in mouse endometrial stromal cells by suppressing the expression and enzymatic activity of matrix metalloproteinase-9 in the trophoblast. Our data indicated that OPN expression in the mouse uterus during early pregnancy is essential for blastocyst hatching and adhesion and that the knockdown of OPN in mouse endometrial stroma cells could lead to a restrained in vitro trophoblast invasion.     

Plasma Progesterone in Gilts During the Prepuberal Period and the First Oestrous Cycle

The progesterone concentration of peripheral plasma in pigs has previously been studied during the oestrous cycle and in early pregnancy (Tillson & Erb 1967, Stabenfeldt et al 1969). The present study was undertaken to obtain some information on the concentration of plasma progesterone during the prepuberal period and during the first oestrous cycle in the pig.     

卵泡刺激素和表皮生长因子对小鼠精原细胞增殖的影响

利用生殖细胞-体细胞体外无血清共培养模型研究了卵泡刺激素(FSH)和表皮生长因子(EGF)对小鼠A型精原细胞增殖的影响。精原细胞在ITS培养液(添加胰岛素、转铁蛋白和亚硒酸钠的DMEM)中培养24h后进行c-kit免疫细胞化学鉴定和EGF及其受体(EGFR)免疫细胞化学检测,72h后测定其形成集落数的情况。结果表明:ITS培养液能维持生殖细胞的活性,增殖细胞核抗原(PCNA)的表达增高。A型精原细胞呈c-kit阳性,EGF和EGFR主要表达于精原细胞。单独的FSH(1～100ng/ml)或EGF(1～10ng/ml)显著促进精原细胞集落数的增加。此外,EGF(0.1ng/ml)联合FSH(10ng/ml)具有加性效应,但更高剂量的EGF(1～10ng/ml)则降低了FSH的刺激作用。结果说明FSH可联合适量的EGF促进精原细胞的增殖。     

The DNA Replication Licensing Factor Miniature Chromosome Maintenance 7 Is Essential for RNA Splicing of Epidermal Growth Factor Receptor,c-Met,and Platelet-derived Growth Factor Receptor

Miniature chromosome maintenance 7 (MCM7) is an essential component of DNA replication licensing complex. Recent studies indicate that MCM7 is amplified and overexpressed in a variety of human malignancies. In this report, we show that MCM7 binds SF3B3. The binding motif is located in the N terminus (amino acids 221–248) of MCM7. Knockdown of MCM7 or SF3B3 significantly increased unspliced RNA of epidermal growth factor receptor, platelet-derived growth factor receptor, and c-Met. A dramatic drop of reporter gene expression of the oxytocin exon 1-intron-exon 2-EGFP construct was also identified in SF3B3 and MCM7 knockdown PC3 and DU145 cells. The MCM7 or SF3B3 depleted cell extract failed to splice reporter RNA in in vitro RNA splicing analyses. Knockdown of SF3B3 and MCM7 leads to an increase of cell death of both PC3 and DU145 cells. Such cell death induction is partially rescued by expressing spliced c-Met. To our knowledge, this is the first report suggesting that MCM7 is a critical RNA splicing factor, thus giving significant new insight into the oncogenic activity of this protein.     

表皮生长因子和前列腺素E1对小鼠精原细胞增殖的促进作用

利用生殖细胞-体细胞无血清共培养模型研究了表皮生长因(EGF)和前列腺素E1(PGE1)对小鼠A型精原细胞增殖的影响.A型精原细胞在ITS培养液(添加胰岛素、转铁蛋白和亚硒酸钠的DMEM)中培养24 h后进行c-kit、EGF、表皮生长因子受体(EGFR)、环氧化酶-1(COX-1)及环氧化酶-2(COX-2)的免疫细胞化学检测,72 h后测定其形成集落教的情况.结果显示,A型精原细胞呈c-kit阳性,EGF、EGFR、COX-1及COX-2主要表达于精原细胞.EGF(10-7～10-6mol/L)或PGE,(10-8.10一mol/I_.)均可显著促进精原细胞集落的形成.此外,前列腺素(PG)受体拮抗剂SCl9220(10-6～10-5mol/L)可抑制PGE1对精原细胞的促增殖作用,COX-1抑制剂SC560(10-7～10-5mol/L)和COX-2抑制剂NS398(10-7～10-5mol/L)能抑制EGF促进精原细胞增殖的作用.因此,EGF可通过促进局部PG的产生而刺激精原细胞的增殖.     

Pattern of Replication of a Colicin Factor During the Cell Cycle of Escherichia coli

The ColBM, trp, lac episome was transferred to a lacZ derivative of Escherichia coli B/r, and the manner of replication of this colicinogenic factor was followed through the cell cycle. The results suggest a pattern of replication not connected with any particular stage in the cell cycle.     

钙离子对鼠角质细胞生长和分化的影响

用无血清培养基培养角质细胞,研究了Ca²⁺对鼠角质细胞生长和分化的影响。实验结果表明,培养基中钙离子最佳浓度为0.2mmol/L。在此浓度下,细胞克隆形成率达到10.8%,细胞的贴壁率达到28.7%,细胞的分化比例和老化比例分别为5.4%和26.3%;当Ca²⁺浓度达到0.6mmol/L以上时,则会引起角质细胞显著的分化和老化。