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1.
Penkov L. I. Platonov E. S. Kondrakhina M. S. Konyukhov B. V. 《Russian Journal of Developmental Biology》2003,34(4):249-253
We studied the effect of the growth factor LIF on the development of parthenogenetic mouse embryos (CBA × C57BL/6)F1. LIF was added to the culture medium at 10, 50, 100, and 250 ng/ml at the morula stage and parthenogenetic embryos were cultured in vitro until the late blastocyst stage and then transplanted in the uterus of pseudopregnant females, which were then sacrificed on day 12 of pregnancy. All the LIF doses used improved the development of parthenogenetic mouse embryos at the preimplantation stages and increased the amount of blastocysts by 15%, on average, as compared to the control. LIF at 50 and 100 ng/ml increased approximately twice the number of embryos that reached the somite stages. Some of them reached the stage of 32–45 somites and had fore and hind limb buds. No such embryos were found in the control. Well formed placenta was observed in 6% of the embryos treated with LIF and the most pronounced effect was recorded at 100 ng/ml. The data we obtained suggest that exogenous LIF can improve pre- and postimplantation development of parthenogenetic mouse embryos due, possibly, to increased survival rate of embryonic stem cells derived from the inner cell mass of blastocysts. LIF improves not only the development of the parthenogenetic embryoper se, but also the formation of its extraembryonic envelopes, which leads to the development of a larger placenta in LIF-treated parthenogenetic embryos, as compared to the control. 相似文献
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E. S. Lobanok L. M. Mezhevikina L. M. Belyanovich R. R. Petrova I. B. Vasilevich I. D. Volotovsky E. E. Fesenko 《Biochemistry (Moscow) Supplemental Series B: Biomedical Chemistry》2009,3(1):54-57
The influence of cytokine LIF (Leukemia Inhibitory Factor) on the viability, and proliferation of mouse embryonic stem cells (ESC) (R1 cell line) and their distribution by cell cycle stages has been investigated. LIF (5–20 ng/ml) increased growth of colonies and maintained high proliferative and pluripotent properties of R1 cells. LIF was also involved into the inhibition of spontaneous cell differentiation and apoptotic cell death; it also decreased the rations of S/G2 + M cell cycle and doubling-time of cell population. 相似文献
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人白血病抑制因子基因LIF的克隆、真核表达及活性检测 总被引:1,自引:0,他引:1
白血病抑制因子 (LIF)是一种多功能细胞因子 ,在生物发育及维持正常生理功能中发挥着重要的作用。以成人血细胞的总RNA为模板 ,利用RT PCR方法从成人外周血细胞中扩增人白血病抑制因子 ,而后将其克隆到真核表达载体pcDNA3,在哺乳动物细胞中成功表达。将分泌到细胞培养基中的LIF因子收集 ,通过LIF信号转导通路下游蛋白质STAT3磷酸化水平的检测、EMSA实验以及荧光酶报告系统检测 ,发现其具有激活STAT3信号通路的生物学活性。同时 ,[3H] TdR参入实验的结果也表明 ,所表达LIF因子能显著抑制鼠骨髓白血病细胞系M1的增殖。 相似文献
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目的:研究FLT4(VEGFR3)基因在斑马鱼早期发育过程中的表达。方法:提取斑马鱼胚胎的总RNA,制备地高辛标记的FLT4RNA反义探针,WISH(整胚胎原位杂交)研究FLT4在斑马鱼早期发育过程中的表达。结果:成功合成FLT4基因探针,获得FLT4基因在斑马鱼早期发育过程中的表达情况:FlT4在2-cell、32-cell、oblong、shield期前普遍性表达(0.75h、1.7h、3.7h、6h);24h在头部表达较多,特别是在ICM(intermediate cell mass,中间细胞群)区处有特异性表达;48h、72h在头部表达均较高表达。结论:FLT4在早期参与了血管的形成和造血的发生,在脑部和肾小管的发育中可能起到了重要作用。 相似文献
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目的:研究sema(semaphorin)4d基因在斑马鱼早期发育过程中的表达.方法:提取斑马鱼胚胎的总RNA,制备地高辛标记的sema4d RNA反义探针,WISH(整胚胎原位杂交)研究sema4d在斑马鱼早期发育过程中的表达.结果:成功合成sema4d基因探针,获得sema4d基因在斑马鱼早期发育过程中的表达情况:sema4d在0.75 hpf(hours post fertilization)、1.0 hpf、1.5 hpf、12 hpf前普遍性表达;17 hpf开始至24 hpf在头部表达较多,在脊髓、肌肉、中间细胞群ICM(intermediate cell mass)区处有特异性表达区处有特异性表达;48 hpf在头部和躯干肌肉持续表达.结论:Sema4d在早期参与了造血的发生,在脑部,脊髓,肌肉的发育中可能起到了重要作用. 相似文献
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Abdel-Majid Khatib Rachid Lahlil Nathalie Scamuffa Marie-Andrée Akimenko Sylvain Ernest Abdderahim Lomri Claude Lalou Nabil G. Seidah Bruno O. Villoutreix Fabien Calvo Geraldine Siegfried 《PloS one》2010,5(7)
Background
In zebrafish, vascular endothelial growth factor-C precursor (proVEGF-C) processing occurs within the dibasic motif HSIIRR214 suggesting the involvement of one or more basic amino acid-specific proprotein convertases (PCs) in this process. In the present study, we examined zebrafish proVEGF-C expression and processing and the effect of unprocessed proVEGF-C on caudal fin regeneration.Methodology/Principal Findings
Cell transfection assays revealed that the cleavage of proVEGF-C, mainly mediated by the proprotein convertases Furin and PC5 and to a less degree by PACE4 and PC7, is abolished by PCs inhibitors or by mutation of its cleavage site (HSIIRR214 into HSIISS214). In vitro, unprocessed proVEGF-C failed to activate its signaling proteins Akt and ERK and to induce cell proliferation. In vivo, following caudal fin amputation, the induction of VEGF-C, Furin and PC5 expression occurs as early as 2 days post-amputation (dpa) with a maximum levels at 4–7 dpa. Using immunofluorescence staining we localized high expression of VEGF-C and the convertases Furin and PC5 surrounding the apical growth zone of the regenerating fin. While expression of wild-type proVEGF-C in this area had no effect, unprocessed proVEGF-C inhibited fin regeneration.Conclusions/Significances
Taken together, these data indicate that zebrafish fin regeneration is associated with up-regulation of VEGF-C and the convertases Furin and PC5 and highlight the inhibitory effect of unprocessed proVEGF-C on fin regeneration. 相似文献8.
白血病抑制因子促进胚泡植入的研究进展 总被引:1,自引:0,他引:1
白血病抑制因子是一多效性细胞因子,能促进哺乳动物的早期胚胎发育和启动胚泡植入,其基因表达和生物合成受母体因素(如甾体激素和其他细胞因子)的控制.gp130是白血病抑制因子家族受体的亲和力转化亚基,其同源/异源亚基的二聚体能够激活酪氨酸激酶,通过不同途径调节靶基因的表达. 相似文献
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Changes in Protein Content of Goldfish Optic Nerve During Degeneration and Regeneration Following Nerve Crush 总被引:10,自引:8,他引:2
Abstract: After the goldfish optic nerve was crushed, the total amount of protein in the nerve decreased by about 45% within 1 week as the axons degenerated, began to recover between 2 and 5 weeks as axonal regeneration occurred, and had returned to nearly normal by 12 weeks. Corresponding changes in the relative amounts of some individual proteins were investigated by separating the proteins by two-dimensional gel electrophoresis and performing a quantitative analysis of the Coomassie Brilliant Blue staining patterns of the gels. In addition, labelling patterns showing incorporation of [3H]proline into individual proteins were examined to differentiate between locally synthesized proteins (presumably produced mainly by the glial cells) and axonal proteins carried by fast or slow axonal transport. Some prominent nerve proteins, ON1 and ON2 (50–55 kD, pI ~6), decreased to almost undetectable levels and then reappeared with a time course corresponding to the changes in total protein content of the nerve. Similar changes were seen in a protein we have designated NF (~130 kD, pI ~5.2). These three proteins, which were labelled in association with slow axonal transport, may be neurofilament constituents. Large decreases following optic nerve crush were also seen in the relative amounts of α- and β-tubulin, which suggests that they are localized mainly in the optic axons rather than the glial cells. Another group of proteins, W2, W3, and W4 (35–45 kD, pI 6.5–7.0), which showed a somewhat slower time course of disappearance and were intensely labelled in the local synthesis pattern, may be associated with myelin. A small number of proteins increased in relative amount following nerve crush. These included some, P1 and P2 (35–40 kD, pIs 6.1–6.2) and NT (~50 kD, pI ~5.5), that appeared to be synthesized by the glial cells. Increases were also seen in one axonal protein, B (~45 kD, pI ~4.5), that is carried by fast axonal transport, as well as in two axonal proteins, HA1 and HA2 (~60 and 65 kD respectively, pIs 4.5–5.0), that are carried mainly by slow axonal transport. Other proteins, including actin, that showed no net changes in relative amount (but presumably changed in absolute amount in direct proportion to the changes in total protein content of the nerve), are apparently distributed in both the neuronal and nonneuronal compartments of the nerve. 相似文献
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In Vivo Phosphorylation of Axonal Proteins in Goldfish Optic Nerve During Regeneration 总被引:4,自引:4,他引:0
In vivo phosphorylation of axonal proteins was investigated in normal and regenerating optic nerves of goldfish by two-dimensional gel electrophoresis. By 6-24 h after intraocular injection of H3(32)PO4, approximately 20 optic nerve proteins ranging in size from 19 to 180 kilodaltons and in pI from 4.4 to 6.8 were seen to have incorporated radiolabel. Five of these proteins showed a robust increase in incorporation of phosphate during regeneration. Among the latter was an acidic (pI 4.5) 45-kilodalton protein, which has previously been shown to be conveyed by fast axonal transport and to increase dramatically in its rate of synthesis during regeneration of goldfish optic axons. 相似文献
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目的考察神经生长因子对视神经损害的疗效。方法腹腔注射二硫化碳致视神经损害的大鼠模型,给药组球后注射或肌内注射神经生长因子,每天一次,每周6d,共计3周,空白对照组球后注射等量生理盐水,于治疗前后测定大鼠模式翻转(FREP)和闪光视觉诱发电位。结果球后注射高剂量组大鼠在治疗10d和20d时各波潜伏时比对照组有显著缩短,而球后注射低剂量组和肌内注射高剂量组大鼠于治疗10d时FVEP各波潜伏时也显著缩短,肌内注射低剂量组大鼠于治疗20d时FEP的P1和P2波潜伏时也显著或非常显著缩短。结论神经生长因子对大鼠视神经损害有明显的治疗作用。 相似文献
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Uridine Metabolism in the Goldfish Retina During Optic Nerve Regeneration: Cell-Free Preparations 总被引:3,自引:2,他引:1
The activities of uridine kinase (EC 2.7.1.48), uridine monophosphate (UMP) kinase (EC 2.7.1.3.14), and uridine diphosphate (UDP) kinase (EC 2.7.4.6) were measured in retinal high-speed supernatant fractions following unilateral optic nerve crush in the goldfish. The enzyme activities followed a similar time course, with initial increases 2-3 days following nerve crush, peak activity at 4 days, and a gradual return to basal levels by day 21. The magnitude of the stimulation on day 4 was about 35% in each case. Activities of two enzymes of intermediary metabolism, pyruvate kinase (EC 2.7.1.40) and lactic dehydrogenase (EC 1.1.1.27), were not altered, indicating that the coordinate increases in nucleoside and nucleotide kinase activities were specific responses to the nerve injury. The increased labeling could not be explained by altered phosphohydrolytic activities. The nature of the enhancement was further studied in UDP kinase, the most active of the kinases examined. Neither low-molecular-weight components nor substrate availability could account for the observed increase in UDP kinase in the 4 day post-crush retinas. The Km for UDP was unaltered, and a mixing experiment did not support the possibility that stimulatory or inhibitory factors played a role. The enhancement of UDP kinase activity was blocked by injection of actinomycin D following nerve crush. The results suggest that the observed increases in enzymes of uridine metabolism result from their increased formation following nerve crush. 相似文献
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Uridine Metabolism in the Goldfish Retina During Optic Nerve Regeneration: Whole Retina Studies 总被引:3,自引:2,他引:1
Linda A. Dokas Shinichi Kohsaka Harry R. Burrell Bernard W. Agranoff 《Journal of neurochemistry》1981,36(3):1160-1165
Accumulation of radioactivity from [3H]uridine in incubations of whole goldfish retinas is increased in the ipsilateral retina during a period of regeneration that follows unilateral optic nerve crush. Brief incubations to investigate the nature of enhanced labeling of the acid-soluble fraction showed a peak uptake 4 days following crush, with a gradual decrease to control levels by 21 days following crush. That nucleoside uptake may not mediate the effect is supported by the observation that the rate of uptake of 5'-deoxyadenosine, a nonmetabolizable nucleoside analog, is the same in post-crush (PC) and normal (N) retinal incubations. Following brief incubations of PC and N retinas with [3H]uridine, there is enhanced labeling in PC retinas relative to N retinas of recovered UMP, UDP, UTP, and uridine nucleotide sugars, whereas recovery of labeled uridine itself is slightly decreased. The results suggest that the increased accumulation of radioactivity in PC retinas following incubation with uridine reflects an increase in the activities of retinal uridine kinase and uridine nucleotide kinases. 相似文献
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Hiroyuki Nawa† Shigetada Nakanishi† Paul H. Patterson 《Journal of neurochemistry》1991,56(6):2147-2150
The cholinergic differentiation factor (CDF) in heart cells is identical to leukemia inhibitory factor (LIF). Recombinant CDF/LIF was shown to alter dramatically neurotransmitter production as well as the levels of several neuropeptides in cultured rat sympathetic neurons. Here it is shown that these changes are likely to be caused by alterations in the mRNA for these proteins and peptides. Growth in 1 nM recombinant CDF/LIF induces mRNA for acetyl CoA: choline-O-acetyltransferase [EC 2.3.1.6; choline acetyltransferase (ChAT)], somatostatin (SOM), substance P, and vasoactive intestinal polypeptide while lowering mRNA levels of tyrosine hydroxylase (EC 1.14.16.2) and neuropeptide Y (NPY). In addition, the sizes of the mRNAs for ChAT, SOM, and NPY are larger after recombinant CDF/LIF treatment. 相似文献
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目的:原核表达重组hLIF融合蛋白并进行诱导表达、纯化及活性鉴定.方法:将hLIF基因克隆至pThioHisA载体,构建融合表达载体pThioHisA-hLIF,转化大肠杆菌BL21(DE3),IPTG诱导表达.表达产物经亲和层析后,Western blot检测目的蛋白的特异性.用小鼠胚胎干细胞脱饲养层培养对纯化后的重组hLIF融合蛋白进行生物活性的鉴定.结果:降低诱导温度和延长诱导时间能增加hLIF融合蛋白的可溶性表达,纯化后的重组蛋白纯度大于95%,Western blot检测显示了良好的特异性.在脱饲养层细胞培养条件下,添加纯化的hLIF融合蛋白能够有效的维持小鼠胚胎干细胞的未分化状态.结论:重组hLIF融合蛋白可在大肠杆菌中高效表达,具有良好的特异性,为干细胞研究及hLIF蛋白的其他功能研究奠定了基础. 相似文献
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Leukemia inhibitory factor (LIF) improves and prolongs the development of parthenogenetic mouse embryos 总被引:2,自引:0,他引:2
We studied the effect of the growth factor LIF on the development of parthenogenetic mouse embryos (CBA x C57BL/6)F1. LIF was added to the culture medium at 10, 50, 100, and 250 ng/ml at the morula stage and parthenogenetic embryos were cultivated in vitro until the late blastocysts stage and then transplanted in the uterus of pseudopregnant females, which were then sacrificed on day 12 of pregnancy. All the LIF doses used improved the development of parthenogenetic mouse embryos at the preimplantation stages and increased the amount of blastocysts by 16%, on average, as compared to the control. LIF at 50 and 100 ng/ml increased approximately twice the number of embryos that reached the somatic stages. Some of them reached the stage of 32-45 somites and had fore and hind limb buds. No such embryos were found in the control. Well formed placenta was observed in 6% of the embryos treated with LIF and the most pronounced effect was recorded at 100 ng/ml. The data we obtained suggest that exogenous LIF can improve pre- and postimplantation development of parthenogenetic mouse embryos due, possibly, to increased survival rate of embryonic stem cells derived from the inner cell mass of blastocysts. LIF improves not only the development of the parthenogenetic embryo per se, but also the formation of its extraembryonic envelopes, which leads to the development of a larger placenta in LIF-treated parthenogenetic embryos, as compared to the control. 相似文献
18.
Diabetic retinopathy is a leading cause of reduced visual acuity and acquired blindness. Axoglial alterations of the distal (close to the chiasm) optic nerve (ON) could be the first structural change of the visual pathway in streptozotocin (STZ)-induced diabetes in rats. We analyzed the effect of environmental enrichment on axoglial alterations of the ON provoked by experimental diabetes. For this purpose, three days after vehicle or STZ injection, animals were housed in enriched environment (EE) or remained in a standard environment (SE) for 6 weeks. Anterograde transport, retinal morphology, optic nerve axons (toluidine blue staining and phosphorylated neurofilament heavy immunoreactivity), microglia/macrophages (ionized calcium binding adaptor molecule 1 (Iba-1) immunoreactivity), astrocyte reactivity (glial fibrillary acid protein-immunostaining), myelin (myelin basic protein immunoreactivity), ultrastructure, and brain derived neurotrophic factor (BDNF) levels were assessed in non-diabetic and diabetic animals housed in SE or EE. No differences in retinal morphology or retinal ganglion cell number were observed among groups. EE housing which did not affect the STZ-induced weight loss and hyperglycemia, prevented a decrease in the anterograde transport from the retina to the superior colliculus, ON axon number, and phosphorylated neurofilament heavy immunoreactivity. Moreover, EE housing prevented an increase in Iba-1 immunoreactivity, and astrocyte reactivity, as well as ultrastructural myelin alterations in the ON distal portion at early stages of diabetes. In addition, EE housing avoided a decrease in BDNF levels induced by experimental diabetes. These results suggest that EE induced neuroprotection in the diabetic visual pathway. 相似文献
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Peter Luedike Christos Rammos Julia Pohl Martin Heisler Matthias Totzeck Werner Kleophas Gerd R. Hetzel Malte Kelm Ulrike Hendgen-Cotta Tienush Rassaf 《PloS one》2015,10(10)
BackgroundEnd stage renal disease (ESRD) patients are characterized by increased morbidity and mortality due to highest prevalence of cardiovascular disease. Macrophage migration inhibitory factor (MIF) is an inflammatory cytokine that controls cellular signaling in human physiology, pathophysiology, and diseases. Increased MIF plasma levels promote vascular inflammation and development of atherosclerosis. We have shown that MIF is associated with vascular dysfunction in ESRD patients. Whether hemodialysis (HD) affects circulating MIF plasma levels is unknown. We here aimed to investigate whether HD influences the circulating MIF pool in ESRD patients.ConclusionMIF is a dialyzable plasma component that is effectively filtrated during HD from the patient blood pool in large amounts. After removal of remarkable amounts of MIF during a single HD session, MIF plasma pool is early reconstituted after termination of HD from unknown sources. 相似文献
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二恶英诱发斑马鱼初期胚的循环系统障碍 总被引:4,自引:0,他引:4
目的 观察二恶英 (2 ,3,7,8 tetrachlorodibenzo p dioxin ,以下略为TCDD)对斑马鱼 (Zebrafish)初期胚胎的毒性作用。方法 斑马鱼胚从受精后 2 4h直至观察放在 2 4孔培养皿内水浴染毒。采用形态学、组织学观察法及细胞色素P4 5 0 1A(CYP1A)抗体染色法。结果 0 1μg LTCDD未观察到明显的变化 ,但是 0 3~ 10 μg L浓度的TCDD染毒时 ,首先观察到后主静脉的血流减缓与停滞为共同特征的毒性症状。在染毒群同时还观察到心囊、卵黄囊、头部、躯体等不同程度的水肿以及头部的畸形。在 180h ,摄食对照群 ,0 1μg L、0 3μg L、0 5 μg L、1μg L和 10 μg L各自的死亡率分别是 0、0、5 %、6 0 2 %、6 1 8%、10 0 %。CYP1A抗体染色发现对照群无阳性反应 ,而染毒群在血管上皮看到极强的阳性反应。结论 本试验表明TCDD对斑马鱼胚胎的循环系统有极强的损坏作用 ,而且这种损坏与CYP1A相关联 相似文献