Reelin expression is upregulated following ocular tissue injury

Purpose Reelin is important in the guidance of neuronal stem cells in the central nervous system during normal development. We wished to determine whether reelin is expressed in the retina and cornea after injury. Methods Mice underwent laceration of their retina as well as corneal epithelial debridement. The mice were sacrificed at 3 days, and eyes were fixed and stained for reelin expression and reelin messenger ribonucleic acid (mRNA). Results In normal eyes, reelin was expressed only at very low levels in the ganglion cell layer of the retina and the endothelial cell layer of the cornea. In injured eyes, there was marked expression in reelin immunoreactivity in the retina and cornea. Reelin gene expression was seen in the retina and cornea. Conclusions Reelin is expressed during normal retinogenesis. This study shows that reelin is also upregulated following injury to the retina and cornea. The expression of reelin following injury suggests that reelin may play an important role in regulating stem cell trafficking in neuronal and nonneuronal tissues following injury similar to its role in normal organogenesis. For consideration of publication in Graefe’s Archive for Clinical and Experimental Ophthalmology.     

The role of pioneer neurons in the development of mouse visual cortex and corpus callosum

The primordial plexiform neuropil is very critical to neocortical development. The pioneer neurons, mainly Cajal-Retzius cells in the marginal zone, and subplate neurons in the subplate, differentiate from the primordial plexiform neuropil. In this study, the development of corpus callosum, visual cortex, and subcortical pathways has been observed in C57BL/6 mice with various methods, such as DiI labeling in vitro and in vivo, DiI and DiA in vitro double labeling, immunocytochemistry, and in vivo BrdU and Fast Blue labeling. As early as E14, the primordial plexiform neuropil can be found in the telencephalic wall, and it contains many pioneer neurons. On E15 the primordial plexiform neuropil differentiates into the marginal zone and the subplate. Cajal-Retzius cells exist in the marginal zone, and subplate neurons are in the subplate. Either Cajal-Retzius cells or subplate neurons have long projections toward the ganglionic eminence, suggesting that they migrate tangentially from the ganglionic eminence. Cajal-Retzius cells are involved in radial migration, and subplate neurons participate to guide pathfinding of subcortical pathways. This study reveals how the pioneer neurons, through radial and tangential migration, play an important role in neocortical formation and in the pathfinding of the corpus callosum and subcortical pathways. Furthermore, DiI labeling in vivo has demonstrated the presence of pioneer neurons all along the corpus callosum pathway, especially in the midline. This suggests that pioneer neurons may also play a role in guiding the pathfinding of the corpus callosum. Accepted: 31 July 2001     

Existing in-between two worlds: supporting asylum seeking women living in temporary accommodation through a creative movement and art intervention

ABSTRACT

In this reflective article we introduce Moving Space, a creative movement and art project supporting female Asylum Seekers as they move through the transient space of temporary accommodation. We explore how this cross-modal approach supports women to anchor experiences of displacement, loss and trauma through the use of embodied and visual creative process. Moreover, we argue that the transient nature of the therapeutic space brings into focus women’s resourcefulness and resilience despite the adversity and uncertainty they are experiencing.     

Migration and distribution of circumpharyngeal crest cells in the chick embryo

The cardiac neural crest is located in a transitional area on the neuraxis between trunk and cephalic regions and gives rise to both the dorsolateral and ventrolateral crest cell populations. Around stage 18 of chick development, a mass of E/C8⁺ cells surrounds the postotic pharyngeal arches and forms a crescent-shaped arch, termed the circumpharyngeal ridge. Using immunohistochemistry and quail-chick chimeras, it was determined that the E/C8⁺ cell mass located in the circumpharyngeal ridge derives from the dorsolateral component of the cardiac neural crest. The ventrolateral cell population of the cardiac crest is located more medially and shows long-persistent HNK-1 immunoreactivity dorsolateral to the foregut. The crest cells that populate the gut arise from the caudal portion of the circumpharyngeal crest and are always located caudal to the caudalmost pharyngeal ectomesenchyme. Circumpharyngeal crest cells continuously populate the pharyngeal arch ectomesenchyme and enteric nervous system on the lateral side of the foregut wall, as well as the hypoglossal pathway which develops within the ventral portion of the circumpharyngeal ridge. E/C8 and HNK-1 immunoreactivity are associated with the cells migrating via the dorsolateral (circumpharyngeal) and ventrolateral pathways, respectively, with one exception: there is a population of putative crest cells along the proximal course of the vagal intestinal branch that shows both immunoreactivities around stage 20. Dil labeling of the cells in the circumpharyngeal ridge suggests that the cells are contributed from the circumpharyngeal ridge to this population. Thus, the distribution of the circumpharyngeal crest cells and their derivatives coincides with the peripheral branch distribution of the cranial nerves IX, X, and XII, whose development is selectively affected in the absence of the cardiac neural crest, the source of the circumpharyngeal crest.© Willey-Liss, Inc.     

Immunohistochemical study about the Flt-1/VEGFR1 expression in the gastrointestinal tract of mouse, rat, dog, swine and monkey

Fms-like tyrosine kinase 1 (Flt-1) performs a subordinate effector role in mesenchymal angiogenesis and potentially serves an equally important functional role as a self-contained receptor in epithelial cells. In both endothelial cells and epithelial cells, Flt-1/vascular endothelial growth factor receptor 1 (VEGFR1) downstream signalling is involved in regulating cellular processes such as cytoskeletal changes and cellular survival protection. Cellular renewal of the gastrointestinal mucosa is based on these processes and might involve Flt-1/VEGFR1 pathway activities; the molecular mechanisms regulating these cellular dynamics remain unclear. This study was performed to investigate the presence and distribution of Flt-1/VEGFR1 in epithelial cells of the gastrointestinal tract by immunohistochemistry (IHC). Gastrointestinal tissues were taken from eight anatomical sites from mouse, rat, dog, swine and monkey. Present results revealed a cytosolic Flt-1/VEGFR1 staining pattern in mucosal epithelial cells for all investigated species. Non-epithelial structures also displayed a distinct Flt-1/VEGFR1 positivity and included vascular smooth muscle walls, enteric smooth muscle layers, the enteric nervous system and capillary endothelial cells. Diverse intensities of the Flt-1/VEGFR1 binding reaction within each species were observed in the intestinal mucosa with a strong immunoreaction in enterocytes and with a low protein expression in the ileum in most species. Crypt cells in the large intestine were mostly negative for Flt-1/VEGFR1. A peculiar and mainly intranuclear antibody binding reaction was found in Brunner's gland epithelial cells of mouse and rat whereas Brunner's glands of dog, swine and monkey remained completely negative. These results indicate a potential involvement of Flt-1/VEGFR1 in normal restitution of gastrointestinal structures in the species studied. Additionally, intranuclear Flt-1/VEGFR1 antibody binding in Brunner's glands of rodents may suggest a nuclear translocation of the transmembrane VEGFR1 which has not previously been described.     

Extracellular detection of K+ release during migration of transformed Madin-Darby canine kidney cells

 Madin Darby canine kidney cells transformed by alkaline stress (MDCK-F cells) constitutively migrate at a rate of about 1 μm·min^–1. Migration depends on the intermittent activity of a Ca²⁺-stimulated, 53-pS K⁺ channel (K_Ca channel) that is inhibitable by charybdotoxin. In the present study we examined whether this intermittent K_Ca channel activity results in a significant K⁺ loss across the plasma membrane. K⁺ efflux from MDCK-F cells should result in a transient increase of extracellular K⁺ ([K⁺]_e) in the close vicinity of a migrating cell. However, due to the rapid diffusion of K⁺ ions into the virtually infinite extracellular space, such a transient increase in [K⁺]_e was too small to be detected by conventional K⁺-selective electrodes. Therefore, we developed a ”shielded ion-sensitive microelectrode” (SIM) that limited diffusion to a small compartment, formed by a shielding pipette which surrounded the tip of the K⁺-sensitive microelectrode. The SIM improved the signal to noise ratio by a factor of at least three, thus transient increases of [K⁺]_e in the vicinity of MDCK-F cells became detectable. They occurred at a rate of 1.3 min^–1. The cell releases 40 fmol K⁺ during each burst of intermittent K_Ca channel activity, which corresponds to about 15% of the total cellular K⁺ content. Since transmembrane K⁺ loss must be accompanied by anion loss and therefore leads to a decrease of cell volume, these findings support the hypothesis that intermittent volume changes are a prerequisite for the migration of MDCK-F cells. Received: 15 April 1996 / Received after revision: 18 June 1996 / Accepted: 23 July 1996     

长链非编码RNA反义RNA 1靶向微小RNA-501-3p对食管癌细胞恶性生物学行为影响的实验研究

目的探讨长链非编码RNA(lncRNA)胸腺生成素反义RNA 1(TMPO-AS1)对食管癌细胞恶性生物学行为的影响及其机制。方法2016年6月至2019年12月,实时荧光定量聚合酶链反应(RT-qPCR)检测20例食管癌组织和癌旁组织中lncRNA TMPO-AS1和微小RNA(miR)-501-3p的水平,食管癌Eca109细胞分为lncRNA TMPO-AS1干扰组(转染si-NC和si-TMPO-AS1);miR-501-3p过表达组(转染miR-NC和miR-501-3p);lncRNA TMPO-AS1过表达组(转染pcDNA和pcDNA-TMPO-AS1);lncRNA TMPO-AS1和miR-501-3p共抑制组(转染si-TMPO-AS1+anti-miR-NC和si-TMPO-AS1+anti-miR-501-3p)。噻唑蓝(MTT)法、流式细胞术和Transwell实验分别检测食管癌Eca109细胞增殖、凋亡率、细胞迁移和侵袭能力,蛋白质印迹法(Western blot)检测相关蛋白水平,双荧光素酶报告系统验证lncRNA TMPO-AS1与miR-501-3p的调控关系。两组间比较采用独立样本t检验,多组间比较采用单因素方差进行分析。结果食管癌组织组中的lncRNA TMPO-AS1水平(2.64±0.26)高于癌旁组织(1.00±0.09,t=26.657,P<0.05),miR-501-3p水平(0.44±0.04)低于癌旁组织(1.00±0.07,t=31.063,P<0.05);干扰lncRNA TMPO-AS1的Eca109细胞24 h增殖(0.36±0.03)低于si-NC组(0.39±0.03,t=2.121,P<0.05),48 h增殖(0.45±0.04)低于si-NC组(0.75±0.07,t=11.163,P<0.05),72 h增殖(0.57±0.05)低于si-NC组(1.16±0.09,t=17.191,P<0.05),迁移细胞数[(54.14±5.65)个]低于si-NC组[(113.02±9.87)个,t=15.531,P<0.05],侵袭细胞数[(47.69±5.01)个]低于si-NC组[(96.32±9.88)个,t=13.169,P<0.05],细胞凋亡率[(22.15±2.22)%]高于si-NC组[(6.98±0.69)%,t=19.576,P<0.05];过表达miR-501-3p的Eca109细胞48 h增殖(0.51±0.05)低于miR-NC组(0.77±0.07,t=9.067,P<0.05),72 h增殖(0.65±0.06)低于miR-NC组(1.15±0.09,t=13.867,P<0.05),迁移细胞数[(61.23±6.33)个]低于miR-NC组[(115.25±9.25)个,t=14.458,P<0.05],侵袭细胞数[(53.14±5.33)]低于miR-NC组[(98.32±8.47)个,t=13.543,P<0.05],细胞凋亡[(19.58±1.74)%]高于miR-NC组[(7.23±0.77)%,t=19.471,P<0.05],差异均有统计学意义。结论lncRNA TMPO-AS1通过靶向miR-501-3p促进食管癌细胞的恶性生物学行为,可被视为食管癌患者的潜在治疗靶标。     

熊果酸对卵巢癌细胞转移的抑制作用

目的 熊果酸(UA)对人卵巢癌细胞株HO-8910PM黏附、侵袭及趋化运动的影响,并探讨其可能的作用机制。方法 采用MTT法方法检测熊果酸对HO-8910PM细胞黏附能力的影响;Transwell小室进行人工重组基底膜侵袭实验检测熊果酸对HO-8910PM细胞侵袭及运动的影响;采用RT-PCR和Western blot方法检测熊果酸对HO-8910PM细胞中MMP-2 mRNA、MMP-9 mRNA和蛋白的表达的影响。结果 在黏附试验中,不同浓度熊果酸作用细胞24h后,抑制率分别为44.03%、46.62%和55.11%,与对照组比较具有统计学意义(P＜0.05)。不同浓度的熊果酸在体外作用24h后可以显著抑制HO-8910PM细胞的侵袭能力,穿膜细胞数与对照组比较具有统计学意义(P＜0.01)。熊果酸处理后细胞趋化运动能力降低与对照组比较差异有统计学意义(P＜0.05)。不同浓度熊果酸作用24小时后,HO-8910PM细胞MMP-2 mRNA、MMP-9 mRNA表达量明显低于正常对照组(P＜0.05),Western blot结果显示:HO-8910PM细胞MMP-2、MMP-9蛋白表达量受到明显抑制(P＜0.05)。结论 熊果酸抑制人卵巢癌细胞株HO-8910PM黏附、侵袭和转移的生物学行为,其作用机制可能与熊果酸显著抑制MMP-2、MMP-9表达水平有密切关系。