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人睫状神经营养因子的原核表达,纯化及其生物效应 总被引:2,自引:0,他引:2
人睫状神经营养因子(hCNTF)克隆入pBV220中,在DH5α菌株中表达,重组蛋白以包含体的形式存在,表达量为菌体总蛋白的50%左右。经比较发现用2mol/L脲洗涤包含体可溶解大量可溶性细菌蛋白,且包含体损失较小。在高浓度变性剂条件下进行sepharcylS-200凝胶过滤,解决了纯化中hCNTF易聚合的问题,在低浓度变性剂条件下进行DEAE离子交换,有利于蛋白活性的保持。经两步纯化后得到均一性hCNTF,纯度达95%以上。在自然状态下使hCNTF复性。纯化复性后的hCNTF对无血清培养的鸡胚背根节神经元和脊髓腹角运动神经元有明显的维持存活和促进生长发育的生物效应。 相似文献
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Zhan Wang Yuxin Song Xingwen Han Peng Qu Wenji Wang 《Journal of cellular physiology》2020,235(4):3634-3645
Exosomes derived from differentiated P12 cells and MSCs were proved to suppress apoptosis of neuron cells, and phosphatase and tensin homolog pseudogene 1 (PTENP1) was reported to inhibit cell proliferation. In this study, we aimed to investigate the role of PTENP1 in the process of post-spinal cord injury (SCI) recovery, so as to evaluate the therapeutic effects of exosomes derived from MSCs transfected with PTENP1 short hairpin RNA (shRNA), as a type of novel biomarkers in the treatment of SCI. Electron microscopy was used to observe the morphology of different exosomes. Real-time polymerase chain reaction and western blot, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, flow cytometry, Nissl staining, immunohistochemistry assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay were conducted to investigate and validate the underlying molecular signaling pathway. PTENP1-shRNA downregulated PTENP1 and PTEN while upregulating miR-21 and miR-19b. PTENP1-shRNA also accelerated cell apoptosis and reduced cell viability. In addition, PTENP1 reduced the miR-21 and miR-19b expression by directly targeting miR-21 and miR-19b. Meanwhile, both miR-21 and miR-19b reduced the expression of PTEN by directly targeting the 3′-untranslated region of PTEN. Furthermore, PTEN level and apoptosis index of neuron cells was the highest in the SCI group, while the treatment with exosomes+PTENP1-shRNA reduced the PTEN expression to a level similar to that in the sham group. Finally, PTENP1 inhibited miR-21 and miR-19b expression but upregulated PTEN expression. The upregulation of miR-21/miR-19b also suppressed the apoptosis of neuron cells by downregulating the PTEN expression. PTENP1 is involved in the recovery of SCI by regulating the expression of miR-19b and miR-21, and exosomes from PTENP1-shRNA-transfected cells may be used as a novel biomarker in SCI treatment. 相似文献
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S. Jennings 《Journal of fish biology》1991,38(3):349-357
Relationships between live and preserved lengths of larval and juvenile bass, Dicentrarchus labrax , subjected to simulated capture, net retention and preservation were investigated. Bass larvae and juveniles shrank during the period of net retention following death. Preserved lengths of larvae, but not of juveniles, decreased significantly when increased periods of net retention preceded preservation. The time which larvae or juveniles remained in 4% formaldehyde in distilled water buffered with 3gl−1 sodium acetate trihydrate or in 70% ethanol in distilled water neutralized with 1gl−1 calcium carbonate powder, had no significant effect upon preserved length. Appropriate data were pooled to calculate transformation factors for estimating live from preserved length. Shrinkage phenomena, and their effects, are discussed. It is suggested that larval bass captured and preserved during standard ichthyoplankton surveys are unsuitable for determination of live length. 相似文献
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《Cell》2021,184(17):4564-4578.e18
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The development of spinal cord supports (bony thickenings which extend into the vertebral canal of vertebrae) in primitive (Salamandrella keyserlingii) and derived (Lissotriton vulgaris) salamanders were described. The spinal cord supports develop as the protuberances of periostal bone of the neural arches in the anteroproximal part of the septal collagenous fibers which connect a transverse myoseptum with the notochord and spinal cord, in the septal bundle inside the vertebral canal. Spinal cord supports were also found in some teleostean (Salmo salar, Oncorhynchus mykiss) and dipnoan (Protopterus sp.) fishes. The absence of the spinal cord supports in vertebrates with cartilaginous vertebrae (lampreys, chondrichthyan, and chondrostean fishes) corresponds to the fact that the spinal cord supports are bone structures. The absence of the spinal cord supports in frogs correlates with the lack of the well developed septal bundles inside the vertebral canal. The spinal cord supports are, presumably, a synapomorphic character for salamanders which originated independently of those observed in teleostean and dipnoan fishes. J. Morphol. 2012. © 2012 Wiley Periodicals, Inc. 相似文献
8.
Postsynaptic currents and action potentials recorded from neurons in a mixed culture of rat dorsal root ganglion and spinal
cord cells are described. The existence of mutual synaptic connections between the above two types of neurons is demonstrated.
Neirofiziologiya/Neurophysiology, Vol. 38, No. 4, pp. 358–360, July–August, 2006. 相似文献
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Spinal muscular atrophy (SMA) is the most common genetic disease that causes infant mortality. Its treatment and prevention represent the paradigmatic example of the ethical dilemmas of 21st-century medicine. New therapies (nusinersen and AVXS-101) hold the promise of being able to treat, but not cure, the condition. Alternatively, genomic analysis could identify carriers, and carriers could be offered in vitro fertilization and preimplantation genetic diagnosis. In the future, gene editing could prevent the condition at the embryonic stage. How should these different options be evaluated and compared within a health system? In this paper, we discuss the ethical considerations that bear on the question of how to prioritize the different treatments and preventive options for SMA, at a policy level. We argue that despite the tremendous value of what we call ‘ex-post’ approaches to treating SMA (such as using pharmacological agents or gene therapy), there is a moral imperative to pursue ‘ex-ante’ interventions (such as carrier screening in combination with prenatal testing and preimplantation genetic diagnosis, or gene editing) to reduce the incidence of SMA. There are moral reasons relating to autonomy, beneficence and justice to prioritize ex-ante methods over ex-post methods. 相似文献