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Transforming growth factor-β2 induces morphological alteration of human corneal endothelial cells in vitro

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International Journal of Ophthalmology(English edition) 2014年第5期7卷 759-763页

作者： Jing Wang Ting-Jun Fan Xiu-Xia Yang Shi-Min Chang College of Life Sciences Langfang Normal University Key Laboratory for Corneal Tissue Engineering College of Marine Life SciencesOcean University of China

AIM:To investigate the morphological altering effect of transforming growth factor-β2(TGF-β2) on untransfected human corneal endothelial cells(HCECs)in vitro.METHODS:After untransfected HCECs were treated with TGF-β2 at different concentrations, the morphology,cytoskeleton distribution, and type IV collagen expression of the cells were examined with inverted contrast light microscopy, fluorescence microscopy,immunofluorescence or Western Blot.RESULTS:TGF-β2 at the concentration of 3-15 μg/L had obviously alterative effects on HCECs morphology in dose and time-dependent manner, and 9 μg/L was the peak concentration. TGF-β2(9 μg/L) altered HCE cell morphology after treatment for 36 h, increased the mean optical density(P microtubule in cytoplasm and collagen type IV in ECM after treatment for 72 h.·CONCLUTION: TGF-β2 has obviously alterative effect on the morphology of HCECs from polygonal phenotype to enlarged spindle-shaped phenotype, in dose and time-dependence manner by inducing more, elongation and alignment of F-actin, rearrangement of microtubule and larger spread area of collagen type IV.

关键词： human corneal endothelial cell transforming growth factor-β 2 F-actin microtubule collagen type IV

Analysis of a single soybean microtubule’s persistence length

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Advances in Bioscience and Biotechnology 2013年第10期4卷 930-936页

作者： Mitra Shojania Feizabadi Jimmy Barrientos Carly Winton Department of Physics Seton Hall University South Orange USA

In this study, we have reported for the first time the persistence length of a single soybean microtubule in vitro. The measurement is based on analysis of thermal bending of a single soybean microtubule. Similar to mammalian microtubules, the length dependency of the persistence length was also observed for this species of plant microtubule. The identification of two intrinsic aspects of plant microtubules: the persistence length and the compositional structure in terms of variety of beta tubulin isotypes, and the potential correlation between these two factors, provides evidence for the functionality mechanism of plant microtubules. In this work, these two fundamental factors have been further discussed for our case study: single soybean microtubule.

关键词： microtubule Soybean Persistence Length β Tubulin Isotype

MAP2c confers drug stability to microtubules in vivo

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Chinese Science Bulletin 1999年第23期44卷 2162-2168页

作者： Xiaoyong Yang Junlin Teng Yang Jin Jianguo Chen Peking Univ Coll Life Sci Dept Cell Biol & Genet Beijing 100871 Peoples R China

microtubules in the axons of nerve cells have unusual stability. To investigate the role of MAP2c in the stabilization of microtubules in neurites, a baculovirus vector has been used to express high levels of MAP2c in insect ovarian Sf9 cells. The cells respond by extending neu-rite-like processes that contain dense bundles of microtubules. The infected cells have

关键词： MAP2c microtubule neuron neurite.

Stabilizing Effects of Ethanolic Extract of Mastic Gum on microtubule Polymers:an In Vitro Study

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Journal of Human Physiology 2019年第2期1卷 18-25页

作者： Ali Dadras Fatemeh Shahrokhi Asl Gholam Hossein Riazi Shahin Ahmadian Tahereh Javdani Khalife Neuro-organic Lab Institute of Biochemistry and Biophysics(IBB)University of TehranTehranIran

Terpenoids are novel natural products isolated from mastic gum.Mastic gum was obtained from the Pistacia Lentiscus tree.Scientific investigations have documented medical and pharmacological properties of mastic gum such as memory enhancement,antifungal,and antibacterial activities.It was astonishing to study the possible interaction of mastic gum extract with microtubule proteins which are involved in memory and consciousness since the administration of mastic gum is evidenced in the improvement of brain functions.Since a number of studies have demonstrated the effect of microtubule dynamics on mammals'memory,in this study,we investigated the effect of Oxygenated Sesquiterpenes(OST)on microtubule polymerization in vitro.OST was purified from the ethanolic extract of mastic gum.The results revealed that OST induces microtubule polymerization;however,microtubule depolymerization was not affected and fluorometric assays showed conformational changes of tubulin in the presence of OST.We interestingly found that colchicine was unable to inhibit MT assembly in the presence of OST and OST was solely more efficient than the combination of OST with paclitaxel for elevating microtubule polymerization rate.We hope that OST could be a promising agent for memory enhancement and the treatment of neurodegenerative diseases as a novel tubulin-binding compound.

关键词： Pistacia lentiscus Mastic gum Oxygenated Sesquitrepene(OST) Tubulin microtubule

Mechanical Properties of Self-Assembled microtubule Curved Protofilaments

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Journal of Biomedical Science and Engineering 2020年第3期13卷 37-44页

作者： Ramana Pidaparti Divya Jakkam School of Environmental Civil Agriculture and Mechanical Engineering College of Engineering University of Georgia Athens GA USA

The microtubule self-assembly process involves the basic building blocks, alpha and beta tubulins which spontaneously bind to one another through polymerization and under controlled intracellular conditions form protofilaments which in turn assemble into microtubules. The mechanical properties of the self-assembled protofilaments play an important role in formation of the microtubules. In this study, we investigate the mechanical properties of the experimentally self-assembled protofilaments (straight and curved) for under different loadings through 3D finite element analysis. Results of force-deformation and stiffness values obtained from the finite element analysis are presented. The results indicate that the stiffness and maximum stress properties change with varying protofilamant curvature. These force-deformation behaviors and stress distributions should help further understand the contribution of protofilaments mechanical properties in forming self-assembled microtubules.

关键词： microtubule Protofilament Tubulin Finite Element Method Mechanical Properties

Cytoplasmic dynein-2： from molecules to human diseases

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Frontiers in Biology 2013年第1期8卷 119-126页

作者： Vidyalakshmi RAJAGOPALAN Jonathan P. D'AMICO David E. WILKES Department of Biological Sciences Indiana University South Bend South Bend IN 46634-7111 USA

The dynein motor protein family is involved in a wide variety of functions in eukaryotic cells. The axonemal dynein class and cytoplasmic dynein-1 subclass have been well characterized. However, the cytoplasmic dynein-2 subclass of the family has only recently begun to be understood. We describe the entire dynein family but focus on cytoplasmic dynein-2. Dynein-2 consists of a heavy, an intermediate, a light intermediate, and a light chain. The complex appears to function primarily as the retrograde motor for intraflagellar transport. This process is important for the formation and maintenance of cilia and flagella. Additionally, dynein-2 has roles in the control of ciliary length and in non-ciliary functions. Mutations in the human dynein-2 heavy chain lead to cilia-related diseases.

关键词： cilia dynein flagella intraflagellar transport microtubule

A likely role for a novel PH-domain containing protein,PEPP2,in connecting membrane and cytoskeleton

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BIOCELL 2012年第3期36卷 127-132页

作者： YI ZOU WENPING ZHONG Department of Biology School of Life Science and TechnologyJinan UniversityGuangzhouChina

PH domains(pleckstrin homology)are well known to bind membrane phosphoinositides with different specificities and direct PH domain-containing proteins to discrete subcellular apartments with assistances of alternative binding partners.PH domain-containing proteins are found to be involved in a wide range of cellular events,including signalling,cytoskeleton rearrangement and vesicular trafficking.Here we showed that a novel PH domain-containing protein,PEPP2,displayed moderate phosphoinositide binding specificity.Full length PEPP2 associated with both plasma membrane and microtubules.The membrane-associated PEPP2 nucleated at cell-cell contacts and the leading edge of migrating cells.Overexpression of PEPP2 increased membrane microviscosity,indicating a potential role of PEPP2 in regulating function of membrane and microtubules.

关键词： microtubule membrane phosphoinositide microviscosity wortmannin

THE COMPARTMENTATION OF microtubule PROTEINS IN THE CELLS OF HIGHER ORGANISMS AND THE FORMATION OF KINETOCHORE microtubuleS

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Chinese Science Bulletin 1984年第1期29卷 104-108页

作者：倪祖梅劳为德施履吉 Shanghai Institute of Cell Biology Academia Silica

The mitotic and meiotic spindle apparatus consists of two groups of microtubules,the chromosomal or kinetochore microtubules and the interpolar or pole-to-pole microtubules. The assembly of kinetochore microtubules constitutes an essential and important process in the later part of spindle development. In the cells of higher organisms,the spindle fiber attachments (SFAs) acting as microtubule organizing centers

关键词： tubulin microtubule spindle chromosomal organizing apparatus assembly Proteins breakdown details

Regulation of blood-testis barrier dynamics by the mT0RCl/rpS6 signaling complex:an in vitro study

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Asian Journal of Andrology 2019年第4期21卷 365-374页

作者： Lin-Xi Li Si-Wen Wu Ming Yan Qing-Quan Lian Ren-Shan Ge C Yan Cheng The Second Affiliated Hospital and Yuying Children's Hospital Wenzhou Medical University Wenzhou 325027 China The Mary M. Wohlford Laboratory for Male Contraceptive Research Center for Biomedical Research Population Council New York NY 10065 USA Jiangsu Key Laboratory of Drug Screening China Pharmaceutical University Nanjing 210009 China

During spermatogenesis, developi ng germ cells that lack the cellular ultrastructures of filopodia and lamellipodia gen erally found in migrating cells, such as macrophages and fibroblasts, rely on Sertoli cells to support their transport across the seminiferous epithelium. These in elude the transport of preleptote ne spermatocytes across the blood-testis barrier (BTB), but also the transport of germ cells, in particular developing haploid spermatids, across the seminiferous epithelium, that is to and away from the tubule lumen, depending on the stages of the epithelial cycle. On the other hand, cell junctions at the Sertoli cell-cell and Sertoli-germ cell in terface also un dergo rapid remodeli ng, invo Iving disassembly and reassembly of cell j un ctions, which, in turn, are supported by actin- and microtubule-based cytoskeletal remodeling. Interestingly, the underlying mechanism(s) and the invoIving biomolecule(s) that regulate or support cytoskeletal remodeling remain largely unknown. Herein, we used an in vitro model of primary Sertoli cell cultures that mimicked the Sertoli BTB in vivo overexpressed with the ribosomal protei n S6 (rpS6, the down stream signali ng protein of mammalian target of rapamycin complex 1 [mTORCl]) cloned into the mammalian expression vector pCI-neo, namely, quadruple phosphomimetic and constitutively active mutant of rpS6 (pCI-neo/p-rpS6-MT) versus pCI-neo/rpS6-WT (wild-type) and empty vector (pCI-neo/Ctrl) for studies. These findings provide compelling evidence that the mT0RCl/rpS6 signal pathway exerted its effects to promote Sertoli cell BTB remodeling. This was mediated through changes in the organization of actin- and microtubulebased cytoskeletons, involving changes in the distribution and/or spatial expression of actin- and microtubule-regulatory proteins.

关键词： blood-testis barrier F-actin microtubule mTORCl rpS6 Sertoli cells testis