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Urban Lendahl - One of the best experts on this subject based on the ideXlab platform.
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immunohistochemical detection of Nestin in pediatric brain tumors
Journal of Histochemistry and Cytochemistry, 2002Co-Authors: Per Almqvist, Björn Jacobsson, Urban Lendahl, Glenda HendsonAbstract:SUMMARY Nestin is an intermediate filament protein (IFP) expressed in undifferentiated cells during CNS development and in CNS tumors. Previous studies have arrived at different conclusions in terms of which types of CNS tumors express Nestin. In this report we establish an immunohistochemical protocol using antigen retrieval, which significantly enhances staining with two polyclonal anti-Nestin antisera, #130 and #4350. The staining pattern was identical for the two Nestin antisera and very similar to that of vimentin, while glial fibrillary acidic protein (GFAP), immunoreactivity was absent from 9.5-week-old forebrain. The current study of 20 primary CNS tumors from pediatric patients included seven ependymomas, seven primitive neuroectodermal tumors (PNETs), five pilocytic astrocytomas, and one glioblastoma multiforme (GBM). All these tumors expressed Nestin to various extents, in contrast to five brain metastases tested. Strong Nestin immunoreactivity was found in malignant primary CNS tumors, whereas benign pilocytic astrocytomas showed low but consistent Nestin expression. In all tumors Nestin immunoreactivity was confined to the cytoplasm of tumor cells and was co-expressed with astrocyte markers vimentin, GFAP, and S-100. Vascular endothelial cells of all neoplasms also showed marked immunoreactivity for Nestin and vimentin, whereas they were negative for GFAP and S-100. In conclusion, antiserum #4350 detected Nestin in formalin-fixed, paraffin-embedded tissue sections by heatinduced antigen retrieval immunohistochemistry. Nestin was expressed in both highly malignant and low malignant gliomas, indicating the potential use of Nestin as a diagnostic tumor marker in surgical pathology. (J Histochem Cytochem 50:147‐158, 2002) The advent of immunohistochemical (IHC) techniques to identify the expression of specific proteins has significantly advanced the typing of CNS tumors. These techniques have been of particular value in identification of intermediate filament proteins (IFPs) (Herrman and Aebi 2000) because these proteins are invaluable markers of cell origin within the CNS. Vimentin, glial fibrillary acidic protein (GFAP), and S-100 � are the markers most often used for identifying astrocytic cells in the intact CNS and in primary CNS neoplasms (Ridet et al. 1997). GFAP is the most specific marker
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mitotic reorganization of the intermediate filament protein Nestin involves phosphorylation by cdc2 kinase
Journal of Biological Chemistry, 2001Co-Authors: Cecilia Sahlgren, Andrey Mikhailov, Yinghao Chou, Jukka Hellman, Robert D. Goldman, Urban Lendahl, John E. ErikssonAbstract:Abstract The intermediate filament protein Nestin is expressed during early stages of development in the central nervous system and in muscle tissues. Nestin expression is associated with morphologically dynamic cells, such as dividing and migrating cells. However, little is known about regulation of Nestin during these cellular processes. We have characterized the phosphorylation-based regulation of Nestin during different stages of the cell cycle in a neuronal progenitor cell line, ST15A. Confocal microscopy of Nestin organization and 32P in vivo labeling studies show that the mitotic reorganization of Nestin is accompanied by elevated phosphorylation of Nestin. The phosphorylation-induced alterations in Nestin organization during mitosis in ST15A cells are associated with partial disassembly of Nestin filaments. Comparative in vitro and in vivo phosphorylation studies identified cdc2 as the primary mitotic kinase and Thr316 as a cdc2-specific phosphorylation site on Nestin. We generated a phosphospecific Nestin antibody recognizing the phosphorylated form of this site. By using this antibody we observed that Nestin shows constitutive phosphorylation at Thr316, which is increased during mitosis. This study shows that Nestin is reorganized during mitosis and that cdc2-mediated phosphorylation is an important regulator of Nestin organization and dynamics during mitosis.
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Nestin expression in embryonic and adult human teeth under normal and pathological conditions
American Journal of Pathology, 2000Co-Authors: Imad About, Dominique Laurentmaquin, Urban Lendahl, Thimios A. MitsiadisAbstract:Nestin is an intermediate filament most related to neurofilaments and expressed predominantly in the developing nervous system and muscles. In the present study we examined the in vivo distribution of Nestin in human teeth during embryonic development and in permanent teeth under normal and pathological conditions. The results show that Nestin is first expressed at the bell stage and that its distribution is restricted in pulpal cells located at the cusp area of the fetal teeth. In young permanent teeth, Nestin is found only in functional odontoblasts, which produce the hard tissue matrix of dentin. Expression is progressively down-regulated and Nestin is absent from older permanent teeth. In carious and injured teeth, Nestin expression is up-regulated in a selective manner in odontoblasts surrounding the injury site, showing a link between tissue repair competence and Nestin up-regulation under pathological conditions. In an in vitro assay system of human dental pulp explants, Nestin is up-regulated after local application of bone morphogenic protein-4. A similar effect is seen in cultures of primary pulp cells during their differentiation into odontoblasts. Taken together, these results suggest that Nestin plays a potential role in odontoblast differentiation during normal and pathological conditions and that bone morphogenic protein-4 is involved in Nestin up-regulation.
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specific and innervation regulated expression of the intermediate filament protein Nestin at neuromuscular and myotendinous junctions in skeletal muscle
American Journal of Pathology, 1999Co-Authors: Samuli Vaittinen, Cecilia Sahlgren, John E. Eriksson, Hannu Kalimo, Urban Lendahl, Riitta Lukka, Jussi Rantanen, Timo HurmeAbstract:The intermediate filament proteins Nestin, vimentin, and desmin show a specific temporal expression pattern during the development of myofibers from myogenic precursor cells. Nestin and vimentin are actively expressed during early developmental stages to be later down-regulated, vimentin completely and Nestin to minimal levels, whereas desmin expression begins later and is maintained in mature myofibers, in which desmin participates in maintaining structural integrity. In this study we have analyzed the expression levels and distribution pattern of Nestin in intact and denervated muscle in rat and in human. Nestin immunoreactivity was specifically and focally localized in the sarcoplasm underneath neuromuscular junctions (NMJs) and in the vicinity of the myotendinous junctions (MTJs), ie, in regions associated with acetylcholine receptors (AChRs). This association prompted us to analyze Nestin in neurogenically and myogenically denervated muscle. Immunoblot analysis disclosed a marked overall increase of accumulated Nestin protein. Similar to the extrajunctional redistribution of AChRs in denervated myofibers, Nestin immunoreactivity extended widely beyond the NMJ region. Re-innervation caused complete reversion of these changes. Our study demonstrates that the expression levels and distribution pattern of Nestin are regulated by innervation, ie, signal transduction into myofibers.
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a rod end deletion in the intermediate filament protein Nestin alters its subcellular localization in neuroepithelial cells of transgenic mice
Journal of Cell Science, 1998Co-Authors: Martha J Marvin, Jonas Dahlstrand, Urban Lendahl, Ronald D G MckayAbstract:Neuroepithelial and radial glial cells span between the ventricular and the pial surfaces of the neural tube and express two intermediate filaments (IFs), Nestin and vimentin, which form a filamentous network throughout the length of the cells. In this report we study the polymerization characteristics of Nestin and examine how mutations affect the assembly and localization of the Nestin protein in cultured cells and in the developing CNS of transgenic mice. A wild-type rat Nestin gene transfected into the IF-free SW13 cell line failed to assemble into a filamentous network but was incorporated into the existing IF network of a subclone expressing vimentin, demonstrating that Nestin requires vimentin for proper assembly. In transgenic mice, rat Nestin formed a network indistinguishable from that formed by endogenous Nestin and vimentin, but a mutant form lacking five amino acids at the carboxy terminus of the rod domain was largely restricted to the pial endfeet. Since Nestin mRNA is localized to the pial endfoot region we propose that both transgenes are translated there, but that the wild-type protein is preferentially incorporated into the IF network. These observations provide evidence for hierarchical assembly and a complex organization of the IF network along the ventricular-pial axis in the early CNS.
Robert M. Hoffman - One of the best experts on this subject based on the ideXlab platform.
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comparison of Nestin expressing multipotent stem cells in the tongue fungiform papilla and vibrissa hair follicle
Journal of Cellular Biochemistry, 2014Co-Authors: Yasuyuki Amoh, Kensei Katsuoka, Sumiyuki Mii, Robert M. HoffmanAbstract:We have previously reported that hair follicles contain multipotent stem cells, which express Nestin and participate in follicle growth at anagen as well as in the extension of the follicle sensory nerve. The Nestin-driven green fluorescent protein (ND-GFP) transgenic mouse labels all Nestin-expressing cells with GFP. The hair follicle Nestin-GFP cells can differentiate into neurons, Schwann cells, and other cell types. In this study, we describe Nestin-expressing multipotent stem cells in the fungiform papilla in the tongue. The Nestin-expressing multipotent stem cells in the fungiform papilla are located around a peripheral sensory nerve immediately below the taste bud and co-express the neural crest cell marker p75NTR. The fungiform papilla cells formed spheres in suspension culture in DMEM-F12 medium supplemented with basic fibroblast growth factor (bFGF). The spheres consisted of Nestin-expressing cells that co-expressed the neural crest marker p75NTR and which developed expression of the stem cell marker CD34. P75NTR, CD34 and Nestin co-expression suggested that Nestin-expressing cells comprising the fungiform papilla spheres were in a relatively undifferentiated state. The Nestin-expressing cells of these spheres acquired the following markers: β III tubulin typical of nerve cells; GFAP typical of glial cells; K15 typical of keratinocytes; and smooth-muscle antigen (SMA), after transfer to RPMI 1640 medium with 10% fetal bovine serum (FBS), suggesting they differentiated into multiple cell types. The results of the current study indicate Nestin-expressing fungiform papilla cells and the Nestin-expressing hair follicle stem cells have common features of cell morphology and ability to differentiate into multiple cell types, suggesting their remarkable similarity. J. Cell. Biochem. 115: 1070–1076, 2014. © 2013 Wiley Periodicals, Inc.
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comparison of Nestin expressing multipotent stem cells in the tongue fungiform papilla and vibrissa hair follicle
Journal of Cellular Biochemistry, 2014Co-Authors: Yasuyuki Amoh, Kensei Katsuoka, Sumiyuki Mii, Robert M. HoffmanAbstract:We have previously reported that hair follicles contain multipotent stem cells, which express Nestin and participate in follicle growth at anagen as well as in the extension of the follicle sensory nerve. The Nestin-driven green fluorescent protein (ND-GFP) transgenic mouse labels all Nestin-expressing cells with GFP. The hair follicle Nestin-GFP cells can differentiate into neurons, Schwann cells, and other cell types. In this study, we describe Nestin-expressing multipotent stem cells in the fungiform papilla in the tongue. The Nestin-expressing multipotent stem cells in the fungiform papilla are located around a peripheral sensory nerve immediately below the taste bud and co-express the neural crest cell marker p75(NTR) . The fungiform papilla cells formed spheres in suspension culture in DMEM-F12 medium supplemented with basic fibroblast growth factor (bFGF). The spheres consisted of Nestin-expressing cells that co-expressed the neural crest marker p75(NTR) and which developed expression of the stem cell marker CD34. P75(NTR), CD34 and Nestin co-expression suggested that Nestin-expressing cells comprising the fungiform papilla spheres were in a relatively undifferentiated state. The Nestin-expressing cells of these spheres acquired the following markers: β III tubulin typical of nerve cells; GFAP typical of glial cells; K15 typical of keratinocytes; and smooth-muscle antigen (SMA), after transfer to RPMI 1640 medium with 10% fetal bovine serum (FBS), suggesting they differentiated into multiple cell types. The results of the current study indicate Nestin-expressing fungiform papilla cells and the Nestin-expressing hair follicle stem cells have common features of cell morphology and ability to differentiate into multiple cell types, suggesting their remarkable similarity.
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the bulge area is the origin of Nestin expressing pluripotent stem cells of the hair follicle
Journal of Cellular Biochemistry, 2011Co-Authors: Aisada Uchugonova, Jennifer Duong, Karsten König, Nan Zhang, Robert M. HoffmanAbstract:: Nestin-expressing pluripotent stem cells have been found both in the bulge area (BA) as well as the dermal papilla (DP). Nestin-expressing stem cells of both the BA and DP have been previously shown to be able to form neurons and other non-follicle cell types. The Nestin-expressing stem cells from the DP have been termed skin precursor or SKP cells. Both Nestin-expressing DP and BA cells have been previously shown to effect repair of the injured spinal cord and peripheral nerve, with the BA being the greater and more constant source of the stem cells. The BA contains Nestin-expressing stem cells throughout the hair cycle, whereas Nestin-expressing dermal papillae stem cells were found in early and mid-anagen only. Our previous studies have shown that the Nestin-expressing stem cells in the BA and DP have similar morphological features. The cells from both regions have a small body diameter of approximately 7 µm with long extrusions, as shown by 2-photon imaging. In the present study, using 2-photon imaging of whisker follicles from transgenic mice expressing Nestin-driven green fluorescent protein (ND-GFP), we demonstrate that the BA is the source of the Nestin-expressing stem cells of the hair follicle. The Nestin-expressing stem cells migrate from the BA to the DP as well as into the surrounding skin tissues including the epidermis, and during wound healing, suggesting that the BA may be the source of the stem cells of the skin itself.
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Nestin expression in hair follicle sheath progenitor cells
Proceedings of the National Academy of Sciences of the United States of America, 2003Co-Authors: Lingna Li, Grigori Enikolopov, John Mignone, Sheldon Penman, Maja Matic, Meng Yang, Robert M. HoffmanAbstract:The intermediate filament protein, Nestin, marks progenitor cells of the CNS. Such CNS stem cells are selectively labeled by placing GFP under the control of the Nestin regulatory sequences. During early anagen or growth phase of the hair follicle, Nestin-expressing cells, marked by GFP fluorescence in Nestin-GFP transgenic mice, appear in the permanent upper hair follicle immediately below the sebaceous glands in the follicle bulge. This is where stem cells for the hair follicle outer-root sheath are thought to be located. The relatively small, oval-shaped, Nestin-expressing cells in the bulge area surround the hair shaft and are interconnected by short dendrites. The precise locations of the Nestin-expressing cells in the hair follicle vary with the hair cycle. During telogen or resting phase and in early anagen, the GFP-positive cells are mainly in the bulge area. However, in mid- and late anagen, the GFP-expressing cells are located in the upper outer-root sheath as well as in the bulge area but not in the hair matrix bulb. These observations show that the Nestin-expressing cells form the outer-root sheath. Results of the immunohistochemical staining showed that Nestin, GFP, keratin 5/8, and keratin 15 colocalize in the hair follicle bulge cells, outer-root sheath cells, and basal cells of the sebaceous glands. These data indicate that Nestin-expressing cells, marked by GFP, in the hair follicle bulge are indeed progenitors of the follicle outer-root sheath. The expression of the unique protein, Nestin, in both neural stem cells and hair follicle stem cells suggests their possible relation.
Toshiyuki Ishiwata - One of the best experts on this subject based on the ideXlab platform.
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inhibition of Nestin suppresses stem cell phenotype of glioblastomas through the alteration of post translational modification of heat shock protein hspa8 hsc71
Cancer Letters, 2015Co-Authors: Yoko Matsuda, Masahito Hagio, Toshiyuki Ishiwata, Hisashi Yoshimura, Tomio AraiAbstract:Nestin, a class VI intermediate filament, was first described as a neuronal stem/progenitor cell marker. We previously reported that knockdown of Nestin expression in human glioblastoma cells suppresses cell proliferation, migration, and invasion. In the present study, we examined the effect of Nestin on stemness, and identified molecules involved in modulating Nestin function in glioblastoma cells. Nestin expression was shown to be higher in high-grade gliomas than in low-grade gliomas. Furthermore, compared with control cells, Nestin short hairpin RNA (shRNA)-transfected glioblastoma cells exhibited reduced sphere formation, decreased expression of NANOG, N-cadherin, CD133, and Oct-4, and decreased tumor size in vivo. To examine the proteins regulated by Nestin in glioblastomas, we carried out two-dimensional electrophoresis using Nestin shRNA-transfected glioblastoma cells. As a result, Nestin shRNA-transfected glioblastoma cells exhibited a decrease in the level of phosphorylation of heat shock cognate 71 kDa protein (HSC71; gene HSPA8). From immunoprecipitation experiments, we demonstrated the direct binding of Nestin, HSC71, and cyclin D1 in vitro. Overexpression of Nestin in glioblastoma cells increased cell growth, sphere formation, and cell invasion. Transfection with HSC71 siRNA restored Nestin expression and cell behavior; therefore, HSC71 knockdown will interfere with enhanced tumorigenic properties of glioblastoma cells that ectopically overexpress Nestin. We have demonstrated that HSC71 and Nestin regulate each other's expression levels or patterns, and that cyclin D1 is located downstream of Nestin and HSC71. In conclusion, Nestin regulates stemness, cell growth, and invasion in glioblastoma cells through the alteration of HSC71. Inhibition of Nestin and HSC71 may thus be a useful molecular target in the treatment of glioblastomas.
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Nestin a novel angiogenesis marker and possible target for tumor angiogenesis
World Journal of Gastroenterology, 2013Co-Authors: Yoko Matsuda, Masahito Hagio, Toshiyuki IshiwataAbstract:Abnormal vasculature, termed tumor vessels, is a hallmark of solid tumors. The degree of angiogenesis is associated with tumor aggressiveness and clinical outcome. Therefore, exact quantification of tumor vessels is useful to evaluate prognosis. Furthermore, selective detection of newly formed tumor vessels within cancer tissues using specific markers raises the possibility of molecular targeted therapy via the inhibition of tumor angiogenesis. Nestin, an intermediate filament protein, is reportedly expressed in repair processes, various neoplasms, and proliferating vascular endothelial cells. Nestin expression is detected in endothelial cells of embryonic capillaries, capillaries of the corpus luteum, which replenishes itself by angiogenesis, and proliferating endothelial progenitor cells, but not in mature endothelial cells. Therefore, expression of Nestin is relatively limited to proliferating vascular endothelial cells and endothelial progenitor cells. Nestin expression is also reported in blood vessels within glioblastoma, prostate cancer, colorectal cancer, and pancreatic cancer, and its expression is more specific for newly formed blood vessels than other endothelial cell markers. Nestin-positive blood vessels form smaller vessels with high proliferation activity in tumors. Knockdown of Nestin in vascular endothelial cells suppresses endothelial cell growth and tumor formation ability of pancreatic cancers in vivo. Using Nestin to more accurately evaluate microvessel density in cancer specimens may be a novel prognostic indicator. Furthermore, Nestin-targeted therapy may suppress tumor proliferation via inhibition of angiogenesis in numerous malignancies, including pancreatic cancer. In this review article, we focus on Nestin as a novel angiogenesis marker and possible therapeutic target via inhibition of tumor angiogenesis.
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neuroepithelial stem cell marker Nestin regulates the migration invasion and growth of human gliomas
Oncology Reports, 2011Co-Authors: Toshiyuki Ishiwata, Kiyoko Kawahara, Yoko Matsuda, Kiyoshi Teduka, Tetsushi Yamamoto, Zenya NaitoAbstract:Nestin, a class VI intermediate filament protein, was originally described as a neuronal stem cell marker during central nervous system development. Nestin is expressed in gliomas, and its expression levels are higher in gliomas with high WHO histopathological classification grades than in those with low grades. In the present study, we examined whether Nestin regulates the biological activities of human glioma cells. Immunohistochemically, the Nestin expression patterns in 10 human glioblastoma patients were examined. The expression levels of Nestin in A172, a human high-grade glioma cell line, and KG-1-C, a human low-grade glioma cell line, were examined using real-time PCR, Western blot and immunofluorescence analyses. An expression vector carrying a short hairpin RNA targeting Nestin was stably transfected into A172 (Sh) cells. The effects of decreased expression levels of Nestin in Sh cells on cell growth, migration, invasion, adhesion to extracellular matrices and fibrillar actin expression on three-dimensional culture plates were examined. The Nestin expression vector was transiently transfected into KG-1-C (Nes) cells, and the effects of the Nestin overexpression on cell growth and migration were examined. Nestin was expressed in the cytoplasm of the glioblastoma cells in all cases examined. Sh cells showed marked decreases in the expression levels of Nestin mRNA and protein, and the growth rate of Sh cells was lower than that of sham (Sc) cells. In contrast, the adhesion activity of Sh cells to types I and IV collagens, fibronectin and laminin was higher than that of Sc cells. Fibrillar actin was clearly detected at the periphery of colonies of Sh cells at the attachment sites on three-dimensional culture plates. The migration and invasion of Sh cells were markedly inhibited compared with those of Sc cells. In contrast, the levels of Nestin expression markedly increased in the Nes cells, which were transiently transfected with the Nestin expression vector. The growth rate and motility of Nes cells were higher than those of the mock cells. In conclusion, Nestin plays important roles in cell growth, migration, invasion and adhesion to extra-cellular matrices in glioma cells. Nestin may serve as a novel candidate for molecular-targeted therapy for gliomas, including glioblastomas.
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Nestin is a novel target for suppressing pancreatic cancer cell migration invasion and metastasis
Cancer Biology & Therapy, 2011Co-Authors: Yoko Matsuda, Kiyoko Kawahara, Nand O. Nakazawa, Murray Korc, Zenya Naito, Toshiyuki IshiwataAbstract:Nestin, is a class VI intermediate filament (IF) that is expressed in 30% of pancreatic ductal adenocarcinoma (PDAC) cases, and its expression in PDAC positively correlates with peripancreatic invasion. An expression vector carrying a short hairpin RNA (shRNA) targeting Nestin was stably transfected into PANC-1 and PK-45H human pancreatic cancer cells, which express high Nestin levels. Alterations in morphology and alignment of actin filaments and α-tubulin were examined by phase-contrast and immunocytochemistry. Effects on cell growth, migration in scratch and Boyden chamber assays, invasion, cell adhesion, and in vivo growth were determined. Differences in mRNA levels were examined by arrays. Nestin shRNA-transfected cells exhibited decreased Nestin expression, a sheet-like appearance with tight cell-cell adhesion, increased expression of filamentous F-actin and E-cadherin, and attenuated migration and invasion, both of which were enhanced following Nestin re-expression. Expression of α-tubulin, and in vitro cell growth and adhesion were not altered by Nestin downregulation, whereas hepatic metastases were decreased. Thus, Nestin plays important roles in pancreatic cancer cell migration, invasion and metastasis by selectively modulating the expression of actin and cell adhesion molecules, and may therefore be a novel therapeutic target in PDAC.
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Nestin in gastrointestinal and other cancers effects on cells and tumor angiogenesis
World Journal of Gastroenterology, 2011Co-Authors: Toshiyuki Ishiwata, Yoko Matsuda, Zenya NaitoAbstract:Nestin is a class VI intermediate filament protein that was originally described as a neuronal stem cell marker during central nervous system (CNS) development, and is currently widely used in that capacity. Nestin is also expressed in non-neuronal immature or progenitor cells in normal tissues. Under pathological conditions, Nestin is expressed in repair processes in the CNS, muscle, liver, and infarcted myocardium. Furthermore, increased Nestin expression has been reported in various tumor cells, including CNS tumors, gastrointestinal stromal tumors, pancreatic cancer, prostate cancer, breast cancer, malignant melanoma, dermatofibrosarcoma protuberances, and thyroid tumors. Nestin is reported to correlate with aggressive growth, metastasis, and poor prognosis in some tumors; however, the roles of Nestin in cancer cells have not been well characterized. Furthermore, Nestin is more specifically expressed in proliferating small-sized tumor vessels in glioblastoma and gastric, colorectal, and prostate cancers than are other tumor vessel markers. These findings indicate that Nestin may be a marker for newly synthesized tumor vessels and a therapeutic target for tumor angiogenesis. It has received a lot of attention recently as a cancer stem cell marker in various cancer cells including brain tumors, malignant rhabdoid tumors, and uterine, cervical, prostate, bladder, head and neck, ovarian, testicular, and pancreatic cancers. The purpose of this review is to clarify the roles of Nestin in cancer cells and in tumor angiogenesis, and to examine the association between Nestin and cancer stem cells. Nestin has the potential to serve as a molecular target for cancers with Nestin-positive cancer cells and Nestin-positive tumor vasculature.
Sumiyuki Mii - One of the best experts on this subject based on the ideXlab platform.
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comparison of Nestin expressing multipotent stem cells in the tongue fungiform papilla and vibrissa hair follicle
Journal of Cellular Biochemistry, 2014Co-Authors: Yasuyuki Amoh, Kensei Katsuoka, Sumiyuki Mii, Robert M. HoffmanAbstract:We have previously reported that hair follicles contain multipotent stem cells, which express Nestin and participate in follicle growth at anagen as well as in the extension of the follicle sensory nerve. The Nestin-driven green fluorescent protein (ND-GFP) transgenic mouse labels all Nestin-expressing cells with GFP. The hair follicle Nestin-GFP cells can differentiate into neurons, Schwann cells, and other cell types. In this study, we describe Nestin-expressing multipotent stem cells in the fungiform papilla in the tongue. The Nestin-expressing multipotent stem cells in the fungiform papilla are located around a peripheral sensory nerve immediately below the taste bud and co-express the neural crest cell marker p75NTR. The fungiform papilla cells formed spheres in suspension culture in DMEM-F12 medium supplemented with basic fibroblast growth factor (bFGF). The spheres consisted of Nestin-expressing cells that co-expressed the neural crest marker p75NTR and which developed expression of the stem cell marker CD34. P75NTR, CD34 and Nestin co-expression suggested that Nestin-expressing cells comprising the fungiform papilla spheres were in a relatively undifferentiated state. The Nestin-expressing cells of these spheres acquired the following markers: β III tubulin typical of nerve cells; GFAP typical of glial cells; K15 typical of keratinocytes; and smooth-muscle antigen (SMA), after transfer to RPMI 1640 medium with 10% fetal bovine serum (FBS), suggesting they differentiated into multiple cell types. The results of the current study indicate Nestin-expressing fungiform papilla cells and the Nestin-expressing hair follicle stem cells have common features of cell morphology and ability to differentiate into multiple cell types, suggesting their remarkable similarity. J. Cell. Biochem. 115: 1070–1076, 2014. © 2013 Wiley Periodicals, Inc.
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comparison of Nestin expressing multipotent stem cells in the tongue fungiform papilla and vibrissa hair follicle
Journal of Cellular Biochemistry, 2014Co-Authors: Yasuyuki Amoh, Kensei Katsuoka, Sumiyuki Mii, Robert M. HoffmanAbstract:We have previously reported that hair follicles contain multipotent stem cells, which express Nestin and participate in follicle growth at anagen as well as in the extension of the follicle sensory nerve. The Nestin-driven green fluorescent protein (ND-GFP) transgenic mouse labels all Nestin-expressing cells with GFP. The hair follicle Nestin-GFP cells can differentiate into neurons, Schwann cells, and other cell types. In this study, we describe Nestin-expressing multipotent stem cells in the fungiform papilla in the tongue. The Nestin-expressing multipotent stem cells in the fungiform papilla are located around a peripheral sensory nerve immediately below the taste bud and co-express the neural crest cell marker p75(NTR) . The fungiform papilla cells formed spheres in suspension culture in DMEM-F12 medium supplemented with basic fibroblast growth factor (bFGF). The spheres consisted of Nestin-expressing cells that co-expressed the neural crest marker p75(NTR) and which developed expression of the stem cell marker CD34. P75(NTR), CD34 and Nestin co-expression suggested that Nestin-expressing cells comprising the fungiform papilla spheres were in a relatively undifferentiated state. The Nestin-expressing cells of these spheres acquired the following markers: β III tubulin typical of nerve cells; GFAP typical of glial cells; K15 typical of keratinocytes; and smooth-muscle antigen (SMA), after transfer to RPMI 1640 medium with 10% fetal bovine serum (FBS), suggesting they differentiated into multiple cell types. The results of the current study indicate Nestin-expressing fungiform papilla cells and the Nestin-expressing hair follicle stem cells have common features of cell morphology and ability to differentiate into multiple cell types, suggesting their remarkable similarity.
John E. Eriksson - One of the best experts on this subject based on the ideXlab platform.
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a Nestin scaffold links cdk5 p35 signaling to oxidant induced cell death
The EMBO Journal, 2006Co-Authors: Cecilia Sahlgren, Yinghao Chou, Hanna-mari Pallari, Tao He, Robert D. Goldman, John E. ErikssonAbstract:The intermediate filament protein, Nestin, has been implicated as an organizer of survival-determining signaling molecules. When Nestin expression was related to the sensitivity of neural progenitor cells to oxidant-induced apoptosis, Nestin displayed a distinct cytoprotective effect. Oxidative stress in neuronal precursor cells led to downregulation of Nestin with subsequent activation of cyclin-dependent kinase 5 (Cdk5), a crucial kinase in the nervous system. Nestin downregulation was a prerequisite for the Cdk5-dependent apoptosis, as overexpression of Nestin efficiently inhibited induction of apoptosis, whereas depletion of Nestin by RNA interference had a sensitizing effect. When the underlying link between Nestin and Cdk5 was analyzed, we observed that Nestin serves as a scaffold for Cdk5, with binding restricted to a specific region following the alpha-helical domain of Nestin, and that the presence and organization of Nestin regulated the sequestration and activity of Cdk5, as well as the ubiquitylation and turnover of its regulator, p35. Our data imply that Nestin is a survival determinant whose action is based upon a novel mode of Cdk5 regulation, affecting the targeting, activity, and turnover of the Cdk5/p35 signaling complex.
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cdk5 regulates the organization of Nestin and its association with p35
Molecular and Cellular Biology, 2003Co-Authors: Cecilia Sahlgren, Andrey Mikhailov, Hanna-mari Pallari, Samuli Vaittinen, Harish C. Pant, John E. Eriksson, Hannu KalimoAbstract:The intermediate filament protein Nestin is characterized by its specific expression during the development of neuronal and myogenic tissues. We identify Nestin as a novel in vivo target for cdk5 and p35 kinase, a critical signaling determinant in development. Two cdk5-specific phosphorylation sites on Nestin, Thr-1495 and Thr-316, were established, the latter of which was used as a marker for cdk5-specific phosphorylation in vivo. Ectopic expression of cdk5 and p35 in central nervous system progenitor cells and in myogenic precursor cells induced elevated phosphorylation and reorganization of Nestin. The kinetics of Nestin expression corresponded to elevated expression and activation of cdk5 during differentiation of myoblast cell cultures and during regeneration of skeletal muscle. In the myoblasts, a disassembly-linked phosphorylation of Thr-316 indicated active phosphorylation of Nestin by cdk5. Moreover, cdk5 occurred in physical association with Nestin. Inhibition of cdk5 activity—either by transfection with dominant-negative cdk5 or by using a specific cdk5 inhibitor—blocked myoblast differentiation and phosphorylation of Nestin at Thr-316, and this inhibition markedly disturbed the organization of Nestin. Interestingly, the interaction between p35, the cdk5 activator, and Nestin appeared to be regulated by cdk5. In differentiating myoblasts, p35 was not complexed with Nestin phosphorylated at Thr-316, and inhibition of cdk5 activity during differentiation induced a marked association of p35 with Nestin. These results demonstrate that there is a continuous turnover of cdk5 and p35 activity on a scaffold formed by Nestin. This association is likely to affect the organization and operation of both cdk5 and Nestin during development.
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mitotic reorganization of the intermediate filament protein Nestin involves phosphorylation by cdc2 kinase
Journal of Biological Chemistry, 2001Co-Authors: Cecilia Sahlgren, Andrey Mikhailov, Yinghao Chou, Jukka Hellman, Robert D. Goldman, Urban Lendahl, John E. ErikssonAbstract:Abstract The intermediate filament protein Nestin is expressed during early stages of development in the central nervous system and in muscle tissues. Nestin expression is associated with morphologically dynamic cells, such as dividing and migrating cells. However, little is known about regulation of Nestin during these cellular processes. We have characterized the phosphorylation-based regulation of Nestin during different stages of the cell cycle in a neuronal progenitor cell line, ST15A. Confocal microscopy of Nestin organization and 32P in vivo labeling studies show that the mitotic reorganization of Nestin is accompanied by elevated phosphorylation of Nestin. The phosphorylation-induced alterations in Nestin organization during mitosis in ST15A cells are associated with partial disassembly of Nestin filaments. Comparative in vitro and in vivo phosphorylation studies identified cdc2 as the primary mitotic kinase and Thr316 as a cdc2-specific phosphorylation site on Nestin. We generated a phosphospecific Nestin antibody recognizing the phosphorylated form of this site. By using this antibody we observed that Nestin shows constitutive phosphorylation at Thr316, which is increased during mitosis. This study shows that Nestin is reorganized during mitosis and that cdc2-mediated phosphorylation is an important regulator of Nestin organization and dynamics during mitosis.
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specific and innervation regulated expression of the intermediate filament protein Nestin at neuromuscular and myotendinous junctions in skeletal muscle
American Journal of Pathology, 1999Co-Authors: Samuli Vaittinen, Cecilia Sahlgren, John E. Eriksson, Hannu Kalimo, Urban Lendahl, Riitta Lukka, Jussi Rantanen, Timo HurmeAbstract:The intermediate filament proteins Nestin, vimentin, and desmin show a specific temporal expression pattern during the development of myofibers from myogenic precursor cells. Nestin and vimentin are actively expressed during early developmental stages to be later down-regulated, vimentin completely and Nestin to minimal levels, whereas desmin expression begins later and is maintained in mature myofibers, in which desmin participates in maintaining structural integrity. In this study we have analyzed the expression levels and distribution pattern of Nestin in intact and denervated muscle in rat and in human. Nestin immunoreactivity was specifically and focally localized in the sarcoplasm underneath neuromuscular junctions (NMJs) and in the vicinity of the myotendinous junctions (MTJs), ie, in regions associated with acetylcholine receptors (AChRs). This association prompted us to analyze Nestin in neurogenically and myogenically denervated muscle. Immunoblot analysis disclosed a marked overall increase of accumulated Nestin protein. Similar to the extrajunctional redistribution of AChRs in denervated myofibers, Nestin immunoreactivity extended widely beyond the NMJ region. Re-innervation caused complete reversion of these changes. Our study demonstrates that the expression levels and distribution pattern of Nestin are regulated by innervation, ie, signal transduction into myofibers.
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the intermediate filament protein Nestin occurs transiently in differentiating testis of rat and mouse
Differentiation, 1997Co-Authors: K Frojdman, Urban Lendahl, Lauri J Pelliniemi, Ismo Virtanen, John E. ErikssonAbstract:Nestin is an intermediate filament (IF) protein (IFP) which occurs during early developmental stages and during regenerative processes in muscle and neuronal cells. The spatial and temporal localization of Nestin in the developing testis of rat and mouse was studied by immunolabeling light and electron microscopy and by immunoblotting. Nestin localization was related to the localization of the other major IFPs specific for this tissue, i.e. cytokeratins, vimentin and desmin. Laminin immunocytochemistry and conventional microscopy were used to identify tissues and cells. With the incipient differentiation of the gonadal anlage, the reaction for Nestin was weak in the gonadal ridge, whereas the cells of the mesonephric mesenchyme showed a prominent reaction for this IFP. The Nestin-specific reaction in the epithelial mesonephric duct and tubules was weak and disappeared at an early phase of differentiation. With the development of the testis proper, Nestin was transiently found in several cell types. Nestin was found as well as vimentin and cytokeratins in the Sertoli cells. In the interstitial cells Nestin was found together with vimentin and desmin IFPs, and was most prominent in the differentiating myoid cells. After birth, Nestin gradually disappeared from the testicular cells and in the rat at puberty was found only in the endothelial cells of some blood vessels. The abolished Nestin synthesis in the testis was confirmed by immunoblotting. These results suggest that Nestin is required transiently during the development of the testis and mesonephros. The temporary presence of Nestin, and several other IFPs during these phases, coincides with key phases of urogenital sex differentiation. This may imply that the orchestrated synthesis of the IFPs Nestin, cytokeratins, vimentin and desmin is likely to be linked with the genes regulating sex differentiation.