Immunohistochemical Detection of Intermediate Filament Nestin

Mokrý, Jaroslav; Němeček, S.

doi:10.14712/18059694.2019.168

Cited by 39 publications

(10 citation statements)

References 27 publications

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“…We additionally viewed horizontally oriented nestin + structures within the retina that were identified as blood vessel cells, an observation in accordance with other reports [26] , [50] , [59] . However, a recent study showed that nestin is only expressed in proliferating endothelial cells, and not in the mature vasculature, further indicating that nestin + cells constitute rather immature cells [60] .…”

Section: Discussionsupporting

confidence: 75%

In Situ Dividing and Phagocytosing Retinal Microglia Express Nestin, Vimentin, and NG2 In Vivo

et al. 2011

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BackgroundFollowing injury, microglia become activated with subsets expressing nestin as well as other neural markers. Moreover, cerebral microglia can give rise to neurons in vitro. In a previous study, we analysed the proliferation potential and nestin re-expression of retinal macroglial cells such as astrocytes and Müller cells after optic nerve (ON) lesion. However, we were unable to identify the majority of proliferative nestin+ cells. Thus, the present study evaluates expression of nestin and other neural markers in quiescent and proliferating microglia in naïve retina and following ON transection in adult rats in vivo.Methodology/Principal FindingsFor analysis of cell proliferation and cells fates, rats received BrdU injections. Microglia in retinal sections or isolated cells were characterized using immunofluorescence labeling with markers for microglia (e.g., Iba1, CD11b), cell proliferation, and neural cells (e.g., nestin, vimentin, NG2, GFAP, Doublecortin etc.). Cellular analyses were performed using confocal laser scanning microscopy. In the naïve adult rat retina, about 60% of resting ramified microglia expressed nestin. After ON transection, numbers of nestin+ microglia peaked to a maximum at 7 days, primarily due to in situ cell proliferation of exclusively nestin+ microglia. After 8 weeks, microglia numbers re-attained control levels, but 20% were still BrdU+ and nestin+, although no further local cell proliferation occurred. In addition, nestin+ microglia co-expressed vimentin and NG2, but not GFAP or neuronal markers. Fourteen days after injury and following retrograde labeling of retinal ganglion cells (RGCs) with Fluorogold (FG), nestin+NG2+ microglia were positive for the dye indicating an active involvement of a proliferating cell population in phagocytosing apoptotic retinal neurons.Conclusions/SignificanceThe current study provides evidence that in adult rat retina, a specific resident population of microglia expresses proteins of immature neural cells that are involved in injury-induced cell proliferation and phagocytosis while transdifferentiation was not observed.

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Section: Discussionsupporting

confidence: 75%

In Situ Dividing and Phagocytosing Retinal Microglia Express Nestin, Vimentin, and NG2 In Vivo

et al. 2011

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“…Lens development may require Nestin for similar reasons. Nestin was reported to be expressed in the epithelium of lens vesicle [26] . Nestin MO may induce apoptosis of the epithelial progenitor cells, resulting in defective lens development and organization.…”

Section: Discussionmentioning

confidence: 99%

Nestin Is Essential for Zebrafish Brain and Eye Development through Control of Progenitor Cell Apoptosis

Chen

Yuh

2010

PLoS ONE

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BackgroundNestin is expressed in neural progenitor cells (NPC) of developing brain. Despite its wide use as an NPC marker, the function of nestin in embryo development is unclear.Methodology/Principal FindingsAs nestin is conserved in zebrafish and its predicted sequence is clustered with the mammalian nestin orthologue, we used zebrafish as a model to investigate its role in embryogenesis. Injection of nestin morpholino (MO) into fertilized eggs induced time- and dose-dependent brain and eye developmental defects. Nestin morphants exhibited characteristic morphological changes including small head, small eyes and hydrocephalus. Histological examinations show reduced hind- and mid-brain size, dilated ventricle, poorly organized retina and underdeveloped lens. Injection of control nestin MO did not induce brain or eye changes. Nestin MO injection reduced expression of ascl1b (achaete-scute complex-like 1b), a marker of NPCs, without affecting its distribution. Nestin MO did not influence Elavl3/4 (Embryonic lethal, abnormal vision, Drosophila-like 3/4) (a neuronal marker), or otx2 (a midbrain neuronal marker), but severely perturbed cranial motor nerve development and axon distribution. To determine whether the developmental defects are due to excessive NPC apoptosis and/or reduced NPC proliferation, we analyzed apoptosis by TUNEL assay and acridine orange staining and proliferation by BrdU incorporation, pcna and mcm5 expressions. Excessive apoptosis was noted in hindbrain and midbrain cells. Apoptotic signals were colocalized with ascl1b. Proliferation markers were not significantly altered by nestin MO.Conclusion/SignificanceThese results suggest that nestin is essential for zebrafish brain and eye development probably through control of progenitor cell apoptosis.

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“…Improved detection of several intermediate filament proteins in formalin-fixed brain tissue has been reported, including phosphorylated and non-phosphorylated NF (Evers and Uylings 1996), vimentin (Ashraf Imam et al 1995; Shi et al 1991; Williams et al 1997) and GFAP (McQuaid et al 1995) and, when compared to protease digestion, MW pretreatment was clearly superior with antigens such as vimentin (Shi et al 1991). Antigen retrieval IHC was used to study nestin expression in developing and adult rodent tissue by use of the mouse MAb Rat-401 (Mokry and Nemecek 1998). Following a protocol similar to ours, these authors reported an enhanced signal for nestin in formalin-fixed tissues, with the highest nestin expression in developing CNS and skeletal muscle.…”

Section: Effects Of Microwave Heatingmentioning

confidence: 99%

“…Since the introduction of epitope retrieval methods, the sensitivity of many antibodies has been enhanced and the ability to retrieve a wide range of antigens in formalin-fixed, paraffin-embedded CNS tissue has greatly increased the range of investigations that can be carried out on this type of stored material (McQuaid et al 1995). Marked increase in sensitivity with no loss of specificity was reported for several IFPs, including vimentin (Shi et al 1991), GFAP (McQuaid et al 1995), (Evers and Uylings 1996) and, most recently, for nestin by the use of Rat-401 mouse monoclonal antibody on rodent tissues (Mokry and Nemecek 1998).…”

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confidence: 99%

Immunohistochemical Detection of Nestin in Pediatric Brain Tumors

Almqvist

Mah

Lendahl

et al. 2002

J Histochem Cytochem.

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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 heat-induced 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.

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Immunohistochemical Detection of Intermediate Filament Nestin

Cited by 39 publications

References 27 publications

In Situ Dividing and Phagocytosing Retinal Microglia Express Nestin, Vimentin, and NG2 In Vivo

In Situ Dividing and Phagocytosing Retinal Microglia Express Nestin, Vimentin, and NG2 In Vivo

Nestin Is Essential for Zebrafish Brain and Eye Development through Control of Progenitor Cell Apoptosis

Immunohistochemical Detection of Nestin in Pediatric Brain Tumors

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