Coexpression of Nestin in Neural and Glial Cells in the Developing Human CNS Defined by a Human-Specific Anti-nestin Antibody

Abstract

The presence of the intermediate filament protein nestin has been the predominant marker used to describe stem and progenitor cells in the mammalian CNS. In this study, a 998-bp fragment in the 3' region of the nestin mRNA was cloned from human fetal brain cells (HFBC). The nucleotide sequence of the cloned cDNA revealed 21 differences with the previously published human nestin sequence, resulting in 17 amino acid changes. A 150-amino-acid fragment derived from the cloned nestin cDNA was coupled to glutathione S-transferase and used as an immunogen to generate a rabbit polyclonal antiserum that selectively detects human nestin. HFBC that proliferated in response to basic fibroblast growth factor incorporated 5-bromo-2'-deoxyuridine into their nuclei and immunostained for nestin, indicating nestin expression in proliferating CNS progenitor cells. In all cell cultures, nestin costained with the neuroepithelial cell marker vimentin. A small subset of nestin-stained cells (1-2%) immunostained with neuronal marker MAP-2 during the first week and after 4 weeks in culture. However, during the first week in culture, approximately 10-30% of the total cell population of HFBC stained for the glial cell marker GFAP, and nearly all coimmunostained for nestin. After 4 weeks in culture, a subset of GFAP-positive cells emerged that no longer costained with nestin. These results describe nestin expression not only in CNS progenitor cells but also in the cells which were in transition from a progenitor stage to glial differentiation. Collectively, these data suggest a differential temporal regulation of nestin expression during glial and neuronal cell differentiation.

Tumor Vascular Interaction in Melanomas and Neurogenesis: A Review

“…Nestinis expressed in endothelial cells of CNS tumor tissues and of adult tissues that replenish by angiogenesis and so can be used as a marker protein for neovascularization [26][27][28][29]. This fact is well brought out in the present study, since the endothelial cells lining the angiogenic vessels are positive for Nes as they enter the tumor.…”

“…These were subjected to routine histochemistry, (HE, PAS, modified reticulin+gold impregnation) [21] enzyme histochemistry (Dopa Oxidase) and Immunohistochemistry using the Avidin/Biotin system [22][23][24][25]. monoclonal antibodies (mAb)used: HMB45, NFP (neuro filament protein), GFAP, Synaptophysin (Syn), (BioGenex) nestin (Nes), (Chemicon) a marker of multi-lineage progenitor cells [26][27][28][29]. The indoleamines: serotonin (SER) & melatonin (MLT), and the catecholamines: dopamine (DA) & noradrenalin (NA), by the avidin-biotin method using Dako Pat kits.…”

Angiogenic endothelium-specific nestin expression is enhanced by the first intron of the nestin gene

“…In neural and glial cells, nestin is expressed in BrdU-incorporated neural progenitor cells. 24 Nestin reportedly induces disassembly of vimentin to nonfilamentous aggregates or particles in mitotic Angiogenesis-specific nestin expression M Aihara et al cells for equal distribution of vimentin to daughter cells. 4 Thus, nestin expression appears to be proliferation-dependent in endothelium.…”

Nestin Is a Potential Mediator of Malignancy in Human Neuroblastoma Cells

“…Nuclear and cytoplasmic fractions were isolated as previously described (22). Western blot procedures have been published (22,24). Primary antibodies included polyclonal antibodies to nestin (331b), homing cell adhesion molecule (HCAM) (Santa Cruz Biotechnology, Santa Cruz, CA), secretogranin II (gift of Dr. Jonathan G. Scammell, Department of South Alabama College of Medicine, Mobile, AL), tissue plasminogen activator (tPA) (ICN Pharmaceuticals, Inc., Aurora, OH), and vimentin (Chemicon International, Temecula, CA) and monoclonal antibodies to neurofilaments (NFs) 70 and 160, chromogranin A, ␤-tubulin, and actin (clone C4) (Roche Applied Science).…”