Class III beta-tubulin is constitutively coexpressed with glial fibrillary acidic protein and nestin in midgestational human fetal astrocytes: implications for phenotypic identity
Language English Country England, Great Britain Media print
Document type Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't
Grant support
P01 NS30916
NINDS NIH HHS - United States
- MeSH
- Astrocytes metabolism MeSH
- Phenotype * MeSH
- Gestational Age * MeSH
- Glial Fibrillary Acidic Protein genetics metabolism MeSH
- Cells, Cultured MeSH
- Humans MeSH
- RNA, Messenger metabolism MeSH
- Cerebral Cortex cytology MeSH
- Mice MeSH
- Nestin MeSH
- Fetus cytology MeSH
- Intermediate Filament Proteins genetics metabolism MeSH
- Nerve Tissue Proteins genetics metabolism MeSH
- Tubulin genetics metabolism MeSH
- Vimentin metabolism MeSH
- Gene Expression Regulation, Developmental MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, N.I.H., Extramural MeSH
- Names of Substances
- Glial Fibrillary Acidic Protein MeSH
- RNA, Messenger MeSH
- NES protein, human MeSH Browser
- Nes protein, mouse MeSH Browser
- Nestin MeSH
- Intermediate Filament Proteins MeSH
- Nerve Tissue Proteins MeSH
- TUBB3 protein, human MeSH Browser
- Tubulin MeSH
- Vimentin MeSH
Class III beta-tubulin isotype (betaIII-tubulin) is widely regarded as a neuronal marker in developmental neurobiology and stem cell research. To test the specificity of this marker protein, we determined its expression and distribution in primary cultures of glial fibrillary acidic protein (GFAP)-expressing astrocytes isolated from the cerebral hemispheres of 2 human fetuses at 18 to 20 weeks of gestation. Cells were maintained as monolayer cultures for 1 to 21 days without differentiation induction. By immunofluorescence microscopy, coexpression of betaIII-tubulin and GFAP was detected in cells at all time points but in spatially distinct patterns. The numbers of GFAP+ cells gradually decreased from Days 1 to 21 in vitro, whereas betaIII-tubulin immunoreactivity was present in 100% of cells at all time points. beta-III-tubulin mRNA and protein expression were demonstrated in cultured cells by reverse-transcriptase-polymerase chain reaction and immunoblotting, respectively. Glial fibrillary acidic protein+/beta-III-tubulin-positive cells coexpressed nestin and vimentin but lacked neurofilament proteins, CD133, and glutamate-aspartate transporter. Weak cytoplasmic staining was detected with antibodies against microtubule-associated protein 2 isoforms. Confocal microscopy, performed on autopsy brain samples of human fetuses at 16 to 20 gestational weeks, revealed widespread colocalization of GFAP and betaIII-tubulin in cells of the ventricular/subventricular zones and the cortical plate. Our results indicate that in the midgestational human brain, betaIII-tubulin is not neuron specific because it is constitutively expressed in GFAP+/nestin+ presumptive fetal astrocytes.
References provided by Crossref.org
Plasticity of calcium signaling cascades in human embryonic stem cell-derived neural precursors
