Class III beta-tubulin is constitutively coexpressed with glial fibrillary acidic protein and nestin in midgestational human fetal astrocytes: implications for phenotypic identity
Jazyk angličtina Země Anglie, Velká Británie Médium print
Typ dokumentu časopisecké články, Research Support, N.I.H., Extramural, práce podpořená grantem
Grantová podpora
P01 NS30916
NINDS NIH HHS - United States
- MeSH
- astrocyty metabolismus MeSH
- fenotyp * MeSH
- gestační stáří * MeSH
- gliový fibrilární kyselý protein genetika metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- messenger RNA metabolismus MeSH
- mozková kůra cytologie MeSH
- myši MeSH
- nestin MeSH
- plod cytologie MeSH
- proteiny intermediálních filament genetika metabolismus MeSH
- proteiny nervové tkáně genetika metabolismus MeSH
- tubulin genetika metabolismus MeSH
- vimentin metabolismus MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- gliový fibrilární kyselý protein MeSH
- messenger RNA MeSH
- NES protein, human MeSH Prohlížeč
- Nes protein, mouse MeSH Prohlížeč
- nestin MeSH
- proteiny intermediálních filament MeSH
- proteiny nervové tkáně MeSH
- TUBB3 protein, human MeSH Prohlížeč
- 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.
Citace poskytuje Crossref.org
Plasticity of calcium signaling cascades in human embryonic stem cell-derived neural precursors
