In the adult mammalian brain, multipotent stem or progenitor cells involved in reproduction of neurons and glial cells have been well investigated only in very restricted regions; the subventricular zone of the lateral ventricle and the dentate gyrus in the hippocampal formation. In the neocortex, a series of in vitro studies has suggested the possible existence of neural progenitor cells possessing neurogenic and/or gliogenic potential in adult mammals. However, the cellular properties of the cortical progenitor cells in vivo have not been fully elucidated. Using 5'-bromodeoxyuridine labeling and immunohistochemical analysis of cell differentiation markers, we found that a subpopulation of NG2-immunopositive cells co-expressing doublecortin (DCX), an immature neuron marker, ubiquitously reside in the adult rat neocortex. Furthermore, these cells are the major population of proliferating cells in the region. The DCX(+)/NG2(+) cells reproduced the same daughter cells, or differentiated into DCX(+)/NG2(-) (approximately 1%) or DCX(-)/NG2(+) (approximately 10%) cells within 2 weeks after cell division. The DCX(+)/NG2(-) cells were also immunopositive for TUC-4, a neuronal linage marker, suggesting that these cells were committed to neuronal cell differentiation, whereas the DCX(-)/NG2(+) cells showed faint immunoreactivity for glutathione S-transferase (GST)-pi, an oligodendrocyte lineage marker, in the cytoplasm, suggesting glial cell lineage, and thereafter the cells differentiated into NG2(-)/GST-pi(+) mature oligodendrocytes after a further 2 weeks. These findings indicate that DCX(+)/NG2(+) cells ubiquitously exist as 'multipotent progenitor cells' in the neocortex of adult rats.

Lamins are major structural proteins of the nuclear envelope. Three lamin subtypes, A/C, B1 and B2, predominate in mammalian somatic cells. While the expression levels of lamins in several tissues are known to change during cell differentiation, lamin expression is poorly understood in the nervous system. To investigate the expression of lamins during neuronal differentiation in the mammalian adult brain, we performed immunohistochemical studies on lamins A/C, B1 and B2 in two neurogenic regions of rat brain: the subgranular zone of the dentate gyrus and the subventricular zone of the lateral ventricle. In particular, three types of cells were analysed using confocal microscopy: GFAP-positive cells as primary progenitor (stem) cells, PSA-NCAM-positive cells as subsequent neuronal progenitor cells, and NeuN-positive mature neurons. GFAP-positive cells possesed lamin A/C (++), B1 (++) and B2 (++), PSA-NCAM-positive cells possessed lamin A/C (-), B1 (+++) and B2 (+), and mature neurons possessed lamin A/C (++), B1 (+) and B2 (+++), in both neurogenic regions. These observations showed that the compositions of expressing lamin subtypes are distinct in particular differentiation stages during neurogenesis in the adult rat brain. Our results suggest that the alteration of nuclear lamina structure is coupled with the progression of neuronal differentiation.

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HighlightsThis work demonstrates the expression patterns for lamin subtypes in neural cells in the adult rat cerebral cortex.This work demonstrates suitable fixative conditions of multiple immunostaining systems for lamins and cell-type specific marker proteins.Neural cells in the adult cerebral cortex showed heterogeneous lamin antibody labeling patterns.