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Because of their potential for self-renewal and the ability for generating many differentiated cell types, progenitor cells are a key player in regenerative and repair processes. In the central nervous system, pools of these cells have been identified in two regions: the subgranular zone of hippocampal gyrus dentatus and the subventricular zone. Neural stem cells that reside in these regions are subject to a specific neurogenesis-stimulating and -regulating environment called ‘niche’. Our model of surgical brain injury (SBI) opens the avenues for studying the mechanisms of repair and reconstruction of brain cortex and enables demonstrating the presence of possible vascular niches in the peri-lesion zone. The present studies were aimed at characterizing of the immune phenotype of the cells that populate this region. The peri-lesion area of the brain cortex showed the presence of dying neurons and glial cells since the first postlesion day. Simultaneously, activated microglial cells and astrocytes appeared, and part of the latter formed a scar on the surface of the damaged cortex. Another fractions of the cells that appeared following the SBI in both the lumen and the vicinity of blood vessels expressed either the macrophagal/monocytic marker CD14, or the marker of hematopoietic progenitor cells and small vessel endothelium CD34. Beginning on the first post-SBI day, the peri-lesion area showed also the presence and accumulation of a variety of cells with immature phenotypes. These included immature endothelial cells building new blood vessels (angiogenesis) and cells with phenotypes of other brain parenchyma-forming cell subpopulations: (1) nestin-positive astroglial and non-glial cells, (2) cells expressing the marker of juvenile astrocytes vimentin-positive, and (3) cells showing doublecortin immunoreactivity (the marker of early differentiated neurons). These results clearly indicate that during the early post-SBI period the peri-lesion zone is being populated by a heterogenic pool of morphologically immature cells that most likely herald the advent of reconstruction and/or repair of the injured brain region. Supported by the Polish Ministry of Science and Higher Education grant No N N404 522838
This study shows that an ICP4– replication-deficient herpes simplex virus containing the Moloney murine leukaemia virus LTR fused with the coding sequence for the β-galactosidase gene can be used as a very effective vector for delivering the β-galactosidase reporter gene into the rat brain septum. F344 rats received bilateral stereotaxic injections into the nucleus of the diagonal band and into the medial septum. The X-gal stain was used to detect the activity of the expressed β-galactosidase enzyme. The delivered reporter gene was expressed successfully not only in the neuronal cells of the injected areas but also in cells that project to the injection area such as cortex cells about 6mm away from the injection sites. Expression was visible at 1, 3 and 9 weeks following injection. We conclude that this vector can effectively deliver genes into different regions of the mature mammalian brain and also to areas distant from the injection site.
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