Elsevier

Neuroscience

Volume 115, Issue 1, 15 November 2002, Pages 295-305
Neuroscience

Stromal cell-derived factor 1 is secreted by meningeal cells and acts as chemotactic factor on neuronal stem cells of the cerebellar external granular layer

https://doi.org/10.1016/S0306-4522(02)00307-XGet rights and content

Abstract

The cerebellar external granular layer (EGL) is an unusually long-lasting neural proliferative zone positioned immediately beneath the pial surface. Its position and stability critically depend on meningeal cells, as their selective destruction leads to its rapid dispersal, creating massive cortical ectopia. Similar ectopias have recently been described as a side effect of deficiency for stromal cell-derived factor 1 (SDF-1), a chemoattractant for haematopoietic precursor cell migration. Here we show that SDF-1 is present in meningeal cells in vivo and in vitro, where it is secreted in functionally relevant concentrations into the medium. Correspondingly, the SDF-1 receptor (termed CXCR4) can be demonstrated on stem cells of the external granular layer, but is absent on postmitotic cells commencing their final inward migration. We show that SDF-1 is concentrated by heparan sulphate proteoglycans highly expressed in the EGL in a laminar fashion, which thus might act to locally restrict SDF-1 action to the EGL in a kind of step gradient. In vitro, SDF-1 chemotactically attracts neuronal cells isolated from the external, but not from the internal granular layer, in a Boyden chamber assay in concentrations found in meningeal cell-conditioned medium. Selective removal of SDF-1 from conditioned media by immunoprecipitation abolishes their chemoattractive action, which can be reconstituted again by the addition of recombinant SDF-1. Meningeal cells are thus an important source for the expression of SDF-1 during brain development, which – comparable to its role in haematopoiesis – appears to be a key factor attracting precursor cells to their proliferative compartment.

Section snippets

Animals

Rats (Wistar strain) were kept in a conventional breeding facility at a 12/12-h light/dark cycle and were mated overnight. For the designation of developmental stages, the day following mating was counted as embryonic day 0 (E 0) and the day of birth as postnatal day zero (P 0).

Immunohistochemistry

Animals were killed by an overdose of anaesthetics and fixed by transcardiac perfusion with either 4% paraformaldehyde in phosphate-buffered saline (PBS) at pH 7.2 or Bouin’s solution diluted 1/4 in PBS (pH 2.5). Brains

Expression of CXCR4 in the cerebellar anlage

We have analysed the in vivo expression of the SDF-1 receptor CXCR4 by immunohistochemistry between E 14 and P 30 (Fig. 1). During prenatal development, immunoreactivity for CXCR4 is first seen in the region of the rhombic lip from E 14 onward, being localized (i) on vascular endothelia both within the developing brain parenchyma and the leptomeningeal tissue and (ii) on emigrating cells setting up the EGL. This latter band of cells passes through the entire thickness of the cerebellar

Developmental biology of the EGL

Key aspects of the ontogeny of the EGL depend on interactions with the overlying meningeal cell layer. Selective destruction of these fibroblasts during a critical developmental period (Sievers et al., 1994a, Sievers et al., 1994b, Sievers et al., 1994c) terminates the tangential caudorostral expansion of this stem cell compartment, most cells of which rapidly leave their subsurface position to become ectopically scattered within the cerebellar cortical anlage. Also, development of the

Acknowledgements

The authors are indebted to Monika Grell, Gundel Jopp, Marion Koelln and Marleen Regent for their expert technical assistance. We thank Prof. Guido David, Department of Human Genetics, KU Leuven, for the generous supply of antibodies against HSPG and heparitinase and many helpful discussions. The work was partly funded by a grant from the Faculty of Medicine, CAU Kiel.

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