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The location and phenotype of proliferating neural-crest-derived cells in the developing mouse gut

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Abstract

Neural crest cells that originate in the caudal hindbrain migrate into and along the developing gastrointestinal tract to form the enteric nervous system. While they are migrating, neural-crest-derived cells are also proliferating. Previous studies have shown that the expression of glial-derived neurotrophic factor (GDNF) and endothelin-3 is highest in the embryonic caecum, and that GDNF alone or in combination with endothelin-3 promotes the proliferation of enteric neural-crest-derived cells in vitro. However, whether neural proliferative zones, like those in the central nervous system, are found along the developing gut is unknown. We used a fluorescent nucleic acid stain to identify dividing cells or BrdU labelling (2 h after administration of BrdU to the mother), combined with antibodies specific to neural crest cells to determine the percentage of proliferating crest-derived cells in various gut regions of embryonic day 11.5 (E11.5) and E12.5 mice. The rate of proliferation of crest-derived cells did not vary significantly in different regions of the gut (including the caecum) or at different distances from the migratory wavefront of vagal crest-derived cells. The phenotype of mitotic enteric crest-derived cells was also examined. Cells expressing the pan-neuronal markers, neurofilament-M and Hu, or the glial marker, S100b, were observed undergoing mitosis. However, no evidence was found for proliferation of cells expressing neuron-type-specific markers, such as nitric oxide synthase (at E12.5) or calcitonin gene-related peptide (at E18.5). Thus, for enteric neurons, exit from the cell cycle appears to occur after the expression of pan-neuronal proteins but prior to the expression of markers of terminally differentiated neurons.

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Acknowledgements

We thank Dr. Vicki Hammond for advice on BrdU labelling and Drs. Piers Emson, Jean Francois Brunet and Miles Epstein for the provision of antisera.

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  1. Department of Anatomy and Cell Biology, University of Melbourne, 3010, Melbourne, Victoria, Australia

    H. M. Young, K. N. Turner & A. J. Bergner

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  1. H. M. Young
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  2. K. N. Turner
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Correspondence to H. M. Young.

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This work was supported by the Australian Research Council (DP0345298) and the National Health and Medical Research Council of Australia (Project grant 145628 and Senior Research Fellowship 170224).

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Young, H.M., Turner, K.N. & Bergner, A.J. The location and phenotype of proliferating neural-crest-derived cells in the developing mouse gut. Cell Tissue Res 320, 1–9 (2005). https://doi.org/10.1007/s00441-004-1057-5

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