Abstract
Balanced cell proliferation and cell death determines neural precursor cell numbers in early stages of neural tube (NT) development. We have previously shown that nitric oxide (NO) regulates cell numbers locally in the NT of eight to 12 somite embryos. Here, we demonstrate that bone morphogenetic protein-4 (BMP-4), which is expressed in the ectoderm and dorsal NT at these developmental stages, induces programmed cell death (PCD) and promotes entry into the S-phase, via nitric oxide synthase (NOS) activity. These effects can be reversed by BMP-4 antagonists, such as follistatin and noggin, or by specific NOS inhibitors, resulting in low NO levels that facilitate mitosis and reduce PCD. Ectopic BMP-4 induction of PCD is restricted to the dorsal NT, whereas promotion of the S-phase is evenly observed across the dorsal–ventral (D–V) axis. Prolonged exposure to either BMP-4 or NOS inhibitors, which results in high or low NO levels, respectively, causes NT defects. The results presented here throw new light on the BMP signaling pathway. The local presence of BMP-4 helps to regulate cell numbers in the developing NT by a NO-mediated pathway, which is essential for normal NT formation.
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Abbreviations
- BMP:
-
bone morphogenetic protein
- BrdU:
-
bromodeoxyuridine
- DAF-2DA:
-
4,5-diaminofluorescein diacetate
- L-NMMA:
-
NG-methyl-L-arginine
- NO:
-
nitric oxide
- NOS:
-
nitric oxide synthase
- NT:
-
neural tube
- PCD:
-
programmed cell death
- PI:
-
propidium iodide
- SNAP:
-
S-nitroso-N-acetyl-penicillamine
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Acknowledgements
This research was supported by The Israel Science Foundation Grant Number 6/10-17.2.
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Supplementary information accompanies this paper on the Cell Death and Differentiation website: http://www.nature.com/cdd
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Supplementary File 1
Video 1.mov: The rotating image is a projection series of 136 microns thick longitudinal optical sections of the NT at the site of the control bead implantation of a 12 somite embryo, the bead was carefully removed before scanning. Note the detection of endogenous NO by DAF-2DA fluorescense in the ectoderm and most dorsal NT region only while no fluorescense is detected ventrally (Fig.3A).
Supplementary File 2
Video 2.mov: The rotating image is a projection series of 136 microns thick longitudinal optical sections of the NT at the site of BMP-4 bead implantation of a 12 somite embryo, the bead was carefully removed before scanning. Note that the DAF-2DA green fluorescence appears within the surface of the pocket left by the bead that was in contact with the BMP-4 bead (Fig. 3B).
Supplementary File 3
Video 3.mov: The rotating image is a projection series of 136 microns thick longitudinal optical sections of the NT at the site of BMP-4 bead implantation of a 12 somite embryo treated with the NOS inhibitor L-NMMA, the bead was carefully removed before scanning. Note the significant reduction in DAF-2DA fluorescence within the bead pocket and throughout the projection caused by the NOS inhibitor (Fig. 3C).
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Traister, A., Abashidze, S., Gold, V. et al. BMP controls nitric oxide-mediated regulation of cell numbers in the developing neural tube. Cell Death Differ 11, 832–841 (2004). https://doi.org/10.1038/sj.cdd.4401404
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DOI: https://doi.org/10.1038/sj.cdd.4401404
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