Notch3 is necessary for neuronal differentiation and maturation in the adult spinal cord
Gabriel Rusanescu,
Jianren Mao,
Corresponding Author
Gabriel Rusanescu
MGH Center for Translational Pain Research, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, Charlestown, MA, USA
Correspondence to: Gabriel RUSANESCU,
MGH Center for Translational Pain Research,
Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, 149 13th Street,
Charlestown, MA 02129, USA.
Tel.: 1-781-526-7412
Fax: 617-726 3441
E-mail: [email protected]
Search for more papers by this authorJianren Mao
MGH Center for Translational Pain Research, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, Charlestown, MA, USA
Search for more papers by this authorGabriel Rusanescu,
Jianren Mao,
Corresponding Author
Gabriel Rusanescu
MGH Center for Translational Pain Research, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, Charlestown, MA, USA
Correspondence to: Gabriel RUSANESCU,
MGH Center for Translational Pain Research,
Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, 149 13th Street,
Charlestown, MA 02129, USA.
Tel.: 1-781-526-7412
Fax: 617-726 3441
E-mail: [email protected]
Search for more papers by this authorJianren Mao
MGH Center for Translational Pain Research, Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, Charlestown, MA, USA
Search for more papers by this authorAbstract
Notch receptors are key regulators of nervous system development and promoters of neural stem cells renewal and proliferation. Defects in the expression of Notch genes result in severe, often lethal developmental abnormalities. Notch3 is generally thought to have a similar proliferative, anti-differentiation and gliogenic role to Notch1. However, in some cases, Notch3 has an opposite, pro-differentiation effect. Here, we show that Notch3 segregates from Notch1 and is transiently expressed in adult rat and mouse spinal cord neuron precursors and immature neurons. This suggests that during the differentiation of adult neural progenitor cells, Notch signalling may follow a modified version of the classical lateral inhibition model, involving the segregation of individual Notch receptors. Notch3 knockout mice, otherwise neurologically normal, are characterized by a reduced number of mature inhibitory interneurons and an increased number of highly excitable immature neurons in spinal cord laminae I–II. As a result, these mice have permanently lower nociceptive thresholds, similar to chronic pain. These results suggest that defective neuronal differentiation, for example as a result of reduced Notch3 expression or activation, may underlie human cases of intractable chronic pain, such as fibromyalgia and neuropathic pain.
Supporting Information
| Filename | Description |
|---|---|
| JCMM12362-sup-0001-Supplementary Figure 1.tifimage/tif, 492.4 KB | Figure S1 Immunofluorescence analysis of Notch3 expression in rat spinal cord, relative to neuronal marker NeuN. |
| JCMM12362-sup-0002-Supplementary Figure 2.tifimage/tif, 795.9 KB | Figure S2 Immunofluorescence 3D imaging shows nuclear Notch3 expression in a NeuN-stained cell (rat spinal cord), validating Notch3 expression in neurons. |
| JCMM12362-sup-0022-Supplem Fig 2B.avivideo/avi, 28.7 MB | |
| JCMM12362-sup-0003-Supplementary Figure 3.tifimage/tif, 454.3 KB | Figure S3 Immunofluorescence analysis of Notch3 expression relative to oligodendrocyte markers in rat spinal cord sections. |
| JCMM12362-sup-0004-Supplementary Figure 4.tifimage/tif, 799.8 KB | Figure S4 Immunofluorescence analysis of Notch3 (N3, green) expression pattern in mouse spinal cord shows a similar Notch3 neuronal specificity as in rat. |
| JCMM12362-sup-0005-Supplementary Figure 5.tifimage/tif, 561.4 KB | Figure S5 Immunofluorescence analysis of neuronal marker synapsin I (Syn, red, arrowhead) expression in mouse dorsal horn spinal cord, after 7 days of EdU injection. |
| JCMM12362-sup-0006-Supplementary Figure 6.tifimage/tif, 707.7 KB | Figure S6 Immunofluorescence analysis of Notch ligands Delta and Jagged expression in rat lumbar spinal cord. |
| JCMM12362-sup-0007-Supplementary Figure 7.tifimage/tif, 919.6 KB | Figure S7 Comparative nuclear DAPI staining of WT and N3KO mouse spinal cord shows altered N3KO mouse morphology, similar to Fig. 6A (shown region T12-L1). |
| JCMM12362-sup-0008-Supplementary Figure 8.tifimage/tif, 801.7 KB | Figure S8 Immunofluorescence analysis of N3KO mouse spinal cord with neuron-specific β3 tubulin. |
| JCMM12362-sup-0009-Supplementary Figure 9.tifimage/tif, 517.3 KB | Figure S9 High magnification of mouse spinal cord laminae I–II (LI–II, double arrows), showing reduced NeuN staining (red) and an increased number of CR+ cells (green) in N3KO mouse relative to WT. |
| JCMM12362-sup-0010-Supplementary Fig Legend.docxWord document, 15.2 KB |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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