Identification of noncanonical Wnt receptors required for Wnt-3a-induced early differentiation of human neural stem cells
Wnt proteins preferentially activate either β-catenin-dependent or β-catenin-independent signals, but the activity of a particular Wnt also depends on cellular context and receptor availability. We previously reported that Wnt-3a induces neural differentiation of human embryonic stem cell-derived ne...
| Main Authors: | , , , , , |
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| Format: | Journal article |
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Springer US
2016
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| _version_ | 1797056180683538432 |
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| author | Bengoa-Vergniory, N Gorrono-Etxebarria, I Lopez-Sanchez, I Marra, M Di Chiaro, P Kypta, R |
| author_facet | Bengoa-Vergniory, N Gorrono-Etxebarria, I Lopez-Sanchez, I Marra, M Di Chiaro, P Kypta, R |
| author_sort | Bengoa-Vergniory, N |
| collection | OXFORD |
| description | Wnt proteins preferentially activate either β-catenin-dependent or β-catenin-independent signals, but the activity of a particular Wnt also depends on cellular context and receptor availability. We previously reported that Wnt-3a induces neural differentiation of human embryonic stem cell-derived neural stem cells (NSCs) in a β-catenin-independent manner by activating a signal involving JNK and the AP-1 family member ATF-2. Here, we report the results of a gene silencing approach to identify the Wnt receptors that mediate this response to Wnt-3a. Silencing of ROR2 increased neuronal differentiation, as measured by expression of the genes DCX, NEUROD1, and NGN1, suggesting ROR2 signals normally prevent differentiation. Silencing of the other Wnt receptors singly did not affect Wnt-3a-induced neuronal differentiation. However, pairwise silencing of ROR1 and FZD4 or FZD5 and of LRP6 and FZD4 or FZD5 inhibited neuronal differentiation, as detected by reductions in the expression of neuronal genes and immunocytochemical detection of DCX, NEUROD1 and DCX. Ectopic expression of these receptors in HEK 293 cells increased ATF2-dependent transcription. In addition, ROR1 coimmunoprecipitated with FZD4 and LRP6 in transfected HEK 293 cells and colocalized with FZD4 and with LRP6 at the cell surface of transfected L cells. Wnt-3a did not appear to affect these interactions but did alter the interactions between LRP6 and FZD4/5. Together, these observations highlight roles for ROR1, LRP6, FZD4, and FZD5 in neural stem cell differentiation and provide support for a model in which dynamic interactions among these receptors mediate Wnt-3a activation of ATF2 signaling. |
| first_indexed | 2024-03-06T19:19:45Z |
| format | Journal article |
| id | oxford-uuid:19a968be-0915-41c8-a04d-ede938e7221a |
| institution | University of Oxford |
| last_indexed | 2024-03-06T19:19:45Z |
| publishDate | 2016 |
| publisher | Springer US |
| record_format | dspace |
| spelling | oxford-uuid:19a968be-0915-41c8-a04d-ede938e7221a2022-03-26T10:50:15ZIdentification of noncanonical Wnt receptors required for Wnt-3a-induced early differentiation of human neural stem cellsJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:19a968be-0915-41c8-a04d-ede938e7221aSymplectic Elements at OxfordSpringer US2016Bengoa-Vergniory, NGorrono-Etxebarria, ILopez-Sanchez, IMarra, MDi Chiaro, PKypta, RWnt proteins preferentially activate either β-catenin-dependent or β-catenin-independent signals, but the activity of a particular Wnt also depends on cellular context and receptor availability. We previously reported that Wnt-3a induces neural differentiation of human embryonic stem cell-derived neural stem cells (NSCs) in a β-catenin-independent manner by activating a signal involving JNK and the AP-1 family member ATF-2. Here, we report the results of a gene silencing approach to identify the Wnt receptors that mediate this response to Wnt-3a. Silencing of ROR2 increased neuronal differentiation, as measured by expression of the genes DCX, NEUROD1, and NGN1, suggesting ROR2 signals normally prevent differentiation. Silencing of the other Wnt receptors singly did not affect Wnt-3a-induced neuronal differentiation. However, pairwise silencing of ROR1 and FZD4 or FZD5 and of LRP6 and FZD4 or FZD5 inhibited neuronal differentiation, as detected by reductions in the expression of neuronal genes and immunocytochemical detection of DCX, NEUROD1 and DCX. Ectopic expression of these receptors in HEK 293 cells increased ATF2-dependent transcription. In addition, ROR1 coimmunoprecipitated with FZD4 and LRP6 in transfected HEK 293 cells and colocalized with FZD4 and with LRP6 at the cell surface of transfected L cells. Wnt-3a did not appear to affect these interactions but did alter the interactions between LRP6 and FZD4/5. Together, these observations highlight roles for ROR1, LRP6, FZD4, and FZD5 in neural stem cell differentiation and provide support for a model in which dynamic interactions among these receptors mediate Wnt-3a activation of ATF2 signaling. |
| spellingShingle | Bengoa-Vergniory, N Gorrono-Etxebarria, I Lopez-Sanchez, I Marra, M Di Chiaro, P Kypta, R Identification of noncanonical Wnt receptors required for Wnt-3a-induced early differentiation of human neural stem cells |
| title | Identification of noncanonical Wnt receptors required for Wnt-3a-induced early differentiation of human neural stem cells |
| title_full | Identification of noncanonical Wnt receptors required for Wnt-3a-induced early differentiation of human neural stem cells |
| title_fullStr | Identification of noncanonical Wnt receptors required for Wnt-3a-induced early differentiation of human neural stem cells |
| title_full_unstemmed | Identification of noncanonical Wnt receptors required for Wnt-3a-induced early differentiation of human neural stem cells |
| title_short | Identification of noncanonical Wnt receptors required for Wnt-3a-induced early differentiation of human neural stem cells |
| title_sort | identification of noncanonical wnt receptors required for wnt 3a induced early differentiation of human neural stem cells |
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