Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryos
Cilia are specialized organelles that extend from plasma membrane, functioning as antennas for signal transduction and are involved in embryonic morphogenesis. Dysfunction of cilia lead to many developmental defects, including neural tube defects (NTDs). Heterodimer WDR60-WDR34 (WD repeat domain 60...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-05-01
|
| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2023.1084245/full |
| _version_ | 1827951627466702848 |
|---|---|
| author | Lu Yan Lu Yan Lu Yan Hailing Yin Hailing Yin Yiwei Mi Yiwei Mi Yu Wu Yu Wu Yufang Zheng Yufang Zheng Yufang Zheng |
| author_facet | Lu Yan Lu Yan Lu Yan Hailing Yin Hailing Yin Yiwei Mi Yiwei Mi Yu Wu Yu Wu Yufang Zheng Yufang Zheng Yufang Zheng |
| author_sort | Lu Yan |
| collection | DOAJ |
| description | Cilia are specialized organelles that extend from plasma membrane, functioning as antennas for signal transduction and are involved in embryonic morphogenesis. Dysfunction of cilia lead to many developmental defects, including neural tube defects (NTDs). Heterodimer WDR60-WDR34 (WD repeat domain 60 and 34) are intermediate chains of motor protein dynein-2, which play important roles in ciliary retrograde transport. It has been reported that disruption of Wdr34 in mouse model results in NTDs and defects of Sonic Hedgehog (SHH) signaling. However, no Wdr60 deficiency mouse model has been reported yet. In this study, piggyBac (PB) transposon is used to interfere Wdr60 and Wdr34 expression respectively to establish Wdr60PB/PB and Wdr34PB/PB mouse models. We found that the expression of Wdr60 or Wdr34 is significantly decreased in the homozygote mice. Wdr60 homozygote mice die around E13.5 to E14.5, while Wdr34 homozygote mice die around E10.5 to E11.5. WDR60 is highly expressed in the head region at E10.5 and Wdr60PB/PB embryos have head malformation. RNAseq and qRT-PCR experiments revealed that Sonic Hedgehog signaling is also downregulated in Wdr60PB/PB head tissue, demonstrating that WDR60 is also required for promoting SHH signaling. Further experiments on mouse embryos also revealed that the expression levels of planar cell polarity (PCP) components such as CELSR1 and downstream signal molecule c-Jun were downregulated in WDR34 homozygotes compared to wildtype littermates. Coincidently, we observed much higher ratio of open cranial and caudal neural tube in Wdr34PB/PB mice. CO-IP experiment showed that WDR60 and WDR34 both interact with IFT88, but only WDR34 interacts with IFT140. Taken together, WDR60 and WDR34 play overlapped and distinct functions in modulating neural tube development. |
| first_indexed | 2024-04-09T13:44:20Z |
| format | Article |
| id | doaj.art-ed0d8f1eba924e909a46a488ee18f730 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-04-09T13:44:20Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-ed0d8f1eba924e909a46a488ee18f7302023-05-09T05:45:16ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2023-05-011110.3389/fcell.2023.10842451084245Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryosLu Yan0Lu Yan1Lu Yan2Hailing Yin3Hailing Yin4Yiwei Mi5Yiwei Mi6Yu Wu7Yu Wu8Yufang Zheng9Yufang Zheng10Yufang Zheng11Obstetrics and Gynecology Hospital, The Institute of Obstetrics and Gynecology, Fudan University, Shanghai, ChinaDepartment of Cellular and Developmental Biology, School of Life Sciences, Fudan University, Shanghai, ChinaState Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, ChinaObstetrics and Gynecology Hospital, The Institute of Obstetrics and Gynecology, Fudan University, Shanghai, ChinaObstetrics Department of the First Affiliated Hospital with Nanjing Medical University, Nanjing, Jiangsu, ChinaDepartment of Cellular and Developmental Biology, School of Life Sciences, Fudan University, Shanghai, ChinaState Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, ChinaDepartment of Cellular and Developmental Biology, School of Life Sciences, Fudan University, Shanghai, ChinaState Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, ChinaObstetrics and Gynecology Hospital, The Institute of Obstetrics and Gynecology, Fudan University, Shanghai, ChinaDepartment of Cellular and Developmental Biology, School of Life Sciences, Fudan University, Shanghai, ChinaState Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University, Shanghai, ChinaCilia are specialized organelles that extend from plasma membrane, functioning as antennas for signal transduction and are involved in embryonic morphogenesis. Dysfunction of cilia lead to many developmental defects, including neural tube defects (NTDs). Heterodimer WDR60-WDR34 (WD repeat domain 60 and 34) are intermediate chains of motor protein dynein-2, which play important roles in ciliary retrograde transport. It has been reported that disruption of Wdr34 in mouse model results in NTDs and defects of Sonic Hedgehog (SHH) signaling. However, no Wdr60 deficiency mouse model has been reported yet. In this study, piggyBac (PB) transposon is used to interfere Wdr60 and Wdr34 expression respectively to establish Wdr60PB/PB and Wdr34PB/PB mouse models. We found that the expression of Wdr60 or Wdr34 is significantly decreased in the homozygote mice. Wdr60 homozygote mice die around E13.5 to E14.5, while Wdr34 homozygote mice die around E10.5 to E11.5. WDR60 is highly expressed in the head region at E10.5 and Wdr60PB/PB embryos have head malformation. RNAseq and qRT-PCR experiments revealed that Sonic Hedgehog signaling is also downregulated in Wdr60PB/PB head tissue, demonstrating that WDR60 is also required for promoting SHH signaling. Further experiments on mouse embryos also revealed that the expression levels of planar cell polarity (PCP) components such as CELSR1 and downstream signal molecule c-Jun were downregulated in WDR34 homozygotes compared to wildtype littermates. Coincidently, we observed much higher ratio of open cranial and caudal neural tube in Wdr34PB/PB mice. CO-IP experiment showed that WDR60 and WDR34 both interact with IFT88, but only WDR34 interacts with IFT140. Taken together, WDR60 and WDR34 play overlapped and distinct functions in modulating neural tube development.https://www.frontiersin.org/articles/10.3389/fcell.2023.1084245/fullWdr60WDR34ciliaSHH signalingPCP signaling |
| spellingShingle | Lu Yan Lu Yan Lu Yan Hailing Yin Hailing Yin Yiwei Mi Yiwei Mi Yu Wu Yu Wu Yufang Zheng Yufang Zheng Yufang Zheng Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryos Frontiers in Cell and Developmental Biology Wdr60 WDR34 cilia SHH signaling PCP signaling |
| title | Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryos |
| title_full | Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryos |
| title_fullStr | Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryos |
| title_full_unstemmed | Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryos |
| title_short | Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryos |
| title_sort | deficiency of wdr60 and wdr34 cause distinct neural tube malformation phenotypes in early embryos |
| topic | Wdr60 WDR34 cilia SHH signaling PCP signaling |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2023.1084245/full |
| work_keys_str_mv | AT luyan deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT luyan deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT luyan deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT hailingyin deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT hailingyin deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT yiweimi deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT yiweimi deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT yuwu deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT yuwu deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT yufangzheng deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT yufangzheng deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos AT yufangzheng deficiencyofwdr60andwdr34causedistinctneuraltubemalformationphenotypesinearlyembryos |