AUTHOR=Yan Lu , Yin Hailing , Mi Yiwei , Wu Yu , Zheng Yufang 

TITLE=Deficiency of Wdr60 and Wdr34 cause distinct neural tube malformation phenotypes in early embryos

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=11

YEAR=2023

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2023.1084245

DOI=10.3389/fcell.2023.1084245

ISSN=2296-634X

ABSTRACT=<p>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 <italic>WDR60</italic>-<italic>WDR34</italic> (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 <italic>Wdr34</italic> in mouse model results in NTDs and defects of Sonic Hedgehog (SHH) signaling. However, no <italic>Wdr60</italic> deficiency mouse model has been reported yet. In this study, piggyBac (PB) transposon is used to interfere <italic>Wdr60</italic> and <italic>Wdr34</italic> expression respectively to establish <italic>Wdr60</italic><sup><italic>PB/PB</italic></sup> and <italic>Wdr34</italic><sup><italic>PB/PB</italic></sup> mouse models. We found that the expression of <italic>Wdr60</italic> or <italic>Wdr34</italic> is significantly decreased in the homozygote mice. <italic>Wdr60</italic> homozygote mice die around E13.5 to E14.5, while <italic>Wdr34</italic> homozygote mice die around E10.5 to E11.5. <italic>WDR60</italic> is highly expressed in the head region at E10.5 and <italic>Wdr60</italic><sup><italic>PB/PB</italic></sup> embryos have head malformation. RNAseq and qRT-PCR experiments revealed that Sonic Hedgehog signaling is also downregulated in <italic>Wdr60</italic><sup><italic>PB/PB</italic></sup> 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 <italic>WDR34</italic> homozygotes compared to wildtype littermates. Coincidently, we observed much higher ratio of open cranial and caudal neural tube in <italic>Wdr34</italic><sup><italic>PB/PB</italic></sup> mice. CO-IP experiment showed that <italic>WDR60</italic> and <italic>WDR34</italic> both interact with IFT88, but only <italic>WDR34</italic> interacts with IFT140. Taken together, <italic>WDR60</italic> and <italic>WDR34</italic> play overlapped and distinct functions in modulating neural tube development.</p>