Prenatal alcohol exposure disrupts Sonic hedgehog pathway and primary cilia genes in the mouse neural tube

• Published in: Reproductive toxicology (Elmsford, N.Y.) N.Y.), 2021-10, Vol.105, p.136-147
• Main Authors: Boschen, Karen E., Fish, Eric W., Parnell, Scott E.
• Format: Article
• Language: English
• Subjects: Animals; Behavior, Animal; Cell cycle; Cell Cycle - genetics; Cilia - genetics; Development; Ethanol - adverse effects; Female; Fetal alcohol spectrum disorders; Gene Expression Regulation, Developmental; Hedgehog Proteins - genetics; Kif3a; Kinesins - genetics; Male; Maternal-Fetal Exchange; Mice; Mice, Inbred C57BL; Mice, Transgenic; Neural Tube - metabolism; Neurulation; Pregnancy; Prenatal Exposure Delayed Effects - genetics
• ISSN: 0890-6238; 1873-1708
• DOI: 10.1016/j.reprotox.2021.09.002

•Neurulation-stage alcohol exposure causes midline craniofacial and CNS defects.•We measured the consequences of prenatal alcohol on the neural tube.•Alcohol reduced neural tube volume and cell layer width.•Alcohol also altered Shh pathway, cilia, and cell cycle gene expression.•Knockdown of a cilia gene exacerbated prenatal alcohol effects on adolescent behavior. Neurulation-stage alcohol exposure (NAE; embryonic day [E] 8–10) is associated with midline craniofacial and CNS defects that likely arise from disruption of morphogen pathways, such as Sonic hedgehog (Shh). Notably, midline anomalies are also a hallmark of genetic ciliopathies such as Joubert syndrome. We tested whether NAE alters Shh pathway signaling and the number and function of primary cilia, organelles critical for Shh pathway transduction. Female C57BL/6 J mice were administered two doses of alcohol (2.9 g/kg/dose) or vehicle on E9. Embryos were collected 6, 12, or 24 h later, and changes to Shh, cell cycle genes, and primary cilia were measured in the rostroventral neural tube (RVNT). Within the first 24 h post-NAE, reductions in Shh pathway and cell cycle gene expression and the ratio of Gli3 forms in the full-length activator state were observed. RVNT volume and cell layer width were reduced at 12 h. In addition, altered expression of multiple cilia-related genes was observed at 6 h post-NAE. As a further test of cilia gene-ethanol interaction, mice heterozygous for Kif3a exhibited perturbed behavior during adolescence following NAE compared to vehicle-treated mice, and Kif3a heterozygosity exacerbated the hyperactive effects of NAE on exploratory activity. These data demonstrate that NAE downregulates the Shh pathway in a region of the neural tube that gives rise to alcohol-sensitive brain structures and identifies disruption of primary cilia function, or a “transient ciliopathy”, as a possible cellular mechanism of prenatal alcohol pathogenesis.