Biology subjects

Lovely, C. B.

Publications and source records attributed to Lovely, C. B..

4 recordsLinked to original sources

bioRxiv2025

Interactions between the Bone Morphogenetic Protein and the Planar Cell Polarity Pathways lead to distinctive ethanol-induced facial defects

BackgroundFetal Alcohol Spectrum Disorders (FASD) describes a spectrum of ethanol-induced neural and facial developmental defects. Ethanol susceptibility is modulated by genetics, but their underlying mechanisms remain poorly understood. In all vertebrates, a series of complex cellular events give rise to the body plan, including convergence & extension (C&E) and endoderm/ cranial neural crest (CNC-which gives rise to the facial skeleton) morphogenesis. These events are critical to establish complex signaling interactions, driving embryo development, including the facial skeleton. In zebrafish, C&E occurs between 6-10 hpf while endoderm/CNC morphogenesis occurs 10-24 hpf. Previous work shows that the PCP mutants are sensitive to ethanol from 6-24 hpf, covering both C&E and endoderm/CNC morphogenesis and exhibiting multiple defects to the forming head raising the question whether ethanol during both time windows drives PCP-ethanol defects. We hypothesize that PCP single and double mutants are ethanol sensitive 10-24 hpf, after C&E. We also hypothesize BMP signaling (sensitive 10-18 hpf) interacts with and sensitizes the PCP pathway to ethanol. MethodsHere, we treated PCP/BMP mutants with ethanol from 6-10, 10-18, 10-24 or 24-30 hpf and combined morphometric and linear measurements to examine facial development. ResultsWe show that PCP mutant larvae are ethanol-sensitive from 10-24 hpf, but not 6-10 or 24-30 hpf. We also show that BMP mutants sensitize PCP mutants to ethanol and lead to novel ethanol-independent midline craniofacial defects. Our results suggest that the ethanol-sensitive role of PCP pathway occurs after C&E, during endoderm/CNC morphogenesis and that the PCP and BMP pathways genetically interact during the morphogenesis events. ConclusionsUltimately, our work builds on a mechanistic paradigm of ethanol-induced birth defects we have been developing, connecting conceptual framework with concrete cellular events that could be ethanol-sensitive beyond facial development.

developmental biology↗

bioRxiv2025

Ethanol induces craniofacial defects in Bmp mutants independent of nkx2.3 by elevating cranial neural crest cell apoptosis

BackgroundFetal Alcohol Spectrum Disorders (FASD) describes a wide range of neurological defects and craniofacial malformations associated with prenatal ethanol exposure. While there is growing evidence for a genetic component to FASD, little is known of the cellular mechanisms underlying these ethanol-sensitive loci in facial development. Endoderm morphogenesis to form lateral protrusions called pouches is one key mechanism in facial development. We have previously shown that multiple members of the Bone Morphogenetic Pathway (Bmp) signaling pathway, a key regulator of pouch formation, interacts with ethanol disrupting facial development. However, ethanol does not directly impact Bmp signaling suggesting that downstream effectors, like nkx2.3 may mediate the impact of ethanol on Bmp mutants. MethodsHere we use an ethanol exposure paradigm with nkx2.3 knockdown approaches to test if loss of nkx2.3 sensitizes Bmp mutants to ethanol induced facial defects. We then combine a morphometric approach with Hybridization Chain Reaction and immunofluorescence to examine the cellular mechanisms underlying Bmp-ethanol interactions. ResultsWe show that Bmp-ethanol interactions alter morphology of the endodermal pouches, independent of nkx2.3 gene expression. Morpholino knock down of nkx2.3 does not sensitize wild type or bmp4 mutant larvae to ethanol-induced facial defects. However, we did observe a significant increase CNCC apoptosis in ethanol-treated Bmp mutants. ConclusionsCollectively, our results suggest that ethanols mode of action is independent of downstream Bmp effectors, converging on CNCC cell survival. Ultimately, our work provides a mechanistic paradigm of ethanol-induced facial defects and connects ethanol exposure with concrete cellular events.

developmental biology↗

bioRxiv2024

Bone Morphogenetic Protein signaling pathway - ethanol interactions disrupt palate formation independent of gata3

Fetal Alcohol Spectrum Disorders (FASD) describes a wide array of neurological defects and craniofacial malformations, associated with ethanol teratogenicity. While there is growing evidence for a genetic component to FASD, little is known of the genes underlying these ethanol-induced defects. Along with timing and dosage, genetic predispositions may help explain the variability within FASD. From a screen for gene-ethanol interactions, we found that mutants for Bmp signaling components are ethanol-sensitive leading to defects in the zebrafish palate. Loss of Bmp signaling results in reductions in gata3 expression in the maxillary domain of the neural crest in the 1st pharyngeal arch, leading to palate defects while upregulation of human GATA3 rescues these defects. Here, we show that ethanol-treated Bmp mutants exhibit misshaped and/or broken trabeculae. Surprisingly, up regulation of GATA3 does not rescue ethanol-induced palate defects and gata3 expression was not altered in ethanol-treated Bmp mutants or dorsomorphin-treated larvae. Timing of ethanol sensitivity shows that Bmp mutants are ethanol sensitive from 10-18 hours post-fertilization (hpf), prior to Bmps regulation of gata3 in palate formation. This is consistent with our previous work with dorsomorphin-dependent knock down of Bmp signaling from 10-18 hpf disrupting endoderm formation and subsequent jaw development. Overall, this suggests that ethanol disrupts Bmp-dependent palate development independent of and earlier than the role of gata3 in palate formation by disrupting epithelial development. Ultimately, these data demonstrate that zebrafish is a useful model to identify and characterize gene-ethanol interactions and this work will directly inform our understanding of FASD. HighlightsO_LIBmp pathway mutants are ethanol sensitive resulting in palate defects. C_LIO_LIEthanol disrupts Bmp-dependent palate development independent of gata3. C_LIO_LITiming of ethanol sensitivity suggests ethanol disrupts Bmp-dependent epithelial morphogenesis. C_LI

developmental biology↗

bioRxiv2022

Next-generation plasmids for transgenesis in zebrafish and beyond

Transgenesis is an essential technique for any genetic model. Tol2-based transgenesis paired with Gateway-compatible vector collections has transformed zebrafish transgenesis with an accessible, modular system. Here, we established several next-generation transgenesis tools for zebrafish and other species to expand and enhance transgenic applications. To facilitate gene-regulatory element testing, we generated Gateway middle entry vectors harboring the small mouse beta-globin minimal promoter coupled to several fluorophores, CreERT2, and Gal4. To extend the color spectrum for transgenic applications, we established middle entry vectors encoding the bright, blue-fluorescent protein mCerulean and mApple as an alternative red fluorophore. We present a series of p2A peptide-based 3 vectors with different fluorophores and subcellular localizations to co-label cells expressing proteins of interest. Lastly, we established Tol2 destination vectors carrying the zebrafish exorh promoter driving different fluorophores as a pineal gland-specific transgenesis marker active prior to hatching and through adulthood. exorh-based reporters and transgenesis markers also drive specific pineal gland expression in the eye-less cavefish (Astyanax). Together, our vectors provide versatile reagents for transgenesis applications in zebrafish, cavefish, and other models.

genetics↗