Ethanol exposure perturbs sea urchin development and disrupts developmental timing

Rodriguez-Sastre, Nahomie; Shapiro, Nicholas; Hawkins, Dakota Y.; Lion, Alexandra T.; Peyreau, Monique; Correa, Andrea E.; Dionne, Kristin; Bradham, Cynthia A.

doi:10.1016/j.ydbio.2022.11.001

Cited by 5 publications

(2 citation statements)

References 73 publications

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“…The stable knockout of ABCB1 in L. pictus (Vyas et al, 2022) presented us with a unique opportunity to develop a powerful drug and toxicant screen at scale. Sea urchins are U.S. Environmental Protection Agency test organisms employed for toxicant screens (Anderson & Phillips, 2016; EPA, 2002) and are also used for studying biochemical and developmental pathways using numerous drugs and inhibitors (Cavalieri & Spinelli, 2015; Duboc et al, 2010; Gupta et al, 2012; Hardin et al, 1992; Patel et al, 1997; Rodríguez‐Sastre et al, 2023; Warner et al, 2016; Wessel & McClay, 1987; Yuh et al, 1998). Until this study it has been difficult to conceive of methods that could be used to systematically study sea urchin embryos at the scale necessary to screen through hundreds or thousands of substrates.…”

Section: Discussionmentioning

confidence: 99%

Semi‐automated, high‐content imaging of drug transporter knockout sea urchin (Lytechinus pictus) embryos

Tjeerdema,

Lee,

Metry

et al. 2023

J Exp Zool Pt B

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A defining feature of sea urchins is their extreme fecundity. Urchins produce millions of transparent, synchronously developing embryos, ideal for spatial and temporal analysis of development. This biological feature has been effectively utilized for ensemble measurement of biochemical changes. However, it has been underutilized in imaging studies, where single embryo measurements are used. Here we present an example of how stable genetics and high content imaging, along with machine learning‐based image analysis, can be used to exploit the fecundity and synchrony of sea urchins in imaging‐based drug screens. Building upon our recently created sea urchin ABCB1 knockout line, we developed a high‐throughput assay to probe the role of this drug transporter in embryos. We used high content imaging to compare accumulation and toxicity of canonical substrates and inhibitors of the transporter, including fluorescent molecules and antimitotic cancer drugs, in homozygous knockout and wildtype embryos. To measure responses from the resulting image data, we used a nested convolutional neural network, which rapidly classified embryos according to fluorescence or cell division. This approach identified sea urchin embryos with 99.8% accuracy and determined two‐cell and aberrant embryos with 96.3% and 89.1% accuracy, respectively. The results revealed that ABCB1 knockout embryos accumulated the transporter substrate calcein 3.09 times faster than wildtypes. Similarly, knockouts were 4.71 and 3.07 times more sensitive to the mitotic poisons vinblastine and taxol. This study paves the way for large scale pharmacological screens in the sea urchin embryo.

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Section: Discussionmentioning

confidence: 99%

Semi‐automated, high‐content imaging of drug transporter knockout sea urchin (Lytechinus pictus) embryos

Tjeerdema,

Lee,

Metry

et al. 2023

J Exp Zool Pt B

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“…Orientation defects about each axis score the large-scale relationships between the skeletal components and are illustrated in Fig. 1 and in our prior work (Piacentino et al, 2016b;Rodriguez-Sastre et al, 2023). Briefly, AP defects reflect abnormally wide or narrow angles between left and right ARs (and BRs), DV defects are reflected by DVCs that are not parallel to each other or perpendicular to the AP axis, and LR defects are indicated by ARs that are oppositely directed.…”

Section: Morphological Imaging and Skeletal Scoringmentioning

confidence: 94%

Voltage-gated sodium channel activity mediates sea urchin larval skeletal patterning through spatial regulation of Wnt5 expression

Thomas

Hawkins

Skidanova

et al. 2022

Preprint

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Defining pattern formation mechanisms during embryonic development is important for understanding the etiology of birth defects and to inform tissue engineering approaches. In this study, we used tricaine, a voltage-gated sodium channel (VGSC) inhibitor, to show that VGSC activity is required for normal skeletal patterning in Lytechinus variegatus sea urchin larvae. We demonstrate that tricaine-mediated patterning defects are rescued by an anesthetic-insensitive version of the VGSC LvScn5a. This channel is expressed in the posterior ventrolateral ectoderm where it spatially overlaps with Wnt5. We show that VGSC activity is required to spatially restrict Wnt5 expression to this ectodermal region that is adjacent and instructive to clusters of primary mesenchymal cells (PMCs) where secretion of the larval skeleton is initiated as triradiates. Tricaine-mediated Wnt5 spatial expansion correlates with the formation of ectopic PMC clusters and triradiates. These tricaine-mediated defects are rescued by Wnt5 knock down, indicating that the spatial expansion Wnt5 is responsible for the patterning defects induced by VGSC inhibition. These results demonstrate a novel connection between bioelectrical status and the spatial control of patterning cue expression during embryonic pattern formation.

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Toxicity of organic solvents and surfactants to the sea urchin embryos

Semenova,

Kuptsova,

Semenov

2024

Chemosphere

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Ethanol exposure perturbs sea urchin development and disrupts developmental timing

Cited by 5 publications

References 73 publications

Semi‐automated, high‐content imaging of drug transporter knockout sea urchin (Lytechinus pictus) embryos

Semi‐automated, high‐content imaging of drug transporter knockout sea urchin (Lytechinus pictus) embryos

Voltage-gated sodium channel activity mediates sea urchin larval skeletal patterning through spatial regulation of Wnt5 expression

Toxicity of organic solvents and surfactants to the sea urchin embryos

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