DOI: 10.5061/dryad.j3tx95xs9
title: Developmental exposures to three mammalian teratogens produces dysmorphic phenotypes in adult Caenorhabditis elegans.

description: Adult morphology was assessed at sublethal concentrations in C. elegans
 using three mammalian teratogens, 5-fluorouracil (5FU), hydroxyurea (HU),
 or ribavirin (RV). C. elegans develop from egg to adult with internal eggs
 in about 70h at 20°C in C. elegans Habitation Medium (CeHM). Cohorts were
 exposed from first feeding after hatching for either 24h (early-only) or
 continuously for four days. Significant increases in the percentage of
 prolapsed individuals were seen on the first day of adulthood in both
 continuous and early-only 5FU and HU exposure groups. In contrast,
 continuous RV exposures induced gonad arm and vulval anomalies that were
 largely but not completely reversed in the early-only exposure groups.
 Reductions in reproductive output with continuous exposures recovered,
 though not fully, with early-only exposures for all three chemicals. To
 our knowledge, this is the first systematic evaluation of C. elegans
 morphology changes induced by mammalian teratogens. These data suggest
 that C. elegans may be a useful model for detecting chemicals with human
 teratogenic potential, but further studies using larger chemical test
 panels are needed.
description: The <em>C. elegans</em> wild-type N2 strain was
 obtained from the <em>Caenorhabditis</em> Genetics Center,
 which is funded by NIH Office of Research Infrastructure Programs (P40
 OD010440). Continuously well-fed cultures were maintained and exposed in
 CeHM as previously described (DOI: 10.1002/jat.4525). Sublethal
 concentrations that allowed for at least semi-synchronous developmental
 timing within exposed cohorts were previously determined (DOI:
 10.1016/j.taap.2024.117108). For continuously exposed <em>C.
 elegans</em>, synchronized newly hatched cohorts were counted and
 diluted to approximately 1,000 first larval stage (L1) worms per mL CeHM,
 then 900 µL worms in CeHM were transferred to 12-well plates and 100 µL of
 10x aqueous dosing solution was added per well within 30 min
 post-first-feeding after hatching. Plates were maintained in 20°C
 incubators for four days. For early-only exposures, the following
 alterations were made: after 24.5h +/-30min exposures, <em>C.
 elegans</em> cohorts were quickly washed first in 14mL water, then
 in 15mL M9 buffer, for a total of no more than 10 min without food, then
 resuspended in 1mL fresh CeHM per well and returned incubators for an
 additional 3 days. Four independent experiments were conducted, each
 consisting of a fresh preparation of dosing solutions and <em>C.
 elegans</em> L1 cohorts. On day four post-L1 feeding and dosing when
 control populations were adults with early-stage progeny, <em>C.
 elegans</em> populations were immobilized for imaging with sodium
 azide directly in CeHM. Washing and pipetting onto slides was not done due
 to the fragility of some phenotypes. Plates were transferred to a Keyence
 (Itasca, IL, USA) automated stage microscope for image capture with a 4x
 objective of the whole well and 10x objective of the center of the
 well.
description: # Developmental exposures to three mammalian teratogens produces
 dysmorphic phenotypes in adult Caenorhabditis elegans Dataset DOI:
 [10.5061/dryad.j3tx95xs9](10.5061/dryad.j3tx95xs9) ## Description of the
 data and file structure As required by Dryad, this image data set is
 submitted to the public domain without restrictions or conditions for
 reuse, modification, or redistribution. 5-fluorouracil (5FU), hydroxyurea
 (HU), and ribavirin (RV) are mammalian teratogens that also cause growth
 retardation in mammals and toxicological model organisms, including *C.
 elegans*. The effects of 5FU, HU, and RV on *C. elegans* adult morphology
 were evaluated after continuous and 24h (early-only) exposures beginning
 from first feeding after hatching.  Data: the images are .jpg files,
 organized in folders by chemical, experiment number, exposure duration,
 and exposure concentration. Data description:* C. elegans* cohorts were
 exposed to sublethal concentrations of three mammalian teratogens from
 first feeding after hatching for either 24h (early-only, wDATe) or four
 days (continuous, wDATc). Culture maintenance and exposures were in *C.
 elegans* Habitation Medium (CeHM) in 12-well plates in 20°C incubators
 without shaking. Four independent experiments were conducted with
 randomized well placement in 12-well plates. Microscopy images for later
 phenotypic analysis were captured on day four post-initiation of L1
 feeding when controls were adults with young progeny. Worms were
 immobilized with sodium azide and imaged directly in CeHM nutrient medium
 to avoid breakup of fragile phenotypes. A Keyence (Itasca, IL, USA)
 automated stage microscope was used for image capture of one well per
 condition per experiment with a 4x objective, and the center of each well
 with a 10x objective. The images are .jpg files, organized in folders
 (5-fluorouracil.zip, hydroxyurea.zip and ribavirin.zip) by chemical
 (5-fluorouracil, hydroxyurea, or ribavirin), experiment number (1-4), and
 exposure concentration in micrograms per mL. The
 Example_Images_Originals.zip folder contains the original full images used
 as examples in figures in manuscript toxics-3687520. These images were
 edited and cropped for clarity in the manuscript and made available here
 as originals for full transparency ### Files and variables #### File:
 5FU_ex1_wDATc_4x **Description:** *C. elegans* continuously exposed for 4
 days from first feeding after hatching (ffah) to 5-fluorouracil.
 Experiment 1, 4x images captured on day 4 post-ffah. #### File:
 5FU_ex1_wDATc_10x **Description:** *C. elegans* continuously exposed for 4
 days from first feeding after hatching (ffah) to 5-fluorouracil.
 Experiment 1, 10x images captured on day 4 post-ffah. #### File:
 5FU_ex1_wDATe_4x **Description:** *C. elegans* exposed for 24h from first
 feeding after hatching (ffah) to 5-fluorouracil. Experiment 1, 4x images
 captured on day 4 post-ffah. #### File: 5FU_ex1_wDATe_10x
 **Description:** *C. elegans* exposed for 24h from first feeding after
 hatching (ffah) to 5-fluorouracil. Experiment 1, 10x images captured on
 day 4 post-ffah. #### File: 5FU_ex2_wDATc **Description:** *C. elegans*
 continuously exposed for 4 days from first feeding after hatching (ffah)
 to 5-fluorouracil. Experiment 2, 4x & 10x images captured on day 4
 post-ffah. #### File: 5FU_ex2_wDATe **Description:** *C. elegans* exposed
 for 24h from first feeding after hatching (ffah) to 5-fluorouracil.
 Experiment 2, 4x & 10x images captured on day 4 post-ffah. #### File:
 5FU_ex3_wDATc **Description:** *C. elegans* continuously exposed for 4
 days from first feeding after hatching (ffah) to 5-fluorouracil.
 Experiment 3, 4x & 10x images captured on day 4 post-ffah. #### File:
 5FU_ex3_wDATe **Description:** *C. elegans* exposed for 24h from first
 feeding after hatching (ffah) to 5-fluorouracil. Experiment 3, 4x &
 10x images captured on day 4 post-ffah. #### File: 5FU_ex4_wDATc
 **Description:** *C. elegans* continuously exposed for 4 days from first
 feeding after hatching (ffah) to 5-fluorouracil. Experiment 4, 4x &
 10x images captured on day 4 post-ffah. #### File: 5FU_ex4_wDATe
 **Description:** *C. elegans* exposed for 24h from first feeding after
 hatching (ffah) to 5-fluorouracil. Experiment 4, 4x & 10x images
 captured on day 4 post-ffah. #### File: HU_ex1_wDATc **Description:** *C.
 elegans* continuously exposed for 4 days from first feeding after hatching
 (ffah) to hydroxyurea. Experiment 1, 4x & 10x images captured on day 4
 post-ffah. #### File: HU_ex1_wDATe **Description:** *C. elegans* exposed
 for 24h from first feeding after hatching (ffah) to hydroxyurea.
 Experiment 1, 4x & 10x images captured on day 4 post-ffah. #### File:
 HU_ex2_wDATc **Description:** *C. elegans* continuously exposed for 4 days
 from first feeding after hatching (ffah) to hydroxyurea. Experiment 2, 4x
 & 10x images captured on day 4 post-ffah. #### File: HU_ex2_wDATe
 **Description:** *C. elegans* exposed for 24h from first feeding after
 hatching (ffah) to hydroxyurea. Experiment 2, 4x & 10x images captured
 on day 4 post-ffah. #### File: HU_ex3_wDATc **Description:** *C. elegans*
 continuously exposed for 4 days from first feeding after hatching (ffah)
 to hydroxyurea. Experiment 3, 4x & 10x images captured on day 4
 post-ffah. #### File: HU_ex3_wDATe **Description:** *C. elegans* exposed
 for 24h from first feeding after hatching (ffah) to hydroxyurea.
 Experiment 3, 4x & 10x images captured on day 4 post-ffah. #### File:
 HU_ex4_wDATc **Description:** *C. elegans* continuously exposed for 4 days
 from first feeding after hatching (ffah) to hydroxyurea. Experiment 4, 4x
 & 10x images captured on day 4 post-ffah. #### File: HU_ex4_wDATe
 **Description:** *C. elegans* exposed for 24h from first feeding after
 hatching (ffah) to hydroxyurea. Experiment 4, 4x & 10x images captured
 on day 4 post-ffah. #### File: RV_ex1_wDATc **Description:** *C. elegans*
 continuously exposed for 4 days from first feeding after hatching (ffah)
 to ribavirin. Experiment 1, 4x & 10x images captured on day 4
 post-ffah. #### File: RV_ex1_wDATe **Description:** *C. elegans* exposed
 for 24h from first feeding after hatching (ffah) to ribavirin. Experiment
 1, 4x & 10x images captured on day 4 post-ffah. #### File:
 RV_ex2_wDATc **Description:** *C. elegans* continuously exposed for 4 days
 from first feeding after hatching (ffah) to ribavirin. Experiment 2, 4x
 & 10x images captured on day 4 post-ffah. #### File: RV_ex2_wDATe
 **Description:** *C. elegans* exposed for 24h from first feeding after
 hatching (ffah) to ribavirin. Experiment 2, 4x & 10x images captured
 on day 4 post-ffah. #### File: RV_ex3_wDATc **Description:** *C. elegans*
 continuously exposed for 4 days from first feeding after hatching (ffah)
 to ribavirin. Experiment 3, 4x & 10x images captured on day 4
 post-ffah. #### File: RV_ex3_wDATe **Description:** *C. elegans* exposed
 for 24h from first feeding after hatching (ffah) to ribavirin. Experiment
 3, 4x & 10x images captured on day 4 post-ffah. #### File:
 RV_ex4_wDATc **Description:** *C. elegans* continuously exposed for 4 days
 from first feeding after hatching (ffah) to ribavirin. Experiment 4, 4x
 & 10x images captured on day 4 post-ffah. #### File: RV_ex4_wDATe
 **Description:** *C. elegans* exposed for 24h from first feeding after
 hatching (ffah) to ribavirin. Experiment 4, 4x & 10x images captured
 on day 4 post-ffah. ## Code/software Files contain images. Any software
 program compatible with .jpg files will allow viewing of images. ## Access
 information Other publicly accessible locations of the data: * NA Data was
 derived from the following sources: * NA