Vandenberg et al. Low Frequency Vibrations Induce Malformations in Aquatic Species
Low Frequency Vibrations Induce Malformations in Two Aquatic Species in a Frequency-, Waveform-, and Direction-Specific Manner
Laura N. Vandenberg, Claire Stevenson, Michael Levin
Abstract
Environmental toxicants such as industrial wastes, air particulates from machinery and transportation vehicles, and pesticide run-offs, as well as many chemicals, have been widely studied for their effects on human and wildlife populations. Yet other potentially harmful environmental pollutants such as electromagnetic pulses, noise and vibrations have remained incompletely understood.
Because developing embryos undergo complex morphological changes that can be affected detrimentally by alterations in physical forces, they may be particularly susceptible to exposure to these types of pollutants.
We investigated the effects of low frequency vibrations on early embryonic development of two aquatic species, Xenopus laevis (frogs) and Danio rerio (zebrafish), specifically focusing on the effects of varying frequencies, waveforms, and applied direction. We observed treatment-specific effects on the incidence of neural tube defects, left-right patterning defects and abnormal tail morphogenesis in Xenopus tadpoles. Additionally, we found that low frequency vibrations latered left-right patterning and tail morphogenesis, but did not induce neural tube defects, in zebrafish.
The results of this study support the conclusion that low frequency vibrations are toxic to aquatic vertebrates, with detrimental effects observed in two important model species with different embryonoic architectures.
Introduction
For several decades, the field of environmental toxicology has been expanding its focus from identifying toxicants and determining their sources of exposure, to assessing the effects of these toxicants on target and non-target species, and determining their mechanisms of action. A large amount of attention has been given to chemicals found in the environment that may be affecting wildlife species including mutagens, carcinogens, and reproductive/developmental toxicants; some of these chemicals have been proposed as contributors to the decline in amphibian populations that has been observed [1,2]. However, there are other environmental perturbations that have received less attention including the effects of noise, electromagnetic fields, and low frequency vibrations. Should these environmental factors be considered developmental toxicants?
In many species, the period encompassing organogenesis is most susceptible to disruption by environmental toxicants because alterations in developmental processes during these stages can have permanent results [3]. Embryogenesis is marked by many complex morphological changes, regardless of whether embryos develop externally (i.e. fish, frogs), in ovo (i.e. chicks, reptiles), or in utero (i.e. mammals) [4,5,6,7]. We propose that because low frequency vibrations can disrupt the cytoskeleton of treated cells [8,9], they alter morphogenesis of the developing embryo and are therefore toxic. We previously examined the effects of low frequency vibrations on Xenopus laevis frog embryos. Xenopus
embryos exposed to low frequency vibrations (,250 Hz) displayed increased rates of heterotaxia, the randomized placement of visceral organs along the left-right (LR) axis [10]. We found that these patterning defects were due to altered cytoplasmic/ cytoskeletal dynamics and tight junctional connections between cells of the early embryo. We also noted that one frequency (15 Hz) additionally produced neural tube defects, suggesting that these two phenotypes (heterotaxia and spina bifida) may have a common intracellular etiology. We thus hypothesized that low frequency vibrations could produce a range of other developmen- tal defects, and could thus be used as a convenient experimental perturbation targeting developmental processes that depend on the cytoskeleton and cell:cell communication.
Here, we have examined the effects of low frequency vibrations on two aquatic species, Xenopus laevis and Danio rerio (zebrafish). These two species were selected because they have different early embryonic architectures and different types of early cell cleavages, and are both widely used as important laboratory models for understanding environmental contaminants, pattern formation and embryogenesis [5,7,11,12,13]. Additional advantages are that they are transparent at early stages, allowing for easy scoring of phenotypes; they develop quickly and require relatively little care; and their embryos are available in large numbers allowing for many variables and endpoints to be tested. We focused our analyses on the effects of vibration on LR patterning, neural tube defects, and tail morphogenesis. These endpoints were chosen because prior data suggested that these examples of large-scale pattern formation were especially sensitive to vibration [10], they are early developmental events that are thought to require the cytoskeleton, intracellular communication, and cell-cell contact [14,15,16,17], and they may have common molecular etiologies, as human patients with ZIC3 mutations often have both abnormal LR patterning and neural tube defects [18]. Because vibrations in the environment are likely to vary, we specifically examined the effects of different frequencies, waveforms, and the direction of applied vibration on these endpoints. In Xenopus embryos, we found different effects of vibration frequency, waveform, and applied direction on all three endpoints examined. In zebrafish embryos, we found similar effects of vibration on two endpoints, LR patterning and tail morphogenesis, suggesting that these patterning events may be susceptible to vibrations across a wider range of species.
Citation: Vandenberg LN, Stevenson C, Levin M (2012) Low Frequency Vibrations Induce Malformations in Two Aquatic Species in a Frequency-, Waveform-, and Direction-Specific Manner. PLoS ONE 7(12): e51473. doi:10.1371/journal.pone.0051473
Editor: Yann Gibert, Deakin School of Medicine, Australia
Received June 14, 2012; Accepted November 6, 2012; Published December 10, 2012
Copyright: © 2012 Vandenberg et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by American Heart Association Established Investigator Grant 0740088N and NIH grant R01-GM077425 to ML, and NRSA grant 1F32GM087107 to LNV. The funders had no additional role in the study design, data analysis, or decision to submit this manuscript.
Competing Interests: The authors have declared that no competing interests exist.
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