On-chip immobilization of planarians for in vivo imaging

Dexter, Joseph P.; Tamme, Mary B.; Lind, Christine H.; Collins, Eva‐Maria S.

doi:10.1038/srep06388

Cited by 21 publications

(15 citation statements)

References 45 publications

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“…A stock solution (1.9 mM) was diluted 1:1000 (0.19 uM) in Poland Springs water, and worms were soaked in the DiBAC solution for >30 min before imaging. Worms were then immobilized in 2% low-meting point agarose, using custom-fabricated Planarian Immobilization Chips as in [ 174 ]. Images of the ventral side of immobilized planaria were captured with the Nikon AZ100 Stereomicroscope, Melville, NY, USA, using epifluorescence optics, and NIS-Elements imaging software (Melville, NY, USA).…”

Section: Methodsmentioning

confidence: 99%

Gap Junctional Blockade Stochastically Induces Different Species-Specific Head Anatomies in Genetically Wild-Type Girardia dorotocephala Flatworms

Emmons-Bell

Durant

Hammelman

et al. 2015

IJMS

117

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The shape of an animal body plan is constructed from protein components encoded by the genome. However, bioelectric networks composed of many cell types have their own intrinsic dynamics, and can drive distinct morphological outcomes during embryogenesis and regeneration. Planarian flatworms are a popular system for exploring body plan patterning due to their regenerative capacity, but despite considerable molecular information regarding stem cell differentiation and basic axial patterning, very little is known about how distinct head shapes are produced. Here, we show that after decapitation in G. dorotocephala, a transient perturbation of physiological connectivity among cells (using the gap junction blocker octanol) can result in regenerated heads with quite different shapes, stochastically matching other known species of planaria (S. mediterranea, D. japonica, and P. felina). We use morphometric analysis to quantify the ability of physiological network perturbations to induce different species-specific head shapes from the same genome. Moreover, we present a computational agent-based model of cell and physical dynamics during regeneration that quantitatively reproduces the observed shape changes. Morphological alterations induced in a genomically wild-type G. dorotocephala during regeneration include not only the shape of the head but also the morphology of the brain, the characteristic distribution of adult stem cells (neoblasts), and the bioelectric gradients of resting potential within the anterior tissues. Interestingly, the shape change is not permanent; after regeneration is complete, intact animals remodel back to G. dorotocephala-appropriate head shape within several weeks in a secondary phase of remodeling following initial complete regeneration. We present a conceptual model to guide future work to delineate the molecular mechanisms by which bioelectric networks stochastically select among a small set of discrete head morphologies. Taken together, these data and analyses shed light on important physiological modifiers of morphological information in dictating species-specific shape, and reveal them to be a novel instructive input into head patterning in regenerating planaria.

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Section: Methodsmentioning

confidence: 99%

Gap Junctional Blockade Stochastically Induces Different Species-Specific Head Anatomies in Genetically Wild-Type Girardia dorotocephala Flatworms

Emmons-Bell

Durant

Hammelman

et al. 2015

IJMS

117

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“…Regeneration in planarians proceeds through activation of cell proliferation and application of bioelectric fields are known to modulate repair and tissue polarity in flatworms [127, 128]. Membrane potential across the whole planarian body could be monitored using DiBAC (4) (3) staining, and the newly developed approach termed Planarian Immobilization Chips (PICs) to visualize bioelectrical changes in real time while minimizing tissue damage [13, 122, 129, 130]. …”

Section: 0 Schmidtea Mediterranea and Tmpmentioning

confidence: 99%

Bioelectrical regulation of cell cycle and the planarian model system

Barghouth

Thiruvalluvan

Oviedo

2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Cell cycle regulation through the manipulation of endogenous membrane potentials offers tremendous opportunities to control cellular processes during tissue repair and cancer formation. However, the molecular mechanisms by which biophysical signals modulate the cell cycle remain underappreciated and poorly understood. Cells in complex organisms generate and maintain a constant voltage gradient across the plasma membrane known as the transmembrane potential. This potential, generated through the combined efforts of various ion transporters, pumps and channels, is known to drive a wide range of cellular processes such as cellular proliferation, migration and tissue regeneration while its deregulation can lead to tumorigenesis. These cellular regulatory events, coordinated by ionic flow, correspond to a new and exciting field termed molecular bioelectricity. We aim to present a brief discussion on the biophysical machinery involving membrane potential and the mechanisms mediating cell cycle progression and cancer transformation. Furthermore, we present the planarian Schmidtea mediterranea as a tractable model system for understanding principles behind molecular bioelectricity at both the cellular and organismal level.

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“…) and immobilization techniques (Dexter et al. ) may offer a way around the current impasse. (3) Much more work is needed to unify bioelectric and biochemical signaling.…”

Section: Discussionmentioning

confidence: 99%

Physiological controls of large‐scale patterning in planarian regeneration: a molecular and computational perspective on growth and form

et al. 2016

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Planaria are complex metazoans that repair damage to their bodies and cease remodeling when a correct anatomy has been achieved. This model system offers a unique opportunity to understand how large‐scale anatomical homeostasis emerges from the activities of individual cells. Much progress has been made on the molecular genetics of stem cell activity in planaria. However, recent data also indicate that the global pattern is regulated by physiological circuits composed of ionic and neurotransmitter signaling. Here, we overview the multi‐scale problem of understanding pattern regulation in planaria, with specific focus on bioelectric signaling via ion channels and gap junctions (electrical synapses), and computational efforts to extract explanatory models from functional and molecular data on regeneration. We present a perspective that interprets results in this fascinating field using concepts from dynamical systems theory and computational neuroscience. Serving as a tractable nexus between genetic, physiological, and computational approaches to pattern regulation, planarian pattern homeostasis harbors many deep insights for regenerative medicine, evolutionary biology, and engineering.

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On-chip immobilization of planarians for in vivo imaging

Cited by 21 publications

References 45 publications

Gap Junctional Blockade Stochastically Induces Different Species-Specific Head Anatomies in Genetically Wild-Type Girardia dorotocephala Flatworms

Gap Junctional Blockade Stochastically Induces Different Species-Specific Head Anatomies in Genetically Wild-Type Girardia dorotocephala Flatworms

Bioelectrical regulation of cell cycle and the planarian model system

Physiological controls of large‐scale patterning in planarian regeneration: a molecular and computational perspective on growth and form

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