Live Imaging and Analysis of Muscle Contractions in &lt;em&gt;Drosophila&lt;/em&gt; Embryo

Chandel, Ishita; Baker, Ryan; Nakamura, Naosuke; Panin, V. Y.

doi:10.3791/59404

Cited by 2 publications

(2 citation statements)

References 15 publications

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“…LGC1 RNAi knockdown causes a significant increase in motoneuron stimulationevoked NMJ transmission, but it was not at all clear how this might translate to changes in the overall animal movement function. Muscle contraction can be accurately quantified (Chandel et al, 2019). Drosophila larval body segments contain a defined motor circuit and muscle pattern, with contraction and relaxation generating peristaltic waves that propel the larva through the environment (Fushiki et al, 2016;Gjorgjieva et al, 2013).…”

Section: Resultsmentioning

confidence: 99%

Secreted C-type lectin regulation of neuromuscular junction synaptic vesicle dynamics modulates coordinated movement

Bhimreddy

Rushton

Kopke

et al. 2021

Journal of Cell Science

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The synaptic cleft manifests enriched glycosylation, with structured glycans coordinating signaling between presynaptic and postsynaptic cells. Glycosylated signaling ligands orchestrating communication are tightly regulated by secreted glycan-binding lectins. Using the Drosophila neuromuscular junction (NMJ) as a model glutamatergic synapse, we identify a new Ca2+-binding (C-type) lectin, Lectin-galC1 (LGC1), which modulates presynaptic function and neurotransmission strength. We find that LGC1 is enriched in motoneuron presynaptic boutons and secreted into the NMJ extracellular synaptomatrix. We show that LGC1 limits locomotor peristalsis and coordinated movement speed, with a specific requirement for synaptic function, but not NMJ architecture. LGC1 controls neurotransmission strength by limiting presynaptic active zone (AZ) and postsynaptic glutamate receptor (GluR) aligned synapse number, reducing both spontaneous and stimulation-evoked synaptic vesicle (SV) release, and capping SV cycling rate. During high-frequency stimulation (HFS) mutants have faster synaptic depression and impaired recovery while replenishing depleted SV pools. Although LGC1 removal increases the number of glutamatergic synapses, we find LGC1 null mutants exhibit decreased SV density within presynaptic boutons, particularly SV pools at presynaptic active zones. Thus, LGC1 regulates NMJ neurotransmission to modulate coordinated movement.

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Section: Resultsmentioning

confidence: 99%

Secreted C-type lectin regulation of neuromuscular junction synaptic vesicle dynamics modulates coordinated movement

Bhimreddy

Rushton

Kopke

et al. 2021

Journal of Cell Science

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“…Briefly, in vitro characterization of spontaneous twitching at day 10 in the bulk and 3D bioprinted constructs was obtained by analyzing the single frames of the live-imaging videos. After properly calibrating images, radial displacement during twitching with respect to a reference point of the image was measured using the Straight Line function [25]. Myofibers size (width and length) was calculated on whole-mount immunofluorescence images using the Straight Line function after image calibration, while coherency was calculated by using the Orientation J plugin, as previously described [11].…”

Section: Image Analysesmentioning

confidence: 99%

A novel extrusion-based 3D bioprinting system for skeletal muscle tissue engineering

et al. 2023

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3D bioprinting is an emerging technology, which turned out to be an optimal tool for tissue engineering approaches. To date, different printing systems have been developed. Among them, the extrusion-based approach demonstrated to be the most suitable for skeletal muscle tissue engineering, due to its ability to produce and deposit printing fibers in a parallel pattern that well mimic the native skeletal muscle tissue architecture. In tissue bioengineering, a key role is played by biomaterials, which must possess the key requisite of “printability”. Nevertheless, this feature is not often well correlated with cell requirements, such as motives for cellular adhesion and/or absorbability. To overcome this hurdle, several efforts have been made to obtain an effective bioink by combining two different biomaterials in order to reach a good printability besides a suitable biological activity. However, despite being efficient, this strategy reveals several outcomes limitations. We report here the development and characterization of a novel extrusion-based 3D bioprinting system, and its application for correction of volumetric muscle loss (VML) injury in a mouse model. The developed bioprinting system is based on the use of PEG-Fibrinogen, a unique biomaterial with excellent biocompatibility, well-suited for skeletal muscle tissue engineering. With this approach, we obtained highly organized 3D constructs, in which murine muscle progenitors were able to differentiate into muscle fibers arranged in aligned bundles and capable of spontaneously contracting when cultured in vitro. Furthermore, to evaluate the potential of the developed system in future regenerative medicine applications, bioprinted constructs laden with either murine or human muscle progenitors were transplanted to regenerate the Tibialis Anterior muscle of a VML murine model, one month after grafting.

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Live Imaging and Analysis of Muscle Contractions in <em>Drosophila</em> Embryo

Cited by 2 publications

References 15 publications

Secreted C-type lectin regulation of neuromuscular junction synaptic vesicle dynamics modulates coordinated movement

Secreted C-type lectin regulation of neuromuscular junction synaptic vesicle dynamics modulates coordinated movement

A novel extrusion-based 3D bioprinting system for skeletal muscle tissue engineering

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