Collective behaviors of Drosophila-derived retinal progenitors in controlled microenvironments

Pena, Caroline D.; Zhang, Stephanie; Markey, Miles; Venkatesh, Tadmiri; Vázquez, Maribel

doi:10.1371/journal.pone.0226250

Cited by 4 publications

(16 citation statements)

References 75 publications

(98 reference statements)

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“…Experiments utilized the GAL4-UAS system [45], where retinal neuroblasts (RNBs) with glial markers expressed green fluorescent protein (GFP) and RNBs with neuronal markers expressed red fluorescent protein (RFP). Drosophila Melanogaster stocks of UAS-GFP (CS: Repo) and UAS-mCD8-GFP; elav GAL4 were used because the Elav (neurons) and Repo (Glia) markers are the only markers to specifically stain cells in the developing retinal ganglion [36,46]. Flies were maintained on standard corn meal agar medium and kept at 25 • C. Stocks were transferred once a week to maintain lines of larvae mixed from the two strains.…”

Section: Drosophila Fly Stocksmentioning

confidence: 99%

“…Scanning Electron Microscopy (Zeiss LS704U SEM, Jena, Germany) was used to capture images of the adult Drosophila compound eye at 6 kV and 2.601 A with the stage at a Z plane of 23.372 nm, as described previously by our group [36]…”

Section: Microscopy and Imagingmentioning

confidence: 99%

“…As Fick's law dictates a linear steady-state concentration gradient for one-dimensional flows, one unique benefit of the µOS is the ability to examine RNB behavior within different concentration ranges of the same gradient field. This is highly advantageous because numerous groups, including our own [26,36,41,42], have illustrated the importance of concentration and concentration gradient, separately, in the motile behavior of different types of cells. The linear concentration gradient within the microarray has been divided into three separate concentration regions along the microchannel, which are grossly denoted as High gradient, H ∆C 1 , Medium gradient, M ∆C 2 , and Low gradient, L ∆C 3 , as shown in Figure 4D.…”

Section: Validation Of the µOs Chemical Environmentmentioning

confidence: 99%

“…Microfluidics-based systems have also been developed to examine electrical synapses and neuromodulation in retina, as well as enable the size profiling of retinal stem cells [31][32][33]. However, very few projects have used microfluidics to examine the collective migration within developing visual systems [34,35] central to retinal regeneration therapies, and even fewer works have used invertebrate cells to do so [36][37][38][39]. Ours is the first microfluidic environment designed to represent the developing visual system of Drosophila Melanogaster to anatomical scale and used to examine collective migration therein.…”

Section: Introductionmentioning

confidence: 99%

“…This device thereby provides an important testing system for studying the mechanisms of Drosophila development to advance emerging strategies for retinal regeneration. Previous work from our group [36,37,[40][41][42] has examined the collective migration of clustered retinal progenitors using microfluidic devices with characteristic lengths much larger than the physiological system examined. In this report, we describe a microfluidic device called the microoptic stalk, or μOS, with dimensions approaching those of the developing eye-brain complex of Drosophila.…”

Section: Introductionmentioning

confidence: 99%

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A Micro-Optic Stalk (μOS) System to Model the Collective Migration of Retinal Neuroblasts

et al. 2020

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Contemporary regenerative therapies have introduced stem-like cells to replace damaged neurons in the visual system by recapitulating critical processes of eye development. The collective migration of neural stem cells is fundamental to retinogenesis and has been exceptionally well-studied using the fruit fly model of Drosophila Melanogaster. However, the migratory behavior of its retinal neuroblasts (RNBs) has been surprisingly understudied, despite being critical to retinal development in this invertebrate model. The current project developed a new microfluidic system to examine the collective migration of RNBs extracted from the developing visual system of Drosophila as a model for the collective motile processes of replacement neural stem cells. The system scales with the microstructure of the Drosophila optic stalk, which is a pre-cursor to the optic nerve, to produce signaling fields spatially comparable to in vivo RNB stimuli. Experiments used the micro-optic stalk system, or μOS, to demonstrate the preferred sizing and directional migration of collective, motile RNB groups in response to changes in exogenous concentrations of fibroblast growth factor (FGF), which is a key factor in development. Our data highlight the importance of cell-to-cell contacts in enabling cell cohesion during collective RNB migration and point to the unexplored synergy of invertebrate cell study and microfluidic platforms to advance regenerative strategies.

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Section: Drosophila Fly Stocksmentioning

confidence: 99%

Section: Microscopy and Imagingmentioning

confidence: 99%

Section: Validation Of the µOs Chemical Environmentmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Micro-Optic Stalk (μOS) System to Model the Collective Migration of Retinal Neuroblasts

et al. 2020

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Retinal Progenitor Cells Exhibit Cadherin-Dependent Chemotaxis across Transplantable Extracellular Matrix of In Vitro Developmental and Adult Models

Markey,

Pena,

Venkatesh

et al. 2023

Journal of Tissue Engineering and Regenerative Medicine

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Retinal degeneration is an escalating public health challenge, as diseases such as age-related macular degeneration, diabetic retinopathy, and retinitis pigmentosa cause irreversible vision loss in millions of adults each year. Regenerative medicine has pioneered the development of stem cell replacement therapies, which treat degeneration by replacing damaged retinal neurons with transplanted stem-like cells (SCs). While the collective migration of SCs plays critical roles during retinal development, our understanding of collective SC behaviors within biomaterials transplanted into adult tissue remains understudied. This project examines the potential therapeutic impacts of collective SC migration during transplantation by correlating the expression of cadherin, cell-cell cohesion molecules that maintain intercellular communication during development, with receptor proteins of chemoattractant molecules prevalent in degenerated adult tissue. Experiments examine these well-conserved biomechanisms by using two different model organisms: Drosophila melanogaster, a seminal model for retinal development, and Mus, an important preclinical model for transplantation. Results indicate that SCs from both animal models significantly upregulate cadherin expression to achieve more directed collective migration towards species-specific chemoattractants and exhibit longer distance motility upon different extracellular matrix substrates.

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VEGF Upregulates EGFR Expression to Stimulate Chemotactic Behaviors in the rMC-1 Model of Müller Glia

Peña

Vázquez

2020

Brain Sciences

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Progressive vision loss in adults has become increasingly prevalent worldwide due to retinopathies associated with aging, genetics, and epigenetic factors that damage the retinal microvasculature. Insufficient supply of oxygen and/or nutrients upregulates factors such as vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), which can induce abnormal angiogenesis and damage the structural arrangement of the retinal blood barrier (BRB). Müller glia (MG) regulate the diffusion of essential compounds across the BRB and respond to retinal insults via reactive gliosis, which includes cell hypertrophy, migration, and/or proliferation near areas of elevated VEGF concentration. Increasing concentrations of exogenous VEGF, upregulated by retinal pigmented epithelium cells, and endogenous epidermal growth factor receptor (EGF-R) stimulation in MG, implicated in MG proliferative and migratory behavior, often lead to progressive and permanent vision loss. Our project examined the chemotactic responses of the rMC-1 cell line, a mammalian MG model, toward VEGF and EGF signaling fields in transwell assays, and within respective concentration gradient fields produced in the glia line (gLL) microfluidic system previously described by our group. rMC-1 receptor expression in defined ligand fields was also evaluated using quantitative polymerase chain reaction (qPCR) and immunocytochemical staining. Results illustrate dramatic increases in rMC-1 chemotactic responses towards EGF gradient fields after pre-treatment with VEGF. In addition, qPCR illustrated significant upregulation of EGF-R upon VEGF pre-treatment, which was higher than that induced by its cognate ligand, EGF. These results suggest interplay of molecular pathways between VEGF and EGF-R that have remained understudied in MG but are significant to the development of effective anti-VEGF treatments needed for a variety of retinopathies.

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Collective behaviors of Drosophila-derived retinal progenitors in controlled microenvironments

Cited by 4 publications

References 75 publications

A Micro-Optic Stalk (μOS) System to Model the Collective Migration of Retinal Neuroblasts

A Micro-Optic Stalk (μOS) System to Model the Collective Migration of Retinal Neuroblasts

Retinal Progenitor Cells Exhibit Cadherin-Dependent Chemotaxis across Transplantable Extracellular Matrix of In Vitro Developmental and Adult Models

VEGF Upregulates EGFR Expression to Stimulate Chemotactic Behaviors in the rMC-1 Model of Müller Glia

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