Pten associates with important gene regulatory network to fine‐tune Müller glia‐mediated zebrafish retina regeneration

Gupta, Shivangi; Sharma, Poonam; Chaudhary, Mansi; Premraj, Sharanya; Kaur, Simran; Vijayan, Vijithkumar; Arun, Manas Geeta; Prasad, Nagaraj Guru; Ramachandran, Rajesh

doi:10.1002/glia.24270

Cited by 8 publications

(3 citation statements)

References 127 publications

(221 reference statements)

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“… 37 , 92 In addition, Gupta and his colleagues have reported that downregulation of Pten activates Akt and enhances MGPCs proliferation in zebrafish. 93 Gao and his colleagues have reviewed the current knowledge regarding the roles of Wnt/β-Catenin and Hedgehog Pathway in RMGC reprogramming. 36 , 94 , 95 …”

Section: Regulators Mediating the Dedifferentiation And Proliferation...mentioning

confidence: 99%

Genetic and epigenetic regulators of retinal Müller glial cell reprogramming

Xiao

Liao

Zou

2023

Advances in Ophthalmology Practice and Research

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Section: Regulators Mediating the Dedifferentiation And Proliferation...mentioning

confidence: 99%

Genetic and epigenetic regulators of retinal Müller glial cell reprogramming

Xiao

Liao

Zou

2023

Advances in Ophthalmology Practice and Research

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“…However, use of these tools in adult zebrafish is more difficult. Many morpholinos used to target genes in adult retina have been designed with a positively charged lissamine fluorescent molecule as a 3′ end modification ( Craig et al, 2010 ; Thummel et al, 2010 , 2011 ; Gupta et al, 2023 ), and require electroporation after injection of the morpholino for successful penetrance into cells. Many zebrafish labs do not have an electroporation device readily available, which cost from US $16,000 to $19,000 in 2021, and must rely on alternative strategies.…”

Section: Introductionmentioning

confidence: 99%

Cost-effective strategies to knock down genes of interest in the retinas of adult zebrafish

Shihabeddin,

Santhanam,

Aronowitz

et al. 2024

Front. Cell. Neurosci.

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High throughput sequencing has generated an enormous amount of information about the genes expressed in various cell types and tissues throughout the body, and about how gene expression changes over time and in diseased conditions. This knowledge has made targeted gene knockdowns an important tool in screening and identifying the roles of genes that are differentially expressed among specific cells of interest. While many approaches are available and optimized in mammalian models, there are still several limitations in the zebrafish model. In this article, we describe two approaches to target specific genes in the retina for knockdown: cell-penetrating, translation-blocking Vivo-Morpholino oligonucleotides and commercially available lipid nanoparticle reagents to deliver siRNA. We targeted expression of the PCNA gene in the retina of a P23H rhodopsin transgenic zebrafish model, in which rapidly proliferating progenitor cells replace degenerated rod photoreceptors. Retinas collected 48 h after intravitreal injections in adult zebrafish reveal that both Vivo-Morpholinos and lipid encapsulated siRNAs were able to successfully knock down expression of PCNA. However, only retinas injected with Vivo-Morpholinos showed a significant decrease in the formation of P23H rhodopsin-expressing rods, a downstream effect of PCNA inhibition. Surprisingly, Vivo-Morpholinos were able to exit the injected eye and enter the contralateral non-injected eye to inhibit PCNA expression. In this article we describe the techniques, concentrations, and considerations we found necessary to successfully target and inhibit genes through Vivo-Morpholinos and lipid encapsulated siRNAs.

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“…Due to the remarkable regenerative capacity of zebrafish, many studies have used this model to uncover the molecular underpinnings of retinal regeneration. Furthermore, a variety of signaling pathways, epigenomic remodeling factors, and immune response components have been identified to be required for Müller glia reprogramming and/or proliferation (e.g., Boyd et al, 2023; Campbell et al, 2021; Conner et al, 2014; Elsaeidi et al, 2018; Fausett et al, 2008; Gupta et al, 2023; Hamon et al, 2016; Kara et al, 2019; Kaur et al, 2018; Lee et al, 2020; Lenkowski et al, 2013; Lourenço et al, 2021; Mitchell et al, 2018; Mitra et al, 2022; Nagashima et al, 2020; Nelson et al, 2012, 2013; Powell et al, 2012, 2013; Rajaram et al, 2014; Ramachandran et al, 2010, 2011; 2012; Sahu et al, 2021; Sharma et al, 2019; Sifuentes et al, 2016; Silva et al, 2020; Song et al, 2023; White et al, 2017; Zhao et al, 2014). Despite these advances, we know little about the transcriptional regulation of pro‐regenerative genes or the genomic and epigenetic landscapes affecting their expression during the injury and regenerative responses.…”

Section: Introductionmentioning

confidence: 99%

Brd activity regulates Müller glia‐dependent retinal regeneration in zebrafish

Lee

Gross

2023

Glia

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The zebrafish retina possesses tremendous regenerative potential. Müller glia underlie retinal regeneration through their ability to reprogram and generate multipotent neuronal progenitors that re‐differentiate into lost neurons. Many factors required for Müller glia reprogramming and proliferation have been identified; however, we know little about the epigenetic and transcriptional regulation of these genes during regeneration. Here, we determined whether transcriptional regulation by members of the Bromodomain (Brd) family is required for Müller glia‐dependent retinal regeneration. Our data demonstrate that three brd genes were expressed in Müller glia upon injury. brd2a and brd2b were expressed in all Müller glia and brd4 was expressed only in reprogramming Müller glia. Utilizing (+)‐JQ1, a pharmacological inhibitor of Brd function, we demonstrate that transcriptional regulation by Brds plays a critical role in Müller glia reprogramming and regeneration. (+)‐JQ1 treatment prevented cell cycle re‐entry of Müller glia and the generation of neurogenic progenitors. Modulating the (+)‐JQ1 exposure window, we identified the first 48 h post‐injury as the time‐period during which Müller glia reprogramming occurs. (+)‐JQ1 treatments after 48 h post‐injury had no effect on the re‐differentiation of UV cones, indicating that Brd function is required only for Müller glia reprogramming and not subsequent specification/differentiation events. Brd inhibition also prevented the expression of reprogramming genes like ascl1a and lepb in Müller glia, but not effector genes like mmp9, nor did it affect microglial recruitment after injury. These results demonstrate that transcriptional regulation by Brds plays a critical role during Müller glia‐dependent retinal regeneration in zebrafish.

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Pten associates with important gene regulatory network to fine‐tune Müller glia‐mediated zebrafish retina regeneration

Cited by 8 publications

References 127 publications

Genetic and epigenetic regulators of retinal Müller glial cell reprogramming

Genetic and epigenetic regulators of retinal Müller glial cell reprogramming

Cost-effective strategies to knock down genes of interest in the retinas of adult zebrafish

Brd activity regulates Müller glia‐dependent retinal regeneration in zebrafish

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