Remarkable Regenerative Abilities Research Articles

Effect of Hydroxyurea on Morphology, Proliferation, and Protein Expression on Taenia crassiceps WFU Strain.

Flatworms are known for their remarkable regenerative ability, one which depends on totipotent cells known as germinative cells in cestodes. Depletion of germinative cells with hydroxyurea (HU) affects the regeneration of the parasite. Here, we studied the reduction and recovery of germinative cells in T. crassiceps cysticerci after HU treatment (25 mM and 40 mM of HU for 6 days) through in vitro assays. Viability and morphological changes were evaluated. The recovery of cysticerci's mobility and morphology was evaluated at 3 and 6 days, after 6 days of treatment. The number of proliferative cells was evaluated using EdU. Our results show morphological changes in the size, shape, and number of evaginated cysticerci at the 40 mM dose. The mobility of cysticerci was lower after 6 days of HU treatment at both concentrations. On days 3 and 6 of recovery after 25 mM of HU treatment, a partial recovery of the proliferative cells was observed. Proteomic and Gene Ontology analyses identified modifications in protein groups related to DNA binding, DNA damage, glycolytic enzymes, cytoskeleton, skeletal muscle, and RNA binding.

Regeneration in the absence of canonical neoblasts in an early branching flatworm.

The remarkable regenerative abilities of flatworms are closely linked to neoblasts - adult pluripotent stem cells that are the only division-competent cell type outside of the reproductive system. Although the presence of neoblast-like cells and whole-body regeneration in other animals has led to the idea that these features may represent the ancestral metazoan state, the evolutionary origin of both remains unclear. Here we show that the catenulid Stenostomum brevipharyngium, a member of the earliest-branching flatworm lineage, lacks conventional neoblasts despite being capable of whole-body regeneration and asexual reproduction. Using a combination of single-nuclei transcriptomics, in situ gene expression analysis, and functional experiments, we find that cell divisions are not restricted to a single cell type and are associated with multiple fully differentiated somatic tissues. Furthermore, the cohort of germline multipotency genes, which are considered canonical neoblast markers, are not expressed in dividing cells, but in the germline instead, and we experimentally show that they are neither necessary for proliferation nor regeneration. Overall, our results challenge the notion that canonical neoblasts are necessary for flatworm regeneration and open up the possibility that neoblast-like cells may have evolved convergently in different animals, independent of their regenerative capacity.

Conserved Genes in Highly Regenerative Metazoans Are Associated with Planarian Regeneration.

Metazoan species depict a wide spectrum of regeneration ability which calls into question the evolutionary origins of the underlying processes. Since species with high regeneration ability are widely distributed throughout metazoans, there is a possibility that the metazoan ancestor had an underlying common molecular mechanism. Early metazoans like sponges possess high regenerative ability, but, due to the large differences they have with Cnidaria and Bilateria regarding symmetry and neuronal systems, it can be inferred that this regenerative ability is different. We hypothesized that the last common ancestor of Cnidaria and Bilateria possessed remarkable regenerative ability which was lost during evolution. We separated Cnidaria and Bilateria into three classes possessing whole-body regenerating, high regenerative ability, and low regenerative ability. Using a multiway BLAST and gene phylogeny approach, we identified genes conserved in whole-body regenerating species and lost in low regenerative ability species and labeled them Cnidaria and Bilaterian regeneration genes. Through transcription factor analysis, we identified that Cnidaria and Bilaterian regeneration genes were associated with an overabundance of homeodomain regulatory elements. RNA interference of Cnidaria and Bilaterian regeneration genes resulted in loss of regeneration phenotype for HRJDa, HRJDb, DUF21, DISP3, and ARMR genes. We observed that DUF21 knockdown was highly lethal in the early stages of regeneration indicating a potential role in wound response. Also, HRJDa, HRJDb, DISP3, and ARMR knockdown showed loss of regeneration phenotype after second amputation. The results strongly correlate with their respective RNA-seq profiles. We propose that Cnidaria and Bilaterian regeneration genes play a major role in regeneration across highly regenerative Cnidaria and Bilateria.

Translational frontiers: insight from lymphatics in skin regeneration.

The remarkable regenerative ability of the skin, governed by complex molecular mechanisms, offers profound insights into the skin repair processes and the pathogenesis of various dermatological conditions. This understanding, derived from studies in human skin and various model systems, has not only deepened our knowledge of skin regeneration but also facilitated the development of skin substitutes in clinical practice. Recent research highlights the crucial role of lymphatic vessels in skin regeneration. Traditionally associated with fluid dynamics and immune modulation, these vessels are now recognized for interacting with skin stem cells and coordinating regeneration. This Mini Review provides an overview of recent advancements in basic and translational research related to skin regeneration, focusing on the dynamic interplay between lymphatic vessels and skin biology. Key highlights include the critical role of stem cell-lymphatic vessel crosstalk in orchestrating skin regeneration, emerging translational approaches, and their implications for skin diseases. Additionally, the review identifies research gaps and proposes potential future directions, underscoring the significance of this rapidly evolving research arena.

Future Liver Remnant Regeneration Post Hepatectomy: Histologic Changes in an Animal Model

Background: The liver possesses remarkable regenerative abilities following parenchymal injury. This study aimed to assess and document the microscopic regenerative changes in the rat liver using a partial hepatectomy model. Methodology: Adult male Sprague Dawley rats weighing approximately 200-300 grams were subjected to partial hepatectomy, leaving approximately 70% future liver remnant. The rats were euthanized at designated time intervals (1/2 hour, 48 hours, 72 hours, 7 days, 11 days, and 14 days) for histological analysis of the liver. Results: Liver regeneration was observed within 1 week of partial hepatectomy. Cytoplasmic vacuolization increased until day 7, but decreased significantly during the regenerative process. Disruption of cytoplasmic membranes and blurring of cell-to-cell and cell-to-sinusoidal space boundaries decreased notably from day 11. Necrosis involving central veins and portal tracts was observed on the first day after hepatectomy. By day 7, the portal tract exhibited granulomatous inflammation with conglomerates of epithelioid macrophages forming giant cells. The sinusoidal spaces displayed increased lymphocyte volume on day 7. Patchy portal inflammation consisting of lymphocytes, plasma cells, occasional eosinophils, and monocytes was observed from day 7 post-hepatectomy. Conclusion: In this partial hepatectomy animal model, the inflammatory cascade was evident through confluent necrosis affecting approximately 10% of the hepatic parenchyma, along with subcapsular infarction. Cytoplasmic vacuolization affected approximately 50% of the cells. The study documented various histological changes during the post-hepatectomy period, demonstrating the initiation of hepatocyte regeneration within one week.

VEGFA mRNA-LNP promotes biliary epithelial cell-to-hepatocyte conversion in acute and chronic liver diseases and reverses steatosis and fibrosis

The liver is known for its remarkable regenerative ability through proliferation of hepatocytes. Yet, during chronic injury or severe hepatocyte death, proliferation of hepatocytes is exhausted. To overcome this hurdle, we propose vascular-endothelial-growth-factor A (VEGFA) as a therapeutic means to accelerate biliary epithelial-cell (BEC)-to-hepatocyte conversion. Investigation in zebrafish establishes that blocking VEGF receptors abrogates BEC-driven liver repair, while VEGFA overexpression promotes it. Delivery of VEGFA via nonintegrative and safe nucleoside-modified mRNA encapsulated into lipid nanoparticles (mRNA-LNPs) in acutely or chronically injured mouse livers induces robust BEC-to-hepatocyte conversion and elimination of steatosis and fibrosis. In human and murine diseased livers, we further identified VEGFA-receptor KDR-expressing BECs associated with KDR-expressing cell-derived hepatocytes. This work defines KDR-expressing cells, most likely being BECs, as facultative progenitors. This study reveals unexpected therapeutic benefits of VEGFA delivered via nucleoside-modified mRNA-LNP, whose safety is widely validated with COVID-19 vaccines, for harnessing BEC-driven repair to potentially treat liver diseases.

In Vivo Stem Cell Discoveries: Promising Implications in Cancer Therapy

The remarkable regenerative abilities and versatility of stem cells have long attracted researchers. Recently, in vivo studies have revealed exciting results related to stem cells, particularly their use in cancer treatment. This review will provide an overview of these discoveries and their broader implications for the future. There is growing in vivo evidence that stem cells have immense therapeutic potential in treating various diseases, including cancer, because of their self-renewal and differentiation capabilities. As a result of in vivo research, critical aspects of stem cell behavior within tumor microenvironments have been clarified, providing a deeper understanding of their potential therapeutic utility. Several in vivo studies have demonstrated the potential of stem cell-engineered tumor-targeting agents or therapeutic payloads for the precise delivery of medicinal drugs when these agents are engineered to express them in tumor cells. Through targeted therapies, off-target effects can be minimized, and the therapeutic index of the anti-cancer agents can be improved. Several stem cell-based delivery systems have shown remarkable efficacy in preclinical in vivo studies, including breast, lung, and pancreatic cancer, indicating their potential as a novel therapeutic strategy. Moreover, in vivo studies have revealed that the immunomodulatory properties of stem cells modulate the immune response and modify the tumor microenvironment to suppress it. In particular, using checkpoint inhibitor therapy with stem cells has paved the way for innovative immunotherapeutic strategies. Research on stem cells in vivo has also provided invaluable insights into stem cell biology and their interaction with cancer cells. Due to these findings, there is an increasing understanding of tumor initiation, progression, and resistance mechanisms, which has opened avenues for improving cancer treatment by developing more effective treatments. As a result of the in vivo studies that have taken place so far, there is a wealth of information regarding the potential of stem cells in cancer treatment. This research opens up exciting prospects for the future of oncology, from the delivery of targeted drugs to immunomodulation and improving our understanding of tumor biology.

Tail regeneration at different ontogenetic stages of the tiger salamander Ambystoma tigrinum suggests possible changes in regeneration between larval and metamorphic individuals

AbstractUrodele amphibians possess remarkable regenerative abilities, allowing them to rebuild lost body parts. Contrary to lizards, salamanders can fully restore their tails, including the neural spine and components of the vertebral column. The axolotl (Ambystoma mexicanum) is the vertebrate model organism for regeneration research due to its ease of breeding in captivity. However, axolotls are paedomorphic, retaining larval somatic features throughout adulthood and do not naturally undergo metamorphosis, a transition phase from aquatic larvae to terrestrial adults with profound morphological and physiological changes. We investigate the influence of metamorphosis on salamander tail regeneration after conspecific biting in the metamorphosing sister taxon Ambystoma tigrinum using histological analysis to answer two key questions: (1) Does regeneration continue during metamorphosis, or is it halted? (2) How does regeneration differ histologically among larval, metamorphosing and postmetamorphic individuals? Our findings demonstrate that regeneration continues even during metamorphic climax, indicating the simultaneous coordination of metamorphosis and regeneration. Additionally, notable distinctions were observed between developmental stages concerning the speed of regeneration and structural differences in the formation of an apical epithelial cap (AEC). While the approach taken in this study necessarily restricts sample size, it offers valuable insights into regeneration in a metamorphosing species under natural conditions.

Regenerative Potential of Injured Spinal Cord in the Light of Epigenetic Regulation and Modulation.

A spinal cord injury is a form of physical harm imposed on the spinal cord that causes disability and, in many cases, leads to permanent mammalian paralysis, which causes a disastrous global issue. Because of its non-regenerative aspect, restoring the spinal cord's role remains one of the most daunting tasks. By comparison, the remarkable regenerative ability of some regeneration-competent species, such as some Urodeles (Axolotl), Xenopus, and some teleost fishes, enables maximum functional recovery, even after complete spinal cord transection. During the last two decades of intensive research, significant progress has been made in understanding both regenerative cells' origins and the molecular signaling mechanisms underlying the regeneration and reconstruction of damaged spinal cords in regenerating organisms and mammals, respectively. Epigenetic control has gradually moved into the center stage of this research field, which has been helped by comprehensive work demonstrating that DNA methylation, histone modifications, and microRNAs are important for the regeneration of the spinal cord. In this review, we concentrate primarily on providing a comparison of the epigenetic mechanisms in spinal cord injuries between non-regenerating and regenerating species. In addition, we further discuss the epigenetic mediators that underlie the development of a regeneration-permissive environment following injury in regeneration-competent animals and how such mediators may be implicated in optimizing treatment outcomes for spinal cord injurie in higher-order mammals. Finally, we briefly discuss the role of extracellular vesicles (EVs) in the context of spinal cord injury and their potential as targets for therapeutic intervention.

Macrophage–Neuroglia Interactions in Promoting Neuronal Regeneration in Zebrafish

The human nervous system exhibits limited regenerative capabilities following damage to the central nervous system (CNS), leading to a scarcity of effective treatments for nerve function recovery. In contrast, zebrafish demonstrate remarkable regenerative abilities, making them an ideal model for studying the modulation of inflammatory processes after injury. Such research holds significant translational potential to enhance our understanding of recovery from damage and disease. Macrophages play a crucial role in tissue repair and regeneration, with their subpopulations indirectly promoting axonal regeneration through developmental signals. The AP-1 signaling pathway, mediated by TNF/Tnfrsf1a, can elevate HDAC1 expression and facilitate regeneration. Furthermore, following spinal cord injury (SCI), pMN progenitors have been observed to switch between oligodendrocyte and motor neuron fates, with macrophage-secreted TNF-α potentially regulating the differentiation of ependymal-radial glia progenitors and oligodendrocytes. Radial glial cells (RGs) are also essential for CNS regeneration in zebrafish, as they perform neurogenesis and gliogenesis, with specific RG subpopulations potentially existing for the generation of neurons and oligodendrocytes. This review article underscores the critical role of macrophages and their subpopulations in tissue repair and regeneration, focusing on their secretion of TNF-α, which promotes axonal regeneration in zebrafish. We also offer insights into the molecular mechanisms underlying TNF-α's ability to facilitate axonal regeneration and explore the potential of pMN progenitor cells and RGs following SCI in zebrafish. The review concludes with a discussion of various unresolved questions in the field, and ideas are suggested for future research. Studying innate immune cell interactions with neuroglia following injury may lead to the development of novel strategies for treating the inflammatory processes associated with regenerative medicine, which are commonly observed in injury and disease.

A CRISPR-Cas9-mediated F0 screen to identify pro-regenerative genes in the zebrafish retinal pigment epithelium

Ocular diseases resulting in death of the retinal pigment epithelium (RPE) lead to vision loss and blindness. There are currently no FDA-approved strategies to restore damaged RPE cells. Stimulating intrinsic regenerative responses within damaged tissues has gained traction as a possible mechanism for tissue repair. Zebrafish possess remarkable regenerative abilities, including within the RPE; however, our understanding of the underlying mechanisms remains limited. Here, we conducted an F0 in vivo CRISPR-Cas9-mediated screen of 27 candidate RPE regeneration genes. The screen involved injection of a ribonucleoprotein complex containing three highly mutagenic guide RNAs per target gene followed by PCR-based genotyping to identify large intragenic deletions and MATLAB-based automated quantification of RPE regeneration. Through this F0 screening pipeline, eight positive and seven negative regulators of RPE regeneration were identified. Further characterization of one candidate, cldn7b, revealed novel roles in regulating macrophage/microglia infiltration after RPE injury and in clearing RPE/pigment debris during late-phase RPE regeneration. Taken together, these data support the utility of targeted F0 screens for validating pro-regenerative factors and reveal novel factors that could regulate regenerative responses within the zebrafish RPE.

Plant <i>in situ</i> tissue regeneration: dynamics, mechanisms and implications for forestry research

Plants possess remarkable developmental plasticity and regenerative ability to reshape themselves in response to external stimulations. After local injuries, they can initiate cellular reprogramming at the wound sites to repair or regrow structures that could substitute the damaged or lost parts functionally. This way of regeneration in plants is called plant <italic>in situ</italic> tissue regeneration. Upon wounding like excision, incision or girdling, the original tissue patterns are completely or partially destroyed, the remanent tissues could perceive the wounding signals and thereby initiate cell de-differentiation, trans-differentiation or re-differentiation to reconstruct the lost or damaged tissues. In this review, we summarize the regenerative dynamics and regulatory mechanisms during the major <italic>in situ</italic> tissue regeneration processes in plants, including secondary vascular tissue (SVT) regeneration after girdling, apex regeneration after excision and tissue reunion after incision. In addition, we compare the features of SVT regeneration, the most relevant system for forestry, with other plant <italic>in situ</italic> tissue regeneration systems. We further discuss the unsolved issues and the potential applications of plant <italic>in situ</italic> regeneration for forestry research, aiming to provide new insights for the study of woody plant development.

Twist2-driven chromatin remodeling governs the postnatal maturation of dermal fibroblasts.

Dermal fibroblasts lose stem cell potency after birth, which prevents regenerative healing. However, the underlying intracellular mechanisms are largely unknown. We uncover the postnatal maturation of papillary fibroblasts (PFs) driven by the extensive Twist2-mediated remodeling of chromatin accessibility. A loss of the regenerative ability of postnatal PFs occurs with decreased H3K27ac levels. Single-cell transcriptomics, assay for transposase-accessible chromatin sequencing (ATAC-seq), and chromatin immunoprecipitation sequencing (ChIP-seq) reveal the postnatal maturation trajectory associated with the loss of the regenerative trajectory in PFs, which is characterized by a marked decrease in chromatin accessibility and H3K27ac modifications. Histone deacetylase inhibition delays spontaneous chromatin remodeling, thus maintaining the regenerative ability of postnatal PFs. Genomic analysis identifies Twist2 as a major regulator within chromatin regions with decreased accessibility during the postnatal period. When Twist2 is genetically deleted in dermal fibroblasts, the intracellular cascade of postnatal maturation is significantly delayed. Our findings reveal the comprehensive intracellular mechanisms underlying intrinsic postnatal changes in dermal fibroblasts.

An Identification and Characterization of the Axolotl (Ambystoma mexicanum, Amex) Telomerase Reverse Transcriptase (Amex TERT).

The Mexican axolotl is one of the few vertebrates that is able to replace its lost body parts during lifespan. Due to its remarkable regenerative abilities, the axolotl emerged as a model organism especially for limb regeneration. Telomeres and the telomerase enzyme are crucial for regeneration and protection against aging processes and degenerating diseases. Despite its relevance for regeneration, the axolotl telomerase and telomere length have not yet been investigated. Therefore, in the present paper, we reveal the sequence of the axolotl telomerase reverse transcriptase gene (Tert) and protein (TERT). Multiple sequence alignment (MSA) showed the known conserved RT- and TERT-specific motifs and residues found in other TERTs. In addition, we establish methods to determine the Tert expression (RT-PCR) and telomerase activity (Q-TRAP) of adult axolotl and blastema tissues. We found that both differentiated forelimb tissue and regenerating blastema tissue express Tert and show telomerase activity. Furthermore, blastema tissue appears to exhibit a higher Tert expression and telomerase activity. The presence of active telomerase in adult somatic cells is a decisive difference to somatic cells of non-regenerating vertebrates, such as humans. These findings indicate that telomere biology may play a key role in the regenerative abilities of cells.

Eye Removal in Living Zebrafish Larvae to Examine Innervation-dependent Growth and Development of the Visual System

Zebrafish exhibit remarkable life-long growth and regenerative abilities. For example, specialized stem cell niches established during embryogenesis support continuous growth of the entire visual system, both in the eye and the brain. Coordinated growth between the retinae and the optic tectum ensures accurate retinotopic mapping as new neurons are added in the eyes and brain. To address whether retinal axons provide crucial information for regulating tectal stem and progenitor cell behaviors such as survival, proliferation, and/or differentiation, it is necessary to be able to compare innervated and denervated tectal lobes within the same animal and across animals. Surgical removal of one eye from living larval zebrafish followed by observation of the optic tectum achieves this goal. The accompanying video demonstrates how to anesthetize larvae, electrolytically sharpen tungsten needles, and use them to remove one eye. It next shows how to dissect brains from fixed zebrafish larvae. Finally, the video provides an overview of the protocol for immunohistochemistry and a demonstration of how to mount stained embryos in low-melting-point agarose for microscopy.

Validation of three reference genes for quantitative RT-PCR analyses in regenerating Lumbriculus variegatus

Regeneration, the ability of organisms to replace body structures lost to injury, varies widely among and within animal taxa. The annelid Lumbriculus variegatus exhibits remarkable regeneration, growing both new head and tail regions from as few as three segments. Identifying orthologous genes upregulated early in regeneration in a variety of species may help pinpoint proteins that are essential for regeneration. Gene expression is frequently assayed with reverse transcription polymerase chain reaction (RT-PCR), a method that requires a validated reference gene. Therefore, we analyzed three common housekeeping genes to determine their suitability as reference genes during early L. variegatus regeneration. We found that all genes tested—18S ribosomal RNA, actin, and glyceraldehyde 3-phosphate dehydrogenase—showed stable RNA levels between intact worms and worms undergoing regeneration at 24 h post-amputation. Our validation of these reference genes will allow for more accurate RT-PCR studies in L. variegatus, and contribute to uncovering the mechanisms behind their remarkable regenerative abilities.

Schmidtea happens: Re-establishing the planarian as a model for studying the mechanisms of regeneration.

Understanding the remarkable regenerative abilities of freshwater planarians was a classic problem of developmental biology. These animals were widely studied until the late 1960s, when their use as experimental subjects declined precipitously after some infamous experiments on memory transfer. By the mid-1990s, only a handful of laboratories worldwide were investigating the mechanisms of planarian regeneration. Here, we provide the personal stories behind our work to reinvigorate studies of these fascinating animals. We recount many of the challenges that had to be overcome and reflect on some of the fortuitous events that helped launch the planarian Schmidtea mediterranea as a model organism for studying the molecular basis of regeneration.

Quantification of planarian behaviors.

Research on individual behaviors can help to reveal the processes and mechanisms that mediate an animal's habits and interactions with the environment. Importantly, individual behaviors arise as outcomes of genetic programs, morphogenesis, physiological processes, and neural functions; thus, behavioral analyses can be used to detect disorders in these processes. Planarians belong to an early branching bilateral group of organisms that possess a simple central nervous system. Furthermore, planarians display various behavioral responses to the environment via their nervous system. Planarians also have remarkable regenerative abilities, including whole-brain regeneration. Therefore, the combination of planarians' phylogenetic position, behavioral properties, regenerative ability, and genetic accessibility provides a unique opportunity to understand the basic mechanisms underlying the anatomical properties of neural morphogenesis and the dynamic physiological processes and neural function. Here, we describe a step-by-step protocol for conducting simple behavioral analyses in planarians with the aim of helping to introduce researchers to the utility of performing behavioral analyses in planarians. Since the conditions of planarians impact experimental results and reproducibility, this protocol begins with a method for maintaining planarians. Next, we introduce the behavioral tests as well as the methods for quantifying them using minimal and cost-effective equipment and materials. Finally, we present a unique RNAi technique that enables conditional silencing of neural activity in the brain of planarians.

Chemical Exposure-Induced Developmental Neurotoxicity in Head-Regenerating Schmidtea mediterranea.

The growing number of commercially used chemicals that are under-evaluated for developmental neurotoxicity (DNT) combined with the difficulty in describing the etiology of exposure-related neurodevelopmental toxicity has created a reticent threat to human health. Current means of screening chemicals for DNT are limited to expensive, time-consuming, and labor-intensive traditional laboratory animal models. In this study, we hypothesize that exposed head-regenerating planarian flatworms can effectively and efficiently categorize DNT in known developmental neurotoxins (ethanol and bisphenol A [BPA]). Planarian flatworms are an established alternative animal model for neurodevelopmental studies and have remarkable regenerative abilities allowing neurodevelopment to be induced via head resection. Here, we observed changes in photophobic behavior and central nervous system (CNS) morphology to evaluate the impact of exposure to low concentrations of ethanol, BPA, and BPA industry alternatives bisphenol F, and bisguaiacol on neurodevelopment. Our studies show that exposure to 1% v/v ethanol during regeneration induces a recoverable 48-h delay in the development of proper CNS integrity, which aligns with behavioral assessments of cognitive ability. Exposure to BPA and its alternatives induced deviations to neurodevelopment in a range of severities, distinguished by suppressions, delays, or a combination of the 2. These results suggest that quick and inexpensive behavioral assessments are a viable surrogate for tedious and costly immunostaining studies, equipping more utility and resolution to the planarian model for neurodevelopmental toxicity in the future of mass chemical screening. These studies demonstrate that behavioral phenotypes observed following chemical exposure are classifiable and also temporally correlated to the anatomical development of the CNS in planaria. This will facilitate and accelerate toxicological screening assays with this alternative animal model.

Retinal ganglion cell survival after severe optic nerve injury is modulated by crosstalk between Jak/Stat signaling and innate immune responses in the zebrafish retina.

Visual information is transmitted from the eye to the brain along the optic nerve, a structure composed of retinal ganglion cell (RGC) axons. The optic nerve is highly vulnerable to damage in neurodegenerative diseases, such as glaucoma, and there are currently no FDA-approved drugs or therapies to protect RGCs from death. Zebrafish possess remarkable neuroprotective and regenerative abilities. Here, utilizing an optic nerve transection (ONT) injury and an RNA-seq-based approach, we identify genes and pathways active in RGCs that may modulate their survival. Through pharmacological perturbation, we demonstrate that Jak/Stat pathway activity is required for RGC survival after ONT. Furthermore, we show that immune responses directly contribute to RGC death after ONT; macrophages/microglia are recruited to the retina and blocking neuroinflammation or depleting these cells after ONT rescues survival of RGCs. Taken together, these data support a model in which crosstalk between macrophages/microglia and RGCs, mediated by Jak/Stat pathway activity, regulates RGC survival after optic nerve injury.