Biology subjects

Palakodeti, D.

Publications and source records attributed to Palakodeti, D..

At least 19 recordsLinked to original sources

bioRxiv2026

Angiogenin activity regulates RNA remodeling during the maternal-to-zygotic transition in early mouse embryogenesis

Angiogenin (ANG), a member of the RNase A superfamily, regulates multiple aspects of RNA metabolism, including rRNA processing and the generation of tRNA-derived small RNAs. However, its function during early mammalian embryogenesis remains unknown. Here, we identify a previously unrecognized requirement for ANG activity during early mouse embryonic development. Pharmacological inhibition of ANG during the first embryonic cell cycle did not prevent progression to the 2-cell stage but markedly impaired subsequent development to the 4-cell stage, indicating a critical requirement for ANG activity during the maternal-to-zygotic transition. Transcriptome-wide analysis revealed that ANG inhibition disrupted the normal remodeling of the embryonic transcriptome, characterized by persistent accumulation of a subset of transcripts that are normally reduced during the 2-cell transition, including many maternally deposited RNAs, together with altered expression of transcripts associated with zygotic genome activation. Small-RNA profiling further revealed widespread alterations in miRNA and tRNA-derived small-RNA populations. Notably, a large proportion of transcripts retained following ANG inhibition contained predicted target sites for miRNAs that were concomitantly reduced, suggesting a potential link between ANG-dependent small-RNA regulation and maternal transcript clearance. ANG inhibition was also associated with altered abundance and organization of components of RNA regulatory granules, including Tudor, TIAR, Dcp1a and Ago2, and increased association of selected retained transcripts Reep4 and Esrrb with Tudor-positive structures. Together, these findings identify ANG activity as an important regulator of RNA remodeling during the maternal-to-zygotic transition and suggest that ANG coordinates maternal transcript clearance, small-RNA dynamics and mRNP organization to support early mouse embryonic development.

developmental biology↗

bioRxiv2026

An oxo-mechanical coupling determines cell state

Across myriad habitats and diverse lifeforms, oxygen and mechanics are the most ubiquitously varying and profoundly influential environmental regulators. While physiological niches feature both varying oxygen levels and heterogeneous mechanics, laboratory experiments typically interrogate these as independent variables. Here, we show that combinatorial regimes defined by varying oxygen partial pressures and environmental mechanics--an oxo-mechanical cue--induce functionally-distinct cellular states in 3D ECM-like contexts. Single-cell morphometrics combined with multi-omics reveal that cellular response to oxygen deprivation depends on external mechanical milieus, whereas, cellular engagement with different mechanical microenvironments depends on oxygen availability. Independently perturbing both hypoxic signaling and cytoskeletal activity further reveals a reciprocal oxo-mechanical regulatory coupling, which operates by differentially altering the global chromatin accessibility for transcriptional regulation in response to specific combinations of oxygen partial pressures and external mechanical milieus. Together, our findings establish that a coupling between oxygen and mechanics drives the emergence of microenvironmentally-defined cell states.

cell biology↗

bioRxiv2026

Actin-dependent mechanotransduction controls nucleocytoplasmic partitioning of DNMT3a through ERK1/2 signaling during cutaneous wound healing

Cutaneous wound healing is a multifaceted physiological process that requires a cell state transition from homeostasis to tissue repair. An important contributor to this reprogramming is the wound-induced mechanical cues that are perceived by epidermal keratinocytes. Previously we identified that the nuclear translocation of the de novo DNA methyltransferase 3A (DNMT3a) upon wounding as an important regulator of this cell-state transition. However, the molecular mechanisms linking mechanotransduction to epigenetic regulation remain incompletely understood. Here we show that under homeostasis, active ERK1/2 phosphorylates DNMT3a, intramolecularly masking its nuclear localization signal (NLS), resulting in its cytoplasmic sequestration. Upon wounding actin cytoskeleton remodeling leads to the downregulation of the Extracellular Signal-Related Kinase 1/2 (ERK 1/2) pathway. Wound-induced ERK1/2 inactivation unmasks the NLS, enabling DNMT3a nuclear translocation. Collectively, these findings define a mechanotransduction-driven signaling axis linking cytoskeletal dynamics to epigenetic regulation and confirms an active role for differentiated keratinocytes in initiating early wound repair programs.

cell biology↗

bioRxiv2025

Calcium: Modulator of Post-transcriptional and post-translational process in mESCs

Calcium ion (Ca2+) is a ubiquitous intracellular and extracellular messenger that regulates several cellular activities. Previous findings have reported the presence of active Ca2+ receptors in mouse embryonic stem cells (mESCs) but the fundamental requirement of Ca2+ signalling remains unclear. We have noted the presence of high Ca2+ in undifferentiated mESCs and showed that its depletion exerts G2/M cell cycle arrest, spontaneous differentiation of mESCs towards mesoderm lineage and mitochondrial biogenesis. Further, our data demonstrates that Ca2+ regulates the homeostasis and stability of Oct4 and Nanog at the post-translational level through pCamkII dependent mechanism independent of polyubiquitin system, JAK-STAT3 pathway, transcriptional and translational control. Our data also signifies the role of Ca2+ at the post-transcriptional level in regulating p-bodies and stress granule markers (Dcp1a, XRN1, Tudor and EDC4), splicing-dependent and 3UTR-dependent NMD activity. Together, this study identifies the broad role of Ca2+ in modulating key processes in mESCs. HighlightsCa2+ is present at elevated levels in mESCs and is important for pluripotency Ca2+ depletion accelerates G2/M cell cycle arrest and mesoderm differentiation Ca2+ regulates pluripotency and p-bodies markers at the post-translational level Ca2+ regulates Oct4, Nanog, dcp1a through pCaMKII dependent mechanism

cell biology↗

bioRxiv2025

TIP60, a key lysine acetyltransferase, acts as a wound-induced factor essential for efficient wound response in planaria

Chromatin modifiers are essential regulators of gene expression, DNA repair, replication, and cell division. Among them, histone acetyltransferases (HATs) such as TIP60 play a central role in modulating chromatin dynamics through acetylation of histone and non-histone proteins. TIP60, a member of the MYST family of HATs, is known to regulate key cellular processes, including transcriptional activation, DNA damage response, and cell cycle progression. Although TIP60s role in stem cell maintenance and differentiation is well established, its function in stem cell-driven regeneration has remained unexplored. In this study, we characterize the role of the TIP60 homolog, SMED-TIP60, in the planarian Schmidtea mediterranea, a model organism renowned for its regenerative capacity. Biochemical assays confirmed SMED-TIP60s acetyltransferase and auto-acetylation activity. RNAi-mediated knockdown of Smed-tip60 resulted in severe defects in tissue homeostasis, survival, and regeneration, including impaired blastema formation and failure to regenerate tissues. In situ hybridization and immunofluorescence analyses revealed a marked reduction in stem cell populations and mitotic activity. Western blotting showed a peak in SMED-TIP60 expression at 5 days post-amputation, suggesting a role during regeneration. RNA-seq analysis revealed widespread dysregulation of gene expression at both anterior and posterior wounds, correlating with increased TIP60 expression post-injury. Notably, wound-response gene expression was aberrant in Smed-tip60 RNAi animals, indicating TIP60s essential role in initiating wound responses and resetting positional cues. Together, these findings establish TIP60 as a critical regulator of stem cell-mediated regeneration and wound healing in planarians.

cell biology↗

bioRxiv2025

Single-cell transcriptomics reveal ddx43+ cells as regulators of slow-cycling neoblasts in planaria

Cell cycle dynamics are fundamental to stem cell maintenance and differentiation. While the G1 phase is known to influence stem cell fate decisions, the mechanisms regulating its duration remain poorly understood. Using Schmidtea mediterranea, a highly tractable model for studying regeneration, we uncovered how the interplay of systemic and local cues from specialized cell types regulates G1 phase progression, thereby priming the cells in a slow-cycling state poised to differentiate. Single-cell transcriptomics identified G1-enriched, non-committed neoblasts that give rise to a previously uncharacterized ddx43 cell type, localized near sub-epidermal muscle and closely associated with surrounding neoblasts. Functional analyses revealed that ddx43 cells act as gatekeepers, maintaining adjacent neoblasts in an extended G1 phase and a differentiation-primed state. ddx43 knockdown led to neoblast hyperproliferation, highlighting their niche-like regulatory role. Following injury, extracellular signal-regulated kinase (Erk) signalling that propagates to distal regions via subepidermal muscle cells is critical for suppression of ddx43 cells by downregulating Notch signalling. This establishes a critical cross-talk between sub-epidermal muscle and ddx43 cells, mediated by the Erk-Notch axis, essential for maintaining G1 phase extension. In summary, we reveal a ddx43-dependent, non-cell autonomous mechanism that regulates a transient, slow-cycling neoblast population, offering new insights into how extrinsic cues orchestrate stem cell cycling during tissue homeostasis and regeneration.

developmental biology↗

bioRxiv2025

Temporal integration and active sensing of light in planarians

Animal light sensing and processing are intimately connected to the design of the sensory apparatus, with eye-design a subject of longstanding interest. Understanding animal response to stimuli can elucidate the nature of neural-processing encoded within the photosensory system, tuned to the specific eye-design. Planarians show exquisite photosensitivity and remarkably diverse light-induced behaviors. Further, they are bilaterally-symmetric and possess prototypic cup-shaped eyes, considered an important landmark in eye evolution. Here, using quantitative analysis of behaviour, we address planarian photo-response to distinct presentation of light gradients. Notably, we find that as the nature and presentation of gradients are altered, planarians switch their navigation strategies. When subjected to side-illumination, planarians display negative-phototaxis with a predominantly angular trajectory which presumably allows the animal to maintain the intensity difference between their two eyes for orientation during taxis. However, when subjected to top illumination where light intensity difference between the two eyes would be minimal, animals are still negatively phototactic but the dominant trajectories tend to be directly away from light. Apart from spatial light-intensity sensing, planarians also appear capable of temporal light sensing; confirmed by their response to a spatially-uniform light source with intensity modified over time. Crucially data show that planarians may be using spatial and temporal light sensing to differing degrees depending on the stimuli. Further, when subjected to light illumination from below, planarians fully switch orientation and go belly-up, indicating an active sampling of light distinct from photo-aversive behaviour. Overall, our results show planarian light-sensing to be multilayered, dependent on distinct, highly plastic sensory strategies and beyond basic photo-aversive behaviour.

animal behavior and cognition↗

bioRxiv2024

Analysis of ribosomes from the Wild-type and FMR1 knockout human embryonic stem cells

Fragile X Messenger Ribonucleoprotein 1 (FMRP) is a multifunctional, multidomain RNA-binding protein whose loss causes Fragile X syndrome. It is also known to associate with ribosomes and modulate translation. In human embryonic stem cells (hESCs), knockout (KO) of the FMR1 gene results in significantly increased protein translation rates and alterations in the 2-O-methylation patterns of rRNA. To understand the structural underpinnings of the process, we performed electron cryomicroscopy analysis of ribosomes isolated from both wild-type (WT) and FMR1 KO hESCs that revealed a subpopulation of dormant ribosomes in the FMR1 KO cells, in addition to ribosomes with tRNAs. This dormant subpopulation, absent in the WT hESCs, is characterized by the binding of SERPINE-mRNA binding protein 1 in the mRNA tunnel and eukaryotic elongation factor 2 near the A-site, preventing translation. The presence of elevated protein translation in FMR1 KO cells, alongside a subpopulation of inactive ribosomes, suggests that FMRP can function as a translational brake. However, due to the high cost of ribosome recycling, the cell appears to adopt a strategy of maintaining a subset of dormant ribosomes. Additionally, we analysed the 2-O-methylation patterns in the 28S rRNA, in both the WT and FMR1 KO hESCs, identifying few potential differentially methylated sites. Thus, these findings provide insights into the mechanisms of ribosome dormancy in the absence of FMRP and lay the groundwork for understanding the role of rRNA methylation in translational regulation.

biochemistry↗

bioRxiv2024

Altering rRNA 2'O-methylation pattern during neuronal differentiation is regulated by FMRP

The Fragile X Messenger Ribonucleoprotein (FMRP) is a selective RNA-binding protein that localizes to the cytoplasm and the nucleus. The loss of FMRP results in Fragile X Syndrome (FXS), an Autism Spectrum Disorder. FMRP interacts with ribosomes and regulates the translation of mRNAs essential for neuronal development and synaptic plasticity. However, the biochemical nature of this translation regulation is unknown. Here we report that a key feature of FMRP-mediated translation regulation during neuronal differentiation is modulating the 2O-methylation of ribosomal RNA. 2O-methylation, facilitated by C/D box snoRNAs in the nucleus, is a major epitranscriptome mark on rRNA, essential for ribosome assembly and function. We found that FMRP influences a distinct rRNA 2O-Methylation pattern across neuronal differentiation. We show that in H9 ESCs, FMRP interacts with a selected set of C/D box snoRNA in the nucleus resulting in the generation of ribosomes with a distinct pattern of rRNA 2O-Methylation. This epitranscriptome pattern on rRNA undergoes a significant change during the differentiation of ESCs to neuronal precursors and cortical neurons. ESCs display maximum hypomethylated residues on rRNA, which is eventually reduced in neuronal precursors and post-mitotic cortical neurons and this is correlated to the change in global protein synthesis among the states of differentiation. Importantly, this gradual change in the 2O-methylation pattern during neuronal differentiation is altered in the absence of FMRP, which could impact neuronal development and contribute to dysregulated protein synthesis observed in Fragile X Syndrome. This also suggests the need for diversity in functional ribosomes during the early stages of development.

molecular biology↗

bioRxiv2024

Dietary stress induced macrophage metabolic reprogramming, a determinant of animal growth

Nutrient sensing and signaling play pivotal roles in animal growth. However, under dietary stress, this system falters, leading to growth defects. While immune cells are increasingly recognized as key nutrient sensors, their impact on animal growth remains poorly understood. In this study, we investigate how Drosophila larval macrophages respond to excessive dietary sugar and identify a reconfiguration of their metabolic state. They undergo a glycolytic shift, intensify TCA activity, and elevate TAG synthesis. While typical of sugarinduced nutrient stress, these changes interestingly exert contrasting effects on animal growth: glycolysis and increased TCA activity inhibit growth, while the lipogenic shift promotes it. However, the lipogenic response is insufficient to counteract the metabolic events suppressing growth, resulting in an overall reduction in adult fly size under high sugar conditions. Stimulating a pro-lipogenic immune state facilitates growth recovery, suggesting a growth paradigm governed by immune-metabolic transitions. This study unveils the unexpected influence of macrophage metabolic reprogramming on organismal growth homeostasis during Drosophila development, highlighting immune cell states as central determinants of growth, particularly under dietary stress.

developmental biology↗

bioRxiv2024

Poly (A) Binding Protein 2 is critical for stem-progenitor differentiation during regeneration in the planarian Schmidtea mediterranea.

Post-transcriptional regulation has emerged as a key mechanism to regulate stem cell renewal and differentiation, which is essential for understanding tissue regeneration and homeostasis. Poly(A)-binding proteins are a family of RNA-binding proteins that play a vital role in post-transcriptional regulation by controlling mRNA stability and protein synthesis. The involvement of poly(A) binding proteins in a wide range of cellular functions is increasingly being investigated. In this study, we used the regenerative model organism planarian Schmidtea mediterranea, to demonstrate the critical role of poly(A)-binding protein 2 (PABP2) in regulating neoblast maintenance and differentiation. A deficit in PABP2 blocks the transition of neoblasts towards immediate early progenitors, leading to an enhanced pool of non-committed neoblasts and a decreased progenitor population. This is reflected in variations in the transcriptome profile, providing evidence of downregulation in multiple lineages. Thus, insufficiency of PABP2 resulted in defective formation and organization of tissue leading to abnormal regeneration. Our study reveals the essential role of PABP2 in regulating genes that mediate stem cell commitment to early progenitors during tissue regeneration.

cell biology↗

bioRxiv2023

Transcriptomic analysis identifies synapse-enriched lncRNAs required for glutamatergic synapse development and fear memory consolidation.

Regulatory functions of lncRNAs in neurons have been majorly limited to the nucleus. The identity of synaptic lncRNAs and their functional roles associated with synapse development and plasticity are poorly understood. Transcriptomic analysis from synaptoneurosomes identified 94 synapse-enriched lncRNAs from the mouse hippocampus. We find Pvt1 to be a specific regulator of excitatory, not inhibitory, synapse development. Synapse-specific loss of Pvt1 regulates synaptic activity and the downregulation of mRNAs pertinent to synapse development. We also report a synapse-centric role for an uncharacterized lncRNA; 2410006H16Rik (SynLAMP); in translation and memory formation. SynLAMP demonstrates enhanced localization to the synapse following fear conditioning and regulates dendritic translation by sequestering the translation repressor FUS; acting as a molecular decoy. Synapse-specific knockdown of SynLAMP inhibits the localised, activity-dependent translation of CamK2a, a FUS target. SynLAMP RNAi occludes fear-memory formation. Comprehensively, our study highlights that de novo activity of lncRNAs are involved in diverse synaptic functions.

neuroscience↗

bioRxiv2022

ES30L: A large subunit rRNA expansion segment enriched in mammals and birds

Ribosomes, the molecular machines that are central to protein synthesis, have gradually been gaining prominence for their regulatory role in translation. Eukaryotic cytosolic ribosomes are typically larger than bacterial ones, partly due to multi-nucleotide insertions at specific conserved positions in the ribosomal RNAs (rRNAs). Such insertions called expansion segments (ESs) are present primarily on the ribosomal surface, with their role in translation and its regulation remaining under-explored. One such ES in the ribosomal large subunit (LSU) is ES30L, which is present only in mammals and birds among eukaryotes. In this study, we show that ES30L possesses complementarity to many protein-coding transcripts in humans and that the complementarity is enriched around the start codon, hinting at a possible role in translation regulation. Further, our in silico analysis analyses and pull-down assays indicate that ES30L may bind to secondary structures in the 5 UTR of several transcripts and RNA binding proteins (RBPs) that are essential for translation. Thus, we have identified a potential regulatory role for ES30L in translation.

molecular biology↗

bioRxiv2021

Function of FMRP domains in regulating distinct roles of neuronal protein synthesis

The Fragile X Mental Retardation Protein (FMRP) is an RNA Binding Protein that regulates translation of mRNAs, essential for synaptic development and plasticity. FMRP interacts with a specific set of mRNAs and aids in their microtubule dependent transport and regulates their translation through its association with ribosomes. However, the biochemical role of individual domains of FMRP in forming neuronal granules and associating with microtubules and ribosomes is currently undefined. Here, we report that the C-terminus domain of FMRP is sufficient to bind to ribosomes as well as polysomes akin to the full-length protein. Furthermore, the C-terminus domain alone is essential and responsible for FMRP-mediated translation repression in neurons. However, FMRP-mediated puncta formation and microtubule association is favored by the synergistic combination of FMRP domains and not by individual domains. Interestingly, we show that the phosphorylation of hFMRP at Serine-500 is important in modulating the dynamics of translation by controlling ribosome/polysome association. This is a fundamental mechanism governing the size and number of FMRP puncta, which appear to contain actively translating ribosomes. Finally through the use of pathogenic mutations, we emphasize the hierarchy of the domains of FMRP in their contribution to translation regulation.

neuroscience↗

bioRxiv2021

DDX24, a D-E-A-D box RNA helicase, is required for muscle fiber organization and anterior pole specification essential for head regeneration in planarians.

Planarians have a remarkable ability to undergo whole-body regeneration. The timely establishment of polarity at the wound site followed by the specification of the organizing centers- the anterior pole and the posterior pole, are indispensable for successful regeneration. In planarians, polarity, pole, and positional-information determinants are predominantly expressed by muscles. The molecular toolkit that enables this functionality of planarian muscles however remains poorly understood. Here we report that SMED_DDX24, a D-E-A-D Box RNA helicase and the homolog of human DDX24, is critical for planarian head regeneration. DDX24 is enriched in muscles and its knockdown leads to defective muscle-fiber organization and failure to re-specify anterior pole/organizer. Overall, loss of DDX24 manifests into gross misregulation of many well-characterized positional-control genes and patterning-control genes, necessary for organogenesis and tissue positioning and tissue patterning. In addition, wound-induced Wnt signalling was also upregulated in ddx24 RNAi animals. Canonical WNT-{beta}CATENIN signalling is known to suppress head identity throughout bilateria, including planarians. Modulating this Wnt activity by {beta}-catenin-1 RNAi, the effector molecule of this pathway, partially rescues the ddx24 RNAi phenotype, implying that a high Wnt environment in ddx24 knockdown animals likely impedes their normal head regeneration. Furthermore, at a sub-cellular level, RNA helicases are known to regulate muscle mass and function by regulating their translational landscape. ddx24 knockdown leads to the downregulation of large subunit ribosomal RNA and the 80S ribosome peak, implying its role in ribosome biogenesis and thereby influencing the translational output. This aspect seems to be an evolutionarily conserved role of DDX24. In summary, our work demonstrates the role of a D-E-A-D box RNA helicase in whole-body regeneration through muscle fiber organization, and pole and positional-information re-specification, likely mediated through translation regulation.

developmental biology↗

bioRxiv2020

Comprehensive annotation and characterization of planarian tRNA and tRNA-derived fragments (tRFs)

tRNA-derived fragments (tRFs) have recently gained a lot of scientific interest due to their diverse regulatory roles in several cellular processes. However, their function in dynamic biological process such as development and regeneration remains unexplored. Here, we identify and characterise a role for tRFs in planarian regeneration. In order to characterise planarian tRFs, we first annotated 457 tRNA loci in S.mediterranea combining two tRNA prediction programs. Annotation of tRNAs facilitated the identification of three main species of tRFs in planarians - the shorter tRF-5s and itRFs, and the abundantly expressed 5-tsRNAs. Spatial profiling of tRFs in sequential transverse sections of planarians revealed diverse expression patterns of these small RNAs, including those that are enriched in the head and pharyngeal regions. Expression analysis of these tRF species revealed dynamic expression of these small RNAs over the course of regeneration suggesting an important role in planarian anterior and posterior regeneration. Finally, we show that 5-tsRNA in planaria interact with all three SMEDWI proteins while sequence analysis revealed a possible involvement of Dicers in the processing of itRFs. In summary, our findings implicate a novel role for tRFs in planarian regeneration, highlighting their importance in regulating complex systemic processes. Our study adds to the catalogue of post-transcriptional regulatory systems in planarian, providing valuable insights on the biogenesis and the function of tRFs in neoblasts and planarian regeneration.

molecular biology↗

bioRxiv2020

PAN-INDIA 1000 SARS-CoV-2 RNA Genome Sequencing Reveals Important Insights into the Outbreak

The PAN-INDIA 1000 SARS-CoV-2 RNA Genome Sequencing Consortium has achieved its initial goal of completing the sequencing of 1000 SARS-CoV-2 genomes from nasopharyngeal and oropharyngeal swabs collected from individuals testing positive for COVID-19 by Real Time PCR. The samples were collected across 10 states covering different zones within India. Given the importance of this information for public health response initiatives investigating transmission of COVID-19, the sequence data is being released in GISAID database. This information will improve our understanding on how the virus is spreading, ultimately helping to interrupt the transmission chains, prevent new cases of infection, and provide impetus to research on intervention measures. This will also provide us with information on evolution of the virus, genetic predisposition (if any) and adaptation to human hosts. One thousand and fifty two sequences were used for phylodynamic, temporal and geographic mutation patterns and haplotype network analyses. Initial results indicate that multiple lineages of SARS-CoV-2 are circulating in India, probably introduced by travel from Europe, USA and East Asia. A2a (20A/B/C) was found to be predominant, along with few parental haplotypes 19A/B. In particular, there is a predominance of the D614G mutation, which is found to be emerging in almost all regions of the country. Additionally, mutations in important regions of the viral genome with significant geographical clustering have also been observed. The temporal haplotype diversities landscape in each region appears to be similar pan India, with haplotype diversities peaking between March-May, while by June A2a (20A/B/C) emerged as the predominant one. Within haplotypes, different states appear to have different proportions. Temporal and geographic patterns in the sequences obtained reveal interesting clustering of mutations. Some mutations are present at particularly high frequencies in one state as compared to others. The negative estimate Tajimas D (D = -2.26817) is consistent with the rapid expansion of SARS-CoV-2 population in India. Detailed mutational analysis across India to understand the gradual emergence of mutants at different regions of the country and its possible implication will help in better disease management.

genomics↗

bioRxiv2020

Mitochondrial state determines functionally divergent stem cell population in planaria

Mitochondrial state changes were shown to be critical for stem cell function. However, variation in the mitochondrial content in stem cells and the implication, if any, on differentiation is poorly understood. Here, using cellular and molecular studies, we show that the planarian pluripotent stem cells (PSCs) have low mitochondrial mass compared to its progenitors. Further, the mitochondrial mass correlated with OxPhos and inhibiting the transition to OxPhos dependent metabolism in cultured cells resulted in higher PIWI-1High neoblasts. Transplantation experiments provided functional validation that neoblasts with low mitochondrial mass are the true PSCs. In summary, we show that low mitochondrial mass is a hallmark of PSCs in planaria and provide a mechanism to isolate live, functionally active, PSCs from different cell cycle stages (G0/G1 and S, G2/M). Our study demonstrates that the change in mitochondrial metabolism, a feature of PSCs is conserved in planaria and highlights its role in organismal regeneration.

cell biology↗