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27 pages, 4385 KB  
Article
Environmental Concentrations of PFOS Accumulate in the Euglena Eyespot and Impair Chloroplast ATP Synthase Activity: A Dual Impairment of Phototaxis and Photosynthetic Light Reactions
by Peirui Liu, Junfeng Wang, Yan Hong, Zilin Chen, Xiaoya Liu, Huayi Chen, Ganning Zeng and Xiangliang Pan
Toxics 2026, 14(6), 540; https://doi.org/10.3390/toxics14060540 (registering DOI) - 22 Jun 2026
Abstract
Perfluorooctane sulfonate (PFOS) is a persistent organic pollutant widely detected in aquatic ecosystems, but its subcellular targets and the mechanisms by which it disrupts light resource utilization in photosynthetic protozoa remain poorly understood at concentrations spanning environmentally typical to supra-environmental levels. Here, Euglena [...] Read more.
Perfluorooctane sulfonate (PFOS) is a persistent organic pollutant widely detected in aquatic ecosystems, but its subcellular targets and the mechanisms by which it disrupts light resource utilization in photosynthetic protozoa remain poorly understood at concentrations spanning environmentally typical to supra-environmental levels. Here, Euglena gracilis G.A. Klebs was exposed to PFOS at concentrations spanning environmentally typical (0.5 µg/L), hotspot-relevant (5 µg/L), and supra-environmental (50 µg/L) levels. Subcellular distribution, phototaxis, photosynthetic light reactions, and energy metabolism were investigated using isolated chloroplast assays, transcriptomics, and proteomics. TEM-EDS mapping revealed pronounced fluorine signal enrichment, attributable to PFOS, in the eyespot and chloroplasts. Eyespot fluorine enrichment was associated with impaired phototactic motility and an altered light perception threshold. PFOS did not acutely inhibit the maximum photochemical efficiency of photosystem II (Fv/Fm); instead, a transient upregulation of photosynthesis-related genes was observed, which weakened with prolonged exposure, whereas the photosynthetic electron transport rate (ETR) was significantly reduced. PFOS significantly reduced ATP levels and ETR, while Fv/Fm remained unchanged and non-photochemical quenching (NPQ) was elevated. Isolated chloroplast assays revealed that PFOS inhibits Mg2+-dependent ATP hydrolytic activity in the chloroplast-enriched fraction and impairs thylakoid electron transport, consistent with impaired chloroplast ATP synthase function, though the specific molecular target and mechanism remain to be conclusively demonstrated. Transcriptomic and proteomic analyses revealed compensatory upregulation of photosynthesis pathways but suppression of ATP synthesis and redox homeostasis. Collectively, our results suggest that PFOS impairs chloroplast ATP synthase function, accompanied by reduced ETR and elevated NPQ. Together with the eyespot-associated phototaxis impairment, these effects suggest that PFOS may dually disrupt light acquisition (behavioral) and light conversion (physiological) in E. gracilis. This dual impairment may compromise the ecological fitness of Euglena in PFOS-contaminated environments, especially under prolonged exposure. It should be noted that the subcellular fluorine mapping is qualitative, the phototaxis assay reflects population-level responses, and the ATP synthase impairment interpretation is indirect; the proposed mechanistic model remains a hypothesis requiring further direct experimental validation. Full article
(This article belongs to the Special Issue Toxic Effects of Emerging Pollutants on Aquatic Organisms and Human)
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15 pages, 3362 KB  
Article
Reduced Glutathione Supplementation Enhances Metamorphosis and Disease Resistance in Larvae of Pacific Oyster Crassostrea gigas
by Xuesong Li, Junlei Cheng, Jiadi Wang, Pan Luo, Chunmei Xiong, Weilin Wang, Lingling Wang and Linsheng Song
Fishes 2026, 11(6), 333; https://doi.org/10.3390/fishes11060333 - 2 Jun 2026
Viewed by 314
Abstract
As an important aquaculture bivalve, the Pacific oyster (Crassostrea gigas) undergoes a metamorphosis process during its embryonic development. The mortality rate of the eye-spot larva during the metamorphosis and settlement stage is relatively high, which urgently needs to be solved. Glutathione [...] Read more.
As an important aquaculture bivalve, the Pacific oyster (Crassostrea gigas) undergoes a metamorphosis process during its embryonic development. The mortality rate of the eye-spot larva during the metamorphosis and settlement stage is relatively high, which urgently needs to be solved. Glutathione (GSH), as a key antioxidant and immune regulator in organisms, plays an important role in the growth, development, and immune defense of marine invertebrates. Analysis of oyster transcriptome data revealed that the expression of key enzyme-producing GSH, Glutathione S-transferase Kappa 1, reached the highest level at the pediveliger larvae (the stage just before eye-spot larvae), suggesting its important role in the larval settlement process. This study investigated the effects of reduced glutathione supplementation in pediveliger larvae on the settlement metamorphosis and immune capacity of eye-spot larvae. The results showed that high concentrations of glutathione (100 μmol/L, 500 μmol/L) significantly reduced the larval eye-spot rate and settlement rate, with larvae in the 500 μmol/L group nearly all dead after 48 h. Meanwhile, low-concentration GSH (7 μmol/L) could significantly increase the settlement rate of larvae, which was 1.17 times (p < 0.05) that of the control group. When the larvae developed to 1-month-old eye-spot larvae, the survival rate of larvae in the low GSH group significantly increased (38.42 ± 15.98%) after Vibrio splendidus stimulation, while they were almost all dead in the control group. Low GSH could also significantly increase lysozyme LZM activity (by 1.39 times, p < 0.05) and Cu/Zn-SOD activity (by 1.15 times, p < 0.05). Gene expression analysis found that low GSH promoted expressions of larval settlement and metamorphosis-related genes, including ecdysone receptor, P450, and calmodulin. Treatment with low GSH also enhanced the expressions of immune genes of SOD, CAT, and LZM. In summary, a low glutathione supplement can promote the settlement process of oyster eye-spot larvae and enhance disease resistance. These findings reveal the role of glutathione in the metamorphosis and settlement of oyster larvae, providing a scientific basis for optimizing techniques for oyster seedling cultivation. Full article
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32 pages, 17226 KB  
Article
Dynamic Physical Distortions of Butterfly Pupal Wings: Potential Mechanical Signals from Eyespot Organizers for Color Pattern Determination
by Yugo Nakazato, Euichi Hirose and Joji M. Otaki
Biology 2026, 15(11), 856; https://doi.org/10.3390/biology15110856 - 29 May 2026
Viewed by 351
Abstract
Butterfly wing color patterns are determined in pupal wing tissues, in which the prospective eyespot focus functions as a developmental organizer. Here, we investigated the microscopic structures of pupal wing tissues containing an eyespot organizer in line with the physical distortion hypothesis. Histochemical [...] Read more.
Butterfly wing color patterns are determined in pupal wing tissues, in which the prospective eyespot focus functions as a developmental organizer. Here, we investigated the microscopic structures of pupal wing tissues containing an eyespot organizer in line with the physical distortion hypothesis. Histochemical staining revealed that the pupal cuticle and epidermis were wavy and thin at 6 h but smooth and thick at 12 h postpupation. The eyespot organizer was associated with the thickest cuticle layer, called the cuticle focal spot. Transmission election microscopy (TEM) revealed that the intervening space (IVS) between the cuticle layer and the cellular apical end was wide at 6 h but narrow at 12 h postpupation. The spatial relationship between cuticle thickness and IVS width was indicative of mechanical buckling of the region adjacent to the cuticle focal spot. Live in vivo imaging revealed that the IVS at and near the eyespot organizer trapped orange fluorescent protein (OFP) injected into hemolymph. Dynamic distortions of the pupal wing epidermis and cuticular surface were detected in live individuals over time. These results suggest that physical distortions of the wing tissue induced by differential cuticle synthesis and subsequent buckling may function as mechanical morphogenic signals from eyespot organizers. Full article
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22 pages, 11713 KB  
Article
Pharmacological Effects of NADPH Oxidase Inhibitors on Butterfly Wing Morphogenesis and Color Pattern Formation in Junonia orithya
by Yugo Nakazato, Momo Ozaki, Ryunosuke Suenaga and Joji M. Otaki
Insects 2026, 17(3), 300; https://doi.org/10.3390/insects17030300 - 10 Mar 2026
Cited by 1 | Viewed by 920
Abstract
During the early pupal stage in butterflies, the peripheral portion of wing tissue undergoes apoptosis to finalize adult wing morphology, and wing color patterns are determined coordinately. We hypothesized that the development of wing morphology and color patterns may involve NADPH oxidase (NOX). [...] Read more.
During the early pupal stage in butterflies, the peripheral portion of wing tissue undergoes apoptosis to finalize adult wing morphology, and wing color patterns are determined coordinately. We hypothesized that the development of wing morphology and color patterns may involve NADPH oxidase (NOX). To test this hypothesis, we treated pupae of the blue pansy butterfly Junonia orithya with NOX inhibitors. When VAS2870, isuzinaxib, or diphenyleneiodonium chloride (DPI) in dimethyl sulfoxide (DMSO) was topically applied to the pupal wing tissue via the sandwich method, wing morphology and color pattern elements, including eyespots, parafocal elements, submarginal bands, and marginal bands, were severely deformed as if the marginal area were surgically removed. The topical application of DMSO alone mildly deformed and enlarged eyespots without affecting other color patterns and wing morphology. When systemically injected into pupae, VAS2870 increased eyespots in males but decreased eyespots in females, likely due to the sexual dimorphism of this species. These results suggest that NOX and probably hydrogen peroxide play important roles in wing morphogenesis and color pattern fate determination in butterfly wings. Sexually dimorphic eyespot size in this species may also be explained by the sexually differential activities of NOX. Full article
(This article belongs to the Section Insect Physiology, Reproduction and Development)
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23 pages, 7830 KB  
Article
TRPA1 for Butterfly Eyespot Formation
by Momo Ozaki and Joji M. Otaki
Int. J. Mol. Sci. 2026, 27(3), 1420; https://doi.org/10.3390/ijms27031420 - 30 Jan 2026
Cited by 2 | Viewed by 726
Abstract
Butterfly wing color pattern formation is a process of two-dimensional morphogenesis involving long-range lateral signaling in pupal wing tissues. We hypothesized that TRP (transient receptor potential) channels, which are multimodal sensors for various stimuli, are involved in this developmental process. Using the blue [...] Read more.
Butterfly wing color pattern formation is a process of two-dimensional morphogenesis involving long-range lateral signaling in pupal wing tissues. We hypothesized that TRP (transient receptor potential) channels, which are multimodal sensors for various stimuli, are involved in this developmental process. Using the blue pansy butterfly Junonia orithya, we injected the TRPA1 antagonists, AM0902 and AP-18, and an agonist, JT010, into pupae and observed that the eyespot core disk area in adult wings increased and decreased in response to AM0902 and JT010, respectively, although AP-18 did not induce any change. Furthermore, the eyespot outer black ring area increased in response to AM0902, and the orange ring area increased in response to JT010. We detected TRPA1 mRNA via RT-PCR in the pupal wing tissues of this species. An antibody against the J. orithya TRPA1 extracellular site induced unique aberrant color patterns with wing vein defects. These results suggest that TRPA1 is expressed in pupal wing tissue and may integrate signaling information to determine eyespot size and structure in butterfly wings. TRPA1 likely suppresses the black core disk and the outer black ring and enhances the nonblack orange ring in eyespots during development. Full article
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13 pages, 852 KB  
Communication
Maize Diseases in Northeast China: Current Status and Emerging Threats
by Bingbing Liang, Dongyu Li, Lingxi He, Huaiyu Dong, Lijuan Wang, Le Chen, Kejie Liu and Ping Wang
Agriculture 2026, 16(2), 249; https://doi.org/10.3390/agriculture16020249 - 19 Jan 2026
Viewed by 763
Abstract
A comprehensive two-year investigation (2024–2025) was conducted across Northeast China’s crucial grain production base to assess the status of maize diseases. Field surveys spanning three provinces and Inner Mongolia revealed a significant shift in the regional disease profile, with diagnosis performed by experienced [...] Read more.
A comprehensive two-year investigation (2024–2025) was conducted across Northeast China’s crucial grain production base to assess the status of maize diseases. Field surveys spanning three provinces and Inner Mongolia revealed a significant shift in the regional disease profile, with diagnosis performed by experienced personnel based on characteristic field symptoms. The results demonstrated that maize white spot (MWS) has emerged as a severe new threat, recording remarkably high disease severity indices exceeding 80 at multiple locations (e.g., LDD25-1: 86.83). Concurrently, gray leaf spot (GLS) was confirmed as the most prevalent foliar disease, forming stable areas of high severity in the eastern mountainous regions where its disease indices consistently surpassed 60 (e.g., LFS25-1: 65.26), thereby exceeding the impact of northern corn leaf blight. In contrast, stalk rot (SR) maintained a low field incidence rate below 10%, while other diseases such as Curvularia leaf spot and maize eyespot were only observed locally or were absent during the 2025 survey period. These findings underscore the emergence of MWS as a critical threat and affirm the dominant status of GLS, offering a scientific foundation for prioritizing disease management strategies in the region. Full article
(This article belongs to the Section Crop Protection, Diseases, Pests and Weeds)
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16 pages, 1551 KB  
Article
Host Response of Winter Wheat to the Causal Agents of Eyespot and Fungicide Resistance of the Pathogens
by Jana Palicová, Pavel Matušinsky, Simona Čejková, Alena Hanzalová, Veronika Dumalasová, Taťána Militká, Dominik Bleša and Jana Chrpová
Plants 2026, 15(2), 285; https://doi.org/10.3390/plants15020285 - 17 Jan 2026
Viewed by 894
Abstract
Eyespot is one of the most important fungal diseases of wheat in the Czech Republic. As part of a long-term study (2015–2024), the occurrence, population structure, and pathogenic variability of Oculimacula yallundae and Oculimacula acuformis were investigated. In total, 356 O. yallundae, [...] Read more.
Eyespot is one of the most important fungal diseases of wheat in the Czech Republic. As part of a long-term study (2015–2024), the occurrence, population structure, and pathogenic variability of Oculimacula yallundae and Oculimacula acuformis were investigated. In total, 356 O. yallundae, 24 O. acuformis, and 33 mixed cultures were collected and identified using PCR. The study also included small-plot inoculation trials (2022–2023) to assess the response of widely grown winter wheat cultivars. Disease severity was evaluated visually, pathogen DNA was quantified using qPCR, and the presence of the resistance gene Pch1 was determined with the STS marker Xorw1. In addition to these analyses, monitoring of fungicide resistance to two commonly used fungicides (fluxapyroxad and prothioconazole) was performed. The results showed significant differences among cultivars and seasons. Genotypes carrying Pch1—including Annie, Campesino, Illusion, KWS Donovan, LG Absalon, and Pallas—exhibited the lowest levels of infection, whereas Mercedes and Dagmar were the most susceptible. The qPCR reliably detected and distinguished both pathogens, with O. yallundae occurring at higher concentrations. Fungicide sensitivity testing revealed EC50 values (mean ± SD) of 0.09 ± 0.13 μg·mL−1 for fluxapyroxad and 0.30 ± 0.22 μg·mL−1 for prothioconazole, indicating that eyespot pathogens remain largely sensitive, with only minor signs of reduced sensitivity. Full article
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28 pages, 11120 KB  
Article
Pharmacological Intervention of PIEZO1 for Butterfly Eyespot Color Patterns in Junonia orithya
by Momo Ozaki and Joji M. Otaki
Receptors 2025, 4(4), 20; https://doi.org/10.3390/receptors4040020 - 21 Oct 2025
Cited by 3 | Viewed by 1777
Abstract
Background: PIEZO channels are mechanoreceptors expressed in various cells. Their contributions to animal development are not entirely clear. According to the physical distortion hypothesis, developmental organizers for butterfly wing eyespots receive and release mechanical signals in pupal wing tissues during development, initiating [...] Read more.
Background: PIEZO channels are mechanoreceptors expressed in various cells. Their contributions to animal development are not entirely clear. According to the physical distortion hypothesis, developmental organizers for butterfly wing eyespots receive and release mechanical signals in pupal wing tissues during development, initiating a calcium signaling cascade and gene expression changes. Objectives: We tested the possible involvement of PIEZO1 in butterfly wing color pattern formation, according to the physical distortion hypothesis. Methods: We performed a pharmacological intervention of PIEZO1, focusing on the eyespots of Junonia orithya. Chemical modulators of PIEZO1 and the actin cytoskeleton were injected into pupae immediately after pupation during the critical period of color pattern determination, and the eyespot color patterns of the emerging adult wings were analyzed. We also tested dimethyl sulfoxide (DMSO) because it was used as a solvent. Results: DMSO significantly enlarged most eyespots examined. In contrast, the specific PIEZO1 activator Jedi2 induced significant reduction in the dorsal hindwing eyespots. Another specific PIEZO1 activator, Yoda1, also induced similar changes, although less clearly. The mechanosensitive channel blocker GsMTx4 produced compromised eyespots in an individual, although statistical support for modification was weak. The actin polymerization activator phalloidin induced blue foci in the ventral forewing eyespots. PIEZO expression in the pupal wings was demonstrated by RT-PCR. Conclusions: These results suggest that eyespot organizers in butterfly wings may employ a PIEZO-mediated mechanotransduction pathway to regulate eyespot color patterns, supporting the physical distortion hypothesis. These results highlight the importance of PIEZO in developmental organizers in animals. Full article
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16 pages, 2508 KB  
Article
Eyespot Variation in the Meadow Brown Butterfly, Maniola jurtina (Insecta: Lepidoptera) in Diverse Climatic Conditions
by Tina Klenovšek, Predrag Jakšić and Franc Janžekovič
Diversity 2025, 17(10), 675; https://doi.org/10.3390/d17100675 - 26 Sep 2025
Viewed by 1637
Abstract
Eyespots are functionally complex and highly variable elements of butterfly wing patterns. The Meadow Brown, Maniola jurtina, is a classic model species studied for variation in eyespots as an index of evolutionary divergence and adaptation. However, the role of fine-scale ecogeographic conditions [...] Read more.
Eyespots are functionally complex and highly variable elements of butterfly wing patterns. The Meadow Brown, Maniola jurtina, is a classic model species studied for variation in eyespots as an index of evolutionary divergence and adaptation. However, the role of fine-scale ecogeographic conditions on eyespot variation remains poorly understood. In this study, we examined hindwing eyespot number, distribution, and combination patterns in male M. jurtina across climatically and topographically diverse north-western Balkans. Compared to the species average, males in this region displayed greater spottiness and phenotypic diversity. While the typical two-spot phenotype was dominant and stable, in some populations, three-spotted and even four-spotted males occurred at similar frequencies. Rare six-spotted individuals were recorded only at mountain localities above 1200 m. Geographic and climatic factors together influenced this variation: higher altitudes and cooler, thermally stable environments promoted increased eyespot number and greater phenotypic plasticity than warmer, more variable environments. This pattern contrasts with large-scale latitudinal trends previously described for the species, emphasizing the importance of local climatic heterogeneity. Our findings suggest the north-western Balkans as a possible transitional zone where environmental complexity promotes elevated eyespot variability, contributing to the understanding of adaptive morphological plasticity in M. jurtina. Full article
(This article belongs to the Section Animal Diversity)
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22 pages, 7620 KB  
Article
DSTANet: A Lightweight and High-Precision Network for Fine-Grained and Early Identification of Maize Leaf Diseases in Field Environments
by Xinyue Gao, Lili He, Yinchuan Liu, Jiaxin Wu, Yuying Cao, Shoutian Dong and Yinjiang Jia
Sensors 2025, 25(16), 4954; https://doi.org/10.3390/s25164954 - 10 Aug 2025
Cited by 2 | Viewed by 1266
Abstract
Early and accurate identification of maize diseases is crucial for ensuring sustainable agricultural development. However, existing maize disease identification models face challenges including high inter-class similarity, intra-class variability, and limited capability in identifying early-stage symptoms. To address these limitations, we proposed DSTANet (decomposed [...] Read more.
Early and accurate identification of maize diseases is crucial for ensuring sustainable agricultural development. However, existing maize disease identification models face challenges including high inter-class similarity, intra-class variability, and limited capability in identifying early-stage symptoms. To address these limitations, we proposed DSTANet (decomposed spatial token aggregation network), a lightweight and high-performance model for maize leaf disease identification. In this study, we constructed a comprehensive maize leaf image dataset comprising six common disease types and healthy samples, with early and late stages of northern leaf blight and eyespot specifically differentiated. DSTANet employed MobileViT as the backbone architecture, combining the advantages of CNNs for local feature extraction with transformers for global feature modeling. To enhance lesion localization and mitigate interference from complex field backgrounds, DSFM (decomposed spatial fusion module) was introduced. Additionally, the MSTA (multi-scale token aggregator) was designed to leverage hidden-layer feature channels more effectively, improving information flow and preventing gradient vanishing. Experimental results showed that DSTANet achieved an accuracy of 96.11%, precision of 96.17%, recall of 96.11%, and F1-score of 96.14%. With only 1.9M parameters, 0.6 GFLOPs (floating point operations), and an inference speed of 170 images per second, the model meets real-time deployment requirements on edge devices. This study provided a novel and practical approach for fine-grained and early-stage maize disease identification, offering technical support for smart agriculture and precision crop management. Full article
(This article belongs to the Section Smart Agriculture)
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20 pages, 1692 KB  
Article
Molecular Mechanism of Metformin Regulating the Regeneration of Planarian Dugesia japonica Through miR-27b
by Kexin Yang, Minmin Feng, Chunmei Zhang, Zelong Zhao, Dandan Yin, Linxia Song and Zhenbiao Xu
Int. J. Mol. Sci. 2025, 26(15), 7092; https://doi.org/10.3390/ijms26157092 - 23 Jul 2025
Viewed by 1412
Abstract
Metformin is one of the most commonly used medications to treat type 2 diabetes. In addition to lowering blood sugar, it can also promote the regeneration of certain organs or tissues. Planarian Dugesia japonica, with its remarkable regenerative capacity, has become an [...] Read more.
Metformin is one of the most commonly used medications to treat type 2 diabetes. In addition to lowering blood sugar, it can also promote the regeneration of certain organs or tissues. Planarian Dugesia japonica, with its remarkable regenerative capacity, has become an important model organism for studying pharmacology and regenerative medicine. Planarian eyespot regeneration involves precise tissue regeneration via mechanisms like cell proliferation, differentiation, and gene regulation following body damage. Experiments on planarian eyespot regeneration have confirmed that 1 mM metformin significantly promotes regeneration. Through analysis of the regenerating planarian miRNA database and the metformin-treated transcriptome database, combined with target gene prediction by TargetScan, the DjmiR-27b/DjPax6 axis was finally determined as the research focus. qPCR showed that metformin significantly affects the expression levels of DjmiR-27b and DjPax6. DjPax6 was identified as the target gene of DjmiR-27b through dual luciferase reporter gene analysis. Functional experiments revealed that metformin regulates the expression of DjPax6 via DjmiR-27b, thereby influencing the regeneration of planarian eyespots. In situ hybridization showed that both DjmiR-27b and DjPax6 are expressed throughout the entire body. This study reveals the molecular mechanism of metformin regulating planarian regeneration through miRNA, providing further insights into its role in the field of regeneration. Full article
(This article belongs to the Section Molecular Biology)
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26 pages, 19485 KB  
Review
Some Insights into the Inventiveness of Dinoflagellates: Coming Back to the Cell Biology of These Protists
by Marie-Odile Soyer-Gobillard
Microorganisms 2025, 13(5), 969; https://doi.org/10.3390/microorganisms13050969 - 24 Apr 2025
Viewed by 2242
Abstract
In this review dedicated to the great protistologist Edouard Chatton (1883–1947), I wanted to highlight the originality and remarkable diversity of some dinoflagellate protists through the lens of cell biology. Their fossilized traces date back to more than 538 million years (Phanerozoic eon). [...] Read more.
In this review dedicated to the great protistologist Edouard Chatton (1883–1947), I wanted to highlight the originality and remarkable diversity of some dinoflagellate protists through the lens of cell biology. Their fossilized traces date back to more than 538 million years (Phanerozoic eon). However, they may be much older because acritarchs from the (Meso) Proterozoic era (1500 million years ago) could be their most primitive ancestors. Here, I described several representative examples of the various lifestyles of free-living (the autotrophic thecate Prorocentrum micans Ehrenberg and the heterotrophic athecate Noctiluca scintillans McCartney and other “pseudo-noctilucidae”, as well as the thecate Crypthecodinium cohnii Biecheler) and of parasitic dinoflagellates (the mixotroph Syndinium Chatton). Then, I compared the different dinoflagellate mitotic systems and reported observations on the eyespot (ocelloid), an organelle that is present in the binucleated Glenodinium foliaceum Stein and in some Warnowiidae dinoflagellates and can be considered an evolutionary marker. The diversity and innovations observed in mitosis, meiosis, reproduction, sexuality, cell cycle, locomotion, and nutrition allow us to affirm that dinoflagellates are among the most innovative unicells in the Kingdom Protista. Full article
(This article belongs to the Special Issue Research on Biology of Dinoflagellates)
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21 pages, 4402 KB  
Article
Transcriptome Sequencing Analysis of the Effects of Metformin on the Regeneration of Planarian Dugesia japonica
by Zelong Zhao, Dandan Yin, Kexin Yang, Chunmei Zhang, Linxia Song and Zhenbiao Xu
Genes 2025, 16(4), 365; https://doi.org/10.3390/genes16040365 - 22 Mar 2025
Cited by 1 | Viewed by 1332
Abstract
Background: Metformin is a widely used oral hypoglycemic agent for treating type 2 diabetes. Planarians, with their remarkable regenerative abilities, are frequently employed as model organisms in stem cell and regeneration studies. This study aimed to investigate the effects of metformin on planarian [...] Read more.
Background: Metformin is a widely used oral hypoglycemic agent for treating type 2 diabetes. Planarians, with their remarkable regenerative abilities, are frequently employed as model organisms in stem cell and regeneration studies. This study aimed to investigate the effects of metformin on planarian regeneration, focusing on the regeneration of eyespots after amputation. Methods: Regenerating planarians with amputated eyespots were exposed to various concentrations of metformin. The regeneration time of the eyespots was measured to assess the effects of metformin. Subsequently, a 1 mmol/L metformin treatment for 24 h was applied to the planarians, followed by transcriptome analysis to identify differentially expressed genes (DEGs). The gene expression was validated through qPCR. The full-length gene of casein kinase 1α (DjCK1α) was cloned using RACE technology. DjCK1α interference was performed to examine its role in regeneration. Results: Low concentrations of metformin significantly reduced the regeneration time of planarians. Transcriptome analysis identified 113 DEGs, including 61 upregulated and 52 downregulated genes. GO and KEGG enrichment analyses were conducted. Notably, DjCK1α, a key gene involved in regeneration, was selected for further validation. qPCR confirmed that DjCK1α was significantly upregulated. The interference of DjCK1α prolonged the regeneration time of the eyespots of planarians cultured in water, while treatment with metformin did not promote the eyespot regeneration of the DjCK1α-interfered planarians. Conclusions: The results suggest that metformin accelerates planarian eyespot regeneration, potentially through the regulation of DjCK1α. This study provides the first transcriptome-based analysis of drug effects on regeneration in planarians, highlighting the role of metformin in the regeneration process. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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34 pages, 36825 KB  
Article
Wound Healing in Butterfly Pupal Wing Tissues: Real-Time In Vivo Imaging of Long-Range Cell Migration, Cluster Formation, and Calcium Oscillations
by Shuka Nagai and Joji M. Otaki
Insects 2025, 16(2), 124; https://doi.org/10.3390/insects16020124 - 27 Jan 2025
Cited by 4 | Viewed by 3347
Abstract
Insects can repair wounds and regenerate body parts in response to physical damage. Wound healing in butterfly pupal wing tissues is developmentally interesting because ectopic color patterns develop during healing, suggesting that normal and damage-induced color patterns may use similar mechanisms. Here we [...] Read more.
Insects can repair wounds and regenerate body parts in response to physical damage. Wound healing in butterfly pupal wing tissues is developmentally interesting because ectopic color patterns develop during healing, suggesting that normal and damage-induced color patterns may use similar mechanisms. Here we physiologically investigated wound healing and ectopic color pattern formation in butterfly pupal wing tissues using the blue pansy butterfly Junonia orithya. In response to physical puncture damage, various ectopic color patterns are formed around the damage site. After the wounding operation, we observed hemocytes migrating over long distances along the wing veins (lacunae) toward the damage site, where hemocytes and epidermal cells formed cellular clusters. Calcium oscillations were observed in cells at and near the damage site. Calcium oscillations were transiently affected by ruthenium red, an inhibitor of calcium transporters and channels, and ruthenium red caused various abnormalities in the scales of adult wings. These results suggest that cell migration, cluster formation, and calcium oscillations play important roles in wound healing and scale development at and near the damage site. Ectopic color patterns may develop in response to local calcium oscillations as a consequence of the evolutionary co-option of the healing process for normal development. Full article
(This article belongs to the Section Insect Physiology, Reproduction and Development)
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22 pages, 25162 KB  
Article
Antibody-Mediated Protein Knockdown Reveals Distal-less Functions for Eyespots and Parafocal Elements in Butterfly Wing Color Pattern Development
by Yugo Nakazato and Joji M. Otaki
Cells 2024, 13(17), 1476; https://doi.org/10.3390/cells13171476 - 2 Sep 2024
Cited by 4 | Viewed by 2445
Abstract
One of the important genes for eyespot development in butterfly wings is Distal-less. Its function has been evaluated via several methods, including CRISPR/Cas9 genome editing. However, functional inhibition may be performed at the right time at the right place using a different [...] Read more.
One of the important genes for eyespot development in butterfly wings is Distal-less. Its function has been evaluated via several methods, including CRISPR/Cas9 genome editing. However, functional inhibition may be performed at the right time at the right place using a different method. Here, we used a novel protein delivery method for pupal wing tissues in vivo to inactivate a target protein, Distal-less, with a polyclonal anti-Distal-less antibody using the blue pansy butterfly Junonia orithya. We first demonstrated that various antibodies including the anti-Distal-less antibody were delivered to wing epithelial cells in vivo in this species. Treatment with the anti-Distal-less antibody reduced eyespot size, confirming the positive role of Distal-less in eyespot development. The treatment eliminated or deformed a parafocal element, suggesting a positive role of Distal-less in the development of the parafocal element. This result also suggested the integrity of an eyespot and its corresponding parafocal element as the border symmetry system. Taken together, these findings demonstrate that the antibody-mediated protein knockdown method is a useful tool for functional assays of proteins, such as Distal-less, expressed in pupal wing tissues, and that Distal-less functions for eyespots and parafocal elements in butterfly wing color pattern development. Full article
(This article belongs to the Section Reproductive Cells and Development)
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