RESUMO
Leber's hereditary optic neuropathy (LHON) is a debilitating mitochondrial disease associated with mutations in mitochondrial DNA (mtDNA). Unfortunately, the available treatment options for LHON patients are limited due to challenges in mitochondrial replacement. In our study, we reprogramming LHON urine cells into induced pluripotent stem cells (iPSCs) and differentiating them into neural progenitor cells (NPCs) and neurons for disease modeling. Our research revealed that LHON neurons exhibited significantly higher levels of mtDNA mutations and reduced mitochondrial function, confirming the disease phenotype. However, through co-culturing LHON iPSC-derived NPCs with mesenchymal stem cells (MSCs), we observed a remarkable rescue of mutant mtDNA and a significant improvement in mitochondrial metabolic function in LHON neurons. These findings suggest that co-culturing with MSCs can enhance mitochondrial function in LHON NPCs, even after their differentiation into neurons. This discovery holds promise as a potential therapeutic strategy for LHON patients.
RESUMO
Biomolecular condensates or membraneless organelles (MLOs) formed by liquid-liquid phase separation (LLPS) divide intracellular spaces into discrete compartments for specific functions. Dysregulation of LLPS or aberrant phase transition that disturbs the formation or material states of MLOs is closely correlated with neurodegeneration, tumorigenesis, and many other pathological processes. Herein, we summarize the recent progress in development of methods to monitor phase separation and we discuss the biogenesis and function of MLOs formed through phase separation. We then present emerging proof-of-concept examples regarding the disruption of phase separation homeostasis in a diverse array of clinical conditions including neurodegenerative disorders, hearing loss, cancers, and immunological diseases. Finally, we describe the emerging discovery of chemical modulators of phase separation.
RESUMO
Increasing attention is being paid to the toxic physiological effects of nanoplastics (NPs) on aquatic organisms. However, few studies have systematically evaluated the regulatory mechanisms of NPs on immune response in crustaceans. In this study, a 28-day chronic exposure experiment was conducted in which shrimps were exposed to various 80-nm polystyrene NPs concentrations (0, 0.1, 1, 5 and 10 mg/L). Transcriptomic analysis was used to investigate the regulatory mechanisms of NPs in immune response of Litopenaeus vannamei. With increasing NPs concentration, the total hemocyte count (THC) content decreased, while phagocytosis rate (PR) and respiratory burst (RB) showed trends of first rising and then falling. High concentration (10 mg/L) of NPs caused the destruction of hepatopancreas tissue structure, the shedding of microvilli, the increase number of hepatocyte apoptosis and autophagy structure. With increasing NPs concentration, the lysozyme (Lys), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities first increased and then decrease, while contents of lipid peroxidation and malondialdehyde increased; the expression levels of Toll, MyD88, GPx, SOD, proPO, Lys, and ALF generally increased at first and then decreased. Transcriptional sequencing analysis showed that the pathway of differentially expressed genes in KEGG enrichment mainly included lysosome (ko04142), apoptosis (ko04210) pathways, indicating that the NPs mainly affected the immune regulatory mechanism. Further analysis by Gene Set Enrichment Analysis (GSEA) showed that the up-regulation pathways of NPs activation mainly included immune response-related pathways such as mitochondrial autophagy, DNA repair, autophagosomes signaling pathway. Our results indicated that NPs exposure induced oxidative stress, apoptosis and autophagy in shrimps. This study provides a basis for further understanding of the mechanisms of antioxidant immune regulation by NPs in shrimp and may serve as a reference for healthy ecological culture of shrimp.
Assuntos
Apoptose , Autofagia , Penaeidae , Poluentes Químicos da Água , Animais , Penaeidae/efeitos dos fármacos , Penaeidae/imunologia , Penaeidae/fisiologia , Penaeidae/genética , Autofagia/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Perfilação da Expressão Gênica , Transcriptoma/efeitos dos fármacos , Microplásticos/toxicidade , Imunidade Inata/efeitos dos fármacos , Nanopartículas/toxicidadeRESUMO
To assess the impact of different substrates in a recirculating water system on the immune response and antioxidant capacity of Babylonia areolata, we conducted a comparative analysis of the transcriptomes and antioxidant performance of the digestive glands in three substrate environments (sand-S group, ceramic granules-C group, and PVC breeding nest-P group). Transcriptome results revealed that the S group and P group exhibited the highest number of differentially expressed genes (DEGs), with a total of 2218 DEGs, including 928 upregulated and 1290 downregulated DEGs. The C group and P group had 1055 DEGs in common, with 316 upregulated and 739 downregulated DEGs. The C group and S group had the fewest DEGs, with 521 in total, including 303 upregulated and 218 downregulated DEGs. GO enrichment analysis showed that in the S vs P group, terms such as catalytic activity, membrane part, and cellular process were enriched with 287, 262, and 180 DEGs, respectively. In the C vs P group, binding, cellular process, and cell part were enriched with 146, 135, and 127 DEGs, respectively. In the C vs S group, catalytic activity, membrane part, and metabolic process were enriched with 90, 83, and 59 DEGs, respectively. Kegg enrichment analysis revealed significant changes in immune-related pathways in the S vs P group, including lysosome, phagosome, and leukocyte transendothelial migration, with 30, 13, and 10 enriched DEGs, respectively. In the C vs P group, phagosome, drug metabolism-other enzymes, and N-Glycan biosynthesis showed significant changes in immune-related pathways, with 9, 6, and 4 enriched DEGs, respectively. In the C vs S group, lysosome, PPAR signaling pathway, and fatty acid degradation exhibited significant changes in immune-related pathways, with 8, 4, and 3 enriched DEGs, respectively. Regarding antioxidant capacity, the S group showed significantly higher total T-AOC than the other experimental groups, while CAT, SOD, POD, and AKP were lower than in the C and P groups. The ACP level in the Sand group was not significantly different from the P group but significantly lower than the C group. In conclusion, substrate environments significantly influence the immune-related genes and key antioxidant enzyme activities in B. areolata.
Assuntos
Aquicultura , Perfilação da Expressão Gênica , Transcriptoma , Animais , Gastrópodes/genética , Gastrópodes/imunologia , Gastrópodes/metabolismo , Antioxidantes/metabolismoRESUMO
The mitochondrial genome transcribes 13 mRNAs coding for well-known proteins essential for oxidative phosphorylation. We demonstrate here that cytochrome b (CYTB), the only mitochondrial-DNA-encoded transcript among complex III, also encodes an unrecognized 187-amino-acid-long protein, CYTB-187AA, using the standard genetic code of cytosolic ribosomes rather than the mitochondrial genetic code. After validating the existence of this mtDNA-encoded protein arising from cytosolic translation (mPACT) using mass spectrometry and antibodies, we show that CYTB-187AA is mainly localized in the mitochondrial matrix and promotes the pluripotent state in primed-to-naive transition by interacting with solute carrier family 25 member 3 (SLC25A3) to modulate ATP production. We further generated a transgenic knockin mouse model of CYTB-187AA silencing and found that reduction of CYTB-187AA impairs females' fertility by decreasing the number of ovarian follicles. For the first time, we uncovered the novel mPACT pattern of a mitochondrial mRNA and demonstrated the physiological function of this 14th protein encoded by mtDNA.
Assuntos
Citocromos b , Animais , Citocromos b/genética , Citocromos b/metabolismo , Camundongos , Feminino , Camundongos Transgênicos , DNA Mitocondrial/genética , DNA Mitocondrial/metabolismo , Mitocôndrias/metabolismo , Proteínas Mitocondriais/metabolismo , Proteínas Mitocondriais/genética , Humanos , Camundongos Endogâmicos C57BL , Genes Mitocondriais , RNA Mensageiro/metabolismo , RNA Mensageiro/genética , MasculinoRESUMO
The widespread occurrence of nanoplastic (NP) pollution in the environment is a growing concern, and its presence poses a potential threat to cultured aquatic animals. Previously, we found that NPs can significantly affect the lipid metabolism of shrimp. However, relevant reports about the effects of increasing dietary lipid levels on NP toxicity are lacking. Therefore, we explored the effects of dietary supplementation with different lipid levels on the growth and lipid metabolism of Pacific white shrimp (Litopenaeus vannamei). We cultured L. vannamei at three dietary lipid levels (3 %, 6 %, and 9 %) and three NP concentrations (0, 1, and 3 mg/L) for 2 months. We evaluated the effects of lipid levels on growth indexes, hepatopancreas morphological structure, lipid metabolism-related enzyme activity, and gene expression of the shrimp. The results showed that as lipid intake increased, the survival rate, body weight growth rate, and hepatosomatic ratio of the shrimp increased while the feed conversion rate decreased. Additionally, the crude protein and crude lipid contents increased, whereas the moisture and ash contents did not change much. We found that the morphological structure of the hepatopancreas was seriously damaged in the 3 mg/L NPs and 3 % dietary lipid group. Finally, lipid metabolism-related enzyme activities and gene expression levels increased with increased dietary lipid levels. Together, these results suggest that increasing dietary lipid content can improve shrimp growth and alleviate lipid metabolism disorders caused by NPs. This study is the first to show that nutrition regulation can alleviate the toxicity of NPs, and it provides a theoretical basis for the green and healthy culture of L. vannamei.
Assuntos
Suplementos Nutricionais , Hepatopâncreas , Metabolismo dos Lipídeos , Penaeidae , Poliestirenos , Poluentes Químicos da Água , Animais , Penaeidae/efeitos dos fármacos , Penaeidae/crescimento & desenvolvimento , Penaeidae/fisiologia , Poluentes Químicos da Água/toxicidade , Hepatopâncreas/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Poliestirenos/toxicidade , Gorduras na Dieta , Nanopartículas/toxicidadeRESUMO
In the field of biomedical research, organoids represent a remarkable advancement that has the potential to revolutionize our approach to studying human diseases even before clinical trials. Organoids are essentially miniature 3D models of specific organs or tissues, enabling scientists to investigate the causes of diseases, test new drugs, and explore personalized medicine within a controlled laboratory setting. Over the past decade, organoid technology has made substantial progress, allowing researchers to create highly detailed environments that closely mimic the human body. These organoids can be generated from various sources, including pluripotent stem cells, specialized tissue cells, and tumor tissue cells. This versatility enables scientists to replicate a wide range of diseases affecting different organ systems, effectively creating disease replicas in a laboratory dish. This exciting capability has provided us with unprecedented insights into the progression of diseases and how we can develop improved treatments. In this paper, we will provide an overview of the progress made in utilizing organoids as preclinical models, aiding our understanding and providing a more effective approach to addressing various human diseases.
RESUMO
The eutrophication of aquaculture water bodies seriously restricts the healthy development of the aquaculture industry. Among them, microcystins are particularly harmful. Therefore, the development of technologies for degrading microcystins is of great significance for maintaining the healthy development of the aquaculture industry. The feasibility and mechanism of removing microcystins-LR by dielectric barrier discharge (DBD) plasma were studied. DBD discharge power of 49.6 W and a treatment time of 40 min were selected as the more suitable DBD parameters, resulting in microcystin-LR removal efficiency of 90.4%. Meanwhile, the effects of initial microcystin-LR concentration, initial pH value, turbidity, anions on the degradation effect of microcystin-LR were investigated. The removal efficiency of microcystin-LR decreased with the increase of initial microcystin-LR concentration and turbidity. The degradation efficiency of microcystin-LR at pH 4.5 and 6.5 is significantly higher than that at pH 8.5 and 3.5. HCO3- can inhibit the removal efficiency of microcystin-LR. Furthermore, five intermediates products (m/z = 1029.5, 835.3, 829.3, 815.4, 642.1) were identified in this study, and the toxicity analysis of these degradation intermediates indicated that DBD treatment can reduce the toxicity of microcystin-LR. eï¼aq, â¢OH, H2O2, and O3 have been shown to play a major role in the degradation of microcystin-LR, and the contribution ranking of these active species is eï¼aq > â¢OH > H2O2 > O3. The application of DBD plasma technology in microcystin-LR removal and detoxification has certain development potential.
Assuntos
Microcistinas , Água , Microcistinas/análise , Peróxido de Hidrogênio , Temperatura , AquiculturaRESUMO
Mitochondria constantly fuse and divide for mitochondrial inheritance and functions. Here, we identified a distinct type of naturally occurring fission, tail-autotomy fission, wherein a tail-like thin tubule protrudes from the mitochondrial body and disconnects, resembling autotomy. Next, utilizing an optogenetic mitochondria-specific mechanostimulator, we revealed that mechanical tensile force drives tail-autotomy fission. This force-induced fission involves DRP1/MFF and endoplasmic reticulum tubule wrapping. It redistributes mitochondrial DNA, producing mitochondrial fragments with or without mitochondrial DNA for different fates. Moreover, tensile force can decouple outer and inner mitochondrial membranes, pulling out matrix-excluded tubule segments. Subsequent tail-autotomy fission separates the matrix-excluded tubule segments into matrix-excluded mitochondrial-derived vesicles (MDVs) which recruit Parkin and LC3B, indicating the unique role of tail-autotomy fission in segregating only outer membrane components for mitophagy. Sustained force promotes fission and MDV biogenesis more effectively than transient one. Our results uncover a mechanistically and functionally distinct type of fission and unveil the role of tensile forces in modulating fission and MDV biogenesis for quality control, underscoring the heterogeneity of fission and mechanoregulation of mitochondrial dynamics.
Assuntos
Proteínas de Membrana , Dinâmica Mitocondrial , Proteínas de Membrana/genética , Proteínas Mitocondriais/genética , Mitocôndrias/genética , DNA Mitocondrial , Controle de Qualidade , Dinaminas/genéticaRESUMO
Nanoplastics (NPs) are widely distributed environmental pollutants that can disrupt intestinal immunity of crustaceans. In this study, the effects of NPs on gut immune enzyme activities, cell morphology, apoptosis, and microbiota diversity of Litopenaeus vannamei were investigated. L. vannamei was exposed to five concentrations of NPs (0, 0.1, 1, 5, and 10 mg/L) for 28 days. The results showed that higher concentrations of NPs damaged the intestinal villi, promoted formation of autophagosomes, increased intestinal non-specific immunoenzyme activities, and significantly increased apoptosis at 10 mg/L. In response to exposure to NPs, the expression levels of ATG3, ATG4, ATG12, Caspase-3, p53, and TNF initially increased and then decreased. In addition, the concentration of NPs was negatively correlated to the expression levels of the genes of interest and intestinal enzyme activities, suggesting that exposure to NPs inhibited apoptosis and immune function. The five dominant phyla of the gut microbiota (Proteobacteria, Firmicutes, Bacteroidetes, Acidobacteria, and Actinomycetes) were similar among groups exposed to different concentrations of NPs, but the abundances tended to differ. Notably, exposure to NPs increased the abundance of pathogenic bacteria. These results confirm that exposure to NPs negatively impacted intestinal immune function of L. vannamei. These findings provide useful references for efficient breeding of L. vannamei.
Assuntos
Microbioma Gastrointestinal , Microbiota , Penaeidae , Animais , Microplásticos , Poliestirenos , Disbiose , Penaeidae/microbiologia , Autofagia , ApoptoseRESUMO
The effect of hydrodynamic mixing on controlling Microcystis blooms or changing the algal community to diatom dominance has been widely studied; however, the effects of colonial Microcystis biomass on the development of the algal community are poorly known. Here, in order to study the changes in Microcystis blooms under continuous aeration mixing, an experiment was carried out in a greenhouse with factors of varying biomass of Microcystis and inorganic nitrogen and phosphorus enrichment in summer. There were three chlorophyll a (Chl-a) levels in six treatments: low Chl-a level of 68.4 µg L-1 (treatments L, L-E), medium Chl-a level of 468.7 µg L-1 (treatments M, M-E), and high Chl-a level of 924.1 µg L-1 (treatments H, H-E). Treatments L-E, M-E and H-E were enriched with the same inorganic nitrogen and phosphorus nutrients. During the experiment of 30 days, the concentration of Microcystis and Chl-a decreased, and diatom Nitzschia palea cells appeared in all the treatments, which became dominant in treatments M, M-E, H and H-E, with the highest biomass of 9.41 ± 1.96 mg L-1 Nitzschia in treatment H-E on day 30. The rank order of the biomass of Nitzschia from low to high was (L = L-E) < (M = M-E) < H < H-E (P < 0.05). In addition, Nitzschia cells were aggregates attached to Microcystis colonies in all the treatments. The results showed that the initial biomass of colonial Microcystis affected the algal shift from Microcystis dominance to Nitzschia dominance. However, the enriched inorganic nitrogen and phosphorus was beneficial for the Nitzschia increase in the high biomass treatment alone. The shift from Microcystis dominance to diatom dominance under continuous aeration mixing may be caused by low light conditions as well as the nutrients released from Microcystis decay. Moreover, the aerobic condition caused by aeration mixing maintained the colonial mucilaginous sheath to support the growth of Nitzschia cells in aggregation. This study found for the first time that Microcystis blooms could shift to diatom Nitzschia dominance in aggregates. It provided a method to control and manipulate Microcystis blooms to diatom dominance through continuous aeration mixing to proper biomass of Microcystis colonies. The shift to diatoms dominance would provide more high quality food organisms for aquaculture and be beneficial to the material cycling and energy flowing in food web dynamics.
Assuntos
Diatomáceas , Microcystis , Biomassa , Clorofila A , Fósforo/farmacologia , Nitrogênio/farmacologiaRESUMO
Harmful cyanobacterial bloom is one of the serious environmental problems worldwide. Microcystis aeruginosa is a representative harmful alga in cyanobacteria bloom. It is of great significance to develop new technologies for the removal of Microcystis aeruginosa and microcystins. The feasibility and mechanism of removing microcystis aeruginosa and degrading microcystins by dielectric barrier discharge (DBD) plasma were studied. The suitable DBD parameters obtained in this study are DBD (41.5 W, 40 min) and DBD (41.5 W, 50 min), resulting in algae removal efficiency of 77.4% and 80.4%, respectively; scanning electron microscope and LIVE-DEATH analysis demonstrate that DBD treatment can disrupt cell structure and lead to cell death; analysis of elemental composition and chemical state indicated that there are traces of oxidation of organic nitrogen and organic carbon in microcystis aeruginosa; further intracellular ROS concentration and antioxidant enzyme activity analysis confirm that DBD damage microcystis aeruginosa through oxidation. Meanwhile, DBD can effectively degrade the microcystin-LR released after cell lysis, the extracellular microcystin-LR concentration in the DBD (41.5 W) group decreased by 88.7% at 60 min compared to the highest concentration at 20 min; further toxicity analysis of degradation intermediates indicated that DBD can reduce the toxicity of microcystin-LR. The contribution of active substances to the inactivation of microcystis aeruginosa is eaq- > â¢OH > H2O2 > O3 > 1O2 > â¢O2- > ONOO-, while on the degradation of microcystin-LR is eaq- > â¢OH > H2O2 > O3 > â¢O2- > 1O2 > ONOO-. The application of DBD plasma technology in microcystis aeruginosa algae removal and detoxification has certain prospects for promotion and application.
Assuntos
Cianobactérias , Toxinas Marinhas , Microcystis , Microcystis/metabolismo , Proliferação Nociva de Algas , Microcistinas/química , Peróxido de Hidrogênio/metabolismo , Estudos de Viabilidade , Cianobactérias/metabolismo , Antioxidantes/metabolismoRESUMO
Aging in mammals is accompanied by an imbalance of intestinal homeostasis and accumulation of mitochondrial DNA (mtDNA) mutations. However, little is known about how accumulated mtDNA mutations modulate intestinal homeostasis. We observe the accumulation of mtDNA mutations in the small intestine of aged male mice, suggesting an association with physiological intestinal aging. Using polymerase gamma (POLG) mutator mice and wild-type mice, we generate male mice with progressive mtDNA mutation burdens. Investigation utilizing organoid technology and in vivo intestinal stem cell labeling reveals decreased colony formation efficiency of intestinal crypts and LGR5-expressing intestinal stem cells in response to a threshold mtDNA mutation burden. Mechanistically, increased mtDNA mutation burden exacerbates the aging phenotype of the small intestine through ATF5 dependent mitochondrial unfolded protein response (UPRmt) activation. This aging phenotype is reversed by supplementation with the NAD+ precursor, NMN. Thus, we uncover a NAD+ dependent UPRmt triggered by mtDNA mutations that regulates the intestinal aging.
Assuntos
Envelhecimento , NAD , Camundongos , Masculino , Animais , NAD/metabolismo , Envelhecimento/genética , Envelhecimento/metabolismo , Mutação , Mitocôndrias/genética , Mitocôndrias/metabolismo , DNA Mitocondrial/genética , DNA Mitocondrial/metabolismo , DNA Polimerase gama/genética , DNA Polimerase gama/metabolismo , Mamíferos/genéticaRESUMO
Litopenaeus vannamei were exposed to 80-nm polystyrene nanoplastics (NPs) at different concentrations (0, 0.1, 1, 5, and 10 mg/L) for 28 days to study the effects on muscle nutritional quality. Our results showed that with increasing NPs concentrations, the survival rate, specific gain rate, and protein efficiency ratio decreased but the feed conversion ratio increased. There was no significant difference in moisture, ash, and crude lipid content in the muscle, and a general decrease in crude protein content was observed. However, the total amino acid and semi-essential amino acid contents decreased. The spacing between muscle fibers and the melting morphology of muscle increased. The hardness of muscle flesh texture increased, but springiness, cohesiveness, and chewiness decreased. Regarding antioxidant enzyme activity, the activity of catalase decreased, but the total antioxidant capacity, superoxide dismutase activity, and reduced glutathione first increased and then decreased. The expression level of the growth-related genes retinoid X receptor (RXR), chitin synthase (CHS), and calmodulin A (CaM) first increased then decreased, but calcium/calmodulin-dependent protein kinase I (CaMKI), ecdysteroid receptor (EcR), chitinase 5 (CHT5), cell division cycle 2 (Cdc2), and cyclin-dependent kinase 2 (CDK2) decreased. Our results suggest that exposure to NPs can inhibit growth by inducing oxidative stress, which leads to muscle tissue damage and changes in amino acid composition. These results will provide a theoretical reference for the risk assessment of NPs and the ecological health aquaculture of shrimp.
Assuntos
Antioxidantes , Penaeidae , Animais , Antioxidantes/metabolismo , Poliestirenos/toxicidade , Poliestirenos/metabolismo , Microplásticos/metabolismo , Aminoácidos/metabolismo , Valor Nutritivo , Músculos/metabolismoRESUMO
Grass carp (Ctenopharyngodon idellus) is the most productive freshwater fish in China, but its traditional aquaculture model still has problems, such as poor water quality and frequent diseases. We have taken monoculture and 80:20 polyculture grass carp ponds as the research object and used EwE software to build the Ecopath model of two ponds. We analyzed and compared the characteristics of ecological structure and energy flow in two ponds. The result showed the highest effective trophic level in the polyculture pond that was higher than that in the monoculture pond, and fish in polyculture had higher EE values which showed the production of fish in polyculture contributed more to the energy conversion efficiency of the ecosystem. Flows into detritus were the largest component of TST both in the two ponds, which accounted for 49.34% and 50.37%. And the average transfer efficiency in monoculture was 13.07%, while that in polyculture was 15.6%. The ascendency/total development capacity (A/TDC) and overhead/total development capacity (O/TDC) were 0.35 and 0.65 both in the two ponds, respectively, which indicated that both systems had a strong anti-perturbation ability, but the stability could be improved. Finn's cycling index (FCI) in polyculture was higher and showed that the polyculture pond was more mature and stable. Unused energy of functional groups will flow to detritus, and that in the monoculture pond was higher, the energy of C. idellus that flowed to detritus in monoculture was 48.17% higher than that in polyculture; unused energy of bacteria and phytoplankton were also high. The result showed that polyculture could improve energy utilization, increase transfer efficiency, and raise the stability of the ecosystem. Grass carp ponds still need to be improved in the aspects of mixed species and energy consumption. It is necessary to improve the ecological and economic benefits of grass carp ponds by optimizing the aquaculture structure and adjusting the aquaculture proportion.
Assuntos
Carpas , Animais , Lagoas/química , Ecossistema , Água Doce , China , AquiculturaRESUMO
In mammalian cells, besides nuclei, mitochondria are the only semi-autonomous organelles possessing own DNA organized in the form of nucleoids. While eukaryotic nuclear DNA compaction, chromatin compartmentalization and transcription are regulated by phase separation, our recent work proposed a model of mitochondrial nucleoid self-assembly and transcriptional regulation by multi-phase separation. Herein, we summarized the phase separation both in the nucleus and mitochondrial nucleoids, and did a comparison of the organization and activity regulating, which would provide new insight into the understanding of both architecture and genetics of nucleus and mitochondrial nucleoids.
RESUMO
Melatonin (MT) is regarded as an antioxidant and immunostimulant that can efficiently scavenge free radicals and activate antioxidant enzymes. The aim of this study was to investigate the effects of dietary MT on the growth performance and immune function of the Pacific white shrimp (Litopenaeus vannamei). Six groups of L. vannamei were supplemented with dietary MT at 0, 22.5, 41.2, 82.7, 165.1, and 329.2 mg/kg levels for 2 months. RNA-Seq analysis was performed to obtain transcriptome data of the control group and the group supplemented with dietary MT at 82.7 mg/kg BW. In total, 1220 DEGs (799 up-regulated and 421 down-regulated) were identified. Pathways and genes related to growth performance and immune function were verified by real-time quantitative polymerase chain reaction. The total hemocyte count, phagocytosis rate, and respiratory burst were significantly increased in the MT (82.7 mg/kg BW) group as compared to the control group. Analysis of antioxidant-related enzymes in the hepatopancreas showed that dietary MT (82.7 mg/kg BW) significantly increased activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase, while dietary MT at 41.2 mg/kg BW significantly increased activities of glutathione S-transferase, lysozyme (LZM), and phenoloxidase (PO). At the transcriptional level, dietary MT up-regulated expression levels of genes associated with antioxidant immunity and growth, which included PO, SOD, LZM, GPx, chitin synthase, ecdysone receptor, calcium-calmodulin dependent protein kinase I, and retinoid X receptor. In conclusion, dietary MT may improve the growth performance and immune function of L. vannamei to some extent.
Assuntos
Melatonina , Penaeidae , Animais , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Melatonina/metabolismo , Melatonina/farmacologia , Transcriptoma , Dieta , Superóxido Dismutase/metabolismo , Fagocitose , Penaeidae/genética , Imunidade Inata , Ração Animal/análiseRESUMO
This study aimed to investigate the effects of dietary melatonin (MT) levels on the antioxidant capacity, immunomodulatory, and transcriptional regulation of red swamp crayfish. Six experimental diets with different levels of MT (0, 22.5, 41.2, 82.7, 165.1, and 329.2 mg/kg diet) were fed to juvenile crayfish for 60 d. The transcriptome data of the control group and the group supplemented with dietary MT at 165.1 mg/kg were obtained using RNA-seq. In total, 3653 differentially expressed genes (2082 up-regulated and 1571 down-regulated) were identified. Pathways and genes related to antioxidant immune and growth performance were verified by qRT-PCR. The total hemocyte count, phagocytosis rate, and respiratory burst were significantly increased in the MT (165.1 mg/kg) group compared to the control group. Analysis of antioxidant immune-related enzymes in the hepatopancreas demonstrated that dietary MT (165.1 mg/kg) significantly increased activities of catalase, superoxide dismutase, glutathione reductase, and glutathione peroxidase and significantly decreased aspartate aminotransferase and alanine aminotransferase activity. At the transcriptional level, dietary MT up-regulated expression levels of genes associated with antioxidant immune and development, which included toll-like receptors, Crustin, C-type lectin, and so on. To conclude, MT could be used as a supplement in crayfish feed to increase immunity and antioxidant capacity and according to the broken line regression, the ideal MT concentration was the 159.02 mg/kg. Overall, this study demonstrates the role of melatonin in the antioxidant responses and immunomodulatory of Procambarus clarkii, laying the foundation for the development of melatonin as a feed additive in the aquaculture of this species.