Pub Date : 2025-08-30DOI: 10.1016/j.cbpb.2025.111152
Haruna Amano , Juna Kamoshita , Masami Ishimoto , Minami Arai , Mitsuru Jimbo , Ko Yasumoto , Go Suzuki
Three egg proteins (EP1, EP2, and EP3) were detected in ovulated eggs of Acropora aff. tenuis, a reef-building stony coral found in tropical and subtropical areas. The proteins were separated into different fractions by gel filtration chromatography, and different patterns were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting with antiserum against A. aff. tenuis egg extract (anti-AtE). EP2 was purified from ovulated egg extracts using a combination of hydroxylapatite and gel filtration chromatography. The molecular mass of native EP2 was estimated to be 286 kDa. SDS-PAGE revealed bands at 70.5, 67.7, 49.6, and 22.9 kDa, with the 70.5 and 67.7 kDa bands staining positive for lipid (Sudan black B). These results indicate that purified EP2 is a lipoprotein in A. aff. tenuis eggs, resembling lipovitellin, the major yolk protein derived from the egg yolk precursor, vitellogenin (Vtg), in oviparous vertebrates. Furthermore, EP2 and its bands were digested with trypsin and analyzed by liquid chromatography using a quadrupole time-of-flight tandem mass spectrometer (LC-QTOF-MS). All fragments were identified as A. aff. tenuis Vtg. This is the first report of the purification and characterization of a lipovitellin-like egg yolk protein in non-bilaterian oviparous animals.
{"title":"Characterization and purification of lipovitellin-like egg yolk protein in a stony coral, Acropora aff. tenuis","authors":"Haruna Amano , Juna Kamoshita , Masami Ishimoto , Minami Arai , Mitsuru Jimbo , Ko Yasumoto , Go Suzuki","doi":"10.1016/j.cbpb.2025.111152","DOIUrl":"10.1016/j.cbpb.2025.111152","url":null,"abstract":"<div><div>Three egg proteins (EP1, EP2, and EP3) were detected in ovulated eggs of <em>Acropora</em> aff. <em>tenuis</em>, a reef-building stony coral found in tropical and subtropical areas. The proteins were separated into different fractions by gel filtration chromatography, and different patterns were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting with antiserum against <em>A.</em> aff. <em>tenuis</em> egg extract (anti-AtE). EP2 was purified from ovulated egg extracts using a combination of hydroxylapatite and gel filtration chromatography. The molecular mass of native EP2 was estimated to be 286 kDa. SDS-PAGE revealed bands at 70.5, 67.7, 49.6, and 22.9 kDa, with the 70.5 and 67.7 kDa bands staining positive for lipid (Sudan black B). These results indicate that purified EP2 is a lipoprotein in <em>A.</em> aff. <em>tenuis</em> eggs, resembling lipovitellin, the major yolk protein derived from the egg yolk precursor, vitellogenin (Vtg), in oviparous vertebrates. Furthermore, EP2 and its bands were digested with trypsin and analyzed by liquid chromatography using a quadrupole time-of-flight tandem mass spectrometer (LC-QTOF-MS). All fragments were identified as <em>A.</em> aff. <em>tenuis</em> Vtg. This is the first report of the purification and characterization of a lipovitellin-like egg yolk protein in non-bilaterian oviparous animals.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111152"},"PeriodicalIF":1.8,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144921425","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-29DOI: 10.1016/j.cbpb.2025.111151
Rowida E. Ibrahim , Gehad E. Elshopakey , Ahmed S. AlSaqufi , Abdallah Tageldein Mansour , Yousef Alkhamis , Hesham A. Hassanien , Ahmed Abbas , Sameh H. Ismail , Tarek Khamis , Afaf N. Abdel Rahman
This study investigated the potential for chitosan nanogel (CNG) to mitigate adverse consequences of infection by Shewanella spp. in the Nile tilapia, Oreochromis niloticus. A total of 160 fish (average weight = 27.55 ± 1.50 g) were allocated to four treatments, each with four replicates, for 14 days. Fish in the control and CNG only groups were intraperitoneally injected with 0.2 mL of sterile saline and placed in water treated with 0 or 75 μg/L, respectively, of CNG. Fish in the Shewanella and CNG + Shewanella groups were injected with 0.2 mL of Shewanella spp. (0.14 × 105 colony forming units) and placed in water treated with 0 or 75 μg/L, respectively, of CNG. Infection by Shewanella induced hemorrhages on the skin and base of the fins, fin rot, and scale loss. Shewanella infection decreased the activity of glutathione peroxidase and catalase, with increased malondialdehyde concentration. Shewanella decreased the brain neurotransmitters (dopamine and serotonin), total protein, and globulin levels, while it increased serum glucose and cortisol. The hepatic and renal dysfunction indicators alanine aminotransferase, aspartate aminotransferase, and creatinine were significantly increased by Shewanella infection. Histopathological changes were produced in the liver, kidney, gills, and brain of Shewanella-infected fish. Notably, Shewanella-infected fish in CNG-treated water had reduced abnormal signs, stress markers, and hepatic and renal dysfunction indicators. CNG water application elevated the brain levels of neurotransmitters and protein and improved the tissue histology. Overall, CNG as a water application is useful for mitigating the adverse effects of Shewanella spp. in Nile tilapia.
{"title":"Chitosan nanogel mitigates Shewanella-induced oxidative stress, brain neurotransmitter imbalance, biochemical, hepatic, and renal dysfunction, and histopathological changes in Nile tilapia","authors":"Rowida E. Ibrahim , Gehad E. Elshopakey , Ahmed S. AlSaqufi , Abdallah Tageldein Mansour , Yousef Alkhamis , Hesham A. Hassanien , Ahmed Abbas , Sameh H. Ismail , Tarek Khamis , Afaf N. Abdel Rahman","doi":"10.1016/j.cbpb.2025.111151","DOIUrl":"10.1016/j.cbpb.2025.111151","url":null,"abstract":"<div><div>This study investigated the potential for chitosan nanogel (CNG) to mitigate adverse consequences of infection by <em>Shewanella</em> spp. in the Nile tilapia, <em>Oreochromis niloticus</em>. A total of 160 fish (average weight = 27.55 ± 1.50 g) were allocated to four treatments, each with four replicates, for 14 days. Fish in the control and CNG only groups were intraperitoneally injected with 0.2 mL of sterile saline and placed in water treated with 0 or 75 μg/L, respectively, of CNG. Fish in the <em>Shewanella</em> and CNG + <em>Shewanella</em> groups were injected with 0.2 mL of <em>Shewanella</em> spp. (0.14 × 10<sup>5</sup> colony forming units) and placed in water treated with 0 or 75 μg/L, respectively, of CNG. Infection by <em>Shewanella</em> induced hemorrhages on the skin and base of the fins, fin rot, and scale loss. <em>Shewanella</em> infection decreased the activity of glutathione peroxidase and catalase, with increased malondialdehyde concentration. <em>Shewanella</em> decreased the brain neurotransmitters (dopamine and serotonin), total protein, and globulin levels, while it increased serum glucose and cortisol. The hepatic and renal dysfunction indicators alanine aminotransferase, aspartate aminotransferase, and creatinine were significantly increased by <em>Shewanella</em> infection. Histopathological changes were produced in the liver, kidney, gills, and brain of <em>Shewanella</em>-infected fish. Notably, <em>Shewanella</em>-infected fish in CNG-treated water had reduced abnormal signs, stress markers, and hepatic and renal dysfunction indicators. CNG water application elevated the brain levels of neurotransmitters and protein and improved the tissue histology. Overall, CNG as a water application is useful for mitigating the adverse effects of <em>Shewanella</em> spp. in Nile tilapia.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111151"},"PeriodicalIF":1.8,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144921578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-23DOI: 10.1016/j.cbpb.2025.111149
Hui Fu, Qichao Fan, Jingjing Miao, Ya Liu, Yuxin Wen, Luqing Pan
The scallop (Chlamys farreri) is an important aquaculture species, and understanding its reproductive regulation is a major research focus. Previous studies have suggested that the orphan nuclear receptor fushi tarazu factor 1 (FTZF1), a member of the nuclear receptor subfamily 5 group A (NR5A), may play a role in regulating gonadal development and differentiation in scallops. In this study, we demonstrate through yeast one-hybrid assays that FTZ-F1 directly binds to the promoters of the gonadotropin releasing hormone receptor (GnRHR), which is involved in steroidogenesis, and DMRT-like family A2 (Dmrta2), which is involved in sex determination. Molecular docking and sequence alignment identified 11 key amino acids within the DNA-binding domain （DBD） essential for promoter binding, 10 of which resided in the conserved DBD region. These findings provide molecular insights into reproductive regulation in bivalves and establish a foundation for further research on sex differentiation and steroidogenesis.
{"title":"Integrated yeast one-hybrid and molecular docking reveal the binding specificity of FTZ-F1 to GnRHR and Dmrta2 promoters in the scallop, Chlamys farreri","authors":"Hui Fu, Qichao Fan, Jingjing Miao, Ya Liu, Yuxin Wen, Luqing Pan","doi":"10.1016/j.cbpb.2025.111149","DOIUrl":"10.1016/j.cbpb.2025.111149","url":null,"abstract":"<div><div>The scallop (<em>Chlamys farreri</em>) is an important aquaculture species, and understanding its reproductive regulation is a major research focus. Previous studies have suggested that the orphan nuclear receptor fushi tarazu factor 1 (FTZ<img>F1), a member of the nuclear receptor subfamily 5 group A (NR5A), may play a role in regulating gonadal development and differentiation in scallops. In this study, we demonstrate through yeast one-hybrid assays that FTZ-F1 directly binds to the promoters of the gonadotropin releasing hormone receptor (<em>GnRHR</em>), which is involved in steroidogenesis, and DMRT-like family A2 (<em>Dmrta2</em>), which is involved in sex determination. Molecular docking and sequence alignment identified 11 key amino acids within the DNA-binding domain （DBD） essential for promoter binding, 10 of which resided in the conserved DBD region. These findings provide molecular insights into reproductive regulation in bivalves and establish a foundation for further research on sex differentiation and steroidogenesis.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111149"},"PeriodicalIF":1.8,"publicationDate":"2025-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144907657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-23DOI: 10.1016/j.cbpb.2025.111150
Thuany da Silva Nogueira , Andressa de Abreu Mello , Nathany da Silva Nogueira , Cintia Monteiro-de-Barros
Reactive oxygen species (ROS) are metabolically generated by neural cells and are essential for maintaining cellular homeostasis. Due to the high metabolic activity and oxygen consumption of the nervous system, large amounts of ROS are produced. Although ROS are traditionally associated with oxidative stress and degeneration, recent studies suggest that they may also play a beneficial role in neural repair. To explore the role of ROS in central nervous system (CNS) regeneration, we employed the adult ascidian Styela plicata, which can regenerate its entire brain within 10 days, as a model. We aimed to elucidate the functions of ROS and the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) in the formation of synapses and/or cell proliferation in the CNS of the ascidian S. plicata following the systemic injection of the neurotoxin 3-acetylpyridine (3-AP). The data revealed two distinct periods of high ROS production: at 0.5 and 5 days. SOD and CAT activity increased only at 5 days. Lipid peroxidation was highest at 0.5 days and gradually decreased during the 10 day post-injection monitoring period. The proliferation marker Ki-67 and the synapse marker synaptophysin were elevated at 3 and 5 days, respectively. These findings suggest a dual role for ROS: an early phase linked to neurodegeneration and a later phase associated with regeneration. This highlights the complex role of redox signaling in CNS regeneration.
{"title":"Dual roles of reactive oxygen species in ascidian brain regeneration and degeneration","authors":"Thuany da Silva Nogueira , Andressa de Abreu Mello , Nathany da Silva Nogueira , Cintia Monteiro-de-Barros","doi":"10.1016/j.cbpb.2025.111150","DOIUrl":"10.1016/j.cbpb.2025.111150","url":null,"abstract":"<div><div>Reactive oxygen species (ROS) are metabolically generated by neural cells and are essential for maintaining cellular homeostasis. Due to the high metabolic activity and oxygen consumption of the nervous system, large amounts of ROS are produced. Although ROS are traditionally associated with oxidative stress and degeneration, recent studies suggest that they may also play a beneficial role in neural repair. To explore the role of ROS in central nervous system (CNS) regeneration, we employed the adult ascidian <em>Styela plicata</em>, which can regenerate its entire brain within 10 days, as a model. We aimed to elucidate the functions of ROS and the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) in the formation of synapses and/or cell proliferation in the CNS of the ascidian <em>S. plicata</em> following the systemic injection of the neurotoxin 3-acetylpyridine (3-AP). The data revealed two distinct periods of high ROS production: at 0.5 and 5 days. SOD and CAT activity increased only at 5 days. Lipid peroxidation was highest at 0.5 days and gradually decreased during the 10 day post-injection monitoring period. The proliferation marker Ki-67 and the synapse marker synaptophysin were elevated at 3 and 5 days, respectively. These findings suggest a dual role for ROS: an early phase linked to neurodegeneration and a later phase associated with regeneration. This highlights the complex role of redox signaling in CNS regeneration.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111150"},"PeriodicalIF":1.8,"publicationDate":"2025-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144904521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-21DOI: 10.1016/j.cbpb.2025.111148
Mengge Xu , Siting Wang , Chen Liang , Dan Yang , Meimei Liu , Zhiguo Dong
This study investigated the tissue-specific expression and subcellular localization of sex differentiation-related genes of Cyclina sinensis, and then analyzed the effects of hormone treatment (1 and 50 μg/L estradiol treatment (E-1, E-50), 1 and 50 μg/L testosterone (T-1, T-50), and a control group (C)) on the sex ratio, gonadosomatic index, hepatosomatic index, hormone content, and expression of sex differentiation-related genes of juvenile C. sinensis. Vitellogenin (Vg), testis-specific serine kinase 1 (Tssk1), doublesex and mab-3 related transcription factor 1 (Dmrt1), and Wnt family member 4 (Wnt4) were significantly expressed in the gonads of adult C. sinensis, and the in situ hybridization signals of forkhead box L2 (Foxl2) and SRY-box transcription factor 9 (Sox9) were localized in nuclei of oogonia and oocytes and in spermatogonia, spermatocytes, and sperm, respectively. The male to female ratio was 7:3 in group T-1 after 60 days of hormone treatment, but this effect was not statistically significant. In the E-1 group, Vg was highly expressed in the ovary at days 30 and 120. TSSK1 was highly expressed in the testes of the T-1 and T-50 groups at 120 days (60 days after the cessation of hormone treatment). The expression of Foxl2 was significantly higher in the ovaries of the T-1 group at 60 days, relative to the control and other hormone-treated groups. Sox9 was highly expressed in the testes at 120 days. The concentrations of testosterone and estradiol in the gonad of C. sinensis increased initially and then decreased. Overall, these results revealed that hormones caused disruption of sex determination and expression of sex-differentiation genes in juvenile C. sinensis.
{"title":"Molecular expression of genes related to sex differentiation and their response to sex steroid hormones in the clam Chinese cyclina (Cyclina sinensis)","authors":"Mengge Xu , Siting Wang , Chen Liang , Dan Yang , Meimei Liu , Zhiguo Dong","doi":"10.1016/j.cbpb.2025.111148","DOIUrl":"10.1016/j.cbpb.2025.111148","url":null,"abstract":"<div><div>This study investigated the tissue-specific expression and subcellular localization of sex differentiation-related genes of <em>Cyclina sinensis,</em> and then analyzed the effects of hormone treatment (1 and 50 μg/L estradiol treatment (E-1, E-50), 1 and 50 μg/L testosterone (T-1, T-50), and a control group (C)) on the sex ratio, gonadosomatic index, hepatosomatic index, hormone content, and expression of sex differentiation-related genes of juvenile <em>C. sinensis</em>. <em>Vitellogenin</em> (<em>Vg</em>), <em>testis-specific serine kinase 1</em> (<em>Tssk1</em>), <em>doublesex and mab-3 related transcription factor 1</em> (<em>Dmrt1</em>), and <em>Wnt</em> family member 4 (<em>Wnt4</em>) were significantly expressed in the gonads of adult <em>C. sinensis</em>, and the <em>in situ</em> hybridization signals of <em>forkhead box L2</em> (<em>Foxl2</em>) and <em>SRY-box transcription factor 9</em> (<em>Sox9</em>) were localized in nuclei of oogonia and oocytes and in spermatogonia, spermatocytes, and sperm, respectively. The male to female ratio was 7:3 in group T-1 after 60 days of hormone treatment, but this effect was not statistically significant. In the E-1 group, <em>Vg</em> was highly expressed in the ovary at days 30 and 120. <em>TSSK1</em> was highly expressed in the testes of the T-1 and T-50 groups at 120 days (60 days after the cessation of hormone treatment). The expression of <em>Foxl2</em> was significantly higher in the ovaries of the T-1 group at 60 days, relative to the control and other hormone-treated groups. <em>Sox9</em> was highly expressed in the testes at 120 days. The concentrations of testosterone and estradiol in the gonad of <em>C. sinensis</em> increased initially and then decreased. Overall, these results revealed that hormones caused disruption of sex determination and expression of sex-differentiation genes in juvenile <em>C. sinensis</em>.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111148"},"PeriodicalIF":1.8,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144921582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-18DOI: 10.1016/j.cbpb.2025.111146
Xishuang Shan , Xiya Zhang , Binbin Tao , Yanlong Song , Zuoyan Zhu , Wei Hu , Ji Chen
Gonadal development and gamete maturation are essential for fish reproduction. The protein tyrosine phosphatase receptor type Fb (Ptprfb) is a member of the tyrosine phosphatase family. In the present study, we used CRISPR/Cas9 to mutate ptprfb in zebrafish. A significantly reduced natural fertilization rate of sperm from mutant fish was observed. The mutant fish produced fewer sperm with shorter flagella, and a smaller proportion of sperm could be activated. RNA-seq analysis revealed abnormal expression of some genes in testicular cells, such as hemoglobin subunit βA1 (hbba1), myosin heavy chain 11b (myh11b), and transgelin (tagln), as well as some genes involved in focal adhesion formation. These findings demonstrate that ptprfb contributes to spermatogenesis in zebrafish, and its dysfunction can adversely affect both the quantity and quality of sperm.
{"title":"CRISPR/Cas9-mediated editing of ptprfb (protein tyrosine phosphatase receptor type fb) reveals its regulatory role in zebrafish spermatogenesis","authors":"Xishuang Shan , Xiya Zhang , Binbin Tao , Yanlong Song , Zuoyan Zhu , Wei Hu , Ji Chen","doi":"10.1016/j.cbpb.2025.111146","DOIUrl":"10.1016/j.cbpb.2025.111146","url":null,"abstract":"<div><div>Gonadal development and gamete maturation are essential for fish reproduction. The protein tyrosine phosphatase receptor type Fb (Ptprfb) is a member of the tyrosine phosphatase family. In the present study, we used CRISPR/Cas9 to mutate <em>ptprfb</em> in zebrafish. A significantly reduced natural fertilization rate of sperm from mutant fish was observed. The mutant fish produced fewer sperm with shorter flagella, and a smaller proportion of sperm could be activated. RNA-seq analysis revealed abnormal expression of some genes in testicular cells, such as hemoglobin subunit βA1 (<em>hbba1</em>), myosin heavy chain 11b (<em>myh11b</em>), and transgelin (<em>tagln</em>), as well as some genes involved in focal adhesion formation. These findings demonstrate that <em>ptprfb</em> contributes to spermatogenesis in zebrafish, and its dysfunction can adversely affect both the quantity and quality of sperm.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111146"},"PeriodicalIF":1.8,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144889982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-18DOI: 10.1016/j.cbpb.2025.111147
Won Chae Jeong , Kun Woo Kim , Jin A Kim , Jun-Hwan Kim , Cheol Young Choi
Microplastics and trace metals such as cadmium (Cd) are environmental contaminants commonly co-occurring in marine ecosystems. We aimed to evaluate the impact of combined exposure of Pacific oyster (Crassostrea gigas) to microbeads (MBs) and Cd, focusing on the effects of the depuration process on contaminant removal and stress-related biomarkers. Pacific oysters were exposed to MBs, Cd, and their combination for 48 h, followed by a 72 h depuration process using uncontaminated seawater. We measured the levels of accumulated MBs and Cd in the whole soft tissue of the Pacific oysters to evaluate the degree of contaminant removal. Additionally, the concentrations of hydrogen peroxide were measured and the mRNA expression levels of antioxidant enzymes, metallothionein, and the apoptosis-related gene caspase-3 were analyzed in the Pacific oyster hepatopancreas tissue to evaluate oxidative stress and apoptosis. Our results indicated that Cd was eliminated more slowly than MBs, and the Pacific oysters exposed to combined MB and Cd contaminants maintained higher levels of oxidative stress-related gene expression than those exposed to individual contaminants. These findings suggest that Cd may persist longer in oyster tissues than MBs, potentially leading to prolonged toxicity in the Pacific oyster. Furthermore, in environments where both MBs and Cd are present, MBs can enhance the toxic effects of Cd through a synergistic interaction. Overall, we provide a reference for understanding the depuration and physiological responses of marine bivalves exposed to MBs and Cd.
{"title":"Depuration and post-exposure recovery of oxidative stress responses to microplastics and cadmium in Pacific oyster (Crassostrea gigas)","authors":"Won Chae Jeong , Kun Woo Kim , Jin A Kim , Jun-Hwan Kim , Cheol Young Choi","doi":"10.1016/j.cbpb.2025.111147","DOIUrl":"10.1016/j.cbpb.2025.111147","url":null,"abstract":"<div><div>Microplastics and trace metals such as cadmium (Cd) are environmental contaminants commonly co-occurring in marine ecosystems. We aimed to evaluate the impact of combined exposure of Pacific oyster (<em>Crassostrea gigas</em>) to microbeads (MBs) and Cd, focusing on the effects of the depuration process on contaminant removal and stress-related biomarkers. Pacific oysters were exposed to MBs, Cd, and their combination for 48 h, followed by a 72 h depuration process using uncontaminated seawater. We measured the levels of accumulated MBs and Cd in the whole soft tissue of the Pacific oysters to evaluate the degree of contaminant removal. Additionally, the concentrations of hydrogen peroxide were measured and the mRNA expression levels of antioxidant enzymes, metallothionein, and the apoptosis-related gene caspase-3 were analyzed in the Pacific oyster hepatopancreas tissue to evaluate oxidative stress and apoptosis. Our results indicated that Cd was eliminated more slowly than MBs, and the Pacific oysters exposed to combined MB and Cd contaminants maintained higher levels of oxidative stress-related gene expression than those exposed to individual contaminants. These findings suggest that Cd may persist longer in oyster tissues than MBs, potentially leading to prolonged toxicity in the Pacific oyster. Furthermore, in environments where both MBs and Cd are present, MBs can enhance the toxic effects of Cd through a synergistic interaction. Overall, we provide a reference for understanding the depuration and physiological responses of marine bivalves exposed to MBs and Cd.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111147"},"PeriodicalIF":1.8,"publicationDate":"2025-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144886744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kuruma shrimp (Penaeus japonicus) exhibits sand-diving behaviour. The genetic control mechanism of this behaviour is still unclear, although previous single-cell transcriptome sequencing suggests that crustacean cardioactive peptide (CCAP) may be involved. In this study, we cloned the CCAP precursor cDNA (Pj-CCAP) in kuruma shrimp. It was predicted that the gene can be translated into four related peptides and one mature peptide (PFCNAFTGC-NH2), and is highly similar to crustaceans and insects. The predicted Pj-CCAP itself is highly conserved in other crustaceans. RT-qPCR and in situ hybridization analysis showed that Pj-CCAP was most highly expressed in muscle tissue. RNA interference and gene overexpression were used to assess the relationship between Pj-CCAP expression and sand-diving behaviour of shrimp. The results showed that at 6 h post-treatment, the sand-diving rate in the RNA interference group was significantly lower than that in the control group, while the heart rate of shrimp subjected to RNA interference was also significantly, but recovered to baseline levels within 12 h post-treatment. Overexpression resulted in a lower sand diving rate, and a transient increase in heart rate. The results suggest that Pj-CCAP is potentially a negative regulator of sand diving behaviour of kuruma shrimp, and may have an important role in regulating its physiological rhythms and behavioural patterns.
{"title":"Crustacean cardioactive peptide (CCAP) negatively regulates sand-diving behaviour in kuruma shrimp, Penaeus japonicus","authors":"Xinyu Zhou , Jiahan Yu , Huimin Zhang , Yang Zhang , Panpan Wang , Jing Xu , Fei Yu , Jianxin Zhang , Qingqi Zhang , Wazir Ali Baloch , Huan Gao","doi":"10.1016/j.cbpb.2025.111143","DOIUrl":"10.1016/j.cbpb.2025.111143","url":null,"abstract":"<div><div>Kuruma shrimp (<em>Penaeus japonicus</em>) exhibits sand-diving behaviour. The genetic control mechanism of this behaviour is still unclear, although previous single-cell transcriptome sequencing suggests that crustacean cardioactive peptide (CCAP) may be involved. In this study, we cloned the CCAP precursor cDNA (<em>Pj-CCAP</em>) in kuruma shrimp. It was predicted that the gene can be translated into four related peptides and one mature peptide (PFCNAFTGC-NH2), and is highly similar to crustaceans and insects. The predicted <em>Pj-CCAP</em> itself is highly conserved in other crustaceans. RT-qPCR and in situ hybridization analysis showed that <em>Pj-CCAP</em> was most highly expressed in muscle tissue. RNA interference and gene overexpression were used to assess the relationship between <em>Pj-CCAP</em> expression and sand-diving behaviour of shrimp. The results showed that at 6 h post-treatment, the sand-diving rate in the RNA interference group was significantly lower than that in the control group, while the heart rate of shrimp subjected to RNA interference was also significantly, but recovered to baseline levels within 12 h post-treatment. Overexpression resulted in a lower sand diving rate, and a transient increase in heart rate. The results suggest that <em>Pj-CCAP</em> is potentially a negative regulator of sand diving behaviour of kuruma shrimp, and may have an important role in regulating its physiological rhythms and behavioural patterns.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111143"},"PeriodicalIF":1.8,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144857096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-08-11DOI: 10.1016/j.cbpb.2025.111145
Zhang Shen, Yu Hua, Li Ying
The purpose of this study was to investigate the effects of conjugated linoleic acid (CLA) on fatty acid metabolism through the mammalian target of rapamycin (mTOR) signaling pathway in grass carp (Ctenopharyngodon idella) adipocytes. Grass carp preadipocytes were cultured and treated with various concentrations of CLA (0, 50, 100, 150 and 200 μmol/L). Lipid accumulation was assessed using Oil Red O staining. The regulatory effects on mTOR signaling pathway were evaluated using real-time quantitative PCR (RT-qPCR) and Western Blot (WB). CLA treatment significantly inhibited lipid accumulation and downregulated the expression of genes related to de novo fatty acid synthesis, including fatty acid synthase (fas), acetyl-CoA carboxylase (acc), stearoyl-CoA desaturase 1 (scd1), and sterol regulatory element binding protein-1c (srebp-1c). In contrast, the expression of genes related to fatty acid transport and β-oxidation, such as carnitine palmitoyl transferase-1α (cpt-1α) and hormone-sensitive lipase (hsl), was upregulated. These regulatory effects were found to be concentration-dependent. Co-treatment with the mTOR inhibitor rapamycin further suppressed the mRNA expression of srebp-1c, fas and acc. Furthermore, CLA reduced the protein expression levels of mTOR and nuclear sterol regulatory element-binding protein 1 (nSrebp1), while exerting minimal effect on phosphorylated mTOR (p-mTOR). In summary, CLA appears to modulate lipid metabolism in grass carp adipocytes by inhibiting srebp-1c within the mTOR signaling pathway.
本研究旨在探讨共轭亚油酸（CLA）通过哺乳动物雷帕霉素（mTOR）信号通路对草鱼脂肪细胞脂肪酸代谢的影响。用不同浓度的CLA（0、50、100、150和200 μmol/L）培养草鱼前脂肪细胞。油红O染色检测脂质积累。采用实时荧光定量PCR （RT-qPCR）和Western Blot （WB）方法评价其对mTOR信号通路的调控作用。CLA处理显著抑制脂肪积累，下调脂肪酸合成相关基因的表达，包括脂肪酸合成酶（fas）、乙酰辅酶a羧化酶（acc）、硬脂酰辅酶a去饱和酶1 （scd1）和甾醇调节元件结合蛋白-1c （srebp-1c）。相反，与脂肪酸转运和β-氧化相关的基因，如肉碱棕榈酰转移酶-1α (cpt-1α)和激素敏感脂肪酶（hsl）的表达上调。发现这些调节作用是浓度依赖性的。与mTOR抑制剂雷帕霉素联合治疗进一步抑制srebp-1c、fas和acc的mRNA表达。此外，CLA降低了mTOR和核固醇调控元件结合蛋白1 （nSrebp1）的蛋白表达水平，而对磷酸化mTOR （p-mTOR）的影响很小。综上所述，CLA似乎通过抑制mTOR信号通路中的srebp-1c来调节草鱼脂肪细胞的脂质代谢。
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Pub Date : 2025-08-09DOI: 10.1016/j.cbpb.2025.111144
Lizeth C. Flores-Méndez , Victor Hugo Caña-Bozada , Bruno Gómez-Gil , Crisantema Hernández
High-density aquaculture of Nile tilapia (Oreochromis niloticus) affects welfare by altering cortisol levels, antioxidant response (superoxide dismutase, catalase, and malondialdehyde), and gut microbiota. Agavin is a fructose- and glucose-based polysaccharide containing β-(2-1) and β-(2-6) linkages, known for its potential to mitigate the adverse effects of oxidative stress. This study evaluated the impact of agavin on the intestinal antioxidant response and investigated its correlation with gut microbiota composition and cortisol levels. Fish were fed a control diet (DC0, no agavin) or diets supplemented with 20 g kg−1 (D20) or 40 g kg−1 (D40) of agavin for 110 days. On day 90, all fish were subjected to high-density stress (63 kg m−3). Intestinal superoxide activity, catalase activity, and malondialdehyde concentrations were correlated with previously published plasma cortisol and microbiota data from the same study. Under high-density conditions, the D20 diet increased superoxide dismutase and catalase activity and reduced malondialdehyde concentration compared to the control. Plasma cortisol and malondialdehyde concentration showed a positive correlation (r = 0.67), but plasma cortisol was negatively correlated with superoxide dismutase activity (r = − 0.50). Sixty-nine and 81 operational taxonomic units (OTUs) were significantly correlated with catalase and superoxide dismutase activity, respectively, with families from the Order Cytophagales being strongly positively correlated. Cytophagales was also negatively correlated with plasma cortisol and malondialdehyde concentrations. These results suggest that, under stress conditions, agavin promotes the antioxidant response, modulating the intestinal microbiota and reducing cortisol release, which could indicate its utility as a prebiotic in aquaculture.
尼罗罗非鱼（Oreochromis niloticus）的高密度养殖通过改变皮质醇水平、抗氧化反应（超氧化物歧化酶、过氧化氢酶和丙二醛）和肠道微生物群来影响福利。Agavin是一种以果糖和葡萄糖为基础的多糖，含有β-(2-1)和β-(2-6)键，以其减轻氧化应激不良影响的潜力而闻名。本研究评估了agavin对肠道抗氧化反应的影响，并研究了其与肠道微生物群组成和皮质醇水平的相关性。饲喂对照饲料（DC0，不添加）或在饲料中添加20 g kg-1 （D20）或40 g kg-1 (D40) 110 d。在第90天，所有鱼都受到高密度应激（63 kg m-3）。肠道超氧化物活性、过氧化氢酶活性和丙二醛浓度与先前发表的血浆皮质醇和微生物群数据相关。在高密度条件下，与对照组相比，D20饲粮提高了超氧化物歧化酶和过氧化氢酶活性，降低了丙二醛浓度。血浆皮质醇与丙二醛浓度呈正相关（r = 0.67），与超氧化物歧化酶活性呈负相关（r = - 0.50）。69个操作分类单位（otu）和81个操作分类单位（otu）分别与过氧化氢酶和超氧化物歧化酶活性显著相关，与胞噬目家族呈显著正相关。细胞吞噬体也与血浆皮质醇和丙二醛浓度呈负相关。这些结果表明，在应激条件下，agavin促进抗氧化反应，调节肠道微生物群并减少皮质醇释放，这可能表明其作为水产养殖益生元的用途。
{"title":"The effect of agavin on the intestinal antioxidant response and its relationship to gut microbiota and plasma cortisol in Nile tilapia (Oreochromis niloticus) subjected to overcrowding stress","authors":"Lizeth C. Flores-Méndez , Victor Hugo Caña-Bozada , Bruno Gómez-Gil , Crisantema Hernández","doi":"10.1016/j.cbpb.2025.111144","DOIUrl":"10.1016/j.cbpb.2025.111144","url":null,"abstract":"<div><div>High-density aquaculture of Nile tilapia (<em>Oreochromis niloticus</em>) affects welfare by altering cortisol levels, antioxidant response (superoxide dismutase, catalase, and malondialdehyde), and gut microbiota. Agavin is a fructose- and glucose-based polysaccharide containing β-(2-1) and β-(2-6) linkages, known for its potential to mitigate the adverse effects of oxidative stress. This study evaluated the impact of agavin on the intestinal antioxidant response and investigated its correlation with gut microbiota composition and cortisol levels. Fish were fed a control diet (DC0, no agavin) or diets supplemented with 20 g kg<sup>−1</sup> (D20) or 40 g kg<sup>−1</sup> (D40) of agavin for 110 days. On day 90, all fish were subjected to high-density stress (63 kg m<sup>−3</sup>). Intestinal superoxide activity, catalase activity, and malondialdehyde concentrations were correlated with previously published plasma cortisol and microbiota data from the same study. Under high-density conditions, the D20 diet increased superoxide dismutase and catalase activity and reduced malondialdehyde concentration compared to the control. Plasma cortisol and malondialdehyde concentration showed a positive correlation (<em>r</em> = 0.67), but plasma cortisol was negatively correlated with superoxide dismutase activity (<em>r</em> = − 0.50). Sixty-nine and 81 operational taxonomic units (OTUs) were significantly correlated with catalase and superoxide dismutase activity, respectively, with families from the Order Cytophagales being strongly positively correlated. Cytophagales was also negatively correlated with plasma cortisol and malondialdehyde concentrations. These results suggest that, under stress conditions, agavin promotes the antioxidant response, modulating the intestinal microbiota and reducing cortisol release, which could indicate its utility as a prebiotic in aquaculture.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"280 ","pages":"Article 111144"},"PeriodicalIF":1.8,"publicationDate":"2025-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144823274","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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