There is an urgent need to elucidate the pathogenesis of myocardial ischemia(MI)and potential drug treatments.Here,the anti-MI mechanism and material basis of Ginkgo biloba L.extract(GBE)were studied from the perspect...There is an urgent need to elucidate the pathogenesis of myocardial ischemia(MI)and potential drug treatments.Here,the anti-MI mechanism and material basis of Ginkgo biloba L.extract(GBE)were studied from the perspective of energy metabolism flux regulation.Metabolic flux analysis(MFA)was performed to investigate energy metabolism flux disorder and the regulatory nodes of GBE components in isoproterenol(ISO)-induced ischemia-like cardiomyocytes.It showed that[U-13 C]glucose derived m+2 isotopologues from the upstream tricarboxylic acid(TCA)cycle metabolites were markedly accumulated in ISO-injured cardiomyocytes,but the opposite was seen for the downstream metabolites,while their total cellular concentrations were increased.This indicates a blockage of carbon flow from glycolysis and enhanced anaplerosis from other carbon sources.A Seahorse test was used to screen for GBE components with regulatory effects on mitochondrial aerobic respiratory dysfunction.It showed that bilobalide protected against impaired mitochondrial aerobic respiration.MFA also showed that bilobalide significantly modulated the TCA cycle flux,reduced abnormal metabolite accumulation,and balanced the demand of different carbon sources.Western blotting and PCR analysis showed that bilobalide decreased the enhanced expression of key metabolic enzymes in injured cells.Bilobalide’s efficacy was verified by in vivo experiments in rats.This is the first report to show that bilobalide,the active ingredient of GBE,protects against MI by rescuing impaired TCA cycle flux.This provides a new mechanism and potential drug treatment for MI.It also shows the potential of MFA/Seahorse combination as a powerful strategy for pharmacological research on herbal medicine.展开更多
Seahorse embryos are brooded in the enclosed nutrient-rich environment of the male brood pouch,which may be prone to bacterial infection.The immune responses of interleukin(IL)genes in the brood pouch have not been we...Seahorse embryos are brooded in the enclosed nutrient-rich environment of the male brood pouch,which may be prone to bacterial infection.The immune responses of interleukin(IL)genes in the brood pouch have not been well studied.We identifi ed 13 interleukins in the lined seahorse Hippocampus erectus.Tissue-specifi c expression analysis revealed increased mRNA expression levels of il-1β,il-18,and il-8 in the brood pouch.When challenged with lipopolysaccharide or Vibrio parahaemolyticus,il-1βand il-18 were active as part of the acute and chronic infl ammatory responses,respectively.Importantly,il-8 may be involved in powerful antibacterial immune responses and may be induced by il-1βand il-18 via a process involving the nuclear factor-κB signaling pathway.These results suggest that il-1β,il-18,and il-8 may play key roles in the antibacterial immune defense of the brood pouch in male seahorses.展开更多
Seagrass ecosystems worldwide have been declining, leading to a decrease in associated fish populations, especially those with low mobility such as syngnathids (pipefish and seahorses). This two-year pilot study inv...Seagrass ecosystems worldwide have been declining, leading to a decrease in associated fish populations, especially those with low mobility such as syngnathids (pipefish and seahorses). This two-year pilot study investigated seasonal patterns in density, growth, site fidelity, and population dynamics of Tampa Bay (FL) syngnathid fishes at a site adjacent to two marinas under construction. Using a modified mark-recapture technique, fish were collected periodically from three closely located sites that varied in seagrass species (Thalassia spp., Syringodium spp., and mixed-grass sites) and their distance from open water, but had consistent physical/chemical environmental characteristics. Fish were marked, photographed for body size and gender measurements, and released the same day at the capture site. Of the 5695 individuals surveyed, 49 individuals were recaptured, indicating a large, flexible population. Population density peaks were observed in July of both years, with low densities in late winter and late summer. Spatially, syngnathid densities were highest closest to the mouth of the bay and lowest near the shoreline. Seven species of syngnathid fishes were observed, and species-specific patterns of seagrass use emerged during the study. However, only two species, Syngnathus scovelli and Hippoeampus zosterae, were observed at high frequencies. For these two species, body size decreased across the study period, but while S. scovelli's population density decreased, H. zosterae's increased. Across six of the seven species, population size declined over the course of this preliminary study; however, seasonal shifts were impossible to distinguish from potential anthropogenic effects of construction [Current Zoology 56 (1): 118-133, 2010].展开更多
Background:Polycystic ovary syndrome(PCOS)is an endocrine disorder that occurs in women of child-bearing age.Moreover,PCOS patients have decreased pregnancy rates and clomiphene citrate resistance.The traditional Chin...Background:Polycystic ovary syndrome(PCOS)is an endocrine disorder that occurs in women of child-bearing age.Moreover,PCOS patients have decreased pregnancy rates and clomiphene citrate resistance.The traditional Chinese medicine formula Kun-Tai-1S(KT1S),consisting of the seahorse species hippocampus,has been reported to elicit therapeutic effects in patients with PCOS.However,given the limited resources and global demand for wild hippocampus,whether KT1S with or without hippocampus can elicit similar therapeutic effects has not been confirmed.Methods:KT1S and Kun-Tai-1A(KT1A,KT1S without dry hippocampus)were used to treat a letrozole-induced rat model of PCOS with an established disease.The serum levels of testosterone,luteinizing hormone,anti-Müllerian hormone,and estradiol were determined,the luteinizing hormone/follicle-stimulating hormone ratio was determined,and the ovarian pathology was evaluated.Results:Similar to the therapeutic effects of cyproterone acetate,both the KT1S and KT1A treatments reduced the body weight and ovarian and uterine indices in the rats with PCOS.The serum levels of testosterone,anti-Müllerian hormone,and luteinizing hormone and the luteinizing hormone/follicle-stimulating hormone ratio were significantly lower in the KT1S and KT1A treatment groups compared to the model group(P<0.01 and P<0.05,respectively).Moreover,the histopathological assessment results suggested that both the KT1S and KT1A treatments significantly ameliorated the PCOS pathology in the rats with an established disease,with a reduced number of cystic and atretic follicles and an increased number of corpora lutea being observed in the ovaries.Notably,there was no obvious difference in the disease outcomes between the KT1S and KT1A-treated groups.Network pharmacology analysis revealed that 4’,7-dihydroxyflavanone,sinpemine A,quercetin,8-isopentenyl-kaempferol,and luteolin in KT1A may promote estrogen signaling;furthermore,the nitric oxide regulation pathway is also closely involved.Conclusion:KT1A and KT1S treatments both significantly ameliorated the PCOS-related pathology in rats,suggesting that the hippocampus component is dispensable for KT1S-mediated amelioration.Given the limited resources and global demand for wild hippocampus for use in complementary medicines,our findings may help conserve this species.Together,our results suggest that KT1A is a promising approach for treating PCOS.展开更多
Vertebrates have developed various modes of reproduction,some of which are found in Teleosts.Over 300 species of the Syngnathidae(seahorses,pipefishes and seadragons)exhibit male pregnancies;the males have specialized...Vertebrates have developed various modes of reproduction,some of which are found in Teleosts.Over 300 species of the Syngnathidae(seahorses,pipefishes and seadragons)exhibit male pregnancies;the males have specialized brood pouches that provide immune protection,nourishment,and oxygen regulation.Chemokines play a vital role at the mammalian maternal–fetal interface;however,their functions in fish reproduction are unclear.This study revealed the evolutionary traits and potential functions of chemokine genes in 22 oviparous,ovoviviparous,and viviparous fish species through comparative genomic analyses.Our results showed that chemokine gene copy numbers and evolutionary rates vary among species with different modes of reproduction.Syngnathidae lost cxcl13 and cxcr5,which are involved in key receptor–ligand pairs for lymphoid organ development.Notably,Syngnathidae have site-specific mutations in cxcl12b and ccl44,suggesting immune function during gestation.Moreover,transcriptome analysis revealed that chemokine gene expression varies among Syngnathidae species with different types of brood pouches,suggesting adaptive variations in chemokine functions among seahorses and their relatives.Furthermore,challenge experiments on seahorse brood pouches revealed a joint immune function of chemokine genes during male pregnancy.This study provides insights into the evolutionary diversity of chemokine genes associated with different reproductive modes in fish.展开更多
Seahorses are a hallmark of specialized morphological features due to their elongated prehensile tail.However,the underlying genomic grounds of seahorse tail development remain elusive.Herein,we evaluated the roles of...Seahorses are a hallmark of specialized morphological features due to their elongated prehensile tail.However,the underlying genomic grounds of seahorse tail development remain elusive.Herein,we evaluated the roles of essential genes from the Wnt gene family for the tail developmental process in the lined seahorse(Hippocampus erectus).Comparative genomic analysis revealed that the Wnt gene family is conserved in seahorses.The expression profiles and in situ hybridization suggested that Wnt5a,Wnt8a,and Wnt11 may participate in seahorse tail development.Like in other teleosts,Wnt5a and Wntll were found to regulate the development of the tail axial mesoderm and tail somitic mesoderm,respectively.However,a significantly extended expression period of Wnt8a during seahorse tail development was observed.Signaling pathway analysis further showed that Wnt5a up-regulated the expression of the tail axial mesoderm gene(Shh),while interaction analysis indicated that Wnt8a could promote the expression of Wntll.In summary,our results indicate that the special extended expression period of Wnt8a might promote caudal tail axis formation,which contributes to the formation of the elongated tail of the seahorse.展开更多
Subject Code:D06Supported by the National Natural Science Foundation of China and the Strategic Priority Research Program of the Chinese Academy of Sciences,the research team led by Prof.Lin Qiang(林强)at the CAS Key ...Subject Code:D06Supported by the National Natural Science Foundation of China and the Strategic Priority Research Program of the Chinese Academy of Sciences,the research team led by Prof.Lin Qiang(林强)at the CAS Key Laboratory of Tropical Marine Bio-resources and Ecology(LMB),South China Sea Institute of Oceanology,Chinese Academy of Sciences,uncovered the seahorse genome and the evolution of its specialized morphology,which was published in Nature as a cover article(2016,540:395—399).展开更多
Seahorses epitomize the exuberance of evolution.They have the unique characteristic of male pregnancy,which includes the carrying of many embryos in a brood pouch that incubates and nourishes the embryos,similar to th...Seahorses epitomize the exuberance of evolution.They have the unique characteristic of male pregnancy,which includes the carrying of many embryos in a brood pouch that incubates and nourishes the embryos,similar to the mammalian placenta.However,the regulatory networks underlying brood pouch formation and pregnancy remain largely unknown.In this study,comparative transcriptomic and metabolomic profiling on the lined seahorse Hippocampus erectus,with unformed,newly formed,and pregnant brood pouches identified a total of 141 and 2,533 differentially expressed genes together with 73 and 121 significantly differential metabolites related to brood pouch formation and pregnancy,respectively.Specifically,integrative omics analysis revealed that retinoic acid(RA)synthesis and signaling pathway played essential roles in the formation of the brood pouch and pregnancy.RA might function upstream of testosterone and progesterone,thereby directly influencing brood pouch formation by regulating the expression of fshr and cyp7a1.Our results also revealed that RA regulates antioxidant defenses,particularly during male pregnancy.Alternatively,pregnancy caused a consistent decrease in RA,canthaxanthin,astaxanthin,and glutathione synthetase,and an increase in susceptibility to oxidative stress,which may balance brood pouch development and reproduction in seahorses and pave the way to successful gestation.展开更多
基金国家自然科学基金面上项目(81573487):二氢神经酰胺类鞘脂及其代谢通路在慢加急性肝衰竭发生发展中的作用及防治措施研究中国医学科学院医学与健康科技创新工程重大协同创新项目(2017-12M-1-013):中药安全风险预警及防控国际合作项目(Grant ID4084):Applications and core technology university research(ACT-UR)proposal form
文摘目的利用Seahorse XF^e24海马细胞能量代谢分析系统分析对乙酰氨基酚(APAP)对人肝细胞线粒体有氧呼吸的整体影响,为APAP在能量代谢角度的毒性防控及毒性机制研究提供方法学的参考。方法体外培养人肝细胞系(HepG2),分别以1、2、4、8、16 mM APAP处理细胞24 h,MTT法检测细胞增殖情况,倒置显微镜观察细胞形态;BCA法定蛋白含量。采用Seahorse XF^e24海马细胞能量代谢分析系统建立HepG2细胞实验方法,并检测不同浓度APAP对人肝细胞线粒体有氧呼吸的影响和特点。结果 Seahorse XF^e24海马细胞能量代谢分析系统检测HepG2细胞能量代谢的最佳条件为:24孔板细胞密度2×10~4/孔,FCCP浓度为2μM。MTT结果中对人肝细胞增殖有显著抑制作用的APAP(4、8、16 mM)能干扰人肝细胞的线粒体有氧呼吸,其中4、8 mM APAP有降低肝细胞基础呼吸和ATP生成的趋势,16 mM APAP能显著抑制肝细胞的基础呼吸、最大呼吸和ATP合成。结论高剂量APAP(16 mM)在人肝细胞活细胞体系中能够显著抑制细胞能量代谢,对活细胞线粒体有氧呼吸显示显著毒性作用。
基金supported by grants from the National Natural Science Foundation of China(Grant No.:81803496)the CAMS Innovation Fund for Medical Sciences(Grant No.:2016-I2M-3-016)the Applications and Core Technology University Research(ACT-UR,Grant No.:4084)。
文摘There is an urgent need to elucidate the pathogenesis of myocardial ischemia(MI)and potential drug treatments.Here,the anti-MI mechanism and material basis of Ginkgo biloba L.extract(GBE)were studied from the perspective of energy metabolism flux regulation.Metabolic flux analysis(MFA)was performed to investigate energy metabolism flux disorder and the regulatory nodes of GBE components in isoproterenol(ISO)-induced ischemia-like cardiomyocytes.It showed that[U-13 C]glucose derived m+2 isotopologues from the upstream tricarboxylic acid(TCA)cycle metabolites were markedly accumulated in ISO-injured cardiomyocytes,but the opposite was seen for the downstream metabolites,while their total cellular concentrations were increased.This indicates a blockage of carbon flow from glycolysis and enhanced anaplerosis from other carbon sources.A Seahorse test was used to screen for GBE components with regulatory effects on mitochondrial aerobic respiratory dysfunction.It showed that bilobalide protected against impaired mitochondrial aerobic respiration.MFA also showed that bilobalide significantly modulated the TCA cycle flux,reduced abnormal metabolite accumulation,and balanced the demand of different carbon sources.Western blotting and PCR analysis showed that bilobalide decreased the enhanced expression of key metabolic enzymes in injured cells.Bilobalide’s efficacy was verified by in vivo experiments in rats.This is the first report to show that bilobalide,the active ingredient of GBE,protects against MI by rescuing impaired TCA cycle flux.This provides a new mechanism and potential drug treatment for MI.It also shows the potential of MFA/Seahorse combination as a powerful strategy for pharmacological research on herbal medicine.
基金Supported by the National Science&Technology Fundamental Resources Investigation Program of China(No.2018FY100106),the National Natural Science Foundation of China(Nos.41825013,41806189)the Key Special Project for Introduced Talents Team of Southern Marine Science and Engineering Guangdong Laboratory(Guangzhou)(No.GML2019ZD0407)the K.C.Wong Education Foundation,and the Guangdong Special Support Program of Leading Scientifi c and Technological Innovation(No.2017TX04N442)。
文摘Seahorse embryos are brooded in the enclosed nutrient-rich environment of the male brood pouch,which may be prone to bacterial infection.The immune responses of interleukin(IL)genes in the brood pouch have not been well studied.We identifi ed 13 interleukins in the lined seahorse Hippocampus erectus.Tissue-specifi c expression analysis revealed increased mRNA expression levels of il-1β,il-18,and il-8 in the brood pouch.When challenged with lipopolysaccharide or Vibrio parahaemolyticus,il-1βand il-18 were active as part of the acute and chronic infl ammatory responses,respectively.Importantly,il-8 may be involved in powerful antibacterial immune responses and may be induced by il-1βand il-18 via a process involving the nuclear factor-κB signaling pathway.These results suggest that il-1β,il-18,and il-8 may play key roles in the antibacterial immune defense of the brood pouch in male seahorses.
基金Field work for this project was conducted under State of Florida Special Activities License Number 05SR-902provided in part by the Tampa Bay Estuary Program+2 种基金Delo Research Professor Grant (University of Tampa)the Dana Award Program (University of Tampa)the Biology Department (University of Tampa)
文摘Seagrass ecosystems worldwide have been declining, leading to a decrease in associated fish populations, especially those with low mobility such as syngnathids (pipefish and seahorses). This two-year pilot study investigated seasonal patterns in density, growth, site fidelity, and population dynamics of Tampa Bay (FL) syngnathid fishes at a site adjacent to two marinas under construction. Using a modified mark-recapture technique, fish were collected periodically from three closely located sites that varied in seagrass species (Thalassia spp., Syringodium spp., and mixed-grass sites) and their distance from open water, but had consistent physical/chemical environmental characteristics. Fish were marked, photographed for body size and gender measurements, and released the same day at the capture site. Of the 5695 individuals surveyed, 49 individuals were recaptured, indicating a large, flexible population. Population density peaks were observed in July of both years, with low densities in late winter and late summer. Spatially, syngnathid densities were highest closest to the mouth of the bay and lowest near the shoreline. Seven species of syngnathid fishes were observed, and species-specific patterns of seagrass use emerged during the study. However, only two species, Syngnathus scovelli and Hippoeampus zosterae, were observed at high frequencies. For these two species, body size decreased across the study period, but while S. scovelli's population density decreased, H. zosterae's increased. Across six of the seven species, population size declined over the course of this preliminary study; however, seasonal shifts were impossible to distinguish from potential anthropogenic effects of construction [Current Zoology 56 (1): 118-133, 2010].
文摘Background:Polycystic ovary syndrome(PCOS)is an endocrine disorder that occurs in women of child-bearing age.Moreover,PCOS patients have decreased pregnancy rates and clomiphene citrate resistance.The traditional Chinese medicine formula Kun-Tai-1S(KT1S),consisting of the seahorse species hippocampus,has been reported to elicit therapeutic effects in patients with PCOS.However,given the limited resources and global demand for wild hippocampus,whether KT1S with or without hippocampus can elicit similar therapeutic effects has not been confirmed.Methods:KT1S and Kun-Tai-1A(KT1A,KT1S without dry hippocampus)were used to treat a letrozole-induced rat model of PCOS with an established disease.The serum levels of testosterone,luteinizing hormone,anti-Müllerian hormone,and estradiol were determined,the luteinizing hormone/follicle-stimulating hormone ratio was determined,and the ovarian pathology was evaluated.Results:Similar to the therapeutic effects of cyproterone acetate,both the KT1S and KT1A treatments reduced the body weight and ovarian and uterine indices in the rats with PCOS.The serum levels of testosterone,anti-Müllerian hormone,and luteinizing hormone and the luteinizing hormone/follicle-stimulating hormone ratio were significantly lower in the KT1S and KT1A treatment groups compared to the model group(P<0.01 and P<0.05,respectively).Moreover,the histopathological assessment results suggested that both the KT1S and KT1A treatments significantly ameliorated the PCOS pathology in the rats with an established disease,with a reduced number of cystic and atretic follicles and an increased number of corpora lutea being observed in the ovaries.Notably,there was no obvious difference in the disease outcomes between the KT1S and KT1A-treated groups.Network pharmacology analysis revealed that 4’,7-dihydroxyflavanone,sinpemine A,quercetin,8-isopentenyl-kaempferol,and luteolin in KT1A may promote estrogen signaling;furthermore,the nitric oxide regulation pathway is also closely involved.Conclusion:KT1A and KT1S treatments both significantly ameliorated the PCOS-related pathology in rats,suggesting that the hippocampus component is dispensable for KT1S-mediated amelioration.Given the limited resources and global demand for wild hippocampus for use in complementary medicines,our findings may help conserve this species.Together,our results suggest that KT1A is a promising approach for treating PCOS.
基金supported by the National Natural Science Foundation of China(41825013)the Marine Economic Development Project(GDNRC[2022]36)+2 种基金National Key R&D Program of China(2022YFC3102403)National Natural Science Foundation of China(42230409,42276127)Guangdong Basic and Applied Basic Research Foundation(2021A1515011380,2021A1515011393).
文摘Vertebrates have developed various modes of reproduction,some of which are found in Teleosts.Over 300 species of the Syngnathidae(seahorses,pipefishes and seadragons)exhibit male pregnancies;the males have specialized brood pouches that provide immune protection,nourishment,and oxygen regulation.Chemokines play a vital role at the mammalian maternal–fetal interface;however,their functions in fish reproduction are unclear.This study revealed the evolutionary traits and potential functions of chemokine genes in 22 oviparous,ovoviviparous,and viviparous fish species through comparative genomic analyses.Our results showed that chemokine gene copy numbers and evolutionary rates vary among species with different modes of reproduction.Syngnathidae lost cxcl13 and cxcr5,which are involved in key receptor–ligand pairs for lymphoid organ development.Notably,Syngnathidae have site-specific mutations in cxcl12b and ccl44,suggesting immune function during gestation.Moreover,transcriptome analysis revealed that chemokine gene expression varies among Syngnathidae species with different types of brood pouches,suggesting adaptive variations in chemokine functions among seahorses and their relatives.Furthermore,challenge experiments on seahorse brood pouches revealed a joint immune function of chemokine genes during male pregnancy.This study provides insights into the evolutionary diversity of chemokine genes associated with different reproductive modes in fish.
基金This work was supported by the K.C.Wong Education Foundation,the National Natural Science Foundation of China(nos.41825013,41706178,41576145,41806189,32000350)the China postdoctoral science foundation grant(no.2019M663151),the Guangdong Special Support Program of Youth Scientific and Technological Innovation(2017TQ04Z269)We are also grateful to Wenqi Hu for help with cell culture.
文摘Seahorses are a hallmark of specialized morphological features due to their elongated prehensile tail.However,the underlying genomic grounds of seahorse tail development remain elusive.Herein,we evaluated the roles of essential genes from the Wnt gene family for the tail developmental process in the lined seahorse(Hippocampus erectus).Comparative genomic analysis revealed that the Wnt gene family is conserved in seahorses.The expression profiles and in situ hybridization suggested that Wnt5a,Wnt8a,and Wnt11 may participate in seahorse tail development.Like in other teleosts,Wnt5a and Wntll were found to regulate the development of the tail axial mesoderm and tail somitic mesoderm,respectively.However,a significantly extended expression period of Wnt8a during seahorse tail development was observed.Signaling pathway analysis further showed that Wnt5a up-regulated the expression of the tail axial mesoderm gene(Shh),while interaction analysis indicated that Wnt8a could promote the expression of Wntll.In summary,our results indicate that the special extended expression period of Wnt8a might promote caudal tail axis formation,which contributes to the formation of the elongated tail of the seahorse.
文摘Subject Code:D06Supported by the National Natural Science Foundation of China and the Strategic Priority Research Program of the Chinese Academy of Sciences,the research team led by Prof.Lin Qiang(林强)at the CAS Key Laboratory of Tropical Marine Bio-resources and Ecology(LMB),South China Sea Institute of Oceanology,Chinese Academy of Sciences,uncovered the seahorse genome and the evolution of its specialized morphology,which was published in Nature as a cover article(2016,540:395—399).
基金supported by the National Natural Science Foundation of China(41825013,41806189),Chinathe K.C.Wong Education Foundation,China+2 种基金the Key Special Project for Introduced Talents Teamof SouthernMarine Science and EngineeringGuangdong Laboratory(Guangzhou)(GML2019ZD0407),Chinathe Guangdong Special Support Program of Youth Scientific and Technological Innovation(2017TQ04Z269),Chinathe ChinaPostdoctoral Science Foundation grant(2019M663151),China.
文摘Seahorses epitomize the exuberance of evolution.They have the unique characteristic of male pregnancy,which includes the carrying of many embryos in a brood pouch that incubates and nourishes the embryos,similar to the mammalian placenta.However,the regulatory networks underlying brood pouch formation and pregnancy remain largely unknown.In this study,comparative transcriptomic and metabolomic profiling on the lined seahorse Hippocampus erectus,with unformed,newly formed,and pregnant brood pouches identified a total of 141 and 2,533 differentially expressed genes together with 73 and 121 significantly differential metabolites related to brood pouch formation and pregnancy,respectively.Specifically,integrative omics analysis revealed that retinoic acid(RA)synthesis and signaling pathway played essential roles in the formation of the brood pouch and pregnancy.RA might function upstream of testosterone and progesterone,thereby directly influencing brood pouch formation by regulating the expression of fshr and cyp7a1.Our results also revealed that RA regulates antioxidant defenses,particularly during male pregnancy.Alternatively,pregnancy caused a consistent decrease in RA,canthaxanthin,astaxanthin,and glutathione synthetase,and an increase in susceptibility to oxidative stress,which may balance brood pouch development and reproduction in seahorses and pave the way to successful gestation.