Hematological parameters can provide key information to an animal health status.However,this information is usually hard to obtain.Here,we described the hematological parameters of Leptodactylus podicipinus in the Bra...Hematological parameters can provide key information to an animal health status.However,this information is usually hard to obtain.Here,we described the hematological parameters of Leptodactylus podicipinus in the Brazilian Pantanal.We measured red blood cell morphometrics,erythrogram,and leukogram.We also tested for phylogenetic signal in the erythrogram and leukogram of 48 frog species from 15 families,testing if body size explains their variation.Lymphocytes were the most abundant leukocytes(>60%)in L.podicipinus,followed by neutrophils(∼10%).Given that L.podicipinus is an abundant and widely distributed species in central Brazil,knowing its hematological pattern can help establish a baseline and improve its use as a bioindicator of environmental degradation.Mean corpuscular hemoglobin and value contributed more to the phylomorphospace of erythrogram,in which Leptodactylus spp.and Hypsiboas raniceps had lower values of these variables,whereas Bufotes viridis and Hyla arborea had high values.The phylogenetic signal was spread throughout the dimensions of the leukogram phylomorphospace.The variables that most contributed to it were total leukocytes counts,lymphocytes,and neutrophils.We also found a moderate phylogenetic signal for both the erythrogram and leukogram.Accordingly,body size accounted for a low proportion of variation in both the leukogram(4.7%)and erythrogram(0.57%).By applying phylogenetic comparative methods to hematological parameters,our results add a new perspective on the evolution of blood cell physiology in frogs.展开更多
Plasma kallikrein（PK）, a serine protease in the trypsin clan（SA）, plays critical roles in many physiological and pathological pathways. Regulating the abnormal activity of PK has been successfully used in the clin...Plasma kallikrein（PK）, a serine protease in the trypsin clan（SA）, plays critical roles in many physiological and pathological pathways. Regulating the abnormal activity of PK has been successfully used in the clinical therapy of hereditary angioedema. In this study, the serine protease domain of murine plasma kallikrein（m PK） was expressed in the pichia pastoris system. The recombinant protein was a glycosylated active enzyme after purification by the cation exchange and size-exclusion chromatography, and was crystallized at the precipitant condition of 25% PEG 3350, 0.1 M Tris-HCl pH 8.5 and 0.1 M Na Cl. The crystal structure of m PK was determined at 2.6 . This is the first published crystal structure of m PK, showing some distinctive features at S2＇ and S3＇ pockets when compared to its human analogue（human plasma kallikrein, h PK）. In addition, m PK show unique structural features in the non-conservative 67-72 and 76-81 loops when compared to other serine proteases. These results provide insights for the design of potent and selective PK inhibitors.展开更多
Salinity(sodium chloride,NaCl)from anthropogenic sources is a persistent contaminant that negatively affects freshwater taxa.Amphibians can be susceptible to salinity,but some species are innately or adaptively tolera...Salinity(sodium chloride,NaCl)from anthropogenic sources is a persistent contaminant that negatively affects freshwater taxa.Amphibians can be susceptible to salinity,but some species are innately or adaptively tolerant.Physiological mechanisms mediating tolerance to salinity are still unclear,but changes in osmoregulatory hor-mones such as corticosterone(CORT)and aldosterone(ALDO)are prime candidates.We exposed larval barred tiger salamanders(Ambystoma mavortium)to environmentally relevant NaCl treatments(<32–4000 mg·L-1)for 24 days to test effects on growth,survival,and waterborne CORT responses.Of those sampled,we also quantified waterborne ALDO from a subset.Using a glucocorticoid antagonist(RU486),we also experimentally suppressed CORT signaling of some larvae to determine if CORT mediates effects of salinity.There were no strong differ-ences in survival among salinity treatments,but salinity reduced dry mass,snout–vent length,and body condition while increasing water content of larvae.High survival and sublethal effects demonstrated that salamanders were physiologically challenged but could tolerate the experimental concentrations.CORT signaling did not attenuate sublethal effects of salinity.Baseline and stress-induced(after an acute stressor,shaking)CORT were not influ-enced by salinity.ALDO was correlated with baseline CORT,suggesting it could be difficult to decouple the roles of CORT and ALDO.Future studies comparing ALDO and CORT responses of adaptively tolerant and previously unexposed populations could be beneficial to understand the roles of these hormones in tolerance to salinity.Nev-ertheless,our study enhances our understanding of the roles of corticosteroid hormones in mediating effects of a prominent anthropogenic stressor.展开更多
基金SISBIO provided collecting permit(#63297-1)This study was funded in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil(CAPES)–Finance Code 001.CESF received a grant from the Fundação de Apoio ao Desenvolvimento do Ensino,Ciência e Tecnologia do Estado de Mato Grosso do Sul(#71/700.136)+1 种基金has been continuously supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico(grant#310058/2020-1)C.O.has been continuously supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico(grant#304552/2019-4).
文摘Hematological parameters can provide key information to an animal health status.However,this information is usually hard to obtain.Here,we described the hematological parameters of Leptodactylus podicipinus in the Brazilian Pantanal.We measured red blood cell morphometrics,erythrogram,and leukogram.We also tested for phylogenetic signal in the erythrogram and leukogram of 48 frog species from 15 families,testing if body size explains their variation.Lymphocytes were the most abundant leukocytes(>60%)in L.podicipinus,followed by neutrophils(∼10%).Given that L.podicipinus is an abundant and widely distributed species in central Brazil,knowing its hematological pattern can help establish a baseline and improve its use as a bioindicator of environmental degradation.Mean corpuscular hemoglobin and value contributed more to the phylomorphospace of erythrogram,in which Leptodactylus spp.and Hypsiboas raniceps had lower values of these variables,whereas Bufotes viridis and Hyla arborea had high values.The phylogenetic signal was spread throughout the dimensions of the leukogram phylomorphospace.The variables that most contributed to it were total leukocytes counts,lymphocytes,and neutrophils.We also found a moderate phylogenetic signal for both the erythrogram and leukogram.Accordingly,body size accounted for a low proportion of variation in both the leukogram(4.7%)and erythrogram(0.57%).By applying phylogenetic comparative methods to hematological parameters,our results add a new perspective on the evolution of blood cell physiology in frogs.
基金Financially supported by the National Natural Science Foundation of China(31570745,31370737,31400637)the Danish National Research Foundation(26-331-6)
文摘Plasma kallikrein（PK）, a serine protease in the trypsin clan（SA）, plays critical roles in many physiological and pathological pathways. Regulating the abnormal activity of PK has been successfully used in the clinical therapy of hereditary angioedema. In this study, the serine protease domain of murine plasma kallikrein（m PK） was expressed in the pichia pastoris system. The recombinant protein was a glycosylated active enzyme after purification by the cation exchange and size-exclusion chromatography, and was crystallized at the precipitant condition of 25% PEG 3350, 0.1 M Tris-HCl pH 8.5 and 0.1 M Na Cl. The crystal structure of m PK was determined at 2.6 . This is the first published crystal structure of m PK, showing some distinctive features at S2＇ and S3＇ pockets when compared to its human analogue（human plasma kallikrein, h PK）. In addition, m PK show unique structural features in the non-conservative 67-72 and 76-81 loops when compared to other serine proteases. These results provide insights for the design of potent and selective PK inhibitors.
基金funded by University of Montana,U.S.Geological Survey(USGS RWO#103 to CWB),National Science Foundation(NSF-DEB 1754474 to EJC)the Nelson Schwab Family Foundation。
文摘Salinity(sodium chloride,NaCl)from anthropogenic sources is a persistent contaminant that negatively affects freshwater taxa.Amphibians can be susceptible to salinity,but some species are innately or adaptively tolerant.Physiological mechanisms mediating tolerance to salinity are still unclear,but changes in osmoregulatory hor-mones such as corticosterone(CORT)and aldosterone(ALDO)are prime candidates.We exposed larval barred tiger salamanders(Ambystoma mavortium)to environmentally relevant NaCl treatments(<32–4000 mg·L-1)for 24 days to test effects on growth,survival,and waterborne CORT responses.Of those sampled,we also quantified waterborne ALDO from a subset.Using a glucocorticoid antagonist(RU486),we also experimentally suppressed CORT signaling of some larvae to determine if CORT mediates effects of salinity.There were no strong differ-ences in survival among salinity treatments,but salinity reduced dry mass,snout–vent length,and body condition while increasing water content of larvae.High survival and sublethal effects demonstrated that salamanders were physiologically challenged but could tolerate the experimental concentrations.CORT signaling did not attenuate sublethal effects of salinity.Baseline and stress-induced(after an acute stressor,shaking)CORT were not influ-enced by salinity.ALDO was correlated with baseline CORT,suggesting it could be difficult to decouple the roles of CORT and ALDO.Future studies comparing ALDO and CORT responses of adaptively tolerant and previously unexposed populations could be beneficial to understand the roles of these hormones in tolerance to salinity.Nev-ertheless,our study enhances our understanding of the roles of corticosteroid hormones in mediating effects of a prominent anthropogenic stressor.
