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1.
PLoS One ; 15(10): e0233481, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33001997

RESUMO

Pearl millet is a key cereal for food security in arid and semi-arid regions but its yield is increasingly threatened by water stress. Physiological mechanisms relating to conservation of soil water or increased water use efficiency can alleviate that stress. Aquaporins (AQP) are water channels that mediate root water transport, thereby influencing plant hydraulics, transpiration and soil water conservation. However, AQP remain largely uncharacterized in pearl millet. Here, we studied AQP function in root water transport in two pearl millet lines contrasting for water use efficiency (WUE). We observed that these lines also contrasted for root hydraulic conductivity (Lpr) and AQP contribution to Lpr. The line with lower WUE showed significantly higher AQP contribution to Lpr. To investigate AQP isoforms contributing to Lpr, we developed genomic approaches to first identify the entire AQP family in pearl millet and secondly, characterize the plasma membrane intrinsic proteins (PIP) gene expression profile. We identified and annotated 33 AQP genes in pearl millet, among which ten encoded PIP isoforms. PgPIP1-3 and PgPIP1-4 were significantly more expressed in the line showing lower WUE, higher Lpr and higher AQP contribution to Lpr. Overall, our study suggests that the PIP1 AQP family are the main regulators of Lpr in pearl millet and may possibly be associated with mechanisms associated to whole plant water use. This study paves the way for further investigations on AQP functions in pearl millet hydraulics and adaptation to environmental stresses.


Assuntos
Aquaporinas , Pennisetum , Raízes de Plantas/fisiologia , Adaptação Fisiológica , Aquaporinas/genética , Aquaporinas/metabolismo , Genes de Plantas , Genoma de Planta , Pennisetum/genética , Pennisetum/fisiologia , Estresse Fisiológico , Transcriptoma , Água/metabolismo
2.
PLoS One ; 15(9): e0236724, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32956406

RESUMO

Aquaporins (AQP) are a family of plasma membrane proteins responsible for water transport through cell membranes. They are differentially expressed in different parts of the alimentary canal of insects where they regulate water transport. These proteins have been studied in detail in some insects, but few data are available for aquaporins of the honey bee, Apis mellifera. We used quantitative PCR to study the expression of six putative aquaporin genes in forager honey bees. We found differential expression of all putative AQP genes in crop, midgut, ileum, rectum and Malpighian tubules. We found the entomoglyceroporin Am_Eglp 1 expressed at extremely high levels in the midgut. We performed a functional characterization of Am_Eglp 1 using heterologous expression in Xenopus laevis oocyte followed by water uptake assays. Our results confirmed that the Am_Eglp 1 gene encodes a functional water transporter. This study shows that all putative honey bee aquaporin genes have complex expression patterns in the digestive and excretory organs of honey bee workers. Our results suggest that Am_Eglp 1 is the principal water transporter in the midgut of A. mellifera workers.


Assuntos
Aquaporinas/metabolismo , Abelhas/metabolismo , Trato Gastrointestinal/metabolismo , Proteínas de Insetos/metabolismo , Animais , Aquaporinas/genética , Abelhas/genética , Genes de Insetos , Proteínas de Insetos/genética , Oócitos , Xenopus laevis
3.
PLoS One ; 15(8): e0234892, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32817668

RESUMO

The mosquito Aedes aegypti vectors the arboviral diseases yellow fever, dengue, Zika and chikungunya. Larvae are usually found developing in freshwater; however, more recently they have been increasingly found in brackish water, potential habitats which are traditionally ignored by mosquito control programs. Aedes aegypti larvae are osmo-regulators maintaining their hemolymph osmolarity in a range of ~ 250 to 300 mOsmol l-1. In freshwater, the larvae must excrete excess water while conserving ions while in brackish water, they must alleviate an accumulation of salts. The compensatory physiological mechanisms must involve the transport of ions and water but little is known about the water transport mechanisms in the osmoregulatory organs of these larvae. Water traverses cellular membranes predominantly through transmembrane proteins named aquaporins (AQPs) and Aedes aegypti possesses 6 AQP homologues (AaAQP1 to 6). The objective of this study was to determine if larvae that develop in freshwater or brackish water have differential aquaporin expression in osmoregulatory organs, which could inform us about the relative importance and function of aquaporins to mosquito survival under these different osmotic conditions. We found that AaAQP transcript abundance was similar in organs of freshwater and brackish water mosquito larvae. Furthermore, in the Malpighian tubules and hindgut AaAQP protein abundance was unaffected by the rearing conditions, but in the gastric caeca the protein level of one aquaporin, AaAQP1 was elevated in brackish water. We found that AaAQP1 was expressed apically while AaAQP4 and AaAQP5 were found to be apical and/or basal in the epithelia of osmoregulatory organs. Overall, the results suggest that aquaporin expression in the osmoregulatory organs is mostly consistent between larvae that are developing in freshwater and brackish water. This suggests that aquaporins may not have major roles in adapting to longterm survival in brackish water or that aquaporin function may be regulated by other mechanisms like post-translational modifications.


Assuntos
Aedes/genética , Aquaporinas/genética , Osmorregulação/genética , Aedes/fisiologia , Animais , Aquaporinas/metabolismo , Infecções por Arbovirus , Transporte Biológico , Ecossistema , Água Doce , Expressão Gênica/genética , Regulação da Expressão Gênica/genética , Hemolinfa/metabolismo , Proteínas de Insetos/metabolismo , Larva/genética , Larva/metabolismo , Osmorregulação/fisiologia , Osmose , Águas Salinas , Salinidade , Água/metabolismo
4.
Ecotoxicol Environ Saf ; 205: 111131, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32827964

RESUMO

Arsenic (As) is one of the most toxic contaminants to food crops, and as such, decreasing crops uptake and accumulation of As cannot be overemphasized. Here, we characterized a functional wheat NIP2;1 homolog of the As transporter, TaNIP2;1. TaNIP2;1 expression was suppressed by arsenite (As(III)) in wheat. Ectopic expression of TaNIP2;1 in the Δfps1 yeast mutant enhanced yeast sensitivity towards As(III). Conversely, the elevated expression of TaNIP2;1 in Δacr3 mutants decreased yeast sensitivity to arsenate (As(V)), demonstrating that TaNIP2;1 showed both influx and efflux transport activities for As(III) in yeasts. This is further supported by increased As concentration in the yeast cells that overproduce TaNIP2;1 in Δfps1, while As concentration decreased in Δacr3. Furthermore, ectopic expression of TaNIP2;1 in Arabidopsis confirmed that TaNIP2;1 can transport As into plants, as supported by increased sensitivity to and uptake of As(III). No change in plant sensitivity was found to Cu(II), Cd(II), Zn(II) or Ni(II), indicating that transport activity of TaNIP2;1 is specific for As(III). Taken together, our data show that TaNIP2;1 may be involved in As(III) transportation in plants. This finding reveals a functional gene that can be manipulated to reduce As content in wheat.


Assuntos
Aquagliceroporinas/genética , Arabidopsis/efeitos dos fármacos , Arsenitos/toxicidade , Expressão Ectópica do Gene/efeitos dos fármacos , Poluentes do Solo/toxicidade , Triticum/efeitos dos fármacos , Adaptação Fisiológica/efeitos dos fármacos , Aquagliceroporinas/metabolismo , Aquaporinas/genética , Aquaporinas/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arsenitos/metabolismo , Bioacumulação , Transporte Biológico , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/metabolismo , Poluentes do Solo/metabolismo , Triticum/genética , Triticum/metabolismo
5.
Gene ; 761: 145043, 2020 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-32777530

RESUMO

Tonoplast Intrinsic Proteins (TIPs) constitute a significant class of the aquaporins. The TIPs control water trade among cytosolic and vacuolar compartments and can also transport glycerol, ammonia, urea, hydrogen peroxide, metals/metalloids, and so forth. Additionally, TIPs are engaged with different abiotic stress responses and developmental processes like leaf expansion, root elongation and seed germination. In this study, ten TIP genes in the rice genome were identified from Oryza sativa ssp indica. Among these, representative groups of TIP genes were cloned and sequenced whilst some TIP sequences showed stop codons in the coding region. The secondary structure analysis represented six conserved transmembrane helices along with the inter-helical regions having conserved motifs. The representative three-dimensional tetrameric design of protein sequence of TIP1;1 displayed key features like NPA motifs, aromatic/arginine (ar/R) selectivity filters, and Froger's residues. The vacuolar localization, transmembrane topological properties, and conserved motif analysis of the cloned genes altogether supported their identity as TIPs. An unrooted phylogenetic tree delineated the relatedness of TIPs from Oryza with different species and bunched them into five clades. The promoter analysis uncovered key regulons associated with administering abiotic stress responses. Gene expression studies showed thatTIPsare differentially regulated under salt and drought stress at various time points in shoots and roots of rice. Also, the pattern of expression was found to be significantly variable in five different rice tissues. The heat-map based tissue and stress- specific expression analysis supported the experimental findings. In conclusion, the identification and transcript-level expression studies of TIPs significantly contribute towards the comprehension of their utilitarian significance in the abiotic stress response.


Assuntos
Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Oryza/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Aquaporinas/genética , Aquaporinas/metabolismo , Clonagem Molecular/métodos , Secas , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Oryza/metabolismo , Filogenia , Folhas de Planta/metabolismo , Cloreto de Sódio/metabolismo , Estresse Fisiológico/genética , Vacúolos/genética , Água/metabolismo
6.
PLoS One ; 15(7): e0236511, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32722717

RESUMO

The severe side effects of chemosynthetic anti-diarrhea drugs have created an interest in low-toxic alternative plant-derived compounds. FengLiao consists of Polygonum hydropiper Linn. and Daphniphyllum calycinum Bench., and is widely used in China to treat diarrhea due to low levels of toxicity. In this study, the effects of FengLiao were analyzed in a castor oil-induced diarrhea model, using the anti-diarrhea drug, loperamide, as the positive control. The effects were evaluated using stool characteristics and the expression levels of various diarrhea-related factors in the jejunum and liver, as well as changes in the microbiota of the jejunum. The symptoms of diarrhea and stool consistency were improved through FengLiao and loperamide treatment. Furthermore, FengLiao down-regulated alpha 1-acid glycoprotein (AGP) and C-reactive protein (CRP) levels, and up-regulated transferrin (TRF) mRNA levels in the liver, and down-regulated Aquaporin 3 (AQP3) and Na+/H+ exchanger isoform 8 (NHE8) expression in the epithelial cells of the jejunum. It also increased the relative abundance of Bifidobacterium, Aerococcus, Corynebacterium_1 and Pseudomonas, and lowered the Firmicutes/Bacteroidetes (F/B) ratio, which maintained the balance between immunity and intestinal health. Taken together, FengLiao alleviated castor oil-induced diarrhea by altering gut microbiota, and levels of jejunum epithelial transport proteins and acute phase proteins.


Assuntos
Proteínas da Fase Aguda/genética , Aquaporinas/genética , Diarreia/tratamento farmacológico , Medicamentos de Ervas Chinesas/farmacologia , Microbioma Gastrointestinal/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Trocadores de Sódio-Hidrogênio/genética , Animais , Óleo de Rícino/toxicidade , Daphniphyllum/química , Diarreia/genética , Diarreia/microbiologia , Medicamentos de Ervas Chinesas/uso terapêutico , Jejuno/efeitos dos fármacos , Jejuno/metabolismo , Jejuno/microbiologia , Camundongos , Polygonum/química
7.
PLoS One ; 15(5): e0233517, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32437405

RESUMO

Vitreo-retinal (VR) surgeries induce conjunctival changes. However, there are no study reports regarding prevalence and severity of dry eye after these surgeries. This study evaluated dry eye outcome after VR surgery. Patients undergoing VR surgery classified as scleral buckle and microincision vitrectomy surgery (n = 44, mean age: 56.09±10.2 years) were recruited. Dry eye evaluation was done before and 8 weeks after surgery (2 weeks after omitting topical eye drops). Conjunctival imprint cytology for goblet cell count and tear Mucin 5AC (MUC5AC) protein estimation was done. Gene expressions of MUC5AC, MUC4, MUC16, Aquaporin 4 (AQP4) and AQP5 were analyzed in the conjunctival imprint cells by qPCR. None of the patients exhibited clinical signs of dry eye after VR surgery. But the conjunctival goblet cell density (GCD) was significantly lowered post-VR surgery (63% cases, **p = 0.012) with no alterations in the tear MUC5AC protein. Post-VR surgery, the conjunctival cell gene expression of MUC4, MUC16 and AQP4 were significantly increased (*p = 0.025, *p = 0.05 and *p = 0.02 respectively) and AQP5 was significantly lowered (*p = 0.037), with no change in MUC5AC expression. Tear cytokines were significantly increased post-VR surgery (anti-inflammatory: IL1RA, IL4, IL5, IL9, FGF; PDGFbb and pro-inflammatory: IL2, IL6, IL15, GMCSF and IFNg). Though clinical signs of dry eye were not observed after VR surgery, ocular surface changes in the form of reduced GCD, altered MUC5AC, MUC4, MUC16, AQP4, AQP5 and cytokines are suggestive of dry eye outcome at the molecular level especially inpatients aged above 51 years, especially female gender and those who are diabetic.


Assuntos
Aquaporinas/genética , Síndromes do Olho Seco/cirurgia , Mucinas/genética , Aquaporina 4/análise , Aquaporina 4/genética , Aquaporina 5/análise , Aquaporina 5/genética , Aquaporinas/análise , Antígeno Ca-125/análise , Antígeno Ca-125/genética , Túnica Conjuntiva/química , Túnica Conjuntiva/metabolismo , Túnica Conjuntiva/patologia , Síndromes do Olho Seco/genética , Síndromes do Olho Seco/patologia , Feminino , Expressão Gênica , Humanos , Masculino , Proteínas de Membrana/análise , Proteínas de Membrana/genética , Pessoa de Meia-Idade , Mucina-5AC/análise , Mucina-5AC/genética , Mucina-4/análise , Mucina-4/genética , Mucinas/análise , Lágrimas/química , Lágrimas/metabolismo
8.
J Anim Sci ; 98(4)2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32227169

RESUMO

The objectives were to determine the effects of forage level and grain processing on whole-body urea kinetics, N balance, serosal-to-mucosal urea flux (Jsm-urea), and messenger ribonucleic acid (mRNA) abundance of urea transporter-B (UT-B; SLC14a1) and aquaporins (AQP) in ovine ruminal, duodenal, and cecal epithelia. Thirty-two wether lambs were blocked by body weight into groups of four and assigned to one of four diets (n = 8) in a 2 × 2 factorial design. Dietary factors were forage level (30% [LF] vs. 70% [HF]) and corn grain processing (whole-shelled [WS] vs. steam-flaked [SF]). Four blocks of lambs (n = 4) were used to determine urea kinetics and N balance using 4-d [15N15N]-urea infusions with concurrent fecal and urine collections. Lambs were killed after 23 d of dietary adaptation. Ruminal, duodenal, and cecal epithelia were collected to determine Jsm-urea and mRNA abundance of UT-B and AQP. Lambs fed LF had greater intakes of dry matter (DMI; 1.20 vs. 0.86 kg/d) and N (NI; 20.1 vs. 15.0 g/d) than those fed HF (P < 0.01). Lambs fed SF had greater DMI (1.20 vs. 0.86 kg/d) and NI (20.6 vs. 14.5 g/d) than those fed WS (P < 0.01). As a percentage of NI, total N excretion was greater in lambs fed HF compared with those fed LF (103% vs. 63.0%; P < 0.01) and was also greater in lambs fed WS compared with those fed SF (93.6% vs. 72.1%; P = 0.02). Retained N (% of NI) was greater in lambs fed LF compared with those fed HF (37.0% vs. -2.55%; P < 0.01). Lambs fed SF had a greater (P = 0.02) retained N (% of NI; 28.0% vs. 6.50%) compared with those fed WS. Endogenous urea production (UER) tended (P = 0.09) to be greater in lambs fed HF compared with those fed LF. As a proportion of UER, lambs fed HF had a greater urinary urea-N loss (0.38 vs. 0.22) and lower urea-N transferred to the gastrointestinal tract (GIT; 0.62 vs. 0.78) or urea-N used for anabolism (as a proportion of urea-N transferred to the GIT; 0.12 vs. 0.26) compared with lambs fed LF (P < 0.01). Ruminal Jsm-urea was unaffected by diet. Duodenal Jsm-urea was greater (P < 0.01) in lambs fed HF compared with LF (77.5 vs. 57.2 nmol/[cm2 × h]). Lambs fed LF had greater (P = 0.03) mRNA expression of AQP3 in ruminal epithelia and tended (P = 0.06) to have greater mRNA expression of AQP3 in duodenal epithelia compared with lambs fed HF. Expression of UT-B mRNA was unaffected by diet. Our results showed that feeding more ruminally available energy improved N utilization, partly through a greater proportion of UER being transferred to the GIT and being used for anabolic purposes.


Assuntos
Ingestão de Energia , Nitrogênio/metabolismo , Ovinos/fisiologia , Ureia/metabolismo , Ração Animal/análise , Animais , Aquaporinas/genética , Dieta/veterinária , Grão Comestível , Epitélio/metabolismo , Fezes/química , Fermentação , Trato Gastrointestinal/metabolismo , Cinética , Masculino , Proteínas de Membrana Transportadoras/genética , Distribuição Aleatória , Ovinos/genética , Zea mays
9.
World J Microbiol Biotechnol ; 36(3): 49, 2020 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-32157439

RESUMO

Glycerol is a by-product of biodiesel, and it has a great application prospect to be transformed to synthesize high value-added compounds. Pseudomonas chlororaphis GP72 isolated from the green pepper rhizosphere is a plant growth promoting rhizobacteria that can utilize amount of glycerol to synthesize phenazine-1-carboxylic acid (PCA). PCA has been commercially registered as "Shenqinmycin" in China due to its characteristics of preventing pepper blight and rice sheath blight. The aim of this study was to engineer glycerol utilization pathway in P. chlororaphis GP72. First, the two genes glpF and glpK from the glycerol metabolism pathway were overexpressed in GP72ANO separately. Then, the two genes were co-expressed in GP72ANO, improving PCA production from 729.4 mg/L to 993.4 mg/L at 36 h. Moreover, the shunt pathway was blocked to enhance glycerol utilization, resulting in 1493.3 mg/L PCA production. Additionally, we confirmed the inhibition of glpR on glycerol metabolism pathway in P. chlororaphis GP72. This study provides a good example for improving the utilization of glycerol to synthesize high value-added compounds in Pseudomonas.


Assuntos
Glicerol/metabolismo , Engenharia Metabólica/métodos , Pseudomonas chlororaphis/genética , Pseudomonas chlororaphis/metabolismo , Aquaporinas/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Capsicum/microbiologia , China , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Regulação Bacteriana da Expressão Gênica , Técnicas de Inativação de Genes , Glicerolfosfato Desidrogenase/genética , Redes e Vias Metabólicas/genética , Fenazinas/metabolismo , Proteínas Repressoras/genética , Rizosfera
10.
Ecotoxicol Environ Saf ; 195: 110471, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32203773

RESUMO

Rice is the most consumed food crop and essential determinant in global food security program. Currently, arsenic (As) accumulation in rice is a critical concern in terms of both crop productivity and grain quality; therefore, it is an urgent need to reduce As accumulation. Here, we selected a glutaredoxin (OsGrx_C7) gene that plays an essential role in AsIII tolerance in rice. To explore the mechanism, we raised OsGrx_C7 overexpression (OE) rice lines, which showed improved plant AsIII tolerance and lowered its accumulation in grains. Arsenic accumulation in husk, unpolished, and polished rice reduced by ca. 65%, 67%, and 85%, respectively, in OE lines, compared to wild-type (WT) plants. To know the rationale, expression of AsIII transporters (aquaporins) in root and shoot tissues were examined, and revealed that OsGrx_C7 regulates the expression of these genes, which ultimately reduces root to shoot AsIII translocation. Additionally, OsGrx_C7 improves root growth by regulating the expression of oxidative stress-induced root expansion related genes, promote root growth and plant health. Overall, current study suggested that AsIII induced OsGrx_C7 markedly enhanced tolerance to AsIII with reduced accumulation in grains by regulating root expansion and controlling root to shoot As transport by altered expression of AsIII aquaporins.


Assuntos
Aquaporinas/genética , Glutarredoxinas/genética , Oryza/genética , Aquaporinas/metabolismo , Arsênico/farmacocinética , Arsênico/toxicidade , Regulação da Expressão Gênica de Plantas , Glutarredoxinas/metabolismo , Oryza/efeitos dos fármacos , Oryza/metabolismo , Raízes de Plantas/metabolismo
11.
PLoS One ; 15(3): e0229245, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32130259

RESUMO

Aquaporins (AQPs) are membrane channels that provide for transport of water and other small molecules across the lipid bilayer of cells. Their function is essential for physiologic processes such as cell volume regulation, chondrocyte hypertrophy during appendicular skeletal growth, water reabsorption in the kidney tubules, and water excretion by the salivary glands. The ten AQP isoforms show tissue specificity and are involved in different pathologies and inflammatory diseases. This study addresses the hypothesis that arthritis, periodontitis, and temporomandibular joint disorders (TMDs) can be influenced by variation in the AQP genes at 12q13.12 locus. Salivary samples of 688 individuals were obtained from the Dental Registry and DNA Repository project at the University of Pittsburgh. Ten polymorphisms in four AQP genes (AQP1, 2, 5, and 6) were genotyped and correlated to disease status as reported by patients. Associations were found between the single nucleotide polymorphism (SNP) rs467323 in AQP2 and TMD in both genotypic (p = 0.03) and recessive (p = 0.02) models, and between rs1996315 in AQP6 and periodontitis (p = 0.05). Combined analysis of TMD and periodontitis showed an association with rs3741559 in AQP2 (p = 0.02). When conducting haplotype analysis of rs467323 and rs10875989 in AQP2, the haplotype CT showed an association with the TMD phenotype (p = 0.007). Our results suggest that the aquaporin locus at 12q13.12 may contribute to the pathogenesis of inflammatory conditions such as periodontitis and TMD. Thus, oral and skeletal health are correlated and potential susceptibility screening strategies may be developed.


Assuntos
Aquaporinas/genética , Loci Gênicos/genética , Predisposição Genética para Doença/genética , Periodontite/complicações , Transtornos da Articulação Temporomandibular/complicações , Transtornos da Articulação Temporomandibular/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Haplótipos , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Adulto Jovem
12.
Nat Biotechnol ; 38(5): 600-608, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32066956

RESUMO

The western corn rootworm (WCR) decimates maize crops worldwide. One potential way to control this pest is treatment with entomopathogenic nematodes (EPNs) that harbor bacterial symbionts that are pathogenic to insects. However, WCR larvae sequester benzoxazinoid secondary metabolites that are produced by maize and use them to increase their resistance to the nematodes and their symbionts. Here we report that experimental evolution and selection for bacterial symbionts that are resistant to benzoxazinoids improve the ability of a nematode-symbiont pair to kill WCR larvae. We isolated five Photorhabdus symbionts from different nematodes and increased their benzoxazinoid resistance through experimental evolution. Benzoxazinoid resistance evolved through multiple mechanisms, including a mutation in the aquaporin-like channel gene aqpZ. We reintroduced benzoxazinoid-resistant Photorhabdus strains into their original EPN hosts and identified one nematode-symbiont pair that was able to kill benzoxazinoid-sequestering WCR larvae more efficiently. Our results suggest that modification of bacterial symbionts might provide a generalizable strategy to improve biocontrol of agricultural pests.


Assuntos
Aquaporinas/genética , Benzoxazinas/farmacologia , Farmacorresistência Bacteriana , Nematoides/microbiologia , Photorhabdus/fisiologia , Zea mays/crescimento & desenvolvimento , Animais , Proteínas de Bactérias/genética , Evolução Molecular , Engenharia Genética , Mutação , Nematoides/patogenicidade , Controle Biológico de Vetores , Photorhabdus/efeitos dos fármacos , Photorhabdus/genética , Doenças das Plantas/prevenção & controle , Zea mays/parasitologia
13.
Int J Mol Sci ; 21(3)2020 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-32046116

RESUMO

BACKGROUND: Ion channels play a crucial role in many physiological processes. Several subtypes are expressed in the endometrium. Endometriosis is strictly correlated to estrogens and it is evident that expression and functionality of different ion channels are estrogen-dependent, fluctuating between the menstrual phases. However, their relationship with endometriosis is still unclear. OBJECTIVE: To summarize the available literature data about the role of ion channels in the etiopathogenesis of endometriosis. METHODS: A search on PubMed and Medline databases was performed from inception to November 2019. RESULTS: Cystic fibrosis transmembrane conductance regulator (CFTR), transient receptor potentials (TRPs), aquaporins (AQPs), and chloride channel (ClC)-3 expression and activity were analyzed. CFTR expression changed during the menstrual phases and was enhanced in endometriosis samples; its overexpression promoted endometrial cell proliferation, migration, and invasion throughout nuclear factor kappa-light-chain-enhancer of activated B cells-urokinase plasminogen activator receptor (NFκB-uPAR) signaling pathway. No connection between TRPs and the pathogenesis of endometriosis was found. AQP5 activity was estrogen-increased and, through phosphatidylinositol-3-kinase and protein kinase B (PI3K/AKT), helped in vivo implantation of ectopic endometrium. In vitro, AQP9 participated in extracellular signal-regulated kinases/p38 mitogen-activated protein kinase (ERK/p38 MAPK) pathway and helped migration and invasion stimulating matrix metalloproteinase (MMP)2 and MMP9. ClC-3 was also overexpressed in ectopic endometrium and upregulated MMP9. CONCLUSION: Available evidence suggests a pivotal role of CFTR, AQPs, and ClC-3 in endometriosis etiopathogenesis. However, data obtained are not sufficient to establish a direct role of ion channels in the etiology of the disease. Further studies are needed to clarify this relationship.


Assuntos
Aquaporinas/metabolismo , Canais de Cloreto/metabolismo , Endometriose/metabolismo , Animais , Aquaporinas/genética , Canais de Cloreto/genética , Endometriose/etiologia , Feminino , Humanos , Transdução de Sinais
14.
Proc Natl Acad Sci U S A ; 117(3): 1779-1787, 2020 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-31907321

RESUMO

Insects are highly successful, in part through an excellent ability to osmoregulate. The renal (Malpighian) tubules can secrete fluid faster on a per-cell basis than any other epithelium, but the route for these remarkable water fluxes has not been established. In Drosophila melanogaster, we show that 4 genes of the major intrinsic protein family are expressed at a very high level in the fly renal tissue: the aquaporins (AQPs) Drip and Prip and the aquaglyceroporins Eglp2 and Eglp4 As predicted from their structure, and by their transport function by expressing these proteins in Xenopus oocytes, Drip, Prip, and Eglp2 show significant and specific water permeability, whereas Eglp2 and Eglp4 show very high permeability to glycerol and urea. Knockdowns of any of these genes result in impaired hormone-induced fluid secretion. The Drosophila tubule has 2 main secretory cell types: active cation-transporting principal cells, wherein the aquaglyceroporins localize to opposite plasma membranes, and small stellate cells, the site of the chloride shunt conductance, with these AQPs localizing to opposite plasma membranes. This suggests a model in which osmotically obliged water flows through the stellate cells. Consistent with this model, fluorescently labeled dextran, an in vivo marker of membrane water permeability, is trapped in the basal infoldings of the stellate cells after kinin diuretic peptide stimulation, confirming that these cells provide the major route for transepithelial water flux. The spatial segregation of these components of epithelial water transport may help to explain the unique success of the higher insects in regulating their internal environments.


Assuntos
Transporte Biológico/fisiologia , Drosophila melanogaster/fisiologia , Túbulos Renais/metabolismo , Água/metabolismo , Animais , Aquagliceroporinas/genética , Aquagliceroporinas/metabolismo , Aquaporinas/genética , Aquaporinas/metabolismo , Permeabilidade da Membrana Celular , Cloretos/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Feminino , Técnicas de Silenciamento de Genes , Túbulos Renais/citologia , Masculino , Túbulos de Malpighi/metabolismo , Modelos Animais , Oócitos/metabolismo , Osmorregulação , Xenopus
15.
Int J Mol Sci ; 21(2)2020 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-31963923

RESUMO

Plant Nodulin 26-like Intrinsic Proteins (NIPs) are multifunctional membrane channels of the Major Intrinsic Protein (MIP) family. Unlike other homologs, they have low intrinsic water permeability. NIPs possess diverse substrate selectivity, ranging from water to glycerol and to other small solutes, depending on the group-specific amino acid composition at aromatic/Arg (ar/R) constriction. We cloned three NIPs (NIP1;1, NIP5;1, and NIP6;1) from grapevine (cv. Touriga Nacional). Their expression in the membrane of aqy-null Saccharomyces cerevisiae enabled their functional characterization for water and glycerol transport through stopped-flow spectroscopy. VvTnNIP1;1 demonstrated high water as well as glycerol permeability, whereas VvTnNIP6;1 was impermeable to water but presented high glycerol permeability. Their transport activities were declined by cytosolic acidification, implying that internal-pH can regulate NIPs gating. Furthermore, an extension of C-terminal in VvTnNIP6;1M homolog, led to improved channel activity, suggesting that NIPs gating is putatively regulated by C-terminal. Yeast growth assays in the presence of diverse substrates suggest that the transmembrane flux of metalloids (As, B, and Se) and the heavy metal (Cd) are facilitated through grapevine NIPs. This is the first molecular and functional characterization of grapevine NIPs, providing crucial insights into understanding their role for uptake and translocation of small solutes, and extrusion of toxic compounds in grapevine.


Assuntos
Aquaporinas/genética , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Saccharomyces cerevisiae/crescimento & desenvolvimento , Vitis/metabolismo , Clonagem Molecular , Glicerol/metabolismo , Mutação com Perda de Função , Proteínas de Membrana/química , Metaloides/química , Família Multigênica , Permeabilidade , Proteínas de Plantas/química , Domínios Proteicos , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Vitis/genética , Água/metabolismo
16.
Artigo em Inglês | MEDLINE | ID: mdl-31945668

RESUMO

Proper storage prolongs peony market supply. Here, we determined the changes in fresh weight and expression of four aquaporin genes under dry storage (DS) and wet storage (WS). It has showed that after harvesting, the fresh weight change was accompanied with flower opening. After both short- and long-term of storage, the water uptake efficiency in DS group was greater during the first few vase days, providing a direct material basis of DS improved vase quality. The gene expression results showed that PlPIP1;3 and PlTIP2;1 were mainly expressed in petals, whereas PlNIP1;2-like and PlSIP2;1 were mainly expressed in the green tissues. In addition, the expression of PlTIP2;1 in the petals was consistent with the flower opening process, indicating that it may play a major role in facilitating water uptake. During cold storage, the expression of PlPIP1;3 and PlTIP2;1 was higher or more rapidly induced in the DS group, and thus we deduced that they play important roles in improving the vase quality of DS. Furthermore, the expression of PlNIP1;2-like in the early stage of the DS group was more stable than in WS, which may also be partially responsible for the vase quality improvement. In contrast, PlSIP2;1 may not be involved, since no significant change was observed between the DS and WS group. In short, the expression of PlPIP1;3 and PlTIP2;1 in the DS group during storage may improve water uptake efficiency during the vase period and then improving the vase quality of cut peony.


Assuntos
Agricultura/métodos , Aquaporinas , Flores , Paeonia , Água , Aquaporinas/genética , Aquaporinas/metabolismo , Transporte Biológico , Regulação da Expressão Gênica de Plantas , Paeonia/metabolismo , Água/metabolismo
17.
Biomed Res Int ; 2020: 2895795, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31998785

RESUMO

In higher plants, aquaporin proteins (AQPs) play important roles in the uptake of water across cell membranes. However, their functions in halophytes are still largely unknown. In this work, we isolated, cloned, and identified KvTIP3, a tonoplast intrinsic protein gene from Kosteletzkya virginica. Bioinformatic analyses demonstrated that KvTIP3 encoded a tonoplast protein with the common properties of AQPs. Further multiple sequence alignment and phylogenetic analyses showed that KvTIP3 shared 65%-82% homology with other AQPs from Arabidopsis, cotton, polar, and cocoa. Quantitative real-time PCR (qPCR) analyses revealed that KvTIP3 was ubiquitously expressed in various tissues such as leaves, stems, and roots, with a predominant expression in roots. In addition, KvTIP3 transcript was strongly induced by NaCl, low temperature, and ABA in K. virginica. Our findings suggest that KvTIP3 encodes a new AQP possibly involved in multiple abiotic stress responses in K. virginica, and KvTIP3 could be used as a potential candidate gene for the improvement of plants resistant to various abiotic stresses.


Assuntos
Aquaporinas , Regulação da Expressão Gênica de Plantas/fisiologia , Malvaceae , Proteínas de Plantas , Estresse Fisiológico/fisiologia , Aquaporinas/biossíntese , Aquaporinas/genética , Malvaceae/genética , Malvaceae/metabolismo , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética
18.
Arch Biochem Biophys ; 679: 108222, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31816311

RESUMO

Aquaporins (AQPs) are a family of transmembrane channel proteins responsible for the transport of water and small uncharged molecules. Thirteen distinct isoforms of AQPs have been identified in mammals (AQP0-12). Throughout the male reproductive tract, AQPs greatly enhance water transport across all biological barriers, providing a constant and expeditious movement of water and playing an active role in the regulation of water and ion homeostasis. This regulation of fluids is particularly important in the male reproductive tract, where proper fluid composition is directly linked with a healthy and competent spermatozoa production. For instance, in the testis, fluid regulation is essential for spermatogenesis and posterior spermatozoa transport into the epididymal ducts, while maintaining proper ionic conditions for their maturation and storage. Alterations in the expression pattern of AQPs or their dysfunction is linked with male subfertility/infertility. Thus, AQPs are important for male reproductive health. In this review, we will discuss the most recent data on the expression and function of the AQPs isoforms in the human, mouse and rat male reproductive tract. In addition, the regulation of AQPs expression and dysfunction linked with male infertility will be discussed.


Assuntos
Aquaporinas/genética , Aquaporinas/metabolismo , Fertilidade/genética , Regulação da Expressão Gênica , Infertilidade/genética , Infertilidade/metabolismo , Animais , Humanos , Infertilidade/fisiopatologia , Masculino , Reprodução
19.
Physiol Plant ; 168(1): 118-132, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31090074

RESUMO

Mineral nutrient supply can affect the hydraulic property of roots. The aim of the present work on sheepgrass (Leymus chinensis L.) plants was to test whether any changes in root hydraulic conductivity (Lp; exudation analyses) in response to a growth-limiting supply of phosphate (P) are accompanied by changes in (1) cell Lp via measuring the cell pressure, (2) the aquaporin (AQP) gene expression by performing qPCR and (3) the formation of apoplastic barriers, by analyzing suberin lamella and Casparian bands via cross-sectional analyses in roots. Plants were grown hydroponically on complete nutrient solution, containing 250 µM P, until they were 31-36 days old, and then kept for 2-3 weeks on either complete solution, or transferred on solution containing 2.5 µM (low-P) or no added P (no-P). Phosphate treatments caused significant decreases in root and cell-Lp and AQP gene expression, while the formation of apoplastic barriers increased, particularly in lateral roots. Experiments using the AQP inhibitor mercury (Hg) suggested that a significant portion of radial root water uptake in sheepgrass occurs along a path involving AQPs, and that the Lp of this path is reduced under low- and no-P. It is concluded that a growth-limiting supply of phosphate causes parallel changes in (1) cell Lp and aquaporin gene expression (decrease) and (2) apoplastic barrier formation (increase), and that the two may combine to reduce root Lp. The reduction in root Lp, in turn, facilitates an increased root-to-shoot surface area ratio, which allocates resources to the root, sourcing the limiting nutrient.


Assuntos
Aquaporinas/genética , Fosfatos , Proteínas de Plantas/genética , Raízes de Plantas/fisiologia , Poaceae/fisiologia , Transporte Biológico , Estudos Transversais , Regulação da Expressão Gênica de Plantas , Exsudatos de Plantas , Poaceae/genética , Água
20.
Artigo em Inglês | MEDLINE | ID: mdl-31841952

RESUMO

The integral parts of the cell membranes are the functional proteins, which are crucial for cell life. Among them, proton-pumping ATPase and aquaporins appear to be of particular importance. There is some knowledge about the effect of the temperature during plant growth, including stress-inducing temperatures, on the accumulation of the membrane proteins: plasma membrane H+-ATPase and aquaporins, but not much is known about the effect of the phytohormones (i.e. brassinosteroids (BR)) on control of accumulation of these proteins. The aim of our study was to answer the question of how a BR deficit and disturbances in the BR perception/signalling affect the accumulation of plasma membrane H+-ATPase (PM H+-ATPase), the aquaporin HvPIP1 transcript and protein in barley growing at 20 °C and during its acclimation at 5 °C and 27 °C. For the studies, the BR-deficient mutant 522DK (derived from the wild-type Delisa), the BR-deficient mutant BW084 and the BR-signalling mutant BW312 and their wild-type Bowman were used. Generally, temperature of growth was significant factor influencing on the level of the accumulation of the H+-ATPase and HvPIP1 transcript and the PM H+-ATPase and HvPIP1 protein in barley leaves. The level of the accumulation of the HvPIP1 transcript decreased at 5 °C (compared to 20 °C), but was higher at 27 °C than at 20 °C in the analyzed cultivars. In both cultivars the protein HvPIP1 was accumulated in the highest amounts at 27 °C. On the other hand, the barley mutants with a BR deficiency or with BR signalling disturbances were characterised by an altered accumulation level of PM H+-ATPase, the aquaporin HvPIP1 transcript and protein (compared to the wild types), which may suggest the involvement of brassinosteroids in regulating PM H+-ATPase and aquaporin HvPIP1 at the transcriptional and translational levels.


Assuntos
Adenosina Trifosfatases/genética , Aquaporinas/genética , Hordeum/fisiologia , Proteínas de Plantas/genética , Aclimatação , Adenosina Trifosfatases/metabolismo , Aquaporinas/metabolismo , Brassinosteroides/metabolismo , Temperatura Baixa , Hordeum/genética , Temperatura Alta , Mutação , Proteínas de Plantas/metabolismo
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