Effects of Bicarbonate Stress on Serum Ions and Gill Transporters in Alkali and Freshwater Forms of Amur Ide (Leuciscus waleckii)

The Amur ide (Leuciscus waleckii) is a fish in the Cyprinidae family. Compared with other Amur ide living in freshwater ecosystems, the Amur ide population in Lake Dali Nor of China is famous for its high tolerance to the alkaline conditions of 54 mM (pH 9.6). Yet, surprisingly, the ionoregulatory mechanism responsible for this remarkable alkaline adaptation remains unclear. Therefore, this study sought to investigate how bicarbonate affects the acid-base balancing and ionoregulatory responses of this animal. Here, using a comparative approach, the alkali form of Amur ide and its ancestral freshwater form living in other freshwater basins were each exposed to 50 mM (pH 9.59 ± 0.09), a level close to the alkalinity of Lake Dali Nor, and their physiological (AE1) adjustment of ions and acid-base regulation were investigated. This study highlighted differences in blood pH and serum ions (e.g., Na+, K+, Cl−, and Ca2+), Na+/K+ ATPase (NKA) activity and its mRNA level, and mRNA expression of gill transporters (Na+/H+ exchanger member 2 and/or 3, Na+/HCO3- cotransporter (NBC1), Cl−/HCO3- exchanger, Na+/Cl− cotransporter (NCC), Na+/K+/2Cl− (NKCC1), SLC26A5, and SLC26A6) for alkalinity adaptation between the two forms of Amur ide differing in alkalinity tolerance. Specifically, close relationships among the serum Na+ and mRNA levels of NCC, NKCC1, and NHE, and also NKA and NBC1, in addition to serum Cl− and bicarbonate transporters (e.g., SLC26A5 and SLC26A6), characterized the alkali form of Amur ide. We propose that this ecotype can ensure its transepithelial Cl− and Na+ uptake/base secretions are highly functional, by its basolateral NKA with NBC1 and apical ionic transporters, and especially NCC incorporated with other transporters (e.g., SLC26). This suggests an evolved strong ability to maintain an ion osmotic and acid-base balance for more effectively facilitating its adaptability to the high alkaline environment. This study provides new insights into the physiological responses of the alkaline form of the Amur ide fish for adapting to extreme alkaline conditions. This information could be used as a reference to cultivating alkaline-tolerant fish species in abandoned alkaline waters.

Decreased Brain pH and Pathophysiology in Schizophrenia

Postmortem studies reveal that the brain pH in schizophrenia patients is lower than normal. The exact cause of this low pH is unclear, but increased lactate levels due to abnormal energy metabolism appear to be involved. Schizophrenia patients display distinct changes in mitochondria number, morphology, and function, and such changes promote anaerobic glycolysis, elevating lactate levels. pH can affect neuronal activity as H+ binds to numerous proteins in the nervous system and alters the structure and function of the bound proteins. There is growing evidence of pH change associated with cognition, emotion, and psychotic behaviors. Brain has delicate pH regulatory mechanisms to maintain normal pH in neurons/glia and extracellular fluid, and a change in these mechanisms can affect, or be affected by, neuronal activities associated with schizophrenia. In this review, we discuss the current understanding of the cause and effect of decreased brain pH in schizophrenia based on postmortem human brains, animal models, and cellular studies. The topic includes the factors causing decreased brain pH in schizophrenia, mitochondria dysfunction leading to altered energy metabolism, and pH effects on the pathophysiology of schizophrenia. We also review the acid/base transporters regulating pH in the nervous system and discuss the potential contribution of the major transporters, sodium hydrogen exchangers (NHEs), and sodium-coupled bicarbonate transporters (NCBTs), to schizophrenia.

Genes encoding putative bicarbonate transporters as a missing molecular link between molt and mineralization in crustaceans

During their life, crustaceans undergo several molts, which if theoretically compared to the human body would be equivalent to replacing all bones at a single event. Such a dramatic repetitive event is coupled to unique molecular mechanisms of mineralization so far mostly unknown. Unlike human bone mineralized with calcium phosphate, the crustacean exoskeleton is mineralized mainly by calcium carbonate. Crustacean growth thus necessitates well-timed mobilization of bicarbonate to specific extracellular sites of biomineralization at distinct molt cycle stages. Here, by looking at the crayfish Cherax quadricarinatus at different molting stages, we suggest that the mechanisms of bicarbonate ion transport for mineralization in crustaceans involve the SLC4 family of transporters and that these proteins play a key role in the tight coupling between molt cycle events and mineral deposition. This discovery of putative bicarbonate transporters in a pancrustacean with functional genomic evidence from genes encoding the SLC4 family—mostly known for their role in pH control—is discussed in the context of the evolution of calcium carbonate biomineralization.

Regulatory adenylnucleotide-mediated binding of the PII-like protein SbtB to the cyanobacterial bicarbonate transporter SbtA is controlled by the cellular energy state

ABSTRACTCyanobacteria have evolved one of the most powerful CO2 concentrating mechanisms (CCM), supporting high photosynthetic rates with limiting inorganic carbon (Ci), which makes their CCM a desirable system for integration into higher plant chloroplasts to enhance photosynthetic yield. The CCM is driven by active Ci uptake, facilitated by bicarbonate transporters and CO2 pumps, which locally elevates the CO2 concentration and carboxylation rate of the primary CO2 fixing enzyme, Rubisco, inside cytoplasmic micro-compartments (carboxysomes). Ci uptake responds allosterically to Ci supply and light, but the molecular signals and regulators of protein function are unknowns. Functional analyses of sodium-dependent bicarbonate transporters classified as SbtA in E. coli support the hypothesis that SbtA activity is negatively regulated through association with its cognate PII-like SbtB protein. Here, we demonstrate that the association of SbtA with SbtB from two phylogenetically distant species, Cyanobium sp. PCC7001 and Synechococcus elongatus PCC7942, depends on the relative amounts of ATP or cAMP compared to ADP or AMP. Higher ATP over ADP or AMP ratios decreased the formation of SbtA:SbtB complexes, consistent with a sensory response to the cellular adenylate energy charge (AEC=[ATP + 0.5 ADP]/[ATP+ADP+AMP]) and the different binding affinities of these adenylates to SbtB protein trimers. Based on evidence for adenylate ligand-specific conformation changes for the SbtB protein trimer of Cyanobium sp. PCC7001, we propose a role for SbtB as a curfew protein locking SbtA into an inactive state as safe-guard against energetically futile and physiologically disadvantageous activation during prolonged low cellular AEC and photosynthetically unfavourable conditions.

Direct Monitoring of Bicarbonate Transport by Emission Spectroscopy

<p>The transmembrane transport of bicarbonate is a key step in many important biological processes, while problems with bicarbonate transport are at the origin of various diseases. Over the past 10 years, many anionophores that have been developed for the transport of chloride, have also been tested as bicarbonate transporters. However, methodology to directly monitor the kinetics of transport of bicarbonate is lacking, hence indirect methods have been used, which mainly rely on the monitoring of chloride concentrations.</p>Here we present an assay that allows the kinetics of bicarbonate transport into liposomes to be monitored directly, using emission spectroscopy. The assay utilises an encapsulated europium(III) complex, which exhibits a large increase in emission upon binding of bicarbonate. The advantages of this assay over existing methodology are that concentrations of bicarbonate are monitored directly and with a high sensitivity. This allows studies at very low concentrations of anionophores, and for the mechanisms of bicarbonate transport to be unravelled. We have distinguished classical antiport with bicarbonate from mechanisms involving CO₂ diffusion and the dissipation of a pH gradient. Furthermore, the use of a standard fluorescence spectrometer and liposomes with a diameter ~200 nm makes this assay readily and reliably applicable in many laboratories, where it can facilitate the development of bicarbonate transporters for applications in physiological studies or therapies.

Direct Monitoring of Bicarbonate Transport by Emission Spectroscopy

<p>The transmembrane transport of bicarbonate is a key step in many important biological processes, while problems with bicarbonate transport are at the origin of various diseases. Over the past 10 years, many anionophores that have been developed for the transport of chloride, have also been tested as bicarbonate transporters. However, methodology to directly monitor the kinetics of transport of bicarbonate is lacking, hence indirect methods have been used, which mainly rely on the monitoring of chloride concentrations.</p>Here we present an assay that allows the kinetics of bicarbonate transport into liposomes to be monitored directly, using emission spectroscopy. The assay utilises an encapsulated europium(III) complex, which exhibits a large increase in emission upon binding of bicarbonate. The advantages of this assay over existing methodology are that concentrations of bicarbonate are monitored directly and with a high sensitivity. This allows studies at very low concentrations of anionophores, and for the mechanisms of bicarbonate transport to be unravelled. We have distinguished classical antiport with bicarbonate from mechanisms involving CO₂ diffusion and the dissipation of a pH gradient. Furthermore, the use of a standard fluorescence spectrometer and liposomes with a diameter ~200 nm makes this assay readily and reliably applicable in many laboratories, where it can facilitate the development of bicarbonate transporters for applications in physiological studies or therapies.

Direct Monitoring of Bicarbonate Transport by Emission Spectroscopy

<p>The transmembrane transport of bicarbonate is a key step in many important biological processes, while problems with bicarbonate transport are at the origin of various diseases. Over the past 10 years, many anionophores that have been developed for the transport of chloride, have also been tested as bicarbonate transporters. However, methodology to directly monitor the kinetics of transport of bicarbonate is lacking, hence indirect methods have been used, which mainly rely on the monitoring of chloride concentrations.</p>Here we present an assay that allows the kinetics of bicarbonate transport into liposomes to be monitored directly, using emission spectroscopy. The assay utilises an encapsulated europium(III) complex, which exhibits a large increase in emission upon binding of bicarbonate. The advantages of this assay over existing methodology are that concentrations of bicarbonate are monitored directly and with a high sensitivity. This allows studies at very low concentrations of anionophores, and for the mechanisms of bicarbonate transport to be unravelled. We have distinguished classical antiport with bicarbonate from mechanisms involving CO₂ diffusion and the dissipation of a pH gradient. Furthermore, the use of a standard fluorescence spectrometer and liposomes with a diameter ~200 nm makes this assay readily and reliably applicable in many laboratories, where it can facilitate the development of bicarbonate transporters for applications in physiological studies or therapies.

Tetragonal crystal form of the cyanobacterial bicarbonate-transporter regulator SbtB from Synechocystis sp. PCC 6803

The PII-like protein SbtB has been identified as a regulator of SbtA, which is one of the key bicarbonate transporters in cyanobacteria. While SbtB from Synechocystis sp. PCC 6803 has previously been shown to be a trimer, a new crystal form is reported here which crystallizes in what is thought to be a non-native tetramer in the crystal, with the C-terminus in an extended conformation. The crystal structure shows the formation of an intermolecular disulfide bond at Cys94 between SbtB monomers, which may stabilize this conformation in the crystal. This motivates the need for future studies to investigate the potential role that the oxidation and reduction of these cysteines may play in the activation and/or function of SbtB.

Direct Monitoring of Bicarbonate Transport by Emission Spectroscopy

<p>The transmembrane transport of bicarbonate is a key step in many important biological processes, while problems with bicarbonate transport are at the origin of various diseases. Over the past 10 years, many anionophores that have been developed for the transport of chloride, have also been tested as bicarbonate transporters. However, methodology to directly monitor the kinetics of transport of bicarbonate is lacking, hence indirect methods have been used, which mainly rely on the monitoring of chloride concentrations.</p>Here we present an assay that allows the kinetics of bicarbonate transport into liposomes to be monitored directly, using emission spectroscopy. The assay utilises an encapsulated europium(III) complex, which exhibits a large increase in emission upon binding of bicarbonate. The advantages of this assay over existing methodology are that concentrations of bicarbonate are monitored directly and with a high sensitivity. This allows studies at very low concentrations of anionophores, and for the mechanisms of bicarbonate transport to be unravelled. We have distinguished classical antiport with bicarbonate from mechanisms involving CO₂ diffusion and the dissipation of a pH gradient. Furthermore, the use of a standard fluorescence spectrometer and liposomes with a diameter ~200 nm makes this assay readily and reliably applicable in many laboratories, where it can facilitate the development of bicarbonate transporters for applications in physiological studies or therapies.