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1. Molecular and biochemical characterization of the bicarbonate sensing soluble adenylyl cyclase from a bony fish, the rainbow trout Oncorhynchus mykiss

   Salmerón C, Harter TS (April 2021, Interface focus)

   Soluble adenylyl cyclase (sAC) is a HCO3 -stimulated enzyme that produces the ubiquitous signalling molecule cAMP, and deemed an evolutionarily conserved acid–base sensor. However, its presence is not yet confirmed in bony fishes, the most abundant and diverse of vertebrates. Here, we identified sAC genes in various cartilaginous, ray-finned and lobe-finned fish species. Next, we focused on rainbow trout sAC (rtsAC) and identified 20 potential alternative spliced mRNAs coding for protein isoforms ranging in size from 28 to 186 kDa. Biochemical and kinetic analyses on purified recombinant rtsAC protein determined stimulation by HCO3 at physiologically relevant levels for fish internal fluids (EC50∼ 7 mM). rtsAC activity was sensitive to KH7, LRE1, and DIDS (established inhibitors of sAC from other organisms), and insensitive to forskolin and 2,5-dideoxyadenosine (modulators of transmembrane adenylyl cyclases). Western blot and immunocytochemistry revealed high rtsAC expression in gill ion-transporting cells, hepatocytes, red blood cells, myocytes and cardiomyocytes. Analyses in the cell line RTgill-W1 suggested that some of the longer rtsAC isoforms may be preferentially localized in the nucleus, the Golgi apparatus and podosomes. These results indicate that sAC is poised to mediate multiple acid–base homeostatic responses in bony fishes, and provide cues about potential novel functions in mammals. 

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2. Rapid blood acid–base regulation by European sea bass ( Dicentrarchus labrax ) in response to sudden exposure to high environmental CO2

   https://doi.org/10.1242/jeb.242735  

   Montgomery, Daniel W.; Kwan, Garfield T.; Davison, William G.; Finlay, Jennifer; Berry, Alex; Simpson, Stephen D.; Engelhard, Georg H.; Birchenough, Silvana N.; Tresguerres, Martin; Wilson, Rod W. (January 2022, Journal of Experimental Biology)

   ABSTRACT Fish in coastal ecosystems can be exposed to acute variations in CO2 of between 0.2 and 1 kPa CO2 (2000–10,000 µatm). Coping with this environmental challenge will depend on the ability to rapidly compensate for the internal acid–base disturbance caused by sudden exposure to high environmental CO2 (blood and tissue acidosis); however, studies about the speed of acid–base regulatory responses in marine fish are scarce. We observed that upon sudden exposure to ∼1 kPa CO2, European sea bass (Dicentrarchus labrax) completely regulate erythrocyte intracellular pH within ∼40 min, thus restoring haemoglobin–O2 affinity to pre-exposure levels. Moreover, blood pH returned to normal levels within ∼2 h, which is one of the fastest acid–base recoveries documented in any fish. This was achieved via a large upregulation of net acid excretion and accumulation of HCO3− in blood, which increased from ∼4 to ∼22 mmol l−1. While the abundance and intracellular localisation of gill Na+/K+-ATPase (NKA) and Na+/H+ exchanger 3 (NHE3) remained unchanged, the apical surface area of acid-excreting gill ionocytes doubled. This constitutes a novel mechanism for rapidly increasing acid excretion during sudden blood acidosis. Rapid acid–base regulation was completely prevented when the same high CO2 exposure occurred in seawater with experimentally reduced HCO3− and pH, probably because reduced environmental pH inhibited gill H+ excretion via NHE3. The rapid and robust acid–base regulatory responses identified will enable European sea bass to maintain physiological performance during large and sudden CO2 fluctuations that naturally occur in coastal environments. 

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3. Marine heatwaves depress metabolic activity and impair cellular acid–base homeostasis in reef‐building corals regardless of bleaching susceptibility

   https://doi.org/10.1111/gcb.15622  

   Innis, Teegan; Allen‐Waller, Luella; Brown, Kristen T.; Sparagon, Wesley; Carlson, Christopher; Kruse, Elisa; Huffmyer, Ariana S.; Nelson, Craig E.; Putnam, Hollie M.; Barott, Katie L. (April 2021, Global Change Biology)

   Abstract Ocean warming is causing global coral bleaching events to increase in frequency, resulting in widespread coral mortality and disrupting the function of coral reef ecosystems. However, even during mass bleaching events, many corals resist bleaching despite exposure to abnormally high temperatures. While the physiological effects of bleaching have been well documented, the consequences of heat stress for bleaching‐resistant individuals are not well understood. In addition, much remains to be learned about how heat stress affects cellular‐level processes that may be overlooked at the organismal level, yet are crucial for coral performance in the short term and ecological success over the long term. Here we compared the physiological and cellular responses of bleaching‐resistant and bleaching‐susceptible corals throughout the 2019 marine heatwave in Hawai'i, a repeat bleaching event that occurred 4 years after the previous regional event. Relative bleaching susceptibility within species was consistent between the two bleaching events, yet corals of both resistant and susceptible phenotypes exhibited pronounced metabolic depression during the heatwave. At the cellular level, bleaching‐susceptible corals had lower intracellular pH than bleaching‐resistant corals at the peak of bleaching for both symbiont‐hosting and symbiont‐free cells, indicating greater disruption of acid–base homeostasis in bleaching‐susceptible individuals. Notably, cells from both phenotypes were unable to compensate for experimentally induced cellular acidosis, indicating that acid–base regulation was significantly impaired at the cellular level even in bleaching‐resistant corals and in cells containing symbionts. Thermal disturbances may thus have substantial ecological consequences, as even small reallocations in energy budgets to maintain homeostasis during stress can negatively affect fitness. These results suggest concern is warranted for corals coping with ocean acidification alongside ocean warming, as the feedback between temperature stress and acid–base regulation may further exacerbate the physiological effects of climate change. 

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4. Ocean acidification decreases molting but not survival of Antarctic amphipods Djerboa furcipes, Gondogeneia antarctica, and Prostebbingia gracilis

   https://doi.org/10.1007/s00300-025-03393-7  

   Oswalt, Hannah E; Amsler, Margaret O; Amsler, Charles D; Schram, Julie B; McClintock, James B; Baker, Paul A (June 2025, Polar Biology)

   Abstract Ocean acidification refers to a decrease in the pH of the world’s oceans from the oceanic uptake of human-derived atmospheric CO₂. Low pH is known to decrease the calcification and survival of many calcifying invertebrates. Shallow, hard bottom communities along the Western Antarctic Peninsula often have incredibly large numbers of invertebrate mesograzers that shelter on and are mutualists with the dominant brown macroalgae. The common amphipod speciesDjerboa furcipes,Gondogeneia antarctica,andProstebbingia graciliswere collected from the immediate vicinity of Palmer Station, Antarctica (64°46′S, 64°03′W) in January–February 2023 and maintained under three different pH treatments simulating ambient conditions (approximately pH 8.0), near-future conditions for 2100 (pH 7.7), and distant future conditions (pH 7.3) for 8 weeks. Molt number and mortality were monitored throughout the course of the experiment. After the 8 week exposure, amphipods were analyzed for their biochemical compositions including the Mg/Ca ratio of their exoskeletons. There was no significant difference in biochemical composition or survival among the pH treatments for any of the amphipod species. All three species, however, had significantly fewer total numbers of molts in the pH 7.3 treatment than in the ambient treatment. These results suggest that amphipods may be able to maintain their survival in decreased pH by reallocating energy into compensatory behaviors, such as acid–base regulation, and away from energy expensive processes like molting. 

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5. Dynamic single-cell intracellular pH sensing using a SERS-active nanopipette

   https://doi.org/10.1039/D0AN00838A  

   Guo, Jing; Sesena Rubfiaro, Alberto; Lai, Yanhao; Moscoso, Joseph; Chen, Feng; Liu, Yuan; Wang, Xuewen; He, Jin (July 2020, The Analyst)

   null (Ed.)

   Glass nanopipettes have shown promise for applications in single-cell manipulation, analysis, and imaging. In recent years, plasmonic nanopipettes have been developed to enable surface-enhanced Raman spectroscopy (SERS) measurements for single-cell analysis. In this work, we developed a SERS-active nanopipette that can be used to perform long-term and reliable intracellular analysis of single living cells with minimal damage, which is achieved by optimizing the nanopipette geometry and the surface density of the gold nanoparticle (AuNP) layer at the nanopipette tip. To demonstrate its ability in single-cell analysis, we used the nanopipette for intracellular pH sensing. Intracellular pH (pH i ) is vital to cells as it influences cell function and behavior and pathological conditions. The pH sensitivity was realized by simply modifying the AuNP layer with the pH reporter molecule 4-mercaptobenzoic acid. With a response time of less than 5 seconds, the pH sensing range is from 6.0 to 8.0 and the maximum sensitivity is 0.2 pH units. We monitored the pH i change of individual HeLa and fibroblast cells, triggered by the extracellular pH (pH e ) change. The HeLa cancer cells can better resist pH e change and adapt to the weak acidic environment. Plasmonic nanopipettes can be further developed to monitor other intracellular biomarkers. 

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