Life in fresh water is osmotically and energetically challenging for living organisms, requiring increases in ion uptake from dilute environments. However, mechanisms of ion uptake from freshwater environments are still poorly understood and controversial, especially in arthropods, for which several hypothetical models have been proposed based on incomplete data. One compelling model involves the proton pump V-type H+ ATPase (VHA), which energizes the apical membrane, enabling the uptake of Na+ (and other cations) via an unknown Na+ transporter (referred to as the \"Wieczorek Exchanger\" in insects). What evidence exists for this model of ion uptake and what is this mystery exchanger or channel that cooperates with VHA? We present results from studies that explore this question in crustaceans, insects, and teleost fish. We argue that the Na+/H+ antiporter (NHA) is a likely candidate for the Wieczorek Exchanger in many crustaceans and insects; although, there is no evidence that this is the case for fish. NHA was discovered relatively recently in animals and its functions have not been well characterized. Teleost fish exhibit redundancy of Na+ uptake pathways at the gill level, performed by different ion transporter paralogs in diverse cell types, apparently enabling tolerance of low environmental salinity and various pH levels. We argue that much more research is needed on overall mechanisms of ion uptake from freshwater habitats, especially on NHA and other potential Wieczorek Exchangers. Such insights gained would contribute greatly to our general understanding of ionic regulation in diverse species across habitats.

Salinity is a worldwide problem limiting the plant growth and risking food security. This study was conducted to examine exogenous application of silicon (Si), gibberellic acid (GA3) upon the ion transport, growth, yield, and antioxidant enzymes activities of pea plant in saline conditions. Two pea varieties Meteor-FSD and Samrina Zard were pre-treated with GA3 (10-4 M) for 12 h. Plants were allowed to grow with or without silicon in washed silica sand. Ten days old seedlings were shifted in pots with 10 kg soil. Twenty-five days old plants were exposed to 0 and 5 dS m-1 sodium stress. Results showed that exogenous application of GA3 + Si was the best treatment for increasing plant biomass and yield in the presence and absence of NaCl. Furthermore, application of Si or GA3 enhanced chlorophyll content in the leaves, thereby increasing the net assimilation rate of pea varieties under NaCl stress by increasing the antioxidant enzyme activity. Treatment of Si alone or in combination with GA3 significantly reduced Na+ movement in both pea varieties. Results showed that Si has more prominent role than GA3 alone to build-up high plant biomass, yield, soluble protein content and reduction of Na+ transport. Samrina Zard variety showed higher yield, shoot and root dry weight as compared to Meteor-FSD variety in presence and absence of salt. It was concluded that Si can be used as a nutrient for pea under saline or non-saline conditions. Moreover, application of GA3 has a potential role for increasing salinity tolerance, mostly in sensitive pea varieties.

The present study examined the effects of waterborne cadmium (Cd) exposure on ionic balance and ionocyte density in developing zebrafish (Danio rerio) (0-4 days post-fertilization). Fish exposed to 1 or 10 μg Cd/L exhibited an increase in whole body Cd level. Exposure to 10 μg Cd/L also significantly reduced whole body content of Ca2+, but not other major ions (e.g., Na+, K+ and Mg2+). Such reduction was accompanied by a decrease in the density of Ca2+-transporting ionocytes, the Na+/K+-ATPase-rich cells (NaRCs). However, the densities of other ionocyte subtypes (e.g., Na+-transporting ionocytes) remained unchanged after exposure to 10 μg Cd/L. The potential interactive effects between water chemistry and Cd exposure on ionocyte density were examined further in Cd-exposed larvae acclimated to different water NaCl or Ca2+ levels. The results demonstrated that NaRC density increased in fish acclimated to low Ca2+ water, presumably increasing Ca2+ uptake for maintaining Ca2+ homeostasis. However, Cd exposure completely abolished the increased NaRC density in low water Ca2+ environments. The increased NaRCs over development was also reduced in Cd-exposed larvae. In conclusion, our study suggested that Cd exposure reduces the density of NaRCs and suppresses the compensatory regulation of NaRCs during acclimation to low water Ca2+ level. These inhibitory effects by Cd exposure ultimately disrupt Ca2+ balance in the early life stages of zebrafish.

The H+-ATPase-rich (HR) cells of zebrafish larvae are a sub-type of ion-transporting cell located on the yolk sac epithelium that are responsible for Na+ uptake and H+ extrusion. Current models of HR cell ion transport mechanisms in zebrafish larvae are well established, but little is known about the involvement of the various ion transport pathways in regulating intracellular acid-base status. Here, a ratiometric imaging technique was developed and validated to monitor intracellular pH (pHi) continuously in larval zebrafish HR cells in vivo Gene knockdown or CRISPR/Cas9 knockout approaches were used to evaluate the roles of the two principal apical membrane acid excretory pathways, the Na+/H+ exchanger (NHE3b; slc9a3.2) and the H+-ATPase (atpv1aa). Additionally, the role of HR cell cytosolic carbonic anhydrase (CAc) was investigated because of its presumed role in providing H+ for Na+/H+ exchange and H+-ATPase. The temporal pattern and extent of intracellular acidification during exposure of fish to 1% CO2 and the extent of post-CO2 alkalisation were altered markedly in fish experiencing knockdown/knockout of CAc, NHE3b or H+-ATPase. Although there were slight differences among the three knockdown/knockout experiments, the typical response was a greater degree of intracellular acidification during CO2 exposure and a reduced capacity to restore pHi to baseline levels post-hypercapnia. The metabolic alkalosis and subsequent acidification associated with 20 mmol l-1 NH4Cl exposure and its washout were largely unaffected by gene knockdown. Overall, the results suggest markedly different mechanisms of intracellular acid-base regulation in zebrafish HR cells depending on the nature of the acid-base disturbance.

Zebrafish (Danio rerio) have become an important model for integrative physiological research. Zebrafish inhabit a hypo-osmotic environment; to maintain ionic and acid-base homeostasis, they must actively take up ions and secrete acid to the water. The gills in the adult and the skin at larval stage are the primary sites of ionic regulation in zebrafish. The uptake of ions in zebrafish is mediated by specific ion transporting cells termed ionocytes. Similarly, in mammals, ion reabsorption and acid excretion occur in specific cell types in the terminal region of the renal tubules (distal convoluted tubule and collecting duct). Previous studies have suggested that functional regulation of several ion transporters/channels in the zebrafish ionocytes resembles that in the mammalian renal cells. Additionally, several mechanisms involved in regulating the epithelial ion transport during metabolic acidosis are found to be similar between zebrafish and mammals. In this article, we systemically review the similarities and differences in ionic regulation between zebrafish and mammals during metabolic acidosis. We summarize the available information on the regulation of epithelial ion transporters during acidosis, with a focus on epithelial Na⁺, Cl- and Ca2+ transporters in zebrafish ionocytes and mammalian renal cells. We also discuss the neuroendocrine responses to acid exposure, and their potential role in ionic compensation. Finally, we identify several knowledge gaps that would benefit from further study.

Rainbow trout (Oncorhynchus mykiss) exposed to acid-base challenges activate branchial mechanisms for the excretion of acid-base equivalents. Current models of branchial acid-base excretion in freshwater rainbow trout propose two main ionocyte types: the peanut lectin agglutinin-positive (PNA+) mitochondrion-rich cell or ionocyte is believed to secrete HCO3- in exchange for Cl-, whereas H+ secretion is thought to occur across PNA- ionocytes in exchange for Na+ Both HCO3- and H+ are supplied by intracellular hydration of CO2 catalysed by cytosolic carbonic anhydrase (CAc). Immunohistochemical approaches revealed that under control conditions, CAc was detectable in 92.3±1.0% (N=11) of PNA- ionocytes, and the abundance of PNA- ionocytes increased in response to systemic acidosis elicited by 72 h exposure to water of low pH (nominally pH 4.5), hypercapnia (1% CO2, nominally 7.6 Torr) or hyperoxia (achieved by gassing water with pure O2), as did the abundance of PNA- ionocytes that exhibited immunofluorescence for CAc. However, just 4.3±0.6% (N=11) of PNA+ ionocytes expressed detectable CAc under control conditions. Marked increases in the abundance of CAc-positive PNA+ ionocytes were detected following exposure of trout to a base load via recovery from hypercapnia or base infusion (72 h infusion with 140 mmol l-1 NaHCO3). The percentage of CAc-positive PNA+ ionocytes also was increased in trout treated with cortisol (10 mg kg-1 hydrocortisone 21-hemisuccinate daily for 7 days). These results suggest that regulation of CA within PNA+ ionocytes and/or the abundance of CAc-positive PNA+ ionocytes plays a role in activating base secretion in response to systemic alkalosis.

The effects of exogenous trehalose (Tre) on salt tolerance of pharmaceutical plant Catharanthus roseus and the physiological mechanisms were both investigated in this study. The results showed that the supplement of Tre in saline condition (250 mM NaCl) largely alleviated the inhibitory effects of salinity on plant growth, namely biomass accumulation and total leaf area per plant. In this saline condition, the decreased level of relative water content (RWC) and photosynthetic rate were also greatly rescued by exogenous Tre. This improved performance of plants under high salinity induced by Tre could be partly ascribed to its ability to decrease accumulation of sodium, and increase potassium in leaves. The exogenous Tre led to high levels of fructose, glucose, sucrose and Tre inside the salt-stressed plants during whole the three-week treatment. The major free amino acids such as proline, arginine, threonine and glutamate were also largely elevated in the first two-week course of treatment with Tre in saline solution. It was proposed here that Tre might act as signal to make the salt-stressed plants actively increase internal compatible solutes, including soluble sugars and free amino acids, to control water loss, leaf gas exchange and ionic flow at the onset of salt stress. The application of Tre in saline condition also promoted the accumulation of alkaloids. The regulatory role of Tre in improving salt tolerance was optimal with an exogenous concentration of 10 mM Tre. Larger concentrations of Tre were supra-optimum and adversely affected plant growth.

Ionic regulation and acid-base balance are fundamental to the physiology of vertebrates including fish. Acidification of freshwater ecosystems is recognized as a global environmental problem, and the physiological responses to acid exposure in a few fish species are well characterized. However, the underlying mechanisms promoting ionic and acid-base balance for most fish species that have been investigated remain unclear. Zebrafish (Danio rerio) has emerged as a powerful model system to elucidate the molecular basis of ionic and acid-base regulation. The utility of zebrafish is related to the ease with which it can be genetically manipulated, its suitability for state-of-the-art molecular and cellular approaches, and its tolerance to diverse environmental conditions. Recent studies have identified several key regulatory mechanisms enabling acclimation of zebrafish to acidic environments, including activation of the sodium/hydrogen exchanger (NHE) and H(+)-ATPase for acid secretion and Na(+) uptake, cortisol-mediated regulation of transcellular and paracellular Na(+) movements, and ionocyte proliferation controlled by specific cell-fate transcription factors. These integrated physiological responses ultimately contribute to ionic and acid-base homeostasis in zebrafish exposed to acidic water. In the present review, we provide an overview of the general effects of acid exposure on freshwater fish, the adaptive mechanisms promoting extreme acid tolerance in fishes native to acidic environments, and the mechanisms regulating ionic and acid-base balance during acid exposure in zebrafish.