Mechanisms Of Ion Regulation Research Articles

Ammonia excretion by the fish gill: discoveries and ideas that shaped our current understanding.

The fish gill serves many physiological functions, among which is the excretion of ammonia, the primary nitrogenous waste in most fishes. Although it is the end-product of nitrogen metabolism, ammonia serves many physiological functions including acting as an acid equivalent and as a counter-ion in mechanisms of ion regulation. Our current understanding of the mechanisms of ammonia excretion have been influenced by classic experimental work, clever mechanistic approaches, and modern molecular and genetic techniques. In this review, I will overview the history of the study of ammonia excretion by the gills of fishes, highlighting the important advancements that have shaped this field with a nearly 100-year history. The developmental and evolutionary implications of an ammonia and gill-dominated nitrogen regulation strategy in most fishes will also be discussed. Throughout the review, I point to areas in which more work is needed to push forward this field of research that continues to produce novel insights and discoveries that will undoubtedly shape our overall understanding of fish physiology.

Proton-driven sodium secretion in a saline water animal

Aquatic animals residing in saline habitats either allow extracellular sodium concentration to conform to environmental values or regulate sodium to lower levels. The latter strategy requires an energy-driven process to move sodium against a large concentration gradient to eliminate excess sodium that diffuses into the animal. Previous studies of invertebrate and vertebrate species indicate a sodium pump, Na+/K+ ATPase, powers sodium secretion. We provide the first functional evidence of a saline-water animal, Aedes taeniorhynchus mosquito larva, utilizing a proton pump to power this process. Vacuolar-type H+ ATPase (VHA) protein is highly expressed on the apical membrane of the posterior rectal cells, and in situ sodium flux across this epithelium increases significantly in larvae held in higher salinity and is sensitive to Bafilomycin A1, an inhibitor of VHA. We also report the first evidence of splice variants of the sodium/proton exchanger, NHE3, with both high and low molecular weight variants highly expressed on the apical membrane of the posterior rectal cells. Evidence of NHE3 function was indicated with in situ sodium transport significantly inhibited by a NHE3 antagonist, S3226. We propose that the outward proton pumping by VHA establishes a favourable electromotive gradient to drive sodium secretion via NHE3 thus producing a hyperosmotic, sodium-rich urine. This H+- driven Na+ secretion process is the primary mechanism of ion regulation in salt-tolerant culicine mosquito species and was first investigated over 80 years ago.

Impact of Various Levels of Salinity Stress on Growth Attributes of Some Salt-Sensitive and Tolerant Varieties of Mung Bean

Salinity stress poses a significant threat to crop productivity, and understanding the mechanisms of ion uptake and distribution in different cultivars is crucial for developing salt-tolerant varieties. This study investigates the patterns of ion uptake and distribution in salt-sensitive and salt-tolerant cultivars of Vigna radiata, commonly known as mung bean. Hydroponic experiments were conducted under controlled conditions to assess the responses of two contrasting cultivars to varying salt concentrations. The study reveals distinct differences in the ion uptake and distribution between salt-sensitive and salt-tolerant cultivars of V. radiata. Salt-sensitive cultivars exhibited increased sodium (Na+) accumulation in both roots and shoots under saline conditions, leading to disrupted ion homeostasis. In contrast, salt-tolerant cultivars demonstrated efficient ion regulation mechanisms, with a higher capacity for potassium (K+) retention and a lower Na+ to K+ ratio. The study also shed light on the involvement of ion transporters and channels in facilitating differential ion accumulation, highlighting the molecular mechanisms contributing to salt tolerance in Vigna radiata. These findings provide valuable insights into the adaptive strategies of salt-tolerant cultivars, offering a basis for targeted breeding programs aimed at enhancing salinity resilience in this economically significant legume. In summary, this research deepens our understanding of the intricate relationship between ion uptake and distribution in Vigna radiata varieties with varying salt tolerance. It lays the groundwork for developing robust crop varieties capable of thriving in saline environments.

Ion transport in nanofluidics under external fields.

Nanofluidic channels with tailored ion transport dynamics are usually used as channels for ion transport, to enable high-performance ion regulation behaviors. The rational construction of nanofluidics and the introduction of external fields are of vital significance to the advancement and development of these ion transport properties. Focusing on the recent advances of nanofluidics, in this review, various dimensional nanomaterials and their derived homogeneous/heterogeneous nanofluidics are first briefly introduced. Then we discuss the basic principles and properties of ion transport in nanofluidics. As the major part of this review, we focus on recent progress in ion transport in nanofluidics regulated by external physical fields (electric field, light, heat, pressure, etc.) and chemical fields (pH, concentration gradient, chemical reaction, etc.), and reveal the advantages and ion regulation mechanisms of each type. Moreover, the representative applications of these nanofluidic channels in sensing, ionic devices, energy conversion, and other areas are summarized. Finally, the major challenges that need to be addressed in this research field and the future perspective of nanofluidics development and practical applications are briefly illustrated.

Na+/H+ exchangers differentially contribute to midgut fluid sodium and proton concentration in the sea urchin larva.

Regulation of ionic composition and pH is a requisite of all digestive systems in the animal kingdom. Larval stages of the marine superphylum Ambulacraria, including echinoderms and hemichordates, were demonstrated to have highly alkaline conditions in their midgut with the underlying epithelial transport mechanisms being largely unknown. Using ion-selective microelectrodes, the present study demonstrated that pluteus larvae of the purple sea urchin have highly alkaline pH (pH∼9) and low [Na+] (∼120 mmol l-1) in their midgut fluids, compared with the ionic composition of the surrounding seawater. We pharmacologically investigated the role of Na+/H+ exchangers (NHE) in intracellular pH regulation and midgut proton and sodium maintenance using the NHE inhibitor 5-(n-ethyl-n-isopropyl)amiloride (EIPA). Basolateral EIPA application decreased midgut pH while luminal application via micro-injections increased midgut [Na+], without affecting pH. Immunohistochemical analysis demonstrated a luminal localization of NHE-2 (SpSlc9a2) in the midgut epithelium. Specific knockdown of spslc9a2 using Vivo-Morpholinos led to an increase in midgut [Na+] without affecting pH. Acute acidification experiments in combination with quantitative PCR analysis and measurements of midgut pH and [Na+] identified two other NHE isoforms, Spslc9a7 and SpSlc9a8, which potentially contribute to the regulation of [Na+] and pH in midgut fluids. This work provides new insights into ion regulatory mechanisms in the midgut epithelium of sea urchin larvae. The involvement of NHEs in regulating pH and Na+ balance in midgut fluids shows conserved features of insect and vertebrate digestive systems and may contribute to the ability of sea urchin larvae to cope with changes in seawater pH.

Influence of Thermal Pollution on the Physiological Conditions and Bioaccumulation of Metals, Metalloids, and Trace Metals in Whitefish (Coregonus lavaretus L.).

The Kola nuclear power plant, which discharges warm water into one of the bays of subarctic Lake Imandra, significantly changes fish habitats. The temperature gradient of the lake is between 2 and 8 °C, which makes it significantly different from the natural temperature of the lake water. The stenothermal cold-water native species (lake whitefish (Coregonus lavaretus L.)), living for more than 40 years under conditions of thermal pollution, has adapted to this stressor. Moreover, this population differs favorably from the population in the natural-temperature environment in terms of its physiological state. Firstly, the hemoglobin concentrations in the fish blood are in the range of the ecological optimum, and secondly, it has a higher somatic growth, as estimated by Fulton’s condition factor. One of its main adaptive mechanisms of ion regulation is an intense metabolism of Na due to the high respiratory activity of the whitefish in warmer water. An increased accumulation of Rb and excretion of Se, Mo, and Si are associated more or less with that feature. Under conditions of an increased water temperature, the main metabolic need is due to a deficiency of Se in fish. The intensive metabolism of selenoproteins may involve risks of toxic effects and the bioaccumulation of Hg, As, and Cu in cases of increased existing stressors or the appearance of new ones.

Are European sea bass as euryhaline as expected? Intraspecific variation in freshwater tolerance

In teleosts, the regulation of hydromineral balance has a direct impact on several physiological functions, biochemical processes, and can influence behaviour, distribution and survival. As European sea bass Dicentrarchus labrax undertake seasonal migrations from seawater (SW) to brackish, estuarine and fresh water (FW) in their habitat, this study investigates their capacity to tolerate fresh water and explores intraspecific variations in physiological responses. Juvenile D. labrax were transferred from SW to FW at various ages. Freshwater-tolerant and non-tolerant phenotypes were discriminated according to behavioural and morphological characteristics. About 30% of the fish exposed to FW were identified as freshwater intolerant following FW challenges performed at different ages. Interestingly, intolerant fish exhibited the same phenotypic traits: erratic swimming, lower speed, isolation from the shoal and darker colour. Freshwater-intolerant fish were also characterised by a significant lower blood osmolality compared to tolerant fish, and significantly lower Na+/K+-ATPase α1a expression in the posterior kidney. An imbalance in ion regulatory mechanisms was further confirmed by a blood Na+/Cl− ratio imbalance observed in some freshwater-intolerant fish. The analysis of glucocorticoid and mineralocorticoid receptor expression levels in gills and kidney revealed significant differences between freshwater-intolerant and -tolerant fish in both organs, suggesting differential stress-related responses. This study clearly shows an intraspecific difference in the responses following FW transfer with a decreased renal ion uptake capacity as a major cause for freshwater intolerance.

Glial Channels and Transporters that Mediate Excretion of K+ in the Microenvironment between Glia and Neurons Shape Neuronal Output in C. elegans

Isolated microenvironments, such as the tripartite synapse, where the concentration of ions is regulated independently from the surrounding tissues, exist throughout the nervous system. While the regulation of ions in these microenvironments is known to be mainly mediated by glia, the molecular mechanisms of ion regulation and effects on neuronal output and animal behavior are poorly understood. Using the model system C. elegans, our lab published that Na+ channels of the DEG/ENaC family expressed in glia control neuronal Ca2+ transients and animal behavior in response to sensory stimuli. DEG/ENaC Na+ channels are known to establish a favorable driving force for K+ excretion, which occurs via inward rectifier K+ channels, in epithelial tissues across species. We hypothesized that a similar mechanism exists in the nervous system. Using molecular, genetic, in vivo imaging, and behavioral approaches, we showed that expression in glia of inward rectifier K+ channels and cationic channels rescues the sensory deficits caused by knock-out of glial DEG/ENaCs, supporting our hypothesis. Based on this model, Na+/K+-ATPases are also needed to maintain ionic concentrations following influx of Na+ and excretion of K+. To test this prediction, we used RNAi to knock down each of the 5 α-subunits of the Na+/K+-pump in glia and test the effect of the knock-downs on sensory behavior. We identified Na+/K+-pump α-subunits eat-6 and catp-1 as specifically required in glia for sensory perception, further supporting a model in which glia shapes neuronal output and, consequently, animal behavior by excreting K+ in the microenvironment surrounding neurons. Given that DEG/ENaCs, inward rectifier K+ channels, and Na+/K+-pumps are also expressed in mammalian glia, we propose that this mode of regulation of neuronal function in conserved across species.

Sodium provides unique insights into transgenerational effects of ocean acidification on bivalve shell formation

Ocean acidification is likely to have profound impacts on marine bivalves, especially on their early life stages. Therefore, it is imperative to know whether and to what extent bivalves will be able to acclimate or adapt to an acidifying ocean over multiple generations. Here, we show that reduced seawater pH projected for the end of this century (i.e., pH7.7) led to a significant decrease of shell production of newly settled juvenile Manila clams, Ruditapes philippinarum. However, juveniles from parents exposed to low pH grew significantly faster than those from parents grown at ambient pH, exhibiting a rapid transgenerational acclimation to an acidic environment. The sodium composition of the shells may shed new light on the mechanisms responsible for beneficial transgenerational acclimation. Irrespective of parental exposure, the amount of Na incorporated into shells increased with decreasing pH, implying active removal of excessive protons through the Na+/H+ exchanger which is known to depend on the Na+ gradient actively built up by the Na+/K+-ATPase as a driving force. However, the shells with a prior history of transgenerational exposure to low pH recorded significantly lower amounts of Na than those with no history of acidic exposure. It therefore seems very likely that the clams may implement less costly and more ATP-efficient ion regulatory mechanisms to maintain pH homeostasis in the calcifying fluid following transgenerational acclimation. Our results suggest that marine bivalves may have a greater capacity to acclimate or adapt to ocean acidification by the end of this century than currently understood.

Tight Coupling of Astrocyte pH Dynamics to Epileptiform Activity Revealed by Genetically Encoded pH Sensors.

Astrocytes can both sense and shape the evolution of neuronal network activity and are known to possess unique ion regulatory mechanisms. Here we explore the relationship between astrocytic intracellular pH dynamics and the synchronous network activity that occurs during seizure-like activity. By combining confocal and two-photon imaging of genetically encoded pH reporters with simultaneous electrophysiological recordings, we perform pH measurements in defined cell populations and relate these to ongoing network activity. This approach reveals marked differences in the intracellular pH dynamics between hippocampal astrocytes and neighboring pyramidal neurons in rodent in vitro models of epilepsy. With three different genetically encoded pH reporters, astrocytes are observed to alkalinize during epileptiform activity, whereas neurons are observed to acidify. In addition to the direction of pH change, the kinetics of epileptiform-associated intracellular pH transients are found to differ between the two cell types, with astrocytes displaying significantly more rapid changes in pH. The astrocytic alkalinization is shown to be highly correlated with astrocytic membrane potential changes during seizure-like events and mediated by an electrogenic Na(+)/HCO3 (-) cotransporter. Finally, comparisons across different cell-pair combinations reveal that astrocytic pH dynamics are more closely related to network activity than are neuronal pH dynamics. This work demonstrates that astrocytes exhibit distinct pH dynamics during periods of epileptiform activity, which has relevance to multiple processes including neurometabolic coupling and the control of network excitability. Dynamic changes in intracellular ion concentrations are central to the initiation and progression of epileptic seizures. However, it is not known how changes in intracellular H(+) concentration (ie, pH) differ between different cell types during seizures. Using recently developed pH-sensitive proteins, we demonstrate that astrocytes undergo rapid alkalinization during periods of seizure-like activity, which is in stark contrast to the acidification that occurs in neighboring neurons. Rapid astrocytic pH changes are highly temporally correlated with seizure activity, are mediated by an electrogenic Na(+)/HCO3- cotransporter, and are more tightly coupled to network activity than are neuronal pH changes. As pH has profound effects on signaling in the nervous system, this work has implications for our understanding of seizure dynamics.

Ocean warming and acidification modulate energy budget and gill ion regulatory mechanisms in Atlantic cod (Gadus morhua).

Ocean warming and acidification are threatening marine ecosystems. In marine animals, acidification is thought to enhance ion regulatory costs and thereby baseline energy demand, while elevated temperature also increases baseline metabolic rate. Here we investigated standard metabolic rates (SMR) and plasma parameters of Atlantic cod (Gadus morhua) after 3–4 weeks of exposure to ambient and future PCO2 levels (550, 1200 and 2200 µatm) and at two temperatures (10, 18 °C). In vivo branchial ion regulatory costs were studied in isolated, perfused gill preparations. Animals reared at 18 °C responded to increasing CO2 by elevating SMR, in contrast to specimens at 10 °C. Isolated gills at 10 °C and elevated PCO2 (≥1200 µatm) displayed increased soft tissue mass, in parallel to increased gill oxygen demand, indicating an increased fraction of gill in whole animal energy budget. Altered gill size was not found at 18 °C, where a shift in the use of ion regulation mechanisms occurred towards enhanced Na+/H+-exchange and HCO3− transport at high PCO2 (2200 µatm), paralleled by higher Na+/K+-ATPase activities. This shift did not affect total gill energy consumption leaving whole animal energy budget unaffected. Higher Na+/K+-ATPase activities in the warmth might have compensated for enhanced branchial permeability and led to reduced plasma Na+ and/or Cl− concentrations and slightly lowered osmolalities seen at 18 °C and 550 or 2200 µatm PCO2 in vivo. Overall, the gill as a key ion regulation organ seems to be highly effective in supporting the resilience of cod to effects of ocean warming and acidification.

Impact of long-term moderate hypercapnia and elevated temperature on the energy budget of isolated gills of Atlantic cod (Gadus morhua)

Effects of severe hypercapnia have been extensively studied in marine fishes, while knowledge on the impacts of moderately elevated CO2 levels and their combination with warming is scarce. Here we investigate ion regulation mechanisms and energy budget in gills from Atlantic cod acclimated long-term to elevated PCO2 levels (2500μatm) and temperature (18°C). Isolated perfused gill preparations were established to determine gill thermal plasticity during acute exposures (10–22°C) and in vivo costs of Na+/K+-ATPase activity, protein and RNA synthesis. Maximum enzyme capacities of F1Fo-ATPase, H+-ATPase and Na+/K+-ATPase were measured in vitro in crude gill homogenates. After whole animal acclimation to elevated PCO2 and/or warming, branchial oxygen consumption responded more strongly to acute temperature change. The fractions of gill respiration allocated to protein and RNA synthesis remained unchanged. In gills of fish CO2-exposed at both temperatures, energy turnover associated with Na+/K+-ATPase activity was reduced by 30% below rates of control fish. This contrasted in vitro capacities of Na+/K+-ATPase, which remained unchanged under elevated CO2 at 10°C, and earlier studies which had found a strong upregulation under severe hypercapnia. F1Fo-ATPase capacities increased in hypercapnic gills at both temperatures, whereas Na+/K+ATPase and H+-ATPase capacities only increased in response to elevated CO2 and warming indicating the absence of thermal compensation under CO2. We conclude that in vivo ion regulatory energy demand is lowered under moderately elevated CO2 levels despite the stronger thermal response of total gill respiration and the upregulation of F1Fo-ATPase. This effect is maintained at elevated temperature.

Hematological adjustments of the bony tongue Arapaima gigas under influence of amazonian waters

Arapaima gigas, populary known as pirarucu is largest freshwater fish in Amazon river, with great, economic and ecological, interest. Despite being an air breather its gill structure is quite close to water breathers, especially in early stages of development [1]. However, in animals above 100g the pillar cell system are embedded into the filament being the gill, an ion regulation active site. The effects of Amazonian rivers waters is well notices in fish [2], mainly by black water (BW), and white water (WW) where the ion fluxes can be meansured [3]. However information on the ionic regulation patters on this species are scarce as well implication by hematological adjustments. Such data could provide inferences on management conditions as well corroborate with what has been suggested by literature, which suggest this species as a panmitic population [4] able to deal with the hydrographic barrier formed by BW in the Amazon basin [5]. The present study aims was to analyzed A. gigas hematological parameters when exposed to BW and WW providing suitable hematological data concerning physiological responses in different types of water. Fish were acclimated seven days in three separated ponds containing BW, WW and well water (C), respectively. Control (C) fish were placed in the latter pond. Blood samples were taken from the caudal vessel at the end of acclimation period in order to perform measurement assays on levels of hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, as well glucose, cholesterol and protein levels. Our findings corroborate the hypothesis stating that BW does interfere on fish adaptation specialy in small fish (~100g). These lack of adaptation should be due the gill morphology of small fish which is close to other water breather fish. However in large fish (~1000g) the findings cleary showed that neither WW or BW can interfere on plasma profile of analysed fish. The compromisse between gas exchange and ion regulation has been demonstrated previously. As hematological adjustment plays a role on such osmorespiratory compromise, the hematological parameters data presented by this study clearly demonstrate that changes in the hematological features play a crucial role in the water to air breathing transition behavior as well in the A. gigas gill ontogenetic changes. Despite black water systems being considered a barrier constraining the dispersion of several species; this seems not to be a problem for this species which has kept its ion-regulatory mechanisms even when immerse in black waters.

A new model for fish ion regulation: identification of ionocytes in freshwater- and seawater-acclimated medaka (Oryzias latipes).

The ion regulation mechanisms of fishes have been recently studied in zebrafish (Danio rerio), a stenohaline species. However, recent advances using this organism are not necessarily applicable to euryhaline fishes. The euryhaline species medaka (Oryzias latipes), which, like zebrafish, is genetically well categorized and amenable to molecular manipulation, was proposed as an alternative model for studying osmoregulation during acclimation to different salinities. To establish its suitability as an alternative, the present study was conducted to (1) identify different types of ionocytes in the embryonic skin and (2) analyze gene expressions of the transporters during seawater acclimation. Double/triple in situ hybridization and/or immunocytochemistry revealed that freshwater (FW) medaka contain three types of ionocyte: (1) Na(+)/H(+) exchanger 3 (NHE3) cells with apical NHE3 and basolateral Na(+)-K(+)-2Cl(-) cotransporter (NKCC), Na(+)-K(+)-ATPase (NKA) and anion exchanger (AE); (2) Na(+)-Cl(-) cotransporter (NCC) cells with apical NCC and basolateral H(+)-ATPase; and (3) epithelial Ca(2+) channel (ECaC) cells [presumed accessory (AC) cells] with apical ECaC. On the other hand, seawater (SW) medaka has a single predominant ionocyte type, which possesses apical cystic fibrosis transmembrane conductance regulator (CFTR) and NHE3 and basolateral NKCC and NKA and is accompanied by smaller AC cells that express lower levels of basolateral NKA. Reciprocal gene expressions of decreased NHE3, AE, NCC and ECaC and increased CFTR and NKCC in medaka gills during SW were revealed by quantative PCR analysis.

Functional anatomy and ion regulatory mechanisms of the antennal gland in a semi-terrestrial crab, Ocypode stimpsoni.

ABSTRACTBrachyuran crabs from diverse habitats show great differences in their osmoregulatory processes, especially in terms of the structural and physiological characteristics of the osmoregulatory organs. In crustaceans, the antennal glands are known to be important in osmoregulation, and they play a functional role analogous to that of the vertebrate kidney. Nevertheless, the detailed structure and function of the antennal glands in different species have rarely been described. The aim of this study is to investigate the role of the antennal gland in ion regulation by examining the ultrastructure of the cells and the distribution of the ion regulatory proteins in each cell type in the antennal gland of a semi-terrestrial crab. The results showed that Na+, K+-ATPase activity significantly increased in the antennal gland after a 4-day acclimation in dilute seawater and returned to its original (day 0) level after 7 days. Three major types of cells were identified in the antennal gland, including coelomic cells (COEs), labyrinthine cells (LBRs) and end-labyrinthine cells (ELBRs). The proximal tubular region (PT) and distal tubular region (DT) of the antennal gland consist of LBRs and COEs, whereas the end tubular region (ET) consists of all three types of cells, with fewer COEs and more ELBRs. We found a non-uniform distribution of NKA immunoreactivity, with increasing intensity from the proximal to the distal regions of the antennal gland. We summarise our study with a proposed model for the urine reprocessing pathway and the role of each cell type or segment of the antennal gland.

Transcriptional responses in Atlantic salmon (Salmo salar) exposed to deltamethrin, alone or in combination with azamethiphos

Recently, Atlantic salmon (Salmo salar) fish farmers have applied a combination of deltamethrin and azamethiphos in high-concentration and short-duration immersion treatment to improve protection against sea-lice (Lepeophtheirus sp.). In this work we aimed to study the effects of deltamethrin, alone or in combination with azamethiphos, on the transcription of stress and detoxification marker genes. Atlantic salmon kept at 12°C (one group was also kept at 4–5°C) were treated with deltamethrin alone or in combination with azamethiphos for a total of 40min, and gill and liver tissue harvested for transcriptional analysis 2 and 24h post treatment. No lethality was observed during the experiment. The result showed that deltamethrin, alone or in combination with azamethiphos, affected the transcriptional levels of several oxidative stress markers, including MnSOD (SOD2) and HSP70 (HSPA8) in the liver, and GPX1, CAT, MnSOD, HSP70 and GSTP1 in the gills. Significant responses for CASP3B, BCLX, IGFBP1B and ATP1A1 (Na–K–ATPase a1b) by some of the treatments suggest that the pharmaceutical drugs may affect apoptosis, growth and ion regulation mechanisms. In fish kept at 4–5°C, different effects were observed, suggesting a temperature-dependent response. In conclusion, the observed responses indicate that short-term exposure to deltamethrin has a profound effect on transcription of the evaluated markers in gills and liver of fish. Co-treatment with azamethiphos appears to have small mitigating effects on the transcriptional response caused by deltamethrin exposure alone.

Allatostatin A-like immunoreactivity in the nervous system and gut of the larval midge Chironomus riparius: modulation of hindgut motility, rectal K+ transport and implications for exposure to salinity.

Evidence for the presence of allatostatin (AST) A-like neuropeptides in the larval midge Chironomus riparius is reported. Immunohistochemical studies on the nervous system and gut revealed the presence of AST A-like immunoreactive (AST-IR) cells and processes. The nerve cord contained AST-IR processes that originated from cells in the brain and travelled the length of nerve cord to the terminal ganglion. Within each ganglion, these processes gave rise to varicosities, suggesting that they formed synapses with neurons in the ganglia. Endocrine cells containing AST-IR were present in three regions of the midgut: near the attachment of the Malpighian tubules, between the anterior and posterior midgut, and in the vicinity of the gastric caecae. The terminal ganglion also contained four AST-IR cells that gave rise to axons that projected onto the hindgut and posterior midgut. Application of a cockroach AST to the semi-isolated hindgut of larval C. riparius led to dose-dependent inhibition of muscle contractions with an EC50 of ~10 nmol l(-1) and a decrease in rectal K(+) reabsorption resulting from reduced rectal Na(+)/K(+)-ATPase and vacuolar type H(+)-ATPase activities. The results suggest the presence of endogenous AST-like neuropeptides in larval C. riparius, where these factors play a role in the function of the gut. Furthermore, regulation of ion reabsorption by ASTs at the rectum could serve as an ideal mechanism of ion regulation in the face of abrupt and acute elevated salt levels.

Expression of ion transporters in gill mitochondrion-rich cells in Japanese eel acclimated to a wide range of environmental salinity

We examined morphological changes and molecular mechanisms of ion regulation in mitochondrion-rich (MR) cells of Japanese eel acclimated to different environmental salinities. Electron microscopic observations revealed that the apical membrane of MR cells appeared as a flat or slightly projecting disk with a mesh-like structure on its surface in eel acclimated to freshwater (FW). In seawater (SW)-acclimated eel, in contrast, the apical membrane of MR cells showed a slightly concave surface without a mesh-like structure. The mRNA expression of Na+/H+ exchanger-3 (NHE3) in deionized FW and normal SW was higher than that in normal FW and 30%-diluted SW. Expression of Na+/K+/2Cl− cotransporter-1a (NKCC1a) became higher with increasing environmental salinity. Immunofluorescence staining showed that the apical NHE3 immunoreaction was stronger in deionized FW and normal SW than in the other groups. Basolateral NKCC1 immunoreaction was most intense in normal SW. These results indicate that apical NHE3 is involved in ion uptake in fish acclimated to hypotonic environments, and that basolateral NKCC1 is important for acclimation to hypertonic environments. The relatively high expression of NHE3 in SW further indicates a possible role of NHE3 in acid-base regulation in the gills in SW-acclimated fish.

Effect of the pesticide monocrotophos on the osmoregulation of a brackish water oligochaete, Pontodrilus bermudensis (beddard) in relation to salinity variations

The toxicity of a commonly used organophosphate pesticide, Monocrotophos, on Pontodrilus bermudensis (Beddard) in optimum salinity was studied from a tropical brackish water environment, the Southern Lighter Channel at the Visakhapatnam Harbour. Physicochemical characteristics were determined at the habitat of the worm during the period of the study. The LC 50 values obtained for this worm subjected to 24, 48, 72 and 96 hours exposure showed that the toxicity of the pesticide increased with the period of exposure. The high LC 50 values indicated its endurance to the toxicant. The concentration changes found in the body fluid (C - , Na+ & K+) in the worms might suggest the possible influence of the pesticide on ion pumps or ion regulatory mechanisms of the worm. The present study confirms that Monocrotophos has interfered remarkably with ion regulation of the worm.

Egg and early larval stages of Baltic cod, Gadus morhua, are robust to high levels of ocean acidification

The accumulation of carbon dioxide in the atmosphere will lower the pH in ocean waters, a process termed ocean acidification (OA). Despite its potentially detrimental effects on calcifying organisms, experimental studies on the possible impacts on fish remain scarce. While adults will most likely remain relatively unaffected by changes in seawater pH, early life-history stages are potentially more sensitive, due to the lack of gills with specialized ion-regulatory mechanisms. We tested the effects of OA on growth and development of embryos and larvae of eastern Baltic cod, the commercially most important fish stock in the Baltic Sea. Cod were reared from newly fertilized eggs to early non-feeding larvae in 5 different experiments looking at a range of response variables to OA, as well as the combined effect of CO2 and temperature. No effect on hatching, survival, development, and otolith size was found at any stage in the development of Baltic cod. Field data show that in the Bornholm Basin, the main spawning site of eastern Baltic cod, in situ levels of pCO2 are already at levels of 1,100 μatm with a pH of 7.2, mainly due to high eutrophication supporting microbial activity and permanent stratification with little water exchange. Our data show that the eggs and early larval stages of Baltic cod seem to be robust to even high levels of OA (3,200 μatm), indicating an adaptational response to CO2.