The fish gill is a multipurpose organ that, in addition to providing for aquatic gas exchange, plays dominant roles in osmotic and ionic regulation, acid-base regulation, and excretion of nitrogenous wastes Thus, despite the fact that all fish groups have functional kidneys, the gill epithelium is the site of many processes that are mediated by renal epithelia in terrestrial vertebrates Indeed, many of the pathways that mediate these processes in mammalian renal epithelial are expressed in the gill, and many of the extrinsic and intrinsic modulators of these processes are also found in fish endocrine tissues and the gill itself The basic patterns of gill physiology were outlined over a half century ago, but modern immunological and molecular techniques are bringing new insights into this complicated system Nevertheless, substantial questions about the evolution of these mechanisms and control remain

https://people.clas.ufl.edu/devans/files/PRVGill.pdf

The Multifunctional Fish Gill: Dominant Site of Gas Exchange, Osmoregulation, Acid-Base Regulation, and Excretion of Nitrogenous Waste

PRL is an anterior pituitary hormone that, along with GH and PLs, forms a family of hormones that probably resulted from the duplication of an ancestral gene. The PRLR is also a member of a larger family, known as the cytokine class-1 receptor superfamily, which currently has more than 20 different members. PRLRs or binding sites are widely distributed throughout the body. In fact, it is difficult to find a tissue that does not express any PRLR mRNA or protein. In agreement with this wide distribution of receptors is the fact that now more than 300 separate actions of PRL have been reported in various vertebrates, including effects on water and salt balance, growth and development, endocrinology and metabolism, brain and behavior, reproduction, and immune regulation and protection. Clearly, a large proportion of these actions are directly or indirectly associated with the process of reproduction, including many behavioral effects. PRL is also becoming well known as an important regulator of immune function. A number of disease states, including the growth of different forms of cancer as well as various autoimmune diseases, appear to be related to an overproduction of PRL, which may act in an endocrine, autocrine, or paracrine manner, or via an increased sensitivity to the hormone. The first step in the mechanism of action of PRL is the binding to a cell surface receptor. The ligand binds in a two-step process in which site 1 on PRL binds to one receptor molecule, after which a second receptor molecule binds to site 2 on the hormone, forming a homodimer consisting of one molecule of PRL and two molecules of receptor. The PRLR contains no intrinsic tyrosine kinase cytoplasmic domain but associates with a cytoplasmic tyrosine kinase, JAK2. Dimerization of the receptor induces tyrosine phosphorylation and activation of the JAK kinase followed by phosphorylation of the receptor. Other receptor-associated kinases of the Src family have also been shown to be activated by PRL. One major pathway of signaling involves phosphorylation of cytoplasmic State proteins, which themselves dimerize and translocate to nucleus and bind to specific promoter elements on PRL-responsive genes. In addition, the Ras/Raf/MAP kinase pathway is also activated by PRL and may be involved in the proliferative effects of the hormone. Finally, a number of other potential mediators have been identified, including IRS-1, PI-3 kinase, SHP-2, PLC gamma, PKC, and intracellular Ca2+. The technique of gene targeting in mice has been used to develop the first experimental model in which the effect of the complete absence of any lactogen or PRL-mediated effects can be studied. Heterozygous (+/-) females show almost complete failure to lactate after the first, but not subsequent, pregnancies. Homozygous (-/-) females are infertile due to multiple reproductive abnormalities, including ovulation of premeiotic oocytes, reduced fertilization of oocytes, reduced preimplantation oocyte development, lack of embryo implantation, and the absence of pseudopregnancy. Twenty per cent of the homozygous males showed delayed fertility. Other phenotypes, including effects on the immune system and bone, are currently being examined. It is clear that there are multiple actions associated with PRL. It will be important to correlate known effects with local production of PRL to differentiate classic endocrine from autocrine/paracrine effects. The fact that extrapituitary PRL can, under some circumstances, compensate for pituitary PRL raises the interesting possibility that there may be effects of PRL other than those originally observed in hypophysectomized rats. The PRLR knockout mouse model should be an interesting system by which to look for effects activated only by PRL or other lactogenic hormones. On the other hand, many of the effects reported in this review may be shared with other hormones, cytokines, or growth factors and thus will be more difficult to study. (ABSTRACT TRUNCATED)

Prolactin (PRL) and Its Receptor: Actions, Signal Transduction Pathways and Phenotypes Observed in PRL Receptor Knockout Mice

This review focuses on the structure and function of the branchial chloride cell in freshwater fishes. The mitochondria-rich chloride cell is believed to be the principal site of trans-epithelial Ca2+ and Cl- influxes. Though currently debated, there is accruing evidence that the pavement cell is the site of Na+ uptake via channels linked electrically to an apical membrane vacuolar H(+)-ATPase (proton pump). Chloride cells perform an integral role in acid-base regulation. During conditions of alkalosis, the surface area of exposed chloride cells is increased, which serves to enhance base equivalent excretion as the rate of Cl-/HCO3- exchange is increased. Conversely, during acidosis, the chloride cell surface area is diminished by an expansion of the adjacent pavement cells. This response reduces the number of functional Cl-/HCO3- exchangers. Under certain conditions that challenge ion regulation, chloride cells proliferate on the lamellae. This response, while optimizing the Ca2+ and Cl- transport capacity of the gill, causes a thickening of the blood-to-water diffusion barrier and thus impedes respiratory gas transfer.

THE CHLORIDE CELL:Structure and Function in the Gills of Freshwater Fishes

The secondary active Clsecretion in seawater (SW) teleost fish gills and elasmobranch rectal gland involves basolateral Na + ,K + -ATPase and NKCC, apical membrane CFTR anion channels, and a paracellular Na + -selective conductance. In freshwater (FW) teleost gill, the mechanism of NaCl uptake is more controversial and involves apical V-type H + -ATPase linked to an apical Na + channel, apical Cl � -HCO3 exchange and basolateral Na + ,K + -ATPase. Ca 2+ uptake (in FW and SW) is via Ca 2+ channels in the apical membrane and Ca 2+ -ATPase in the basolateral membrane. Mainly this transport occurs in mitochondria rich (MR) chloride cells, but there is a role for the pavement cells also. Future research will likely expand in two major directions, molded by methodology: first in physiological genomics of all the transporters, including their expression, trafficking, operation, and regulation at the molecular level, and second in biotelemetry to examine multivariable components in behavioral physiological ecology, thus widening the integration of physiology from the molecular to the environmental levels while deepening understanding at all levels. J. Exp. Zool. 293:264-283, 2002. r 2002 Wiley-Liss, Inc.

/pdf/na-cl-ca2-and-zn2-transport-by-fish-gills-retrospective-4d21bs1iq8.pdf

Na+, Cl−, Ca2+ and Zn2+ transport by fish gills: retrospective review and prospective synthesis

Compared to terrestrial animals, fish have to cope with more-challenging osmotic and ionic gradients from aquatic environments with diverse salinities, ion compositions, and pH values. Gills, a unique and highly studied organ in research on fish osmoregulation and ionoregulation, provide an excellent model to study the regulatory mechanisms of ion transport. The present review introduces and discusses some recent advances in relevant issues of teleost gill ion transport and functions of gill ionocytes. Based on accumulating evidence, a conclusive model of NaCl secretion in gills of euryhaline teleosts has been established. Interpretations of results of studies on freshwater fish gill Na+/Cl- uptake mechanisms are still being debated compared with those for NaCl secretion. Current models for Na+/Cl- uptake are proposed based on studies in traditionally used model species. Many reported inconsistencies are claimed to be due to differences among species, various experimental designs, or acclimation conditions. Having the benefit of advanced techniques in molecular/cellular biology, functional genomics, and model animals, several new notions have recently been raised concerning relevant issues of Na+/Cl- uptake pathways. Several new windows have been opened particularly in terms of molecular mechanisms of ionocyte differentiation and energy metabolite transport between gill cells during environmental challenge.

New insights into fish ion regulation and mitochondrion-rich cells

Calcium fluxes and internal calcium pools were measured in fed, rapidly growing, male tilapia, Oreochromis mossambicus , acclimated to 0.8 mmol l −1 (FW) and 0.2mmoll −1 (LFW) Ca 2+ . Plasma calcium levels were slightly and significantly higher in the LFW tilapia, but muscle calcium concentrations were independent of ambient Ca 2+ . At the time of the experiments, the LFW fish were growing and accumulated calcium, although the calcium content of their hard tissues was reduced. The LFW fish had higher Ca 2+ influx and efflux rates than the FW fish. The increase in the influx of Ca 2+ in LFW fish was, however, substantially greater than the increase in the efflux of Ca 2+ , giving these fish a more than four-fold increase in net Ca 2+ influx from the water: for a 20-g tilapia net uptake rates of Ca 2+ from the water were 390 and 1620 nmol Ca 2+ h −1 for FW- and LFW-adapted fish, respectively. These values were calculated to represent at least 69% of the total calcium accumulated by these growing fish. This indicates that even in low-calcium water, tilapia absorb a significant amount of their calcium requirement directly from the water. The pool of readily exchangeable calcium in the bone of FW fish was estimated to be about 7 % of the total hard tissue calcium. In the fish acclimated to LFW, this percentage increased to about 15 % as total hard tissue mineralization decreased. This may indicate that tilapia can increase the mobility of their hard tissue calcium during periods of low calcium stress.

/pdf/effects-of-low-ambient-calcium-levels-on-whole-body-ca2-flux-3kv9epoh37.pdf

Effects of low ambient calcium levels on whole-body Ca2+ flux rates and internal calcium pools in the freshwater cichlid teleost, Oreochromis mossambicus

Rainbow trout (Oncorhynchus mykiss) were exposed to ion-poor (soft) water to test the hypothesis that naturally induced proliferation of branchial chloride cells causes a thickening of the blood-to-water diffusion barrier. This was achieved by using a combination of scanning and transmission electron-microscopic techniques. Fish were exposed to soft-water conditions ([Na+]= 0.055 mmol l-1, [Cl–]≈0.029 mmol 1–1, [Ca2+]≈ 0.059 mmol 1–1, and [K+]≈0.007 mmol 1–1) for 1, 2, and 4 weeks. Marked chloride cell proliferation was evident at all sampling times with an approximate doubling of the gill epithelial surface area covered by chloride cells exposed to the water (”chloride cell fractional area”). The increases in chloride cell fractional area resulted from both increased numbers of cells and expanded apical surfaces of exposed individual cells. As a result of chloride cell proliferation, soft-water exposure was associated with a doubling of the lamellar blood-to-water diffusion distance from 3.26±0.08 μm to 6.58±0.43 μm as determined from transmission electron micrographs. These data demonstrated a positive correlation between chloride cell fractional area and blood-to-water diffusion distance. We conclude that, in trout, chloride cell proliferation during soft-water exposure, while presumably benefiting ionic regulation, may impair gas transfer owing to the associated thickening of the blood-to-water diffusion barrier.

The effects of soft-water acclimation on gill structure in the rainbow trout Oncorhynchus mykiss

1. 1. The characteristics of Ca2+-activated ATPase activities previously often postulated as components for the calcium transporting system in fish gills do not fulfil the requirements of a transport Ca2+-ATPase. 2. 2. The chelation of Ca2+- or Mg2+-ions is a prerequisite for the adenosinephosphate esters to serve as substrate for gill plasma membrane phosphatases. 3. 3. Ca2+-activated ATP hydrolysis results from the activity of a heterogeneous pool of phosphatases located in the plasma membranes of the branchial epithelium, as is concluded from substrate specificity tests and the effects of various inhibitors on these hydrolytic activities. 4. 4. In the present study only non-specific phosphatases could be shown.

/pdf/ca2-dependent-phosphatase-and-atpase-activities-in-eel-gill-ri0tuts2l0.pdf

Ca2+-dependent phosphatase and ATPase activities in eel gill plasma membranes—I. Identification of Ca2+-activated ATPase activities with non-specific phosphatase activities

The objectives of this study were to induce chloride cell (CC) proliferation on the gill lamellae of rainbow trout Oncorhynchus mykiss and to evaluate the consequences for respiratory function. Chronic elevation of hormone levels was used to induce CC proliferation; fish were injected with a combination of cortisol (8 mg kg-1 intramuscularly every day for 10 days) and ovine growth hormone (2 mg kg-1 intraperitoneally every second day for 10 days). The extent of CC proliferation was quantified using scanning electron microscopy and a two-dimensional analysis. An extracorporeal preparation in combination with environmental hypoxia was used to assess the effects of CC proliferation on respiratory function. Arterial blood was routed from the coeliac artery through an external circuit in which pH (pHa), partial pressure of oxygen (PaO2) and partial pressure of carbon dioxide (PaCO2) were monitored continuously. Environmental hypoxia was imposed by gassing a water equilibration column supplying the experimental chamber with N2. The hormone treatment increased the average CC surface area by 2.7-fold and CC density by 2.2-fold; the combined effect was a fivefold increase in CC fractional area. While the PaO2 values of hormone-treated and control fish were similar at PwO2>12.0 kPa, the arterial O2 tensions of treated fish were significantly lower than those of the control group for PwO2f12.0 kPa. In comparison with control fish at all environmental O2 tensions, the hormone-treated fish exhibited elevated PaCO2 values and a significant acidosis. The effects of CC proliferation on blood gas variables in hormone-treated fish were accompanied by a significantly elevated ventilation amplitude and a lowered ventilation frequency. The results of this study demonstrated (i) that impairment of respiratory gas transfer coincides with CC proliferation, (ii) that O2 and CO2 transfer are influenced differently and (iii) that partial compensation is achieved through physiological adjustments.

/pdf/the-effects-of-branchial-chloride-cell-proliferation-on-3ecjoaycgs.pdf

The effects of branchial chloride cell proliferation on respiratory function in the rainbow trout oncorhynchus mykiss

The effects of branchial chloride cell proliferation on ion transport capability and gill morphometry were evaluated in the rainbow trout, Oncorhynchus mykiss, to test the hypothesis that chloride cell (CC) proliferation benefits ionic regulation at the expense of efficient gas transfer. The extent of hormone-induced CC proliferation (using ovine growth hormone (oGH), cortisol, or a combination of both) on the gill filament epithelium was assessed by determining the fractional surface area of exposed cells using scanning electron microscopy. Cortisol and oGH were equally effective in increasing CC fractional surface area (~2×), owing to the enlargement of individual CCs. The combined cortisol/oGH treatment resulted in an even greater increase in CC fractional area (~6×), as both the size and number of CCs increased. Sham injections were without effect on CC surface area or number. Significant increases in Na+ (Jin Na+) and Cl− uptake (Jin Cl−) were observed after all hormone treatments and were correlated...

J.C. Fenwick

Papers

Effects of low ambient calcium levels on whole-body Ca2+ flux rates and internal calcium pools in the freshwater cichlid teleost, Oreochromis mossambicus

The effects of soft-water acclimation on gill structure in the rainbow trout Oncorhynchus mykiss

Ca2+-dependent phosphatase and ATPase activities in eel gill plasma membranes—I. Identification of Ca2+-activated ATPase activities with non-specific phosphatase activities

The effects of branchial chloride cell proliferation on respiratory function in the rainbow trout oncorhynchus mykiss

Branchial chloride cell proliferation in the rainbow trout, Oncorhynchus mykiss: implications for gas transfer