Cytoplasmic carbonic anhydrase isozymes in rainbow trout<i>Oncorhynchus mykiss</i>: comparative physiology and molecular evolution

Esbaugh, Andrew J.; Perry, Steve F.; Bayaa, M.; Georgalis, Tina; Nickerson, James G.; Tufts, Bruce L.; Gilmour, K. M.

doi:10.1242/jeb.01551

Cited by 68 publications

(60 citation statements)

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“…Rahim et al (1988) also provided immunological evidence for the presence of distinct branchial and blood CA isoforms in rainbow trout and carp, a finding that was recently confirmed by the cloning of rainbow trout blood (tCAb) and cytosolic (tCAc) CA isoforms (Esbaugh et al, 2004(Esbaugh et al, , 2005. Northern and real-time PCR analyses identified the gills as a site of considerable tCAc mRNA expression (Esbaugh et al, 2005).…”

Section: Introductionmentioning

confidence: 79%

“…The annealing and extension temperatures over 40 cycles were 58°C (30·s) and 72°C (30·s), respectively. The primer pairs used were those designed and reported by Esbaugh et al (2005): ␤-actin forward 5Ј-CCA ACA GAT GTG GAT CAG CAA-3Ј, ␤-actin reverse 5Ј-GGT GGC ACA GAG CTG AAG TGG TA-3Ј, tCAc forward 5Ј-CAG TCT CCC ATT GAC ATC GTA-3Ј, and tCAc reverse 5Ј-CGT TGT CGT CGG TGT AGG T-3Ј. The specificity of the primers was verified by the cloning (TOPO TA cloning kit; Invitrogen, Burlington, ON, Canada) and sequencing of amplified products.…”

Section: Analysis Of Tcac Mrna Expressionmentioning

confidence: 99%

“…10-12·h after the onset of hypercarbia. The acetazolamide dose (30·mg·kg -1 ; delivered as a bolus injection in 1·ml of saline via the dorsal aortic cannula) was calculated so as to elicit an initial circulating concentration of ~450·mol·l -1 , which would be expected to fully inhibit branchial tCAc activity, as the inhibition constant for Az against tCAc is ~1·nmol·l -1 (Esbaugh et al, 2005). An arterial blood sample (600·l) was withdrawn via the dorsal aortic cannula prior to the initiation of hypercarbia and at the beginning of each measurement period; any red cells not used for analysis were resuspended in saline and returned to the fish.…”

Section: Series Ii: Determination Of the Role Of Branchial Ca Activitmentioning

confidence: 99%

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The role of branchial carbonic anhydrase in acid-base regulation in rainbow trout (Oncorhynchus mykiss)

Georgalis

Perry

Gilmour

2006

Journal of Experimental Biology

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SUMMARY The objective of the present study was to examine the branchial distribution of the recently identified rainbow trout cytoplasmic carbonic anhydrase isoform (tCAc) and to investigate its role in the regulation of acid-base disturbances in rainbow trout (Oncorhynchus mykiss). In situ hybridization using an oligonucleotide probe specific to tCAc revealed tCAc mRNA expression in both pavement cells and mitochondria-rich cells (chloride cells). Similarly, using a homologous polyclonal antibody,tCAc immunoreactivity was localized to pavement cells and mitochondria-rich cells in the interlamellar region and along the lamellae of the gills. Exposure of rainbow trout to hypercarbia (∼0.8% CO2) for 24 h resulted in significant increases in tCAc mRNA expression (∼20-fold;quantified by real-time PCR) and protein levels (∼1.3-fold; quantified by western analysis) but not enzyme activity (assessed on crude gill homogenates using the delta-pH CA assay). Inhibition of branchial CA activity in vivo using acetazolamide reduced branchial net acid excretion significantly by 20%. This effect was enhanced to a 36% reduction in branchial net acid excretion by subjecting the trout to hypercarbia (∼0.8%CO2) for 10 h prior to acetazolamide injection, an exposure that significantly increased branchial net acid excretion. The results of the present study support the widely held premise that branchial intracellular CA activity (tCAc) plays a key role in regulating acid-base balance in freshwater teleost fish.

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Section: Introductionmentioning

confidence: 79%

Section: Analysis Of Tcac Mrna Expressionmentioning

confidence: 99%

Section: Series Ii: Determination Of the Role Of Branchial Ca Activitmentioning

confidence: 99%

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The role of branchial carbonic anhydrase in acid-base regulation in rainbow trout (Oncorhynchus mykiss)

Georgalis

Perry

Gilmour

2006

Journal of Experimental Biology

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“…Besides, in vivostudies of deltametrinewere performed on CA activity of rainbow trout gill. Fish gill CA was significantly inhibited at three concentrations (0.25, 1.0 and 2.5 μg/L) at 24 and 48 h (Ceyhun et al, 2010). Söyüt and Beydemir (2011) …”

Section: Discussionmentioning

confidence: 98%

“…Up to now, CA has been purified from many different tissues including human erythrocytes (Bayram et al, 2008;Şentürk et al, 2009), fish gills (Bone et al, 1995;Skaggs and Henry, 2002;Hisar et al, 2004;Ceyhun et al, 2010), fish erythrocytes (Bülbül et al, 2003;Hisar et al, 2005;Doğan, 2006), rainbow trout brain and liver . Although, CA and the inhibitory effects of many metals on CA have been studied in most fish tissues, no study has been done on C. umbla gill CA, yet.…”

Section: Discussionmentioning

confidence: 99%

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Kırıcı¹,

Kırıcı²,

Beydemır³

et al. 2016

Turk. J. Fish. Aquat. Sci.

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In this study, in vitroeffects of some metal ions (Fe 3+ , Cd 2+ , Pb 2+ and Ni 2+ )on cytoplasmic carbonic anhydrase(CA, EC 4.2.1.1) from Capoeta umbla gill was investigated. CA was purified from the gills of C. umbla for the first time. It was purified with the Sepharose-4B-L-Tyrosine Sulphanilamide affinity chromatography method. The overall purification was approx. 31.69-fold with a yield of 53.33%, and a specific activity of 326.73 EU/mg proteins. Sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDS-PAGE) showed a single band corresponding to a molecular weight of approx. 29 kDa. ve Ni +2 ) in vitro etkileri incelendi. Karbonik anhidraz enzimi C. umbla solungaç dokusundan ilk kez saflaştırıldı. Enzim, Sepharose 4B-L-tirozin-sulfanilamid afinite kromatografisi yöntemi ile saflaştırıldı. Bu yöntem sonucunda enzim, spesifik aktivitesi 326,73 EÜ/mg protein ve %53,3 verimle yaklaşık 31,69 kat olarak saflaştırıldı. Sodyum dodesil sülfat-poliakrilamid jel elektroforezinde (SDS-PAGE) yaklaşık 29 kDa'da tek bant olarak görüldü. Metal iyonları için enzim inhibitör kompleks sabitleri (Ki) ve %50 inhibitör değerleri (IC50), sırasıyla Lineweaver-Burk grafikleri ve % aktivite-[I] grafikleri çizilerek hesaplandı. Ki sabitleri ve IC50 değerleri, Fe +3 için 0,012±0,0135 ve 0,136 mM, Cd +2 için 0,019±0,0113 ve 0,191 mM, Pb +2 için 0,041±0,0075 ve 0,289 mM ve Ni +2 için 0,120±0,034 ve 0,924 mM olarak belirlendi. Fe +3 , Cd +2 ve Pb +2 iyonları enzimi, yarışmalı olarak inhibe ederken, Ni +2 iyonu yarışmasız olarak inhibe etmiştir. Sonuç olarak, Fe +3 iyonunun C. umbla solungaç karbonik anhidraz enzimi için güçlü bir inhibitör olduğu belirlendi.Anahtar Kelimeler: Capoeta umbla, karbonik anhidraz, solungaç, metal toksisitesi.

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Toxicological perspective on the osmoregulation and ionoregulation physiology of major ions by freshwater animals: Teleost fish, crustacea, aquatic insects, and Mollusca

Griffith

2016

Enviro Toxic and Chemistry

141

136

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Anthropogenic sources increase freshwater salinity and produce differences in constituent ions compared with natural waters. Moreover, ions differ in physiological roles and concentrations in intracellular and extracellular fluids. Four freshwater taxa groups are compared, to investigate similarities and differences in ion transport processes and what ion transport mechanisms suggest about the toxicity of these or other ions in freshwater. Although differences exist, many ion transporters are functionally similar and may belong to evolutionarily conserved protein families. For example, the Na+/H+-exchanger in teleost fish differs from the H+/2Na+ (or Ca2+)-exchanger in crustaceans. In osmoregulation, Na+ and Cl− predominate. Stenohaline freshwater animals hyperregulate until they are no longer able to maintain hypertonic extracellular Na+ and Cl− concentrations with increasing salinity and become isotonic. Toxic effects of K+ are related to ionoregulation and volume regulation. The ionic balance between intracellular and extracellular fluids is maintained by Na+/K+-adenosine triphosphatase (ATPase), but details are lacking on apical K+ transporters. Elevated H+ affects the maintenance of internal Na+ by Na+/H+ exchange; elevated HCO3− inhibits Cl− uptake. The uptake of Mg2+ occurs by the gills or intestine, but details are lacking on Mg2+ transporters. In unionid gills, SO42− is actively transported, but most epithelia are generally impermeant to SO42−. Transporters of Ca2+ maintain homeostasis of dissolved Ca2+. More integration of physiology with toxicology is needed to fully understand freshwater ion effects.

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Cytoplasmic carbonic anhydrase isozymes in rainbow troutOncorhynchus mykiss: comparative physiology and molecular evolution

Cited by 68 publications

References 74 publications

The role of branchial carbonic anhydrase in acid-base regulation in rainbow trout (Oncorhynchus mykiss)

The role of branchial carbonic anhydrase in acid-base regulation in rainbow trout (Oncorhynchus mykiss)

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Toxicological perspective on the osmoregulation and ionoregulation physiology of major ions by freshwater animals: Teleost fish, crustacea, aquatic insects, and Mollusca

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