RESUMEN
Stenohaline freshwater stingrays (Potamotrygon spp.) are endemic to the very dilute (Na(+), Cl(-), Ca2(+)
Asunto(s)
Calcio/metabolismo , Agua Dulce/química , Rajidae/fisiología , Sodio/metabolismo , Amoníaco/metabolismo , Animales , Brasil , Calcio/análisis , Sustancias Húmicas/metabolismo , Concentración de Iones de Hidrógeno , Iones/análisis , Iones/metabolismo , Cinética , Rajidae/metabolismo , Sodio/análisisRESUMEN
We examined the ion composition of mosquito breeding sites located in the Amazon rain forest and the ion regulatory patterns of larvae from these habitats. We found larvae of Toxorhynchites haemorroidalis, Limatus durhamii, Culex (Carrollia) bonnei, and Culex (Culex) sp. residing in fallen palm bracts, leaves, and tree holes that were filled with water. These breeding sites had micromolar levels of Na(+) (1.6-99 micromol L(-1)), but K(+) and Cl(-) concentrations were higher and varied over a large range (231-17,615 micromol L(-1) K(+); 355-2,700 micromol L(-1) Cl(-)). Despite the variability in environmental ion levels and ratios, all four species maintain high hemolymph NaCl levels (80-120 mmol L(-1) Na(+); 60-80 mmol L(-1) Cl(-)). However, the species differed in the means by which they maintain hemolymph ion balance, as indicated by the range of unidirectional Na(+) and Cl(-) uptake rates. Toxorhynchites haemorroidalis had extremely low rates of Na(+) uptake and undetectable Cl(-) uptake, whereas L. durhamii had high rates of uptake for both ions. This variability in rates of uptake may reflect species differences in rates of diffusive ion loss (i.e., permeability). We observed the same curious pattern of Na(+) inhibition and Cl(-) stimulation by low-pH exposure in all four species of mosquitoes, as has been documented in other mosquitoes and aquatic insects. Kinetic analyses of Na(+) and Cl(-) uptake in C. bonnei larvae revealed an unusual pattern of Na(+) uptake that increases linearly (nonsaturable) to extremely high rates, while Cl(-) uptake is a low-affinity, low-capacity system. This pattern contrasts with L. durhamii and Culex (Culex) sp. larvae, which had large increases in both Na(+) and Cl(-) uptake when external NaCl levels were increased. Our results suggest that although these rain forest mosquito larvae are residing in habitats with similar low Na(+), high Cl(-) composition and maintain similar hemolymph NaCl levels, the underlying mechanisms of ion regulation differ among the species.
Asunto(s)
Cloruros/metabolismo , Culicidae/metabolismo , Larva/metabolismo , Sodio/metabolismo , Clima Tropical , Equilibrio Hidroelectrolítico , Adaptación Fisiológica , Animales , Agua Corporal/metabolismo , Brasil , Hemolinfa/química , Hemolinfa/metabolismo , Concentración de Iones de Hidrógeno , Transporte Iónico , Especificidad de la Especie , ÁrbolesRESUMEN
This study is the first step in characterizing ion uptake mechanisms of mosquito larvae from the Amazon region of Brazil. Hemolymph NaCl levels and rates of unidirectional Na(+) and Cl(-) uptake were measured in larvae of Aedes aegypti and Culex quinquefasciatus in a series of environmental manipulations that are known to challenge ion regulation in other aquatic animals. Despite being reared for numerous generations in dilute media (20 micromol L(-1) NaCl), both species were able to maintain high hemolymph NaCl concentrations, a departure from previous studies. Exposure to distilled water or high-NaCl media did not affect hemolymph ion levels, but pH 3 caused significant decreases in hemolymph Na(+) and Cl(-) levels in both species. Exposure to water from Rio Negro (pH 5.5), an organically rich but ion-poor body of water, did not disturb hemolymph Na(+) and Cl(-) levels or the uptake of these ions. Acute exposure to control media or Rio Negro water titrated to pH 3.5 caused inhibition of Na(+) uptake and stimulation of Cl(-) uptake in C. quinquefasciatus, but A. aegypti larvae experienced only a significant reduction of Na(+) uptake in Rio Negro/pH 3.5 treatment. The stimulation of Cl(-) uptake at low pH has been documented only in aquatic insects and differs from all other invertebrate and vertebrate species. A similar pattern of Na(+) uptake inhibition and Cl(-) uptake stimulation was observed in A. aegypti larvae exposed to bafilomycin A(1), a blocker of V-type H(+) ATPase. Culex quinquefasciatus larvae were unaffected by this drug. Both Na(+) and Cl(-) uptake were reduced when C. quinquefasciatus larvae were exposed to acetazolamide, indicating that H(+) and HCO(3)(-), derived from hydration of CO(2), are involved with Na(+) and Cl(-) uptake. Kinetic analysis of Na(+) and Cl(-) uptake in C. quinquefasciatus, A. aegypti, and Anopheles nuneztovari larvae indicate that these Amazonian species share similar high-capacity and high-affinity mechanisms. Comparison of the Amazonian C. quinquefasciatus with a Californian population provided evidence of both phenotypic plasticity and population disparity in Na(+) and Cl(-) uptake, respectively. When the California population of C. quinquefasciatus was reared in a medium similar to that of the Amazonian group (60 micromol L(-1) NaCl) instead of 4,000 micromol L(-1) NaCl, larvae increased both Na(+) uptake capacity (J(max)) and affinity (i.e., reduced K(m)), yet Cl(-) uptake did not change from its nonsaturating, low-capacity pattern. In the reverse experiment, Amazonian C. quinquefasciatus demonstrated plasticity in both Na(+) and Cl(-) uptake by significantly reducing rates when held in 4,000 micromol L(-1) NaCl for 3 d.
Asunto(s)
Cloruros/metabolismo , Culicidae/metabolismo , Larva/metabolismo , Sodio/metabolismo , Clima Tropical , Equilibrio Hidroelectrolítico , Adaptación Fisiológica , Animales , Agua Corporal/metabolismo , Brasil , Hemolinfa/química , Hemolinfa/metabolismo , Concentración de Iones de Hidrógeno , Transporte Iónico/efectos de los fármacos , Cinética , Fenotipo , Especificidad de la EspecieRESUMEN
Stingrays of the family Potamotrygonidae are the only stenohaline freshwater elasmobranchs. Potomotrygon sp. collected from the ion-poor blackwaters ([Na(+)], [Cl(-)] and [Ca(2+)]=10-30 micro mol l(-1), pH 6.1) of the Rio Negro, Amazonas, Brazil, were ammoniotelic (91% ammonia-N, 9% urea-N excretion) and exhibited blood chemistry (Na(+), Cl(-), urea, ammonia and glucose levels and osmolality) typical of freshwater teleosts. Unidirectional Na(+) and Cl(-) influx rates, measured with radiotracers, displayed saturation kinetics. The relationships for Cl(-) and Na(+) had similar K(m) values (300-500 micro mol l(-1)), but J(max) values for Cl(-) (approximately 950 micro mol kg(-1) h(-1)) were almost twice those for Na(+) (approximately 500 micro mol kg(-1) h(-1)). Cl(-) efflux rates varied with external concentration, but Na(+) efflux rates did not. There were no differences in the kinetic variables (K(m), J(max)) for influx between animals acclimated to their native ion-poor blackwater or to ion-rich hard water, but efflux rates for both Na(+) and Cl(-) were lower in the former, yielding much lower balance points (external Na(+) or Cl(-) levels at which influx and efflux were equal). Na(+), Cl(-) and Ca(2+) uptake were all strongly inhibited by acute exposure to pH 4.0, but efflux rates and Ca(2+) binding to the body surface did not change. Na(+) influx was inhibited by amiloride (10(-4) mol l(-1)) and by two of its analogs, phenamil (4 x 10(-5) mol l(-1)) and HMA (4 x 10(-5) mol l(-1)), with the latter being slightly more potent, while Cl(-) fluxes were unaffected. Cl(-) fluxes were insensitive to DIDS (2 x 10(-5) mol l(-1) or 10(-4) mol l(-1)) and SITS (10(-4) mol l(-1)), but both influx and efflux rates were strongly inhibited by DPC (10(-4) mol l(-1)) and thiocyanate (10(-4) mol l(-1)). Ammonia excretion was unresponsive to large changes in water Na(+) concentration, but was elevated by 70% during acute exposure to pH 4.0 and transiently inhibited by approximately 50% by amiloride and its analogues. The strategy of adaptation to ion-poor blackwater appears similar to that of some Rio Negro teleosts (Cichlidae) in which low-affinity transport systems are relatively sensitive to inhibition by low pH but are complemented by low diffusive loss rates. Ionic transport systems in these freshwater elasmobranchs, although superficially similar to those in some freshwater teleosts, may bear more resemblance to their presumed evolutionary precursors in marine elasmobranchs.