Dopaminergic regulation of ion transport in gills of the euryhaline semiterrestrial crab Chasmagnathus granulatus: Interaction between D1- and D2-like receptors

The effects of dopamine (DA) and dopaminergic agonists and antagonists on ion transport were studied in isolated perfused gills of the crab Chasmagnathus granulatus. DA applied under steady state conditions (perfusion with hemolymph-like saline) produced a transient increase of the transepithelial potential difference (Vtc) from 2.2±0.2 to 4.8±0.3 mV, describing an initial cAMP-dependent stimulating phase followed by an inhibitory phase. Spiperone and domperidone (antagonists of D2-like DA receptors in vertebrates) completely blocked the response to DA, while the D1-like antagonist SCH23390 blocked only the inhibitory phase. Theophylline (phosphodiesterase inhibitor) and okadaic acid (protein phosphatases PP1 and PP2A inhibitor) were also able to block the inhibitory phase, suggesting that it depends on adenylyl cyclase inhibition and on protein phosphatases. When the gills were perfused with hypoosmotic solution, or with the adenylyl cyclase activator forskolin, Vte was increased several-fold. DA applied under these stimulated conditions partially reversed the Vte increase by 54% and 25%, respectively. Similarly, the D1-like agonist, fenoldopam, produced a 33% reduction in the stimulated Vte. We propose that, in C. granulatus gills, DA stimulates adenylyl cyclase and therefore ion transport through D1-like receptors linked to a Gs protein, although they respond to antagonists that interact with D2-like receptors in vertebrates. The inhibitory phase seems to be mediated by D2-like receptors linked to a Gi/o protein, which inhibits adenylyl cyclase, although these receptors can be activated or blocked by agonists or antagonists that interact with D1-like receptors in vertebrates and insects.Fil:Genovese, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Ortiz, N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Regueira, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Tresguerres, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Luquet, C.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Induction of branchial ion transporter mRNA expression during acclimation to salinity change in the euryhaline crab Chasmagnathus granulatus

Using quantitative real-time PCR, the expression of mRNAs encoding three transport-related proteins and one putative housekeeping protein was analyzed in anterior and posterior gills of the euryhaline crab Chasmagnathus granulatus following transfer from isosmotic conditions (30‰ salinity) to either dilute (2‰) or concentrated (45‰) seawater. Modest changes were observed in the abundance of mRNAs encoding the housekeeping protein arginine kinase and the vacuolar-type H+-ATPase B-subunit, both of which were highly expressed under all conditions. By contrast, the expression of Na +/K+-ATPase α-subunit mRNA and Na+/K +/2Cl- cotransporter mRNA was strongly responsive to external salinity. During acclimation to dilute seawater, cotransporter mRNA increased 10-20-fold in posterior gills within the first 24 h while Na +/K+-ATPase α-subunit mRNA increased 35-55-fold. During acclimation to concentrated seawater, cotransporter mRNA increased 60-fold by 96 h and Na+/K+-ATPase α-subunit increased approximately 25-fold in posterior gills. Our results indicate a complex pattern of transcriptional regulation dependent upon the direction of salinity change and the developmental background of the gills.Fil:Luquet, C.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Dopaminergic regulation of ion transport in gills of the euryhaline semiterrestrial crab Chasmagnathus granulatus: Interaction between D1- and D2-like receptors

The effects of dopamine (DA) and dopaminergic agonists and antagonists on ion transport were studied in isolated perfused gills of the crab Chasmagnathus granulatus. DA applied under steady state conditions (perfusion with hemolymph-like saline) produced a transient increase of the transepithelial potential difference (V tc) from 2.2±0.2 to 4.8±0.3 mV, describing an initial cAMP-dependent stimulating phase followed by an inhibitory phase. Spiperone and domperidone (antagonists of D2-like DA receptors in vertebrates) completely blocked the response to DA, while the D1-like antagonist SCH23390 blocked only the inhibitory phase. Theophylline (phosphodiesterase inhibitor) and okadaic acid (protein phosphatases PP1 and PP2A inhibitor) were also able to block the inhibitory phase, suggesting that it depends on adenylyl cyclase inhibition and on protein phosphatases. When the gills were perfused with hypoosmotic solution, or with the adenylyl cyclase activator forskolin, V te was increased several-fold. DA applied under these stimulated conditions partially reversed the Vte increase by 54% and 25%, respectively. Similarly, the D1-like agonist, fenoldopam, produced a 33% reduction in the stimulated V te. We propose that, in C. granulatus gills, DA stimulates adenylyl cyclase and therefore ion transport through D1-like receptors linked to a Gs protein, although they respond to antagonists that interact with D2-like receptors in vertebrates. The inhibitory phase seems to be mediated by D2-like receptors linked to a Gi/o protein, which inhibits adenylyl cyclase, although these receptors can be activated or blocked by agonists or antagonists that interact with D1-like receptors in vertebrates and insects.Fil:Genovese, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Ortiz, N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Regueira, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Tresguerres, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Luquet, C.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina