Effects of injection of biogenic amines on expression of gill related ion transporter mRNA and α-subunit protein in Litopenaeus vannamei

Abstract

We used semi-quantitative reverse transcription and the polymerase chain reaction (RT-PCR) to identify and characterize the mRNA of Na +/K +-ATPase α-subunit and Vacuolar-type H/-ATPase (V-ATPase) β-subunit prepared from gills of the white shrimp Litopenaeus vannamei. Western blot and immunocytochemistry analysis were employeed to investigate the effects of biogenic amines such as dopamine (DA) and 5-hydroxytryptamine (5-HT) injection on expression of protein abundance and immunostained sites. Our results show that the gill Na +/K +-ATPase of 5-HT treatments increased lag to those of DA treatments. However, the DA and 5-HT have no significant effects on the activity of V-ATPase. The biogenic amines could induce the synthesis of Na +/K +-ATPase α-subunit mRNA in the first 12 h. The degree of change in Na +/K +-ATPase α-subunit mRNA expression were different between DA and 5-HT treated groups which depend on the dose. The expression of V-ATPase β-subunit mRNA was not markedly affected by the biogenic amines. Concomitant elevation of Na +/K +-ATPase activity, α-subunit mRNA as well as protein abundance in response to injected biogenic amines was also found in Na +/K +-ATPase immunostained sites within the gill epithelium.