Effects of injection of biogenic amines on expression of gill related ion transporter mRNA and alpha-subunit protein in Litopenaeus vannamei.
We used semi-quantitative reverse transcription and the polymerase chain reaction (RT-PCR) to identify and characterize the mRNA of Na(+)/K(+)-ATPase alpha-subunit and Vacuolar-type H/-ATPase (V-ATPase) beta-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 alpha-subunit mRNA in the first 12 h. The degree of change in Na(+)/K(+)-ATPase alpha-subunit mRNA expression were different between DA and 5-HT treated groups which depend on the dose. The expression of V-ATPase beta-subunit mRNA was not markedly affected by the biogenic amines. Concomitant elevation of Na(+)/K(+)-ATPase activity, alpha-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.
Liu
Hong-Yu
HY
The Key Laboratory of Mariculture of Chinese, Ministry of Education, Ocean University of China, Qingdao 266003, China.
Pan
Lu-Qing
LQ
Zheng
De-Bin
DB
D000818:Animals; D004298:Dopamine / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D005880:Gills / Q000187:drug effects / Q000378:metabolism*; D033561:Penaeidae / Q000235:genetics; D021122:Protein Subunits / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D012701:Serotonin / Q000494:pharmacology*; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics*; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics*
D021122:Protein Subunits; D012333:RNA, Messenger; D012701:Serotonin; D025262:Vacuolar Proton-Translocating ATPases; D000254:Sodium-Potassium-Exchanging ATPase; D004298:Dopamine