TY - JOUR T1 - Ontogeny of osmoregulation in the Pacific blue shrimp, Litopenaeus stylirostris (decapoda, penaeidae): Deciphering the role of the Na+/K+-ATPase A1 - Pham,Dominique A1 - Charmantier,Guy A1 - Boulo,Viviane A1 - Wabete,Nelly A1 - Ansquer,Dominique A1 - Dauga,Clement A1 - Grousset,Evelyse A1 - Labreuche,Yannick A1 - Charmantier-Daures,Mireille AD - IFREMER, Lagon, Environm & Aquaculture Durable, Boulouparis, Nouvelle Caledo, France. AD - Univ Montpellier, IFREMER, Adaptat Ecophysiol & Ontogenese, Marbec,UM,CNRS,IRD,UMR 9190, Pl Eugene Bataillon,CC 092, F-34095 Montpellier 05, France. AD - IFREMER, Unite Physiol Fonct Organismes Marins, ZI Pointe Diable, CS 10070, F-29280 Plouzane, France. AD - Univ Paris 06, Univ Paris 04, Stn Biol Roscoff, Integrat Biol Marine Models,CNRS,UMR 8227, CS 90074, F-29688 Roscoff, France. UR - https://doi.org/10.1016/j.cbpb.2015.12.007 DO - 10.1016/j.cbpb.2015.12.007 KW - Penaeid KW - Litopenaeus stylirostris KW - Ontogeny KW - Osmoregulation KW - Na+/K+-ATPase KW - RNA interference N2 - The role of the main ion transporting enzyme Na +/K + − ATPase in osmoregulation processes was investigated in Litopenaeus stylirostris. The development and localization of the osmoregulation sites were studied during ontogenesis by immunodetection of Na+ K+-ATPase using monoclonal antibodies and transmission electron microscopy (TEM). Osmoregulation sites were identified as the pleurae and branchiostegites in the zoeae and mysis stages. In the subsequent post-metamorphic stages the osmoregulatory function was mainly located in the epipodites and branchiostegites and osmotic regulation was later detected in the gills. The presence of ionocytes and microvilli in these tissues confirmed their role in ionic processes. The complete open reading frame of the mRNA coding for the α-subunit of Na + K + − ATPase was characterized in L. stylirostris. The resulting 3092-bp cDNA (LsNKA) encodes a putative 1011-amino-acid protein with a predicted molecular mass of 112.3 kDa. The inferred amino acid sequence revealed that the putative protein possesses the main structural characteristics of the Na + K + − ATPase α-subunits. Quantitative RT-PCR analyses indicated that LsNKA transcripts did not significantly vary between the different developmental stages. The number of transcripts was about 2.5-fold higher in the epipodites and gills than in any other tissues tested in juveniles. A reverse genetic approach was finally implemented to study the role of LsNKA in vivo. Knock down of LsNKA expression by gene-specific dsRNA injection led to an increase of shrimp mortality following an abrupt salinity change compared to control animals. These data strongly suggest that LsNKA plays an important role in osmoregulation when the shrimp are challenged by changing salinities. Y1 - 2016/06 PB - Elsevier Science Inc JF - Comparative Biochemistry And Physiology B-biochemistry & Molecular Biology SN - 1096-4959 VL - 196 SP - 27 EP - 37 ID - 42176 ER -