In all cases this signal was less intense than that in the PR. An. albimanus: saline tolerant anopheline Protein localization in the recta of An. albimanus was identical to that in An. gambiae when larvae were reared in freshwater: Na K ATPase appeared to be restricted to the basal infoldings of the non DAR cells ; CA9 protein was evident only in the cytoplasm of the DAR cells ; V ATPase localized to the apical infoldings of the non DAR cells and the cytoplasm of the DAR cells . This apparent cytoplasmic localization is better seen at a higher magnification in Fig. 1I. The localization pattern of all three proteins was identical in another saline tolerant anopheline species, An. farauti, when reared in freshwater . The distributions of CA9 and V ATPase proteins remained unchanged in larvae reared in 50% ASW compared with those reared in freshwater, but Na K ATPase underwent a dramatic shift and appeared to localize mainly to the basal infoldings of the DAR cells , presenting a drastic increase of this protein in the DAR cells and a contrasting reduction in the non DAR cells . The change in Na K ATPase distribution is shown graphically in figure 1L.
When reared in freshwater, Na K ATPase 50% ASW, it was significantly STAT inhibitors greater in the DAR cells. To determine if the Na K ATPase protein shift is a reversible event, larvae were reared in either freshwater or 25% ASW to 2nd, 3rd, or 4th instar and were transferred to 25% ASW or freshwater, respectively for 24, 48 or 72 hours. For each image in Fig. 3, the data are presented graphically as the ratio of Na K ATPase peak pixel intensity in the DAR cells versus the non DAR cells . Lowercase bar labels in Fig. 3H correspond to the experimental group indicated by the uppercase letters . When larvae were reared in freshwater and exposed briefly to 25% ASW, the ability for larvae to shift rectal Na K ATPase localization depended on the larval stage at which the exposure occurred. If exposed to ASW during the 2nd or 3rd instar stages, a change in Na K ATPase peak signal intensity from the non DAR cells to the DAR cells was evident within 24 hours .
Fourth instar larvae exposed only for 24 hours expressed Na K ATPase in Sodium valproate both DAR and non DAR cells, as if in an intermediate stage . However, a change in Na K ATPase localization from the non DAR cells to DAR cells was evident after 48 hours . In most cases, Na K ATPase shift was most dramatic in larvae exposed during the 2nd larval stage. Most 3rd and 4th instar larvae retained some Na K ATPase signal in the non DAR after exposure to 25% ASW. Slightly different results were found for larvae reared in 25% ASW and exposed to freshwater . While 2nd instar larvae shifted Na K ATPase localization from DAR to non DAR cells within 24 hours , 3rd instar and 4thinstar larvae did not fully shift Na K ATPase localization after 72 hours or 48 hours, respectively, and expressed the protein in both DAR and non DAR cells.