BUSCADOR RECOLECTA

Peer reviewed

Code and preprocessed dataset used in the work "A data-driven approach for the assessment of the thermal stratification of reservoirs based on readily available data." Original data were obtained from the web page of the Ebro Automatic Water Quality Information System (SAICA Ebro) and the web page of the Ebro Automatic Hydrographic Information System (SAIH Ebro), in particular from the weather station of El Val (code EM71) and the monitoring station of the reservoir storage state (code E071). The data were preprocessed as explained in the article., Peer reviewed

Aquaporin-mediated oocyte hydration is a developmentally regulated adaptive mechanism that co-occurs with meiosis resumption in marine teleosts. It provides the early embryos with vital water until osmoregulatory systems develop, and in the majority of marine teleosts causes their eggs to float. Recent studies have shown that the subdomains of two water channels (Aqp1ab1 and Aqp1ab2) encoded in a teleost-specific aquaporin-1 cluster (TSA1C) co-evolved with duplicated Ywhaz-like (14-3-3ζ-like) binding proteins to differentially control their membrane trafficking for maximal egg hydration. Here, we report that in species that encode the full TSA1C, in-frame intronic splice variants of Aqp1ab1 result in truncated proteins that cause dominant-negative inhibition of the canonical channel trafficking to the plasma membrane. The inhibition likely occurs through hetero-oligomerization and retention in the endoplasmic reticulum (ER) and ultimate degradation. Conversely, in species that only encode the Aqp1ab2 channel we found an in-frame intronic splice variant that results in an intact protein with an extended extracellular loop E, and an out-of frame intronic splice variant with exon readthrough that results in a truncated protein. Both isoforms cause dominant-negative enhancement of the degradation pathway. However, the extended and truncated Aqp1ab2-type variants can also partially escape from the ER to reach the oocyte plasma membrane, where they dominantly-negatively inhibit water flux. The ovarian follicular expression ratios of the Aqp1ab2 isoforms in relation to the canonical channel are lowest during oocyte hydration, but subsequently highest when the canonical channel is recycled, thus leaving the eggs endowed with >90% water. These findings suggest that the expression of inhibitory isoforms of Aqp1ab1 and Aqp1ab2 may represent a new regulatory mechanism through which the cell-surface expression and the activity of the canonical channels can be physiologically modulated during oocyte hydration in marine teleosts., Peer reviewed

Peer reviewed

Oligonucleotide primers employed for the RT-PCR screening of splice forms expression of aqp1ab-type genes in teleosts., Peer reviewed

(A-C) Genomic organization of tandemly arranged aqp1ab2 and aqp1ab1 genes in seabream (A) and Atlantic halibut (B), and of the single aqp1ab2 gene in Senegalese sole and Common sole (C). The four exons (1–4) of both genes are represented by blue and red boxes, and the start codons are indicated by upper arrows. Lower arrows indicate the position of the oligonucleotide primers used for RT-PCR. (D-E) Representative RT-PCR analysis of aqp1ab1 and aqp1ab2 expression in different adult tissues from seabream (D) and halibut (E). A single aqp1ab2 mRNA species is expressed in seabream and halibut, whereas two or one splice variants (aqp1ab1_v1 and aqp1ab1_v2) in addition to aqp1ab1 are detected, respectively, as indicated. (F-G) RT-PCR of aqp1ab2 expression in Senegalese sole (F) and Common sole (G) where two splice forms (aqp1ab2_v1 and aqp1ab2 _v2) in addition to aqp1ab2 are amplified in the ovary and testis. In D-G, the sizes (kb) of molecular markers are indicated on the left., Peer reviewed

(A-C) Schematic diagram of the seabream and halibut aqp1ab1 and sole aqp1ab2 genes, and the different emerging transcript variants identified in each species. The numbers represent the exons, and the initiation and stop codons in each mRNA are indicated. (D-F) Integral membrane views (cartoon renders) of the wild type and variant channels. Canonical transmembrane domains are shown in cyan, loops in magenta and the two conserved Asn-Pro-Ala (NPA) motifs in dark blue. Variant regions are shown in red. In sole Aqp1ab2_v2 (C and F), the splicing of the distal region of intron 2 causes out-of-frame readthrough of exon 3 and the spliced exon 4 resulting in a premature stop codon and the truncated protein., Peer reviewed

(A-C) Pf of X. laevis oocytes injected with water (W, controls) or expressing seabream SaAqp1ab1-WT, SaAqp1ab1_v1 or SaAqp1ab1_v2 (A), Atlantic halibut HhAqp1ab1-WT or HhAqp1ab1_v1 (B), or sole SsAqp1ab2-WT, SsAqp1ab2_v1 or SsAqp1ab2_v2 (C). Oocytes injected with SsAqp1ab2-WT or splice forms were co-injected with halibut YwhazLb and exposed to FSK for 1 h prior to the swelling assay. The data are the mean ± SEM (n indicated above each bar) and were statistically analyzed by an unpaired Student’s t-test (***, P < 0.001; with respect to controls). (D-F) Double immunostaining of oocytes expressing untagged SaAqp1ab1-WT or HA-tagged SaAqp1ab1_v1 or HA-SaAqp1ab1_v2 (D), Flag-tagged HhAqp1ab1-WT or HA-tagged HhAqp1_v1 (E), or Flag-tagged SsAqp1ab2-WT or HA-tagged SsAqp1ab2_v1 or HA-SsAqp1ab2_v2 (F), and the ER marker protein disulfide isomerase (PDI). The plasma membrane is indicated by an arrowhead, whereas the co-localization of aquaporin channels and PDI in the cytoplasm is indicated by arrows. Scale bars, 10 μm (insets 5 μm)., Peer reviewed

(A-C) Changes in Pf of X. laevis oocytes injected with water or expressing SaAqp1ab1-WT (A), HhAqp1ab1-WT (B) or SsAqp1ab2-WT (C) alone or in combination with different amounts of the isoforms of each paralog. For SsAqp1ab2-WT and splice forms, oocytes were co-injected with halibut YwhazLb and exposed to FSK. (D) Effect of the inhibition of the oocyte Pf by the different Aqp1ab1 splice forms, at a concentrations that produced approximately half-maximal reduction (as observed in A-C), in SaAqp1ab1 or HhAqp1ab1 expressing oocytes in the presence or absence of YwhazLa, and exposed to FSK. The percentage of Pf inhibition elicited by each isoform under the two conditions is indicated in each plot. In all panels, data are the mean ± SEM (n indicated above each bar) and were statistically analyzed by an unpaired Student’s t-test (*, P < 0.05: **, P < 0.01; ***, P < 0.001; with respect to oocytes injected with the WT form alone)., Peer reviewed

Figure S1: UV-VIS spectra of the SERS substrates herein studied: Glass_ Ag, Glass_AuNPs, Glass_Ag_AuNPs, 1G_Ag_AuNPs, 3G_Ag_AuNPs; Figure S2: SERS spectra of 6 spots recorded on bottom and top of 1G_Ag_AuNPs sample upon exposure to 1.2 ppmV DMMP in gas phase., Peer reviewed