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Antarctic icefish is the only known vertebrate species that lacks oxygen-carrying hemoglobin and functional erythrocytes. To reveal the unique hematopoietic process of icefish, we used an integrated approach including tandem mass tag (TMT) labeling and liquid chromatography–tandem mass spectrometry (LC–MS/MS) to quantify the dynamic changes in the head kidney whole proteome of a white-blooded icefish, Chionodraco hamatus, compared to those in two other red-blooded Antarctic fish, Trematomus bernacchii and Notothenia coriiceps. Of the 4672 identified proteins, in the Antarctic icefish head kidney, 123 proteins were significantly upregulated and 95 proteins were downregulated. The functional grouping of differentially expressed proteins (DEPs) based on KEGG pathway analysis shows that white blood fish and red blood fish have significant differences in erythropoiesis, heme biogenesis, leucocyte and platelet cell development. The proteins involved in the hematopoietic process in icefish showed a clear trend of downregulation of erythroid lineage marker proteins (β-Spectrin, hemoglobin, CTCF and tfr1a) and upregulation of lymphoid and megakaryocytic lineage marker proteins, including CD9, ITGB2, and mTOR, which suggests a shift in hematopoiesis in the icefish head kidney due to the loss of erythrocytes. Moreover, western blotting analysis showed that a significantly down expression level of CTCF protein was detected in the icefish head kidney, which was in line with the proteomics data. The results of the present study not only provide basic datasets for the head kidney proteins of Antarctic fishes, but also provide important references for studies on immunity and hematopoiesis in various species. [ABSTRACT FROM AUTHOR]

The identification of loci under selection (outliers) is a major challenge in evolutionary biology, being critical to comprehend evolutionary processes leading to population differentiation and speciation, and for conservation purposes, also in light of recent climate change. However, detection of selected loci can be difficult when populations are weakly differentiated. This is the case of marine fish populations, often characterized by high levels of gene flow and connectivity, and particularly of fish living in the Antarctic marine environment, characterized by a complex and strong circulating system promoting individual dispersal all around the continent. With the final aim of identifying outlier loci putatively under selection in the Chionodraco genus, we used 21 microsatellites, including both genomic (Type II) and EST-linked loci (Type I), to investigate the genetic differentiation among the three recently derived Chionodraco species that are endemic to the freezing Antarctic waters. Neutrality tests were applied in interspecific comparisons in order to identify candidate loci showing high levels of genetic differentiation, which might reveal imprints of past selection. Three outlier loci were identified, detecting a higher differentiation between species than did neutral loci. Outliers showed sequence similarity to a calmodulin gene, to an antifreeze glycoprotein/trypsinogen-like protease gene and to nonannotated fish mRNAs. Selective pressures acting on outlier loci identified in this study might reflect past evolutionary processes, which led to species divergence and local adaptation in the Chionodraco genus. Used loci will provide a valuable tool for future population genetic studies in Antarctic notothenioids. [ABSTRACT FROM AUTHOR]

ntarctic icefishes (Family: Channichthyidae) are the only adult vertebrate that do not express the circulatory oxygen-binding protein, hemoglobin. Six species of icefishes, including Dacodraco hunteri, also do not express myoglobin (Mb), an oxygen-binding protein found in ventricular tissue of Antarctic notothenioids. Sequence of the Mb gene from D. hunteri contains a duplicated TATA box in the promoter region of the Mb gene that is absent in Chionodraco rastrospinosus, an icefish species known to express Mb. The 15-base pair insertion in D. hunteri is identical to the duplicated TATA box that is thought to be directly responsible for preventing transcription of the Mb gene in C. aceratus, another icefish species that lacks Mb expression. Sequencing of the Mb promoter from the remaining channichthyid species revealed the duplicated TATA box in 14 of 16 icefish species, including 8 species known to express Mb. Based on the presence of the duplicated TATA box in promoters of both Mb-expressing and Mb-lacking species, loss of Mb expression in D. hunteri and C. aceratus is either independent of the duplicated TATA box, or the duplicated TATA box requires presence or absence of a distant regulatory element to prevent transcription. [ABSTRACT FROM AUTHOR]