Francisca Fábrega-Guerén

Previously, we reported that mating induces an early transcriptional response in the oviductal mucosa of rats. The functional category ‘cell-to-cell signaling and interaction’ was overrepresented in this gene list. Therefore, in the present study, we describe the role of one of these genes, carbohydrate sulfotransferase 10 (Chst10), in the oviductal mucosa. CHST10 participates in the synthesis of the carbohydrate moiety human natural killer-1 (HNK-1), which mediates cell-to-cell interactions. When using one-dimensional Western blot and sulfotransferase analyses, we found that mating increased the protein level and activity of CHST10 in the oviductal mucosa at 3 h after stimulation. A two-dimensional Western blot analysis and mass spectrometry were used to identify the novel HNK-1 glycoproteins aldehyde dehydrogenase 9 family, member A1 (ALDH9A1), fructose bisphosphate aldolase A (ALDOA), and four and a half LIM domains protein 1 (FHL1) in the oviductal mucosa, and we found that mating induces the synthesis of their acidic variants. Interestingly, in the utero-tubal junction (UTJ), acrosome-reacted sperm apparently were interacting with regions in which ALDH9A1 and HNK-1 signals overlap. Furthermore, vaginocervical stimulation applied to unmated rats increased the mRNA level of Chst10 in the oviductal mucosa. In conclusion, mating increases the activity of CHST10 in the oviductal mucosa, which in turn induces the synthesis of acidic variants of ALDH9A1 and FHL1 via HNK-1 glycosylation. ALDH9A1, HNK-1-ALDH9A1, and/or other HNK-1 glycoproteins could participate in the negative selection of sperm in the UTJ, since we detected acrosome-reacted sperm apparently interacting with regions where these proteins are located. Finally, the sensorial component of mating could regulate early events (e.g., sperm transport and selection) occurring in the oviductal mucosa after mating.

Previously, we reported that mating induces an early transcriptional response in the oviductal mucosa of rats. The functional category ‘cell-to-cell signaling and interaction’ was overrepresented in this gene list. Therefore, in the present study, we describe the role of one of these genes, carbohydrate sulfotransferase 10 (Chst10), in the oviductal mucosa. CHST10 participates in the synthesis of the carbohydrate moiety HNK-1, which mediates cell-to-cell interactions. Using 1D western blot and sulfotransferase assays, we showed that mating increased the protein level and activity of CHST10 in the oviductal mucosa at 3 h after stimulation. A 2D western blot assay and mass spectrometry were used to identify the novel HNK-1 glycoproteins ALDH9A1, ALDOA and FHL1 in the oviductal mucosa, and we showed that mating induces the synthesis of their acidic variants. Interestingly, in the utero-tubal junction (UTJ), acrosome-reacted sperm are in close contact with HNK-1-ALDH9A1 regions, and acrosome-intact sperm are in close contact with other regions in which HNK-1 and ALDH9A1 do not colocalize. Furthermore, vaginocervical stimulation applied to unmated rats increases the mRNA level of Chst10 in the oviductal mucosa. In conclusion, mating increases the activity of CHST10 in the oviductal mucosa, which in turn induces the synthesis of acidic variants of ALDH9A1 and FHL1 via HNK-1 glycosylation. In the UTJ, HNK-1-ALDH9A1 can control the quality of the sperm arriving at the fertilization site, since we detected acrosome-reacted sperm in close contact with this glycoprotein, a finding that merits further research to propose the use of HNK-1-ALDH9A1 in the selection of sperm in procedures of assisted reproduction. Finally, the sensorial component of mating could regulate events occurring in the oviductal mucosa early after mating.

The Amuyo Ponds (APs) are a group of three brackish hydrothermal lagoons located at 3700 m above sea level in a pre-Andean setting in the Atacama Desert. Each pond shows a conspicuous green (GP), red (RP), or yellow (YP) coloration, and discharges water rich in arsenic and boron into the Caritaya River (Camarones Basin, northern Chile). Microorganisms are subjected to harsh environmental conditions in these ponds, and the microbial composition and diversity in the Amuyo Ponds’ sediments are unknown. The microbial life colonizing AP sediments was explored by metagenomics analyses, showing a diverse microbial life dominated by members of the bacterial domain, with nearly 800 bacterial genome sequences, and sequences associated with Archaea, Eukarya, and viruses. The genus Pseudomonas was more abundant in GP and YP sediments, while the genera Pseudomonas, Aeromonas, and Shewanella were enriched in RP sediments. Archaeal composition was similar in all sediments, and enriched with methanogens sequences from the Archaeoglobi and Halobacteria classes. Abundant fungi sequences were detected in all sediments from the phyla Blastocladiomycota and Ascomycota. We also report putative functional capabilities related to virulence and defense genes, the biosynthesis of secondary metabolites, and tolerance to arsenic. Thirteen bacterial and fourteen viral metagenome-assembled genomes were reconstructed and informed here. This work expands our knowledge on the richness of the microorganisms in the APs and open further studies on the ecology and genomics of this striking Andean geosite.

Amuyo Ponds (AP) are a group of three hydrothermal lagoons located at 3,700 m above sea level, in a pre-Andean setting at the Atacama Desert. Each pond shows a conspicuous green, red, or yellow coloration, and discharge waters rich in arsenic and boron into the Caritaya River (Camarones Basin, Arica and Parinacota Region, in northern Chile). Microorganisms at these ponds are subjected to harsh environmental conditions, and microbial composition and diversity in Amuyo Ponds sediments are unknown. Then, microbial life colonizing AP sediments was explored by metagenomics analyses. This approach showed a highly diverse microbial life dominated by members of the Bacterial Domain (nearly 800 bacterial genome sequences) and includes sequences associated to Archaea, Eukarya, and viruses. We also report putative functional capabilities related to virulence and defense genes, and biosynthesis of secondary metabolites. Thirteen bacterial and fourteen virus metagenome-assembled genomes were reconstructed and are here informed. This work expands our knowledge on the richness of microorganisms in AP and open further studies on the ecology and genomics of this striking Andean geosite.