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Machado, M. C.

Publications and source records attributed to Machado, M. C..

4 recordsLinked to original sources

Representations in the hippocampal-entorhinal system emerge from learning sensory predictions

The hippocampal formation and adjacent parahippocampal areas are central to intelligent behaviour such as memory and navigation. Understanding how systems in the brain generate structured representations from experience remains a fundamental goal in neuroscience. A central open question is whether a single computational principle can account for the diverse neural responses observed across the hippocampal-entorhinal circuit. Existing models often rely on hand-crafted features or specialized learning mechanisms unrelated to sensory observations, and typically express representations of only a small subset of known cell types. Further, representations learned in such models are often not empirically evaluated against neural representations observed in the navigating brain. Here, we introduce a neurobiologically-inspired and robust computational model in which diverse cell types emerge from a single learning objective with minimal hand-engineered assumptions. Our model applies contrastive graph representation learning to transitions between high-dimensional visual observations, constructing a metric space in which temporally adjacent sensory observations are mapped to nearby states. Inspired by the anatomical information flow of the hippocampal-entorhinal system, and anchored in output representations based on neural coding in the entorhinal cortex, the model gives rise to activity resembling place cells, grid cells, boundary vector cells, band cells, corner cells, and conjunctive cells among others. Across varied environments and sensory streams, the framework captures not only diverse neural response patterns but also the functional dependencies between them, mirroring the proposed sequential representational structure observed in the hippocampal-entorhinal system. Crucially, place-cell-like features of the model quantitatively reproduce remapping dynamics observed in CA1 of freely moving animals, and afford theoretical explanatory power of existing neurobiologically-informed models. This work thus offers a unified computational model of spatial coding in the hippocampal-entorhinal system and a testable framework for generating mechanistic hypotheses in silico, to be evaluated in vivo.

neuroscience↗

The ribonucleoprotein-mediated CRISPR-Cas9 system induces recurrent Aire gene mutations in contrast to the nickase expression vector in murine in vitro or in vivo models

Although in vitro mTEC cultures can help determine the role of the autoimmune regulator (Aire) gene, the mechanisms of Aire mutations were identified using Aire mutant mice. Nevertheless, long-term cultures of mTECs from mice are difficult to establish. To overcome this, we used a CRISPR-Cas9 system to edit Aire in a murine mTEC line in vitro and mouse embryos. Two ribonucleoprotein (RNP) complexes were designed to separately target Aire exons 6 and 8. NHEJ-derived indels or HDR-derived mutations were obtained. Recurrent NHEJ-derived mutations were observed among editions, one in exon 6 (del 3554G) and the other in exon 8 (del 5676_5677TG), i.e., the exon 6 mutation was kept in an mTEC clone edited in vitro and in vivo in a mouse, and the exon 8 mutation was kept in mTECs in vitro. None of the mutations obtained with the nickase system were recurrent, indicating the participation of the RNP complex.

molecular biology↗

miR-155 exerts posttranscriptional control of autoimmune regulator (Aire) and tissue-restricted antigen genes in medullary thymic epithelial cells

BackgroundThe autoimmune regulator (Aire) gene is critical for the appropriate establishment of central immune tolerance. As one of the main controllers of promiscuous gene expression in the thymus, Aire promotes the expression of thousands of downstream tissue-restricted antigen (TRA) genes, cell adhesion genes and transcription factor genes in medullary thymic epithelial cells (mTECs). Despite the increasing knowledge about the role of Aire as an upstream transcriptional controller, little is known about the mechanisms by which this gene could be regulated. ResultsHere, we assessed the posttranscriptional control of Aire by miRNAs. The in silico miRNA-mRNA interaction analysis predicted thermodynamically stable hybridization between the 3' UTR of Aire mRNA and miR-155, which was confirmed to occur within the cellular milieu through a luciferase reporter assay. This finding enabled us to hypothesize that miR-155 might play a role as an intracellular posttranscriptional regulator of Aire mRNA. To test this hypothesis, we transfected a murine mTEC cell line with a miR-155 mimic in vitro, which reduced the mRNA and protein levels of Aire. Moreover, large-scale transcriptome analysis showed the modulation of 311 downstream mRNAs, which included 58 TRA mRNAs. Moreover, miR-155 mimic-transfected cells exhibited a decrease in their chemotaxis property compared with control thymocytes. ConclusionOverall, the results indicate that miR-155 may posttranscriptionally control Aire mRNA as well as a crucial process by which mTECs allow migration of thymocytes through chemotaxis.

molecular biology↗

Development of microsatellite markers for the threatened species Coleocephalocereus purpureus (Cactaceae) using next-generation sequencing

Ten microsatellite loci were developed and validated for the endangered cactus species Coleocephalocereus purpureus. The markers were obtained from sequences generated by whole genome shotgun sequencing approaches. A testing group of 36 specimens of the main grouping were genotyped and all described markers presented suitable outcomes to population genetic studies, showing polymorphic status for C. purpureus testing group with clean and reproducible amplification. No evidence for scoring errors, null alleles or linkage disequilibrium was detected. Number of alleles per locus ranged from 3 to 6 and expected heterozygosity ranged from 0.78 to 0.99. These new microsatellite loci are suitable to be used in future diversity and structure population studies of C. purpureus.

genetics↗