Project description:Aging is associated with progressive decline in lung function and responses to environmental stress culminating in increased incidence of lung diseases. Initial analysis of Atp8b1 (ATPase, Aminophospholipid Transporter, Class I, Type 8B, Member 1) mutant mice (on C57BL/6J background) revealed deficiency in transporter function and was associated with decreased lung function. Therefore, to further study the aging-related genes in the lungs at the molecular level, the transcriptome analysis of Atp8b1 mutant and C57BL/6J lungs were carried out at 14 M and 7_9 wks of age. Global transcriptome analysis revealed 532 differentially expressed genes in Atp8b1 mutant lungs, 157 differentially expressed genes in C57BL/6J mice and 37 overlapping genes. Interestingly, the majority of age-related (350) genes were unique to Atp8b1 mutant lungs in contrast to the 85 genes that were unique to C57BL/6J lungs. Further, Ingenuity Pathway Analysis (IPA) of aged genes in Atp8b1 mutant lungs showed enrichment of canonical signaling pathways such as xenobiotic metabolism signaling and Nrf2-mediated signaling pathway. We validated Adamts2 and Mmp13 in Atp8b1 mutant lungs by quantitative real-time PCR (qRT-PCR). In addition, Col1a1 and CxCr6 were also validated in both Atp8b1 mutant and C57BL/6J lungs, respectively. To summarize, our study provide insights into altered age-related genes in Atp8b1 lungs that are associated with mutation and environmental trigger when compared to C57BL/6J lungs.
Project description:The importance of unanchored Ub in innate immunity has been shown only for a limited number of unanchored Ub-interactors. We investigated what additional cellular factors interact with unanchored Ub and whether unanchored Ub plays a broader role in innate immunity. To identify unanchored Ub-interacting factors from murine lungs, we used His-tagged recombinant poly-Ub chains as bait. These chains were mixed with lung tissue lysates and protein complexes were isolated with Ni-NTA beads. Sample elutions were subjected to mass spectrometry (LC-MSMS) analysis.
Project description:To characterize the genetic basis of hybrid male sterility in detail, we used a systems genetics approach, integrating mapping of gene expression traits with sterility phenotypes and QTL. We measured genome-wide testis expression in 305 male F2s from a cross between wild-derived inbred strains of M. musculus musculus and M. m. domesticus. We identified several thousand cis- and trans-acting QTL contributing to expression variation (eQTL). Many trans eQTL cluster into eleven ‘hotspots,’ seven of which co-localize with QTL for sterility phenotypes identified in the cross. The number and clustering of trans eQTL - but not cis eQTL - were substantially lower when mapping was restricted to a ‘fertile’ subset of mice, providing evidence that trans eQTL hotspots are related to sterility. Functional annotation of transcripts with eQTL provides insights into the biological processes disrupted by sterility loci and guides prioritization of candidate genes. Using a conditional mapping approach, we identified eQTL dependent on interactions between loci, revealing a complex system of epistasis. Our results illuminate established patterns, including the role of the X chromosome in hybrid sterility.
