Project description:Adult Sprague Dawley rats were treated i.g. with 50 mg/kg alpha-naphthylisothiocyanate (ANIT) or vehicle (corn oil). Hepatobiliary liver injury occurred at 24 h postdose in ANIT rats with repair at 120h. Livers were extracted from rats at 24h and 120 h post ANIT exposure. This study investigated differences in mRNA expression between the injury and repair phases in the context of ANIT exposure.
Project description:To determine the LncRNA expression profile in dorsal root ganglia tissues of 7 days after rats received 150 μl of CFA (1 mg/ml Mycobacterium tuberculosis, Sigma, USA) through the patella tendon into the right knee joint. CFA injection rats macthed saline injection rats, we uesed LncRNA microArray analysis form Arraystar to examine the expression of LncRNAs and mRNAs in dorsal root ganglia tissues of 7 days after rats received CFA through the patella tendon into the right knee joint and matched Saline through the patella tendon into the right knee joint.
Project description:Turnover of matrix proteins is essential to maintain tissue homeostasis, and dysregulation of protein turnover has been implicated in a variety of diseases. However, proteome dynamics in the musculoskeletal system, particularly tendon, remain relatively unexplored. The aim of this study was therefore to calculate the turnover rate of individual proteins within tendon and its constituent components. We hypothesised that turnover of proteins in the interfascicular matrix would be greater than in the fascicles. Achilles tendons were harvested from rats fed heavy labelled water over a period of 127 days. Proteins were extracted from one Achilles tendon, while laser capture microdissection was used to isolate regions of interfascicular and fascicular matrix from the other Achilles prior to protein extraction. Samples were analysed using tandem mass spectrometry and the rate of label incorporation used to calculate turnover rate of individual proteins. Results demonstrate complex proteome dynamics within tendon, with differences in protein turnover rates approaching 1000-fold. Collagen turnover was relatively slow, with much more rapid turnover of proteoglycans and glycoproteins. Data support the hypothesis, demonstrating overall faster protein turnover within the interfascicular matrix compared to the fascicles. More rapid turnover of the interfascicular matrix may be a mechanism to repair microdamage to this region which is exposed to high levels of shear during locomotion. The techniques used here provide a powerful approach to interrogate alterations in protein turnover in the musculoskeletal system with ageing and/or disease which are likely to influence injury risk.
Project description:Adult Sprague Dawley rats were treated i.g. with 50 mg/kg alpha-naphthylisothiocyanate (ANIT) or vehicle (corn oil). Hepatobiliary liver injury occurred at 24 h postdose in ANIT rats with repair at 120h. Livers were extracted from rats at 24h and 120 h post ANIT exposure. This study investigated differences in mRNA expression between the injury and repair phases in the context of ANIT exposure. 8 week male Sprague Dawley rats were administered ANIT or vehicle for quantification of hepatobiliary injury via clinical chemistry biomarkers and pathology between 6 and 168 h postdosing. Rats sacrificed at 24 h and 120h postdosing coincided with peak injury and injury resolution, respectively.
Project description:Injury was induced in the left Achilles tendon by needle puncture. Rats were sacrificed 4 and 21 days post injury. Shams, in which the tendon was isolated but not punctured, were included as controls. Serum was collected post-mortem and RNAseq used to identify differentially expressed ncRNAs.
Project description:Heterogeneity in pluripotent cells marks a metastable state where cells may drift between native and lineage-primed populations. While the role for these heterogeneities are unclear, they may reflect the dynamic equilibriums of signaling networks and have a direct effect on differentiation potentialities. Here, we report the role of the cell cycle in establishing heterogeneity of human pluripotent stem cells. By utilizing the FUCCI cell cycle indicator system coupled to fluorescent activated cell sorting (FACS), we have uncovered that the cell cycle drives heterogeneity at the epigenetic, transcriptional and post-transcriptional levels. Our data show widespread dynamics in 5-hydroxymethylcytosine (5hmC) during the cell cycle. Furthermore, transcript profiling by RNA-sequencing identified >500 genes that were cell cycle-regulated, of which the largest cohort of genes were transcriptional regulators. In sum, we demonstrate the role of the cell cycle in coordinating cellular transitions between metastable states in pluripotent stem cells. mRNA sequencing of the cell cycle phases; early & late G1, S and G2/S from human ES cells in triplicate.
Project description:Achilles tendinopathy is often thought to be a consequence of overuse of the Cells within the Achilles tendon of healthy rats undergo a series of changes following physiologic levels of mechanical stimulation after running, and we further explored the transcriptome changes in Achilles tendon cells during post-exercise recovery. Our current experiment reveals RNA-seq analysis of the transcriptome of the rat Achilles tendon after 12 hours of rest following running.