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:We report transcriptomic differences between neonatal and adult Treg responses to tendon jury using NGS sequencing. We performed tendon injuries to neonatal mice at P5 and 4-6 month old adult mice and isolated Tregs by FACS (CD4-PE+, Foxp3-GFP+) from injured tendons and autologus spleens 14 days after inury. We then performed RNA-sequencing with biological triplicates to distinguish unique transcriptomic signatures that can illuminate the role of Tregs in neonatal tendon regeneration. Tregs from neonatal tendon show a type 2 immune signatures with a unique gene signature that is distinc from canonical Treg activation, while adult Tregs demonstrate a type 1 signature.
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:Impaired healing of adult tendons with fibrosis remains clinical challenges while neonatal tendons have full functional restoration. However, age-associated cellular and molecular changes in tendon cells and tendon stem/progenitor cells (TSPCs) remain unknown. Here, comparative single cell transcriptomics of early postnatal (2-week-old) and adult (20-week-old) mouse tendons revealed that adult tendons have reduced number of TSPCs, decreased gene expression in tendon and cartilage development, and a greater population of fibro-tenogenic cells. Notably, adult TSPCs and tenocytes exhibit increased expression of immune-response and oxidative-stress genes with higher EGFR but decreased IGF signaling. Adult tendon cells show increased levels of intracellular reactive oxygen species (ROS) in vivo. In contrast, antioxidant treatment of adult tendons significantly reduces intracellular ROS of TSPCs and improves tendon strength in vivo. Hence, these findings suggest that increased inflammation and ROS during tendon aging deteriorates tendon function and regeneration that can be mitigated by antioxidant treatment.
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.