Project description:Identify differentially expressed microRNAs in mild and severe equine distal interphalangeal joint osteoarthritis plasma and synovial fluid samples Determine the effects of selected osteoarthritis-related miRNAs on equine chondrocytes in monolayer culture through the application of miRNA agomirs and antagomirs
Project description:Equine lameller tissues were collected to compare normal vs laminitis generated differences in transcriptom level. Keywords: Laminitis, Equine, Diseased foot
Project description:The enteric nervous system (ENS), which is derived from enteric neural crest cells (ENCCs) during gut development, represents the neuronal innervation of the gastrointestinal tract and is critical for regulating normal intestinal function. Compromised ENCC migration can lead to Hirschsprung Disease, which is characterized by an aganglionic distal bowel. We find that removal of the ceca, a paired structure present at the midgut-hindgut junction in avian intestine, leads to severe hindgut aganglionosis, suggesting that the ceca are required for ENS development. To test this, we replaced the ceca of embryonic day 6 (E6) wild-type chicks with ceca from transgenic GFP chicks. Interestingly, the entire hindgut ENS arises from the GFP+ ceca-derived ENCC population. Comparative transcriptome profiling of the cecal buds compared to the interceca region shows that the non-canonical Wnt signaling pathway is preferentially expressed within the ceca. Specifically, Wnt11 is highly expressed in the ceca, as confirmed by RNA in situ hybridization, leading us to hypothesize that cecal expression of Wnt11 is important for ENCC colonization of the hindgut. Organ cultures were prepared using E6 avian intestine, when ENCCs are migrating through the ceca, and showed that Wnt11 inhibits enteric neuronal differentiation. These results reveal an essential role for the ceca during hindgut ENS formation and highlight an important function for non-canonical Wnt signaling in regulating ENCC differentiation and thereby promoting their migration into the colon.
Project description:Aedes aegypti mosquitoes are efficient vectors of human disease including Zika, Dengue and Chikungunya. After taking a blood meal, proteins from the blood are broken down into amino acids that are essential for egg development and growth. We have discovered a novel role for a neuropeptide receptor in the hindgut that coordinates blood utilization. The hindgut is not a tissue normally implicated in blood digestion, therefore we conducted RNA-seq profiling of the hindgut in wild type females to obtain a transcriptomic profile of these tissues in non-blood fed conditions.
Project description:Anorectal malformation (ARM), a common congenital anomaly of the digestive tract, is a result of insufficient elongation of the urorectal septum. The cytoplasmic protein Receptor of Activated C-Kinase 1 (Rack1) is involved in embryonic neural development; however, its role in embryonic digestive tract development and ARM formation is unexplored. Our study explored the hindgut development and cell death mechanisms in ARM-affected rats using spatial transcriptome analysis. We induced ARM in rats by administering ethylenethiourea via gavage on gestational day (GD) 10. On GDs 14-16, embryos from both normal and ARM groups underwent spatial transcriptome sequencing, which identified key genes and signalling pathways. Rack1 exhibited significant interactions among differentially expressed genes on GDs 15 and 16. Reduced Rack1 expression in the ARM-affected hindgut, verified by Rack1 silencing in intestinal epithelial cells, led to increased P38 phosphorylation and activation of the MAPK signalling pathway. The suppression of this pathway downregulated Nqo1 and Gpx4 expression, resulting in elevated intracellular levels of ferrous ions, reactive oxygen species (ROS) and lipid peroxides. Downregulation of Gpx4 expression in the ARM hindgut, coupled with Rack1 co-localisation and consistent mitochondrial morphology, indicated ferroptosis. In summary, Rack1, acting as a hub gene, modulates ferrous ions, lipid peroxides, and ROS via the P38-MAPK/Nqo1/Gpx4 axis. This modulation induces ferroptosis in intestinal epithelial cells, potentially influencing hindgut development during ARM onset.
Project description:The aim of the study was to investigate the effects of autologous equine serum (AES) incubated for 24 h and autologous conditioned serum (ACS) on inflamed equine chondrocyte pellets in vitro.
Project description:Equine lameller tissues were collected to compare normal vs laminitis generated differences in transcriptom level. Experiment Overall Design: Three Laminitis generated vs three normal Equine hoof tissues were subjected to comparison analysis in transcriptom level by using the Affymetrix Bovine GeneChip. Experiment Overall Design: The reasons for Bovine chip were; 1) Genetic similarity to Equine. Experiment Overall Design: 2) More transcriptom was search at that Affymetrix platform comparing the Equine GeneChip at the time of the study.