Project description:This study looks at the effect of dietary manipulation on the development of hepatic steatosis and changes in hepatic gene expression in a feline model. We used microarray analysis to examine changes in hepatic gene transcription in response to Trans fat, High Fructose Corn Syrup (HFCS) and/or Monosodium Glutamate (MSG) in the domestic cat. The use of human Affymetrix arrays for the study of feline gene expression has previously been validated by Dowling and Bienzle, 2005, Journal of General Virology. 86(Pt 8), 2239-48 (PMID 16033971).
Project description:Purpose:MicroRNAs (miRNAs) are members of a rapidly growing class of small endogenous non-coding RNAs that play crucial roles in post-transcriptional regulator of gene expression in many biological processes. Feline Panleukopenia Virus (FPV) is a highly infectious pathogen that causes severe disease in pets, economically important animals and wildlife in worldwide. However, the molecular mechanisms underlying the pathogenicity of FPV have not been completely clear. To study the involvement of miRNAs in the FPV infection process, miRNAs expression profiles were identified via deep sequencing in the feline kidney cell line (F81) infected and uninfected with FPV. Methods:miRNA-sequencing analysis was performed on an Illumina Hiseq 2500 (LC Sciences, USA) following the vendor's recommended protocol Results:As a result, 673 known miRNAs belonging to 210 families and 278 novel miRNAs were identified. Then we found 57 significantly differential expression miRNAs by comparing the results between uninfected and FPV-infected groups. Furthermore, stem-loop qRT-PCR was applied to validate and profile the expression of the randomly selected miRNAs; the results were consistent with those by deep sequencing. Furthermore, the potential target genes were predicted. The target genes of differential expression miRNAs were analyzed by GO and KEGG pathway. Conclusions:The identification of miRNAs in feline kidney cell line before and after infection with Feline Panleukopenia Virus will provide new information and enhance our understanding of the functions of miRNAs in regulating biological processes.
Project description:This study looks at the effect of dietary manipulation on the development of hepatic steatosis and changes in hepatic gene expression in a feline model. We used microarray analysis to examine changes in hepatic gene transcription in response to Trans fat, High Fructose Corn Syrup (HFCS) and/or Monosodium Glutamate (MSG) in the domestic cat. The use of human Affymetrix arrays for the study of feline gene expression has previously been validated by Dowling and Bienzle, 2005, Journal of General Virology. 86(Pt 8), 2239-48 (PMID 16033971). Our study animals were bred from female Felis catus previously placed on one of 4 different dietary regimens for a period of 3 weeks prior to mating. The four dietary regimens used in this study were: [1] Standard Chow Control feline diet (Test Diet Purina catalog #5003); [2] MSG diet consisting of Control diet with 1.125% added Monosodium Glutamate (Diet A: Test Diet Purina catalog #5C1J); [3] Trans-fat/HFCS diet, containing 8.6% Trans fat and 24% HFCS (Diet B: Test Diet Purina catalog #5B4K); and [4] Trans-fat/HFCS and MSG diet, containing 8.6% Trans fat, 24% HFCS and 1.125% MSG (Diet C: Test Diet Purina catalog #5C1H). Following mating, the 4 groups of dams were maintained on their respective diets throughout the gestation and nursing period. Male offspring used in the following experiments were weaned onto the same diets and maintained on their respective dietary regimens until they reached 9 months of age. Hepatic tissues (4-5 per diet group) were used for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Through the application of a transcriptome profiling strategy, we were able to ascertain the differentially expressed genes and complicated pathways involved in the interactions between Atractylenolide-I and feline ovarian granulosa cells. Based on the results of our transcriptome profiling study, we found the highest number of DEGs participated in cholesterol metabolism pathways, the activation of which might be a major factor underlying Atractylenolide I promote the luteinization of the feline ovarian granulosa cells.
Project description:Adipose-derived mesenchymal stem cells (ASCs) are a promising cell therapy to treat inflammatory and immune-mediated diseases. Development of appropriate pre-clinical animal models is critical to determine safety and attain early efficacy data for the most promising therapeutic candidates. Naturally occurring diseases in cats already serve as valuable models to inform human clinical trials in oncologic, cardiovascular and genetic diseases. The objective of this study was to complete a comprehensive side-by-side comparison of human and feline ASCs with an emphasis on their immunomodulatory capacity and transcriptome. Similar to human ASCs, feline ASCs were highly proliferative at low passages and fit the minimal criteria of multipotent stem cells including a compatible surface protein phenotype, osteogenic capacity and normal karyotype. Like ASCs from all species, feline ASCs inhibited mitogen activated lymphocyte proliferation in vitro, with or without direct ASC-lymphocyte contact. Feline ASCs mimic human ASCs in their mediator secretion pattern including prostaglandin E2, indoleamine 2,3 dioxygenase, transforming growth factor beta and interleukin-6, all augmented by interferon gamma secretion by lymphocytes. The transcriptome of 3 unactivated feline ASC lines were highly similar. Functional analysis of the most highly expressed genes highlighted processes including: 1) the regulation of apoptosis, 2) cell adhesion, 3) response to oxidative stress, and 4) regulation of cell differentiation. Finally, feline ASCs had a similar gene expression profile to noninduced human ASCs. These data will help inform clinical trials using cats with naturally occurring diseases as surrogate models for human clinical trials in the regenerative medicine arena.
Project description:The purpose of this study was to characterize the transcriptomic alterations accompanying the inflammation involved in feline chronic gingivostomatitis (FCGS). Towards this goal next-generation sequencing (NGS)-based gene expression profiling (RNA-Sequencing; RNA-Seq) was performed on matched pairs of FCGS diseased and healthy tissues obtained from three feline subjects.
Project description:The aim of this study is to discover new genes and cells involved in life-long tooth replacement. Here we study the adult dentition of the leopard gecko (Eublepharis macularius). Bulk RNAseq was used to compare teeth that are in function versus unerupted, developing teeth and single cell RNA-seq was carried out on jaw segments containing the dental forming tissues. In bulk RNAseq data, we found that functional teeth expressed genes involved in bone and tooth resorption. Indeed, we found expression of these markers in multinucleated odontoclasts within resorbing functional teeth. Chemotaxis genes SEMA3A and SEMA3E, were expressed within odontoblasts and in adjacent mesenchyme using RNAscope. Semaphorins could be involved in regulating odontoclast formation, recruitment or repulsion from developing teeth. The scRNA-seq experiment successfully isolated dental mesenchyme and several epithelial clusters. We confirmed that some of these genes are expressed in the earliest tooth buds within the tooth forming field and in erupting teeth. This work will lead to discovery of genes and cell populations that may have been gained or lost during evolution of amniotes. Moreover, gene differences may lead to dental therapies to prevent tooth loss from disease or injury.
Project description:The aim of this study is to discover new genes and cells involved in life-long tooth replacement. Here we study the adult dentition of the leopard gecko (Eublepharis macularius). Bulk RNAseq was used to compare teeth that are in function versus unerupted, developing teeth and single cell RNA-seq was carried out on jaw segments containing the dental forming tissues. In bulk RNAseq data, we found that functional teeth expressed genes involved in bone and tooth resorption. Indeed, we found expression of these markers in multinucleated odontoclasts within resorbing functional teeth. Chemotaxis genes SEMA3A and SEMA3E, were expressed within odontoblasts and in adjacent mesenchyme using RNAscope. Semaphorins could be involved in regulating odontoclast formation, recruitment or repulsion from developing teeth. The scRNA-seq experiment successfully isolated dental mesenchyme and several epithelial clusters. We confirmed that some of these genes are expressed in the earliest tooth buds within the tooth forming field and in erupting teeth. This work will lead to discovery of genes and cell populations that may have been gained or lost during evolution of amniotes. Moreover, gene differences may lead to dental therapies to prevent tooth loss from disease or injury.