Study On Goat And Sheep Pain Response Genetic Mechanism By Transcriptome Sequencing

Chronic pain is a common complex nerve pathology and subjective symptoms, there are significant differences in the feelings of pain between differentspecies.Spinal dorsal root ganglion (dorsal root ganglion, DRG) is the primary center of conduction and maintain of pain, researching on DRG transcriptome is helpful for comprehensive and systematic kowing the chronic pain and regulatory mechanism,and screening pathogeni city-related genes of pain. We choose the CFA inducing pain (experimental group) and untreated (control group) in sheep and goat each two groups, each group of three.We sequence and analysis the transcriptome of their DRG tissues by RNA-seq technology and bioinformatics methods, in order to reveal the gene regulation model in pain response between different species.we obtains the main results as follows:1.The pain is regulated at the level of transcription and alternative splicing.2. The cluster analysis in differential expression genes suggests goats and sheep gene expression patterns changed dramatically in the process of response pain.3. DEG analysis show that 1748 differentially expressed genes were obtain in goat,2441 were obtain in sheep; 2935 DEG were obtain between goat and sheep compared, among of 426 gene were expressed only in sheep DRG,353 genes only in goat DRG.4. GO analysis shows that those genes expression which participate in the inflammatory response (8 up,19 down)、immune response (20 up,3 down)、damage response (13 up,46 down) and defense response (14up,25 down)and the gated ion channels、neurotransmitters and their receptors related geneand so on have significantly changed in goat,such as:CCL27, PRKACA, GABRA4, GRIA2.GRIK3, CACNA1E, SCN3A. KCNK1.Those genes play a key role in the process of response to pain through up or down gene expression.The results suggest the immune system of goats changed dramatically when inflammatory pain occurs.These differences genes involved in the pathway.such as Chemokine signaling pathway. Retrograde endocannabinoid、Calcium signaling pathway-Glutamatergic synapse、MAPK signaling pathway and so on. These pathway activated play a important role in response to pain in the goat.5. GO analysis shows that those inflammatory response related genes were not enrichment in sheep, but these genes expression which encoded inhibitory neurotransmitter and receptor genes was up regualated in sheep, such as GABRG3, GABRB2. GABRB1, GLRA3, CLCN7;in addition, involved in neurotransmitter-gating ion channel ligand binding and neural activity of NMDA receptor ligand receptor interaction, coding and potassium channel related gene,their expression were up regualated, such as GRIK1, GRID2, KCND2, KCNH7, CATSPER3. CACNG2, CHRNA5, etc. Importantly,These gene including SCN9A, SCN4B, SCN11A, SCN8A. SCN7A, SCN10A, TRPV1 were down regulated expression,which code the gated sodium channel sodium ion transport, neurotrophic factors related gene expression and so on, especially the down expression of SCN9A, TRPV1 is associated with pain tolerance.Differences in gene involved in GABAergicsynapse, Neurotrophin signaling pathway, Retrograde endocannabinoid signaling, Morphine addiction, Nicotine addiction, Neurotransmitter-gated ion channel-ligand binding-pathwaysand so on.The results shown that these pathways are involved in response to pain in the sheep.6. According to GO analysis of the DEG between species shows that there is a certain difference in some genes at the transcription regulation background level between sheep and goat, including voltage-gated ion channels, damage response, inflammation response, defense response related genes.7. Alternative splicing events analysis showed that alternative splicing events is reduced in the process of pain response, at the same time we detected abundant novel AS events and SJs from goats and sheep samples. We detected 15576 novel AS events and 107786 novel SJsfrom goats six samples; 17260 novel AS events and 83358 novel SJs From six sheep samples.8. Alternative splicing differences analysis found that 371 genes AS differences in goat,360 genes in sheep.DASG function clusters suggest in the process of pain response in sheep, involved in regulating the Ras GTP enzyme activity and the Ras protein signal transduction related genes made changed significantly in alternative splicing events.including FGD1, RABGAP1. TBC1D14. AGAP1, TBC1D1, SCRIB Moreover, participated in the sodium ions and transport genes, calcium ion channel related genes and ion transport alternative splicing events had altered obviously, including ATP1A2. HCN4. CLCN3. TRPC4. KCNC4. CACNA2D1. SGK1. etc.But in the process of the pain response in goats only GRIK1 and GRIK2 gene significantly changedin AS events.The research found that gene expression patterns of DRG tissues in sheep and goats changed significantly in the process of chronic pain, involved in all kinds of ion channels, neurotransmitters and their receptors, inflammatory mediators, cytokines, signal transduction molecules,their expression were up or down regulated to response pain.We reveals the pain response mechanism between goat and sheep is difference in transcription, transcription regulation and alternative splicing level at the first time. Goats could be increased in Peripheral Sensitization to enhance the pain, instead of sheep may be weakened pain transduction by release lots of inhibitory neurotransmitter, and found a large number of potential analgesic drug targets.The results provides a scientific reference for the genetic basis of pain, response, and symptoms and the regulation mechanism also development for the treatment of drugs.