Finding the Genes Expressed in Female Reproductive Tissues in Pigs

A molecular understanding of porcine reproduction is of biological interest and economic importance. Our Midwest Consortium has produced cDNA libraries containing the majority of genes expressed in major female reproductive tissues, and we have deposited into public databases 21,499 expressed sequence tag (EST) gene sequences from the 3\u27 end of clones from these libraries. These sequences represent 10,574 different genes, based on sequence comparison among these data, and comparison to existing porcine genes indicate as many as 4,652 are novel. Computer analysis identified sequences that are expressed in specific pig tissues or organs, and confirmed the broad expression in pig for many genes ubiquitously expressed in human tissues. Furthermore, we have developed computer software to identify sequence similarity of these pig genes with their human counterparts, and to extract the mapping information of these human homologues from genome databases. We used this software to localize 61 genes on the porcine physical map of chromosomes 5, 10, and 14. Thus our sequence data is useful in accelerating mapping studies and will be useful in understanding pig reproductive biology

Cloning of the Pig Counterpart of a Gene Involved in Resistance to Bacterial Infection

The pig gene corresponding to a mouse protein known to cause susceptibility to infection by several different bacteria (NRAMP1) was cloned and the entire protein coding region sequenced. The pig protein encoded within this gene is highly similar to the mouse and human NRAMP1. A preliminary expression profile of pig NRAMP1 indicates it is expressed in spleen, a rich source of immune cells, and may be expressed in other tissues at low levels. Taken together, these data strongly indicate that the newly cloned gene has a similar physiological function in pigs to that seen for mouse NRAMP1. With this new information, the association of NRAMP1 to Salmonella infection in pigs can be tested

Genetic markers for improved disease resistance in animals (BPI)

A method for determining improved disease resistance in animals is disclosed. The method assays for a novel genetic alleles of the BPI gene of the animal. The alleles are correlated with superior disease resistance. Novel nucleotide sequences, assays and primers are disclosed for the methods of the invention

Heat Stress during Pig Oocyte In Vitro Maturation Impacts Embryonic Development and Gene Expression

Gene expression of heat stressed oocytes matured in vitro was evaluated for potential markers which could be used to characterize the effects of heat stress on developing oocytes in the pig. Three heat stress scenarios were administered during in vitro oocyte maturation. Heat stressed oocytes had reduced maturation rates, decreased developmental competency, and altered expression of heat stress and developmental competency markers

Genetic markers for screening animals for improved disease resistance (NRAMP)

A method for determining improved innate immunity, disease resistance or performance in animals is disclosed. The method involves assays for a genetic differences in the NRAMP1 gene of the animal which is associated with superior disease resistance. Novel NRAMP1 sequence, assays, and compositions for identifying the presence of absence of these alleles are provided

Pig immune response to general stimulus and to porcine reproductive and respiratory syndrome virus infection: a meta-analysis approach

BACKGROUND: The availability of gene expression data that corresponds to pig immune response challenges provides compelling material for the understanding of the host immune system. Meta-analysis offers the opportunity to confirm and expand our knowledge by combining and studying at one time a vast set of independent studies creating large datasets with increased statistical power. In this study, we performed two meta-analyses of porcine transcriptomic data: i) scrutinized the global immune response to different challenges, and ii) determined the specific response to Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) infection. To gain an in-depth knowledge of the pig response to PRRSV infection, we used an original approach comparing and eliminating the common genes from both meta-analyses in order to identify genes and pathways specifically involved in the PRRSV immune response. The software Pointillist was used to cope with the highly disparate data, circumventing the biases generated by the specific responses linked to single studies. Next, we used the Ingenuity Pathways Analysis (IPA) software to survey the canonical pathways, biological functions and transcription factors found to be significantly involved in the pig immune response. We used 779 chips corresponding to 29 datasets for the pig global immune response and 279 chips obtained from 6 datasets for the pig response to PRRSV infection, respectively. RESULTS: The pig global immune response analysis showed interconnected canonical pathways involved in the regulation of translation and mitochondrial energy metabolism. Biological functions revealed in this meta-analysis were centred around translation regulation, which included protein synthesis, RNA-post transcriptional gene expression and cellular growth and proliferation. Furthermore, the oxidative phosphorylation and mitochondria dysfunctions, associated with stress signalling, were highly regulated. Transcription factors such as MYCN, MYC and NFE2L2 were found in this analysis to be potentially involved in the regulation of the immune response. The host specific response to PRRSV infection engendered the activation of well-defined canonical pathways in response to pathogen challenge such as TREM1, toll-like receptor and hyper-cytokinemia/ hyper-chemokinemia signalling. Furthermore, this analysis brought forth the central role of the crosstalk between innate and adaptive immune response and the regulation of anti-inflammatory response. The most significant transcription factor potentially involved in this analysis was HMGB1, which is required for the innate recognition of viral nucleic acids. Other transcription factors like interferon regulatory factors IRF1, IRF3, IRF5 and IRF8 were also involved in the pig specific response to PRRSV infection. CONCLUSIONS: This work reveals key genes, canonical pathways and biological functions involved in the pig global immune response to diverse challenges, including PRRSV infection. The powerful statistical approach led us to consolidate previous findings as well as to gain new insights into the pig immune response either to common stimuli or specifically to PRRSV infection