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1 Immunogenetics Laboratory, Department of Animal Science, Michigan State University, East Lansing, MI, USA
2 Animal Health and Welfare, Teagasc, Grange Research Centre, Dunsany, Co., Meath, Ireland
3 Center for Animal Functional Genomics, Department of Animal Science, Michigan State University, East Lansing, MI, USA
4 Immunogenetics Laboratory, Department of Animal Science, Michigan State University, East Lansing, MI, USA; Center for Animal Functional Genomics, Department of Animal Science, Michigan State University, East Lansing, MI, USA
* To whom correspondence should be addressed. E-mail: burtonj{at}msu.edu.
It is well documented that blood neutrophils from parturient dairy cows do not perform as well as neutrophils from non-parturient cows in laboratory assays of adhesion, migration, or phagocytosis-induced respiratory burst. However, little is known about the possible molecular basis for parturition-induced changes in neutrophils. cDNA microarray analysis was used in the current study to explore parturition-induced changes in gene expression profiles in bovine blood neutrophils. Total RNA from isolated blood neutrophils of 4 parturient Holstein cows was obtained before, during and after parturition, reverse transcribed into cDNA, and sequentially labeled with Cy3 or Cy5 dyes prior to paired hybridizations to 1056 member bovine total leukocyte (BOTL-3) microarrays in a loop design. Resulting gene expression data were LOESS normalized by array and analyzed using a mixed model approach. Results showed that expression profiles for 302 BOTL-3 genes were influenced by parturition. BLASTn analysis and preliminary clustering of affected genes by biological function indicated that the largest proportion (14%) of changed genes encode proteins critical to regulation of apoptosis. Independent confirmation of altered expression for 16 of these genes was achieved using Q-RT-PCR. A predominantly survival phenotype inferred from the microarray and Q-RT-PCR results was substantiated by monitoring apoptosis status of blood neutrophils from castrated male cattle cultured in the presence of sera from parturient cows. Thus, our combined gene expression and apoptosis phenotyping results suggest that bovine parturition may induce prolonged survival in normally short-lived blood neutrophils.
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