Technical Abstract: Transcript profiling was performed by Affymetrix microarray analysis and SAGE to characterize changes in gene expression in the bovine mammary gland in response to 4× versus 2× daily milking during the first week of lactation. These changes in gene expression may contribute to the increased milk production observed in response to increased milking frequency. Our results indicate that increased milking frequency alters expression of genes in the mammary gland related to increased extracellular matrix remodeling, neovascularization, metabolism, cell proliferation and apoptosis. Assession Number GSE9344.