title: Effects of increased milking frequency on gene expression in the bovine mammary gland.
abstract: BACKGROUND
  Previous research has demonstrated that increased milking frequency of dairy cattle during the first few weeks of lactation enhances milk yield, and that the effect persists throughout the entire lactation period. The specific mechanisms controlling this increase in milk production are unknown, but suggested pathways include increased mammary epithelial cell number, secretory capacity, and sensitivity to lactogenic hormones. We used serial analysis of gene expression (SAGE) and microarray analysis to identify changes in gene expression in the bovine mammary gland in response to 4x daily milking beginning at d 4 of lactation (IMF4) relative to glands milked 2x daily (Control) to gain insight into physiological changes occurring within the gland during more frequent milking.
  
  
  RESULTS
  Results indicated changes in gene expression related to cell proliferation and differentiation, extracellular matrix (ECM) remodeling, metabolism, nutrient transport, and immune function in IMF4 versus Control cows. In addition, pathways expected to promote neovascularization within the gland appeared to be up regulated in IMF4 cows. To validate this finding, immunolocalization of Von Willebrandt's factor (VWF), an endothelial cell marker, and its co-localization with the nuclear proliferation antigen Ki67 were evaluated in mammary tissue sections at approximately d 7 and d 14 of lactation in cows milked 4x daily versus Controls to estimate endothelial cell abundance and proliferation within the gland. Consistent with expression of genes related to neovascularization, both abundance of VWF and its co-localization with Ki67 appeared to be elevated in cows milked 4x daily, suggesting persistent increased milk yield in response to increased milking frequency may be mediated or complemented by enhanced mammary ECM remodeling and neovascularization within the gland.
  
  
  CONCLUSIONS
  Additional study is needed to determine whether changes in ECM remodeling and neovascularization of the mammary gland result in increased milk yield during increased milking frequency, or occur in response to an increased demand for milk production. Gene pathways identified by the current study will provide a basis for future investigations to identify factors mediating the effects of milking frequency on milk yield.
authors.affiliation: Bovine Functional Genomics Laboratory, USDA-ARS, Beltsville Agricultural Research Center, 10300 Baltimore Avenue, Beltsville, MD 20705, USA. erin.connor@ars.usda.gov
authors.forename: Erin E
authors.initials: EE
authors.lastname: Connor
authors.forename: Stephen
authors.initials: S
authors.lastname: Siferd
authors.forename: Theodore H
authors.initials: TH
authors.lastname: Elsasser
authors.forename: Christina M
authors.initials: CM
authors.lastname: Evock-Clover
authors.forename: Curtis P
authors.initials: CP
authors.lastname: Van Tassell
authors.forename: Tad S
authors.initials: TS
authors.lastname: Sonstegard
authors.forename: Violet M
authors.initials: VM
authors.lastname: Fernandes
authors.forename: Anthony V
authors.initials: AV
authors.lastname: Capuco
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D002417:Cattle; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation; D003612:Dairying / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism; D005260:Female; D005502:Food; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D016678:Genome; D007150:Immunohistochemistry; D007774:Lactation / Q000235:genetics / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D008892:Milk / Q000378:metabolism*; D018919:Neovascularization, Physiologic / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D013997:Time Factors