title: Proteotyping of mammary tissue from transgenic and gene knockout mice with immunohistochemical markers: a tool to define developmental lesions.
abstract: Through the use of transgenic and gene knockout mice, several studies have identified specific genes required for the functional development of mammary epithelium. Although histological and milk protein gene analyses can provide useful information regarding functional differentiation, they are limited in their ability to precisely define the molecular lesions. For example, mice that carry a mutation in one of the subunits of the IkappaB kinase, IKKalpha, cannot lactate despite the presence of histologically normal alveolar compartment and the expression of milk protein genes. To further define and understand such lesions on a molecular level, we sought evidence for proteins that are differentially expressed during mammary gland development with a view to generating a tissue proteotype. Using database screens and immunohistochemical analyses, we have identified three proteins that exhibit distinct profiles. Here, using mouse models as test biological systems, we demonstrate the development and application of mammary tissue proteotyping and its use in the elucidation of specific developmental lesions. We propose that the technique of proteotyping will have wide applications in the analyses of defects in other mouse models.
authors.affiliation: Laboratory of Genetics and Physiology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA. jonshi@helix.nih.gov
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Shillingford
authors.forename: Keiko
authors.initials: K
authors.lastname: Miyoshi
authors.forename: Gertraud W
authors.initials: GW
authors.lastname: Robinson
authors.forename: Brian
authors.initials: B
authors.lastname: Bierie
authors.forename: Yixue
authors.initials: Y
authors.lastname: Cao
authors.forename: Michael
authors.initials: M
authors.lastname: Karin
authors.forename: Lothar
authors.initials: L
authors.lastname: Hennighausen
mesh_terms: D000818:Animals; D051402:Aquaporin 5; D020346:Aquaporins / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis; D004268:DNA-Binding Proteins / Q000235:genetics; D005260:Female; D051550:I-kappa B Kinase; D007150:Immunohistochemistry; D028322:Inhibin-beta Subunits / Q000235:genetics; D053614:Janus Kinase 2; D008321:Mammary Glands, Animal / Q000254:growth & development / Q000378:metabolism*; D008565:Membrane Proteins; D051379:Mice; D018345:Mice, Knockout; D008819:Mice, Nude; D008822:Mice, Transgenic; D008894:Milk Proteins; D016328:NF-kappa B / Q000235:genetics / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D011518:Proto-Oncogene Proteins; D011981:Receptors, Prolactin / Q000235:genetics; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor; D015398:Signal Transduction; D050608:Sodium-Phosphate Cotransporter Proteins; D028021:Sodium-Potassium-Chloride Symporters / Q000378:metabolism*; D064506:Solute Carrier Family 12, Member 2; D027981:Symporters / Q000378:metabolism*; D015534:Trans-Activators / Q000235:genetics
chemical_list: C496038:Aqp5 protein, mouse; D051402:Aquaporin 5; D020346:Aquaporins; D015415:Biomarkers; D004268:DNA-Binding Proteins; C498905:Inhbb protein, mouse; D008565:Membrane Proteins; D008894:Milk Proteins; D016328:NF-kappa B; D011518:Proto-Oncogene Proteins; D011981:Receptors, Prolactin; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor; C578878:Slc12a2 protein, mouse; D050608:Sodium-Phosphate Cotransporter Proteins; D028021:Sodium-Potassium-Chloride Symporters; D064506:Solute Carrier Family 12, Member 2; C494087:Stat3 protein, mouse; D027981:Symporters; D015534:Trans-Activators; D028322:Inhibin-beta Subunits; D011505:Protein-Tyrosine Kinases; C507926:Jak2 protein, mouse; D053614:Janus Kinase 2; D017346:Protein Serine-Threonine Kinases; C496564:Chuk protein, mouse; D051550:I-kappa B Kinase; C496562:Ikbkb protein, mouse; C496566:Ikbke protein, mouse