title: Insulin, a key regulator of hormone responsive milk protein synthesis during lactogenesis in murine mammary explants.
abstract: Murine milk protein gene expression requires insulin, hydrocortisone, and prolactin; however, the role of insulin is not well understood. This study, therefore, examined the requirement of insulin for milk protein synthesis. Mammary explants were cultured in various combinations of the lactogenic hormones and global changes in gene expression analysed using Affymetrix microarray. The expression of 164 genes was responsive to insulin, and 18 were involved in protein synthesis at the level of transcription and posttranscription, as well as amino acid uptake and metabolism. The folate receptor gene was increased by fivefold, highlighting a potentially important role for the hormone in folate metabolism, a process that is emerging to be central for protein synthesis. Interestingly, gene expression of two milk protein transcription factors, Stat5a and Elf5, previously identified as key components of prolactin signalling, both showed an essential requirement for insulin. Subsequent experiments in HCll cells confirmed that Stat5a and Elf5 gene expression could be induced in the absence of prolactin but in the presence of insulin. Whereas prolactin plays an essential role in phosphorylating and activating Stat5a, gene expression is only induced when insulin is present. This indicates insulin plays a crucial role in the transcription of the milk protein genes.
authors.affiliation: Department of Zoology, University of Melbourne, VIC, Australia. karensa.menzies@deakin.edu.au
authors.forename: Karensa K
authors.initials: KK
authors.lastname: Menzies
authors.forename: Heather J
authors.initials: HJ
authors.lastname: Lee
authors.forename: Christophe
authors.initials: C
authors.lastname: Lefèvre
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Ormandy
authors.forename: Keith L
authors.initials: KL
authors.lastname: Macmillan
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Nicholas
mesh_terms: D000818:Animals; D002417:Cattle; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D007328:Insulin / Q000494:pharmacology*; D007774:Lactation / Q000187:drug effects / Q000378:metabolism*; D008321:Mammary Glands, Animal / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D008894:Milk Proteins / Q000096:biosynthesis*; D008957:Models, Genetic; D020411:Oligonucleotide Array Sequence Analysis; D014176:Protein Biosynthesis / Q000187:drug effects* / Q000235:genetics; D015203:Reproducibility of Results; D050799:STAT5 Transcription Factor / Q000235:genetics / Q000378:metabolism; D046509:Tissue Culture Techniques; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; C500759:Elf5 protein, mouse; D007328:Insulin; D008894:Milk Proteins; D050799:STAT5 Transcription Factor; D014157:Transcription Factors