A quantitative RT-PCR study of the mRNA expression profile of the IGF axis during mammary gland development

M. Boutinaud, J.H. Shand, J. Beattie, M. Park, G.J. Allan, K. Phillips, D.J. Flint

Research output: Contribution to journalArticle

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Abstract

We have used quantitative RT-PCR to analyse the mRNA expression profile of the major components of the IGF axis in different stages of murine mammary gland development, including late pregnancy, lactation and involution. We have shown that all the genes studied, IGF-I, IGF-II, IGF receptor (IGFR) and IGF-binding protein (IGFBP)-1 to -6, were expressed in every stage, albeit at greatly differing levels and displaying unique expression profiles between developmental stages. IGF-I was always expressed at significantly higher levels than either IGF-II or IGFR. This suggests that IGF-I may be the more important IGF during mammary morphogenesis. Overall, IGFBP-3 demonstrated the highest level of expression of any of the IGFBP genes throughout all the developmental stages studied. However, within developmental stages, by far the highest level of expression of any of the IGFBPs was that of IGFBP-5 at day 2 of involution; this was almost an order of magnitude higher than any of the other IGFBP levels recorded. This corroborated our previous findings that the levels of IGFBP-5 protein are highly elevated in the involuting mammary gland, and demonstrated that this up-regulation of IGFBP-5 operates at the level of transcriptional control or message stability. Comparison of the expression profile for these different genes would strongly suggest that they are likely to have differential functions throughout mammary gland development, and also highlights potential interactions and co-regulation between different members of this axis. In addition, our results have identified some similarities and differences in the expression of IGFBPs between the mouse mammary epithelial cell line, HC11, and the normal mammary gland which are worthy of study, most notably the differential regulation of IGFBP-2 and the site of expression of IGFBP-4 and -6. Overall, this study has demonstrated the importance and complexity of the IGF axis during mammary gland development and provides a valuable resource for future research in this area.
LanguageEnglish
Pages195-207
Number of pages12
JournalJournal of Molecular Endocrinology
Volume33
Issue number1
DOIs
Publication statusPublished - Aug 2004

Fingerprint

Human Mammary Glands
Insulin-Like Growth Factor Binding Proteins
Insulin-Like Growth Factor Binding Protein 5
Polymerase Chain Reaction
Messenger RNA
Insulin-Like Growth Factor I
Breast
Insulin-Like Growth Factor Binding Protein 6
Insulin-Like Growth Factor Binding Protein 4
Insulin-Like Growth Factor Binding Protein 2
IGF Type 2 Receptor
Insulin-Like Growth Factor Binding Protein 1
Insulin-Like Growth Factor Binding Protein 3
Insulin-Like Growth Factor II
Morphogenesis
Transcriptome
Lactation
Genes
Up-Regulation
Epithelial Cells

Keywords

  • mammary glands
  • physiology
  • IGF axis

Cite this

Boutinaud, M. ; Shand, J.H. ; Beattie, J. ; Park, M. ; Allan, G.J. ; Phillips, K. ; Flint, D.J. / A quantitative RT-PCR study of the mRNA expression profile of the IGF axis during mammary gland development. In: Journal of Molecular Endocrinology. 2004 ; Vol. 33, No. 1. pp. 195-207.
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A quantitative RT-PCR study of the mRNA expression profile of the IGF axis during mammary gland development. / Boutinaud, M.; Shand, J.H.; Beattie, J.; Park, M.; Allan, G.J.; Phillips, K.; Flint, D.J.

In: Journal of Molecular Endocrinology, Vol. 33, No. 1, 08.2004, p. 195-207.

Research output: Contribution to journalArticle

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AU - Shand, J.H.

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AU - Phillips, K.

AU - Flint, D.J.

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AB - We have used quantitative RT-PCR to analyse the mRNA expression profile of the major components of the IGF axis in different stages of murine mammary gland development, including late pregnancy, lactation and involution. We have shown that all the genes studied, IGF-I, IGF-II, IGF receptor (IGFR) and IGF-binding protein (IGFBP)-1 to -6, were expressed in every stage, albeit at greatly differing levels and displaying unique expression profiles between developmental stages. IGF-I was always expressed at significantly higher levels than either IGF-II or IGFR. This suggests that IGF-I may be the more important IGF during mammary morphogenesis. Overall, IGFBP-3 demonstrated the highest level of expression of any of the IGFBP genes throughout all the developmental stages studied. However, within developmental stages, by far the highest level of expression of any of the IGFBPs was that of IGFBP-5 at day 2 of involution; this was almost an order of magnitude higher than any of the other IGFBP levels recorded. This corroborated our previous findings that the levels of IGFBP-5 protein are highly elevated in the involuting mammary gland, and demonstrated that this up-regulation of IGFBP-5 operates at the level of transcriptional control or message stability. Comparison of the expression profile for these different genes would strongly suggest that they are likely to have differential functions throughout mammary gland development, and also highlights potential interactions and co-regulation between different members of this axis. In addition, our results have identified some similarities and differences in the expression of IGFBPs between the mouse mammary epithelial cell line, HC11, and the normal mammary gland which are worthy of study, most notably the differential regulation of IGFBP-2 and the site of expression of IGFBP-4 and -6. Overall, this study has demonstrated the importance and complexity of the IGF axis during mammary gland development and provides a valuable resource for future research in this area.

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