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Quantifying promoter-specific Insulin-like Growth Factor 1 gene expression by interrogating public databases.

ABSTRACT: The actions of insulin-like growth factor 1 (IGF1), a small, secreted protein, are essential for normal somatic growth in children and are important for tissue regeneration and repair in adults. Similar functions are conserved in other mammalian species. IGF1 gene regulation is complicated in mammals, with transcription being controlled by different hormonal, nutritional, and tissue-specific inputs. Quantifying IGF1 gene expression in different organs and tissues also has been difficult because of the variable contributions of its two promoters and because of the lack of standard platforms for analysis. Here, I have taken advantage of the wealth of information found in publicly accessible RNA-sequencing libraries to measure steady-state levels of IGF1 mRNAs from human and macaque, species chosen because they are not readily tractable experimental organisms, yet retain similar IGF1 gene organization. Results demonstrate that IGF1 transcripts are highly expressed in fat and liver in both species, and are induced during human adipocyte differentiation. IGF1 mRNAs also are increased in macaque skeletal muscle after selected dietary manipulations. In the organs and tissues examined, IGF1 promoter 1 appears to be far more active than promoter 2. Collectively, these observations show that interrogating large-scale public genomic resources is an effective strategy for quantifying gene expression across different tissues and species.

SUBMITTER: Rotwein P

PROVIDER: S-EPMC6317063 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Quantifying promoter-specific Insulin-like Growth Factor 1 gene expression by interrogating public databases.

Rotwein Peter P

Physiological reports 20190101 1

The actions of insulin-like growth factor 1 (IGF1), a small, secreted protein, are essential for normal somatic growth in children and are important for tissue regeneration and repair in adults. Similar functions are conserved in other mammalian species. IGF1 gene regulation is complicated in mammals, with transcription being controlled by different hormonal, nutritional, and tissue-specific inputs. Quantifying IGF1 gene expression in different organs and tissues also has been difficult because ...[more]

PMID: 30604932

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Project description:Previously we showed that CCN family member 2/connective tissue growth factor (CCN2) promotes the proliferation, differentiation, and maturation of growth cartilage cells in vitro. To elucidate the specific role and molecular mechanism of CCN2 in cartilage development in vivo, in the present study we generated transgenic mice overexpressing CCN2 and analyzed them with respect to cartilage and bone development. Transgenic mice were generated expressing a ccn2/lacZ fusion gene in cartilage under the control of the 6 kb-Col2a1-enhancer/promoter. Changes in cartilage and bone development were analyzed histologically and immunohistologically and also by micro CT. Primary chondrocytes as well as limb bud mesenchymal cells were cultured and analyzed for changes in expression of cartilage-related genes, and non-transgenic chondrocytes were treated in culture with recombinant CCN2. Newborn transgenic mice showed extended length of their long bones, increased content of proteoglycans and collagen II accumulation. Micro-CT analysis of transgenic bones indicated increases in bone thickness and mineral density. Chondrocyte proliferation was enhanced in the transgenic cartilage. In in vitro short-term cultures of transgenic chondrocytes, the expression of col2a1, aggrecan and ccn2 genes was substantially enhanced; and in long-term cultures the expression levels of these genes were further enhanced. Also, in vitro chondrogenesis was strongly enhanced. IGF-I and IGF-II mRNA levels were elevated in transgenic chondrocytes, and treatment of non-transgenic chondrocytes with recombinant CCN2 stimulated the expression of these mRNA. The addition of CCN2 to non-transgenic chondrocytes induced the phosphorylation of IGFR, and ccn2-overexpressing chondrocytes showed enhanced phosphorylation of IGFR. Our data indicates that the observed effects of CCN2 may be mediated in part by CCN2-induced overexpression of IGF-I and IGF-II. These findings indicate that CCN2-overexpression in transgenic mice accelerated the endochondral ossification processes, resulting in increased length of their long bones. Our results also indicate the possible involvement of locally enhanced IGF-I or IGF-II in this extended bone growth.

Dataset Information

Quantifying promoter-specific Insulin-like Growth Factor 1 gene expression by interrogating public databases.

Publications

Quantifying promoter-specific Insulin-like Growth Factor 1 gene expression by interrogating public databases.

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