Microarray analysis of insulin-like growth factor-I-induced changes in messenger ribonucleic acid expression in cultured porcine granulosa cells : Possible role of insulin-like growth factor-I in angiogenesis
Articolo
Data di Pubblicazione:
2009
Citazione:
Microarray analysis of insulin-like growth factor-I-induced changes in messenger ribonucleic acid expression in cultured porcine granulosa cells : Possible role of insulin-like growth factor-I in angiogenesis / J.A. Grado-Ahuir, P.Y. Aad, G. Ranzenigo, F. Caloni, F. Cremonesi, L.J. Spicer. - In: JOURNAL OF ANIMAL SCIENCE. - ISSN 0021-8812. - 87:6(2009), pp. 1921-1933.
Abstract:
ABSTRACT: Insulin-like growth factor-I in conjunction
with gonadotropins are important stimulators of
mitosis and ovarian steroid production by granulosa
and thecal cells, which are required for normal oocyte
development and hormonal feedback signaling to the
hypothalamus and pituitary. However, a comprehensive
evaluation of the changes in gene expression induced
by IGF-I has not been conducted. Our objective was to
characterize granulosa cell gene expression in response
to IGF-I treatment. Porcine granulosa cells were pooled
in 4 biological replicates and treated with FSH (baseline)
or FSH+IGF-I for 24 h in vitro. The RNA was collected
and hybridized to 8 Affymetrix Porcine GeneChips
(Affymetrix, Santa Clara, CA) in a paired design. Differentially
regulated gene sequence element sets (P <
0.01) were used as queries in the UniGene database
searching for annotated genes. Abundance of messenger
RNA (mRNA) for genes differentially expressed in the
microarray analysis was determined through multiplex
assays of one-step real-time reverse transcription-PCR
and further analyzed under a statistical model including
the fixed effect of treatment. A total of 388 gene
sequence element sets were differentially expressed, and
42 matched annotated genes in the UniGene database.
Of the 3 upregulated target genes selected for further
quantitative reverse transcription-PCR analysis, only
FGF receptor 2 III c (FGFR2IIIc) mRNA abundance
was significantly increased by IGF-I. Of the 3 downregulated
target genes selected for further analysis, only
thrombospondin-1 (THBS1) mRNA abundance was
significantly decreased by IGF-I. Further study revealed
that neither FSH nor estradiol affected the IGF-I-induced
suppression of THBS1 mRNA abundance. These
results provide the first comprehensive assessment of
IGF-I-induced gene expression in granulosa cells and
will contribute to a better understanding of the molecular
mechanisms of IGF-I regulation of follicular development.
Involvement of FGFR2IIIc and THBS1 in
mediating IGF-I-induced granulosa cell steroidogenesis
and proliferation during follicular development is novel,
but their specific roles will require further elucidation.
Tipologia IRIS:
01 - Articolo su periodico
Elenco autori:
J.A. Grado-Ahuir, P.Y. Aad, G. Ranzenigo, F. Caloni, F. Cremonesi, L.J. Spicer
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