The ovarian expression of the antigonadotropic insulin-like growth factor binding protein-2 is theca-interstitial cell-selective: Evidence for hormonal regulation

Elisabctta Ricciarelli, Elcuterio R. Hernandez, Aryc Hurwitz, Ehud Kokia, Ronald (Ron) Rosenfeld, Eli Y. Adashi

Research output: Contribution to journalArticle

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Abstract

To begin the process of identification and charactization of rat ovarian insulin-like growth factor binding proteins, we have undertaken to explore the ovarian expression, cellular localization, and hormonal regulation of the insulin-like growth factor binding protcin-2 (1GFBP-2) gene lor which an anligonadoiropic potential has recently been demonstrated. To this end, a solution hybridization/RNasc protection assay was employed wherein total ovarian RNA (2(Vg) from immature (21-23 davs old) female rats was hybridized with a 32P-laballed 1GFBP-2 nboprobe. As in liver, a single protected fragment (550 bases long.) corresponding to IGFBP-2 transcripts was identified in whole ovarian material. Cellular localization studies revealed the IGFBP-2 gene to be exclusively expressed in the ihcca-intcrstilial rather than the granulosa cell compartment. To confirm the cellular disinbution of the IGFBP-2 protein, media conditioned by cultured granulosa or theca-intcrsiitial cells weresubjectedtoimmunoprecipation using two IGFBP-2-directedpolyclonalanliscraExpectdly.bothantibodies(butnotnon-immunerabbitserum) readily.mmunoprccip.latcd the 28kDa rat IGFBP-2 species “cncratcd by hepatic BRL-3A cells. Similarly, both antibodies effectively immunoprccipitatcd an IGFBP the size ol rat IGFBP-2 elaborated by theca-micrstiiial cells. In contrast, neither antibody immunoprccipitatcd the 28-29kDa IGFBP species elaborated by granulosa cells otherwise readily apparent in conventional Western ligand blots. Hypophyscctomy resulted in a 3-fold decrease (P<0.05) in the relative (dcnsiiomctncally-quantidcd) abundance ol ovarian IGFBP-2 transcripts, a diametrically opposed effect (P<0.05) being noted at the level of the liver. In contrast, treatment ol immature hypophyscctomized rats with a dicthvlstilbcstrol-containing subcutaneous silastic implant for a total of 5 days resulted in a concordant 3-fold increase (P<0.05) in the relative abundance of IGFBP-2 transcripts in both ovary and liver when compared with untreated hypophyscctomized controls. Taken together, these findings document rat ovarian IGFPB-2 gene expression to be Ihcca-inlcrslitial (rather than granulosa) cell-selective, and subject to uprcgulatory control by pituitary principlc(s) and/or by estrogens. Although equally estrogen-dependent, hepatic IGFBP-2 gene expression proved constitutive in nature and subject to (diametrically opposed) inhibitory control by (potentially distinct) pituitary principlc(s).

Original languageEnglish (US)
Pages (from-to)2266-2268
Number of pages3
JournalEndocrinology
Volume129
Issue number4
DOIs
StatePublished - 1991
Externally publishedYes

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Insulin-Like Growth Factor Binding Protein 2
Theca Cells
Insulin-Like Growth Factor Binding Proteins
Granulosa Cells
Liver
Estrogens
pX Genes
Gene Expression
Antibodies
Somatomedins
Conditioned Culture Medium
Ovary
Western Blotting
RNA
Ligands

ASJC Scopus subject areas

  • Endocrinology

Cite this

The ovarian expression of the antigonadotropic insulin-like growth factor binding protein-2 is theca-interstitial cell-selective : Evidence for hormonal regulation. / Ricciarelli, Elisabctta; Hernandez, Elcuterio R.; Hurwitz, Aryc; Kokia, Ehud; Rosenfeld, Ronald (Ron); Adashi, Eli Y.

In: Endocrinology, Vol. 129, No. 4, 1991, p. 2266-2268.

Research output: Contribution to journalArticle

Ricciarelli, Elisabctta ; Hernandez, Elcuterio R. ; Hurwitz, Aryc ; Kokia, Ehud ; Rosenfeld, Ronald (Ron) ; Adashi, Eli Y. / The ovarian expression of the antigonadotropic insulin-like growth factor binding protein-2 is theca-interstitial cell-selective : Evidence for hormonal regulation. In: Endocrinology. 1991 ; Vol. 129, No. 4. pp. 2266-2268.
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abstract = "To begin the process of identification and charactization of rat ovarian insulin-like growth factor binding proteins, we have undertaken to explore the ovarian expression, cellular localization, and hormonal regulation of the insulin-like growth factor binding protcin-2 (1GFBP-2) gene lor which an anligonadoiropic potential has recently been demonstrated. To this end, a solution hybridization/RNasc protection assay was employed wherein total ovarian RNA (2(Vg) from immature (21-23 davs old) female rats was hybridized with a 32P-laballed 1GFBP-2 nboprobe. As in liver, a single protected fragment (550 bases long.) corresponding to IGFBP-2 transcripts was identified in whole ovarian material. Cellular localization studies revealed the IGFBP-2 gene to be exclusively expressed in the ihcca-intcrstilial rather than the granulosa cell compartment. To confirm the cellular disinbution of the IGFBP-2 protein, media conditioned by cultured granulosa or theca-intcrsiitial cells weresubjectedtoimmunoprecipation using two IGFBP-2-directedpolyclonalanliscraExpectdly.bothantibodies(butnotnon-immunerabbitserum) readily.mmunoprccip.latcd the 28kDa rat IGFBP-2 species “cncratcd by hepatic BRL-3A cells. Similarly, both antibodies effectively immunoprccipitatcd an IGFBP the size ol rat IGFBP-2 elaborated by theca-micrstiiial cells. In contrast, neither antibody immunoprccipitatcd the 28-29kDa IGFBP species elaborated by granulosa cells otherwise readily apparent in conventional Western ligand blots. Hypophyscctomy resulted in a 3-fold decrease (P<0.05) in the relative (dcnsiiomctncally-quantidcd) abundance ol ovarian IGFBP-2 transcripts, a diametrically opposed effect (P<0.05) being noted at the level of the liver. In contrast, treatment ol immature hypophyscctomized rats with a dicthvlstilbcstrol-containing subcutaneous silastic implant for a total of 5 days resulted in a concordant 3-fold increase (P<0.05) in the relative abundance of IGFBP-2 transcripts in both ovary and liver when compared with untreated hypophyscctomized controls. Taken together, these findings document rat ovarian IGFPB-2 gene expression to be Ihcca-inlcrslitial (rather than granulosa) cell-selective, and subject to uprcgulatory control by pituitary principlc(s) and/or by estrogens. Although equally estrogen-dependent, hepatic IGFBP-2 gene expression proved constitutive in nature and subject to (diametrically opposed) inhibitory control by (potentially distinct) pituitary principlc(s).",
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T2 - Evidence for hormonal regulation

AU - Ricciarelli, Elisabctta

AU - Hernandez, Elcuterio R.

AU - Hurwitz, Aryc

AU - Kokia, Ehud

AU - Rosenfeld, Ronald (Ron)

AU - Adashi, Eli Y.

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N2 - To begin the process of identification and charactization of rat ovarian insulin-like growth factor binding proteins, we have undertaken to explore the ovarian expression, cellular localization, and hormonal regulation of the insulin-like growth factor binding protcin-2 (1GFBP-2) gene lor which an anligonadoiropic potential has recently been demonstrated. To this end, a solution hybridization/RNasc protection assay was employed wherein total ovarian RNA (2(Vg) from immature (21-23 davs old) female rats was hybridized with a 32P-laballed 1GFBP-2 nboprobe. As in liver, a single protected fragment (550 bases long.) corresponding to IGFBP-2 transcripts was identified in whole ovarian material. Cellular localization studies revealed the IGFBP-2 gene to be exclusively expressed in the ihcca-intcrstilial rather than the granulosa cell compartment. To confirm the cellular disinbution of the IGFBP-2 protein, media conditioned by cultured granulosa or theca-intcrsiitial cells weresubjectedtoimmunoprecipation using two IGFBP-2-directedpolyclonalanliscraExpectdly.bothantibodies(butnotnon-immunerabbitserum) readily.mmunoprccip.latcd the 28kDa rat IGFBP-2 species “cncratcd by hepatic BRL-3A cells. Similarly, both antibodies effectively immunoprccipitatcd an IGFBP the size ol rat IGFBP-2 elaborated by theca-micrstiiial cells. In contrast, neither antibody immunoprccipitatcd the 28-29kDa IGFBP species elaborated by granulosa cells otherwise readily apparent in conventional Western ligand blots. Hypophyscctomy resulted in a 3-fold decrease (P<0.05) in the relative (dcnsiiomctncally-quantidcd) abundance ol ovarian IGFBP-2 transcripts, a diametrically opposed effect (P<0.05) being noted at the level of the liver. In contrast, treatment ol immature hypophyscctomized rats with a dicthvlstilbcstrol-containing subcutaneous silastic implant for a total of 5 days resulted in a concordant 3-fold increase (P<0.05) in the relative abundance of IGFBP-2 transcripts in both ovary and liver when compared with untreated hypophyscctomized controls. Taken together, these findings document rat ovarian IGFPB-2 gene expression to be Ihcca-inlcrslitial (rather than granulosa) cell-selective, and subject to uprcgulatory control by pituitary principlc(s) and/or by estrogens. Although equally estrogen-dependent, hepatic IGFBP-2 gene expression proved constitutive in nature and subject to (diametrically opposed) inhibitory control by (potentially distinct) pituitary principlc(s).

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