Characterization and hormonal regulation of granulosa cell-derived insulin-like growth factor binding protein-4

DooSeok Choi, Lechoslaw T. Putowski, Paul J. Fielder, Ronald (Ron) Rosenfeld, Richard M. Rohan, Eli Y. Adashi

Research output: Contribution to journalArticle

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Abstract

OBJECTIVE: Because of the potential importance of insulin-like growth factor binding protein-4 (IGFBP-4) to ovarian physiology and the obvious limitations imposed by in vivo-exclusive experimental paradigms, we set out to delineate the characteristics and hormonal regulation of granulosa cell- derived IGFBP-4 under in vitro circumstances. METHODS: Granulosa cells obtained by follicular puncture of the ovaries from diethylstilbestrol- primed intact immature rats were subjected to culture for up to 72 hours. Insulin-like growth factor binding protein-4 mRNA extracted from culture was subjected to Northern blot hybridization. Data normalization was assured by reprobing with the hamster Chinese hamster ovary B(CHOB) dna, and the IGFBP- 4/CHOB ratio was calculated. Conditioned culture media were subjected to Western ligand blot before and after immunoprecipitation with a rat IGFBP-4- directed polyclonal antiserum (α-B104). RESULTS: Immunoprecipitation studies revealed granulosa cell-derived IGFBP-4 to be composed of a major 24-kDa species as well as a relatively minor 27-kDa moiety. Given cultures of untreated granulosa cells from immature estrogen-treated rats, transcripts corresponding to IGFBP-4 displayed an initial temporary decline culminating in a 6-hour nadir (a decrease of 67%; P19 aromatase substrate androstenedione (10-7 mol/L) to culture medium from 72 hours enhanced the inhibitory effect of FSH (100 ng/mL) for a maximal decrement in IGFBP-4 transcript of 49% (P <.05). Treatment with insulin-like growth factor (IGF)- I produced limited inhibition (up to 26%) of the steady-state levels of IGFBP-4 transcripts (P <.05). CONCLUSION: Finding indicate the existence of heterogenously-sized IGFBP-4 species, of which the 27-kDa (as distinct from the 24-kDa) IGFBP-4 moiety constitutes a relatively minor component. The steady-state levels of granulosa cell-derived IGFBP-4 transcripts display relatively limited regulation in response to treatment with either FSH or IGF-L.

Original languageEnglish (US)
Pages (from-to)145-151
Number of pages7
JournalJournal of the Society for Gynecologic Investigation
Volume3
Issue number3
DOIs
StatePublished - May 1996
Externally publishedYes

Fingerprint

Insulin-Like Growth Factor Binding Protein 4
Granulosa Cells
Ovary
Cricetulus
Immunoprecipitation
Diethylstilbestrol
Aromatase
Somatomedins
Conditioned Culture Medium
Insulin-Like Growth Factor I
Punctures
Cricetinae
Northern Blotting

Keywords

  • carrier proteins
  • gonadotropins
  • Insulin-like growth factors

ASJC Scopus subject areas

  • Obstetrics and Gynecology

Cite this

Characterization and hormonal regulation of granulosa cell-derived insulin-like growth factor binding protein-4. / Choi, DooSeok; Putowski, Lechoslaw T.; Fielder, Paul J.; Rosenfeld, Ronald (Ron); Rohan, Richard M.; Adashi, Eli Y.

In: Journal of the Society for Gynecologic Investigation, Vol. 3, No. 3, 05.1996, p. 145-151.

Research output: Contribution to journalArticle

Choi, DooSeok ; Putowski, Lechoslaw T. ; Fielder, Paul J. ; Rosenfeld, Ronald (Ron) ; Rohan, Richard M. ; Adashi, Eli Y. / Characterization and hormonal regulation of granulosa cell-derived insulin-like growth factor binding protein-4. In: Journal of the Society for Gynecologic Investigation. 1996 ; Vol. 3, No. 3. pp. 145-151.
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abstract = "OBJECTIVE: Because of the potential importance of insulin-like growth factor binding protein-4 (IGFBP-4) to ovarian physiology and the obvious limitations imposed by in vivo-exclusive experimental paradigms, we set out to delineate the characteristics and hormonal regulation of granulosa cell- derived IGFBP-4 under in vitro circumstances. METHODS: Granulosa cells obtained by follicular puncture of the ovaries from diethylstilbestrol- primed intact immature rats were subjected to culture for up to 72 hours. Insulin-like growth factor binding protein-4 mRNA extracted from culture was subjected to Northern blot hybridization. Data normalization was assured by reprobing with the hamster Chinese hamster ovary B(CHOB) dna, and the IGFBP- 4/CHOB ratio was calculated. Conditioned culture media were subjected to Western ligand blot before and after immunoprecipitation with a rat IGFBP-4- directed polyclonal antiserum (α-B104). RESULTS: Immunoprecipitation studies revealed granulosa cell-derived IGFBP-4 to be composed of a major 24-kDa species as well as a relatively minor 27-kDa moiety. Given cultures of untreated granulosa cells from immature estrogen-treated rats, transcripts corresponding to IGFBP-4 displayed an initial temporary decline culminating in a 6-hour nadir (a decrease of 67{\%}; P19 aromatase substrate androstenedione (10-7 mol/L) to culture medium from 72 hours enhanced the inhibitory effect of FSH (100 ng/mL) for a maximal decrement in IGFBP-4 transcript of 49{\%} (P <.05). Treatment with insulin-like growth factor (IGF)- I produced limited inhibition (up to 26{\%}) of the steady-state levels of IGFBP-4 transcripts (P <.05). CONCLUSION: Finding indicate the existence of heterogenously-sized IGFBP-4 species, of which the 27-kDa (as distinct from the 24-kDa) IGFBP-4 moiety constitutes a relatively minor component. The steady-state levels of granulosa cell-derived IGFBP-4 transcripts display relatively limited regulation in response to treatment with either FSH or IGF-L.",
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T1 - Characterization and hormonal regulation of granulosa cell-derived insulin-like growth factor binding protein-4

AU - Choi, DooSeok

AU - Putowski, Lechoslaw T.

AU - Fielder, Paul J.

AU - Rosenfeld, Ronald (Ron)

AU - Rohan, Richard M.

AU - Adashi, Eli Y.

PY - 1996/5

Y1 - 1996/5

N2 - OBJECTIVE: Because of the potential importance of insulin-like growth factor binding protein-4 (IGFBP-4) to ovarian physiology and the obvious limitations imposed by in vivo-exclusive experimental paradigms, we set out to delineate the characteristics and hormonal regulation of granulosa cell- derived IGFBP-4 under in vitro circumstances. METHODS: Granulosa cells obtained by follicular puncture of the ovaries from diethylstilbestrol- primed intact immature rats were subjected to culture for up to 72 hours. Insulin-like growth factor binding protein-4 mRNA extracted from culture was subjected to Northern blot hybridization. Data normalization was assured by reprobing with the hamster Chinese hamster ovary B(CHOB) dna, and the IGFBP- 4/CHOB ratio was calculated. Conditioned culture media were subjected to Western ligand blot before and after immunoprecipitation with a rat IGFBP-4- directed polyclonal antiserum (α-B104). RESULTS: Immunoprecipitation studies revealed granulosa cell-derived IGFBP-4 to be composed of a major 24-kDa species as well as a relatively minor 27-kDa moiety. Given cultures of untreated granulosa cells from immature estrogen-treated rats, transcripts corresponding to IGFBP-4 displayed an initial temporary decline culminating in a 6-hour nadir (a decrease of 67%; P19 aromatase substrate androstenedione (10-7 mol/L) to culture medium from 72 hours enhanced the inhibitory effect of FSH (100 ng/mL) for a maximal decrement in IGFBP-4 transcript of 49% (P <.05). Treatment with insulin-like growth factor (IGF)- I produced limited inhibition (up to 26%) of the steady-state levels of IGFBP-4 transcripts (P <.05). CONCLUSION: Finding indicate the existence of heterogenously-sized IGFBP-4 species, of which the 27-kDa (as distinct from the 24-kDa) IGFBP-4 moiety constitutes a relatively minor component. The steady-state levels of granulosa cell-derived IGFBP-4 transcripts display relatively limited regulation in response to treatment with either FSH or IGF-L.

AB - OBJECTIVE: Because of the potential importance of insulin-like growth factor binding protein-4 (IGFBP-4) to ovarian physiology and the obvious limitations imposed by in vivo-exclusive experimental paradigms, we set out to delineate the characteristics and hormonal regulation of granulosa cell- derived IGFBP-4 under in vitro circumstances. METHODS: Granulosa cells obtained by follicular puncture of the ovaries from diethylstilbestrol- primed intact immature rats were subjected to culture for up to 72 hours. Insulin-like growth factor binding protein-4 mRNA extracted from culture was subjected to Northern blot hybridization. Data normalization was assured by reprobing with the hamster Chinese hamster ovary B(CHOB) dna, and the IGFBP- 4/CHOB ratio was calculated. Conditioned culture media were subjected to Western ligand blot before and after immunoprecipitation with a rat IGFBP-4- directed polyclonal antiserum (α-B104). RESULTS: Immunoprecipitation studies revealed granulosa cell-derived IGFBP-4 to be composed of a major 24-kDa species as well as a relatively minor 27-kDa moiety. Given cultures of untreated granulosa cells from immature estrogen-treated rats, transcripts corresponding to IGFBP-4 displayed an initial temporary decline culminating in a 6-hour nadir (a decrease of 67%; P19 aromatase substrate androstenedione (10-7 mol/L) to culture medium from 72 hours enhanced the inhibitory effect of FSH (100 ng/mL) for a maximal decrement in IGFBP-4 transcript of 49% (P <.05). Treatment with insulin-like growth factor (IGF)- I produced limited inhibition (up to 26%) of the steady-state levels of IGFBP-4 transcripts (P <.05). CONCLUSION: Finding indicate the existence of heterogenously-sized IGFBP-4 species, of which the 27-kDa (as distinct from the 24-kDa) IGFBP-4 moiety constitutes a relatively minor component. The steady-state levels of granulosa cell-derived IGFBP-4 transcripts display relatively limited regulation in response to treatment with either FSH or IGF-L.

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KW - gonadotropins

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