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  • Title: Expression and regulation of insulin-like growth factor-binding protein-1, -2, -3, and -4 messenger ribonucleic acids in purified rat Leydig cells and their biological effects.
    Author: Lin T, Wang D, Nagpal ML, Shimasaki S, Ling N.
    Journal: Endocrinology; 1993 May; 132(5):1898-904. PubMed ID: 7682935.
    Abstract:
    The actions of insulin-like growth factor-I (IGF-I) are modified by binding proteins [IGF-binding proteins (IGFBPs)]. Previously, we reported that IGF-I enhances Leydig cell steroidogenesis and that IGF-I mRNA is expressed in Leydig cells. In the present study, we evaluated the expression and regulation of IGFBP-1, -2, -3, and -4 in purified rat Leydig cells and their biological effects. We found that none of the testicular crude interstitial cells, purified Leydig cells, or seminiferous tubules expressed IGFBP-1 mRNA. This indicated that IGFBP-1 mRNA is not expressed in the testis in detectable amounts. In contrast, large amounts of IGFBP-2 with a size of 1.8 kilobases (kb) were expressed in purified Leydig cells, and lesser amounts in crude interstitial cells. Small amounts of IGFBP-2 mRNA were expressed in seminiferous tubules, but none could be detected in liver. IGFBP-3 mRNA was predominantly expressed in purified Leydig cells, crude interstitial cells, and liver, while appreciable amounts were not found in seminiferous tubules. Liver had the highest amounts of IGFBP-4 mRNA, whereas purified Leydig cells and crude interstitial cells had lesser amounts. We next evaluated the pituitary dependency of IGFBP mRNAs in Leydig cells. Purified Leydig cells were isolated from 50-day-old rats 5 days after hypophysectomy. IGFBP-2, -3, and -4 mRNA levels in Leydig cells decreased 22%, 80%, and more than 90%, respectively, after hypophysectomy. In the liver, however, IGFBP-2 mRNA levels increased, and IGFBP-3 mRNA levels decreased, while IGFBP-4 remained unchanged. As expected, hypophysectomy caused decrements in large (7.0- to 7.5-kb; a 75% reduction) and small (0.8- to 1.2-kb; a 90% reduction) IGF-I mRNA levels in Leydig cells. Hypophysectomy also reduced IGF-I mRNA expression in liver. Finally, the effects of IGFBP-2, -3, and -4 on Leydig cell testosterone formation were investigated. hCG in a concentration of 10 ng/ml increased testosterone formation from 0.6 +/- 0.01 to 27.4 +/- 1.01 ng/10(6) cells.h. In the presence of IGF-I (10 ng/ml), testosterone formation was further increased to 58.6 +/- 1.6 ng/10(6) cells.h (P < 0.01). IGFBP-3 (0.1, 1, and 2.5 pmol/ml) caused a dose-dependent inhibition of IGF-I- plus hCG-induced testosterone formation. IGFBP-3 in a concentration of 2.5 pmol/ml completely neutralized the effects of IGF-I on Leydig cell steroidogenesis. IGFBP-4 had a lesser effect, while IGFBP-2 had no effect on IGF-I- plus hCG-induced testosterone formation.(ABSTRACT TRUNCATED AT 400 WORDS)
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