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J Clin Invest. Dec 1994; 94(6): 2475–2480.
PMCID: PMC330080

Estrogen levels in childhood determined by an ultrasensitive recombinant cell bioassay.

Abstract

We hypothesized that estradiol levels are higher in prepubertal girls than in prepubertal boys and that this greater secretion of estradiol might drive the more rapid epiphyseal development and earlier puberty in girls. Since previous estradiol assays have lacked adequate sensitivity to test the hypothesis of higher estradiol levels in girls, we developed a new ultrasensitive assay to measure estrogen levels. The assay uses a strain of Saccharomyces cerevisiae genetically engineered for extreme sensitivity to estrogen. Yeast were transformed with plasmids encoding the human estrogen receptor and an estrogen-responsive promoter fused to the structural gene for beta-galactosidase. Ether extracts of 0.8 ml of serum were incubated with yeast for 8 h and the beta-galactosidase response was used to determine estrogen bioactivity relative to estradiol standards prepared in charcoal-stripped plasma. The assay was highly specific for estradiol with < 3% cross-reactivity with estrone, estriol, or estradiol metabolites. The detection limit was < 0.02 pg/ml estradiol equivalents (100-fold lower than existing assays). Using this assay, we measured estrogen levels in 23 prepubertal boys (9.4 +/- 2.0 yr) and 21 prepubertal girls (7.7 +/- 1.9 [SD] yr). The estrogen level in girls, 0.6 +/- 0.6 pg/ml estradiol equivalents, was significantly greater than the level in boys, 0.08 +/- 0.2 pg/ml estradiol equivalents (P < 0.05). We conclude that the ultrasensitive recombinant cell bioassay for estrogen is approximately 100-fold more sensitive than previous estradiol assays, that estrogen levels are much lower prepubertally, in both sexes, than reported previously, and that prepubertal girls have 8-fold higher estrogen levels than prepubertal boys.

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Selected References

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  • Garibaldi LR, Aceto T, Jr, Weber C. The pattern of gonadotropin and estradiol secretion in exaggerated thelarche. Acta Endocrinol (Copenh) 1993 Apr;128(4):345–350. [PubMed]
  • Marshall WA, Tanner JM. Variations in pattern of pubertal changes in girls. Arch Dis Child. 1969 Jun;44(235):291–303. [PMC free article] [PubMed]
  • Caruso-Nicoletti M, Cassorla F, Skerda M, Ross JL, Loriaux DL, Cutler GB., Jr Short term, low dose estradiol accelerates ulnar growth in boys. J Clin Endocrinol Metab. 1985 Nov;61(5):896–898. [PubMed]
  • Vanderschueren-Lodeweyckx M, Massa G, Maes M, Craen M, van Vliet G, Heinrichs C, Malvaux P. Growth-promoting effect of growth hormone and low dose ethinyl estradiol in girls with Turner's syndrome. J Clin Endocrinol Metab. 1990 Jan;70(1):122–126. [PubMed]
  • Ross JL, Cassorla FG, Skerda MC, Valk IM, Loriaux DL, Cutler GB., Jr A preliminary study of the effect of estrogen dose on growth in Turner's syndrome. N Engl J Med. 1983 Nov 3;309(18):1104–1106. [PubMed]
  • Ross JL, Cassorla F, Carpenter G, Long LM, Royster MS, Loriaux DL, Cutler GB., Jr The effect of short term treatment with growth hormone and ethinyl estradiol on lower leg growth rate in girls with Turner's syndrome. J Clin Endocrinol Metab. 1988 Sep;67(3):515–518. [PubMed]
  • Laue L, Jones J, Barnes KM, Cutler GB., Jr Treatment of familial male precocious puberty with spironolactone, testolactone, and deslorelin. J Clin Endocrinol Metab. 1993 Jan;76(1):151–155. [PubMed]
  • Forest MG, De Peretti E, Bertrand J. Hypothalamic-pituitary-gonadal relationships in man from birth to puberty. Clin Endocrinol (Oxf) 1976 Sep;5(5):551–569. [PubMed]
  • Reyes FI, Boroditsky RS, Winter JS, Faiman C. Studies on human sexual development. II. Fetal and maternal serum gonadotropin and sex steroid concentrations. J Clin Endocrinol Metab. 1974 Apr;38(4):612–617. [PubMed]
  • Shutt DA, Smith ID, Shearman RP. Oestrone, oestradiol-17beta and oestriol levels in human foetal plasma during gestation and at term. J Endocrinol. 1974 Feb;60(2):333–341. [PubMed]
  • Kenny FM, Angsusingha K, Stinson D, Hotchkiss J. Unconjugated estrogens in the perinatal period. Pediatr Res. 1973 Oct;7(10):826–831. [PubMed]
  • Bidlingmaier F, Wagner-Barnack M, Butenandt O, Knorr D. Plasma estrogens in childhood and puberty under physiologic and pathologic conditions. Pediatr Res. 1973 Nov;7(11):901–907. [PubMed]
  • Baker HW, Burger HG, de Kretser DM, Hudson B, O'Connor S, Wang C, Mirovics A, Court J, Dunlop M, Rennie GC. Changes in the pituitary-testicular system with age. Clin Endocrinol (Oxf) 1976 Jul;5(4):349–372. [PubMed]
  • Belgorosky A, Rivarola MA. Progressive increase in nonsex hormone-binding globulin-bound testosterone and estradiol from infancy to late prepuberty in girls. J Clin Endocrinol Metab. 1988 Aug;67(2):234–237. [PubMed]
  • Ducharme JR, Forest MG, De Peretti E, Sempé M, Collu R, Bertrand J. Plasma adrenal and gonadal sex steroids in human pubertal development. J Clin Endocrinol Metab. 1976 Mar;42(3):468–476. [PubMed]
  • Picard D, Khursheed B, Garabedian MJ, Fortin MG, Lindquist S, Yamamoto KR. Reduced levels of hsp90 compromise steroid receptor action in vivo. Nature. 1990 Nov 8;348(6297):166–168. [PubMed]
  • Fuqua SA, Fitzgerald SD, Chamness GC, Tandon AK, McDonnell DP, Nawaz Z, O'Malley BW, McGuire WL. Variant human breast tumor estrogen receptor with constitutive transcriptional activity. Cancer Res. 1991 Jan 1;51(1):105–109. [PubMed]
  • McDonnell DP, Nawaz Z, Densmore C, Weigel NL, Pham TA, Clark JH, O'Malley BW. High level expression of biologically active estrogen receptor in Saccharomyces cerevisiae. J Steroid Biochem Mol Biol. 1991 Sep;39(3):291–297. [PubMed]
  • Pham TA, Hwung YP, Santiso-Mere D, McDonnell DP, O'Malley BW. Ligand-dependent and -independent function of the transactivation regions of the human estrogen receptor in yeast. Mol Endocrinol. 1992 Jul;6(7):1043–1050. [PubMed]
  • Moehle CM, Aynardi MW, Kolodny MR, Park FJ, Jones EW. Protease B of Saccharomyces cerevisiae: isolation and regulation of the PRB1 structural gene. Genetics. 1987 Feb;115(2):255–263. [PMC free article] [PubMed]
  • Menzel R. A microtiter plate-based system for the semiautomated growth and assay of bacterial cells for beta-galactosidase activity. Anal Biochem. 1989 Aug 15;181(1):40–50. [PubMed]
  • Yocum RR, Hanley S, West R, Jr, Ptashne M. Use of lacZ fusions to delimit regulatory elements of the inducible divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Oct;4(10):1985–1998. [PMC free article] [PubMed]
  • MARTIN L, CLARINGBOLD PJ. The mitogenic action of oestrogens in the vaginal epithelium of the ovariectomized mouse. J Endocrinol. 1960 May;20:173–186. [PubMed]
  • MARTIN L. The use of 2-3-5-triphenyltetrazolium chloride in the biological assay of oestrogens. J Endocrinol. 1960 May;20:187–197. [PubMed]
  • DORFMAN RI, DORFMAN AS. Estrogen assays using the rat uterus. Endocrinology. 1954 Jul;55(1):65–69. [PubMed]
  • BATES RW, COHEN H. Experimental basis for selecting the optimal conditions for quantitative fluorometry of natural estrogens. Endocrinology. 1950 Sep;47(3):166–181. [PubMed]
  • Abraham GE. Radioimmunoassay of steroids in biological fluids. J Steroid Biochem. 1975 Mar-Apr;6(3-4):261–270. [PubMed]
  • Loriaux DL, Ruder HJ, Lipsett MB. The measurement of estrone sulfate in plasma. Steroids. 1971 Oct;18(4):463–472. [PubMed]
  • Ruder HJ, Loriaux L, Lipsett MB. Estrone sulfate: production rate and metabolism in man. J Clin Invest. 1972 Apr;51(4):1020–1033. [PMC free article] [PubMed]
  • Nagai N, Longcope C. Estradiol-17-beta and estrone: studies on their binding to rabbit uterine cytosol and their concentration in plasma. Steroids. 1971 Jun;17(6):631–647. [PubMed]
  • Spratt DI, Longcope C, Cox PM, Bigos ST, Wilbur-Welling C. Differential changes in serum concentrations of androgens and estrogens (in relation with cortisol) in postmenopausal women with acute illness. J Clin Endocrinol Metab. 1993 Jun;76(6):1542–1547. [PubMed]
  • Longcope C, Franz C, Morello C, Baker R, Johnston CC., Jr Steroid and gonadotropin levels in women during the peri-menopausal years. Maturitas. 1986 Oct;8(3):189–196. [PubMed]
  • Longcope C, Pratt JH. Relationship between urine and plasma estrogen ratios. Cancer Res. 1978 Nov;38(11 Pt 2):4025–4028. [PubMed]
  • Longcope C, Watson D, Williams KI. The effects of synthetic estrogens on the metabolic clearance and production rates of estrone and estradiol. Steroids. 1974 Jul;24(1):15–30. [PubMed]
  • England BG, Niswender GD, Midgley AR., Jr Radioimmunoassay of estradiol-17beta without chromatography. J Clin Endocrinol Metab. 1974 Jan;38(1):42–50. [PubMed]
  • Korenman SG, Stevens RH, Carpenter LA, Robb M, Niswender GD, Sherman BM. Estradiol radioimmunoassay without chromatography: procedure, validation and normal values. J Clin Endocrinol Metab. 1974 Apr;38(4):718–720. [PubMed]
  • Hansen JW, Hoffman HJ, Ross GT. Monthly gonadotropin cycles in premenarcheal girls. Science. 1975 Oct 10;190(4210):161–163. [PubMed]
  • Demers LM, Lipton A, Harvey HA, Kambic KB, Grossberg H, Brady C, Santen RJ. The efficacy of CGS 20267 in suppressing estrogen biosynthesis in patients with advanced stage breast cancer. J Steroid Biochem Mol Biol. 1993 Mar;44(4-6):687–691. [PubMed]

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