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Sex steroids do not alter sex differences in tyrosine hydroxylase activity of dopaminergic neurons in vitro

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Summary

In order to distinguish the effects of genetic sex from those of sex hormones on the sexual differentiation of dopaminergic neurons, catecholamine synthesis was studied in gender-specific cultures of embryonic day-14 rat diencephalon. In addition to embryos from normal dams, embryos were used whose mothers had been treated with the estrogen antagonist tamoxifen or the testosterone antagonist cyproterone acetate on days 12 and 13 of gestation. Cultures from embryos of untreated dams were fed daily with a medium containing 17β-estradiol or testosterone. After 10 days in vitro, cultures were immunostained for tyrosine hydroxylase and the accumulation of dihydroxyphenylalanine (DOPA) was measured in the presence of the DOPA decarboxylase inhibitor NSD 1015. Rates of DOPA synthesis, unlike the numbers of tyrosine hydroxylase-immunoreactive neurons, were markedly higher in female cultures under all experimental conditions. Treatment of dams with antisteroids prior to removal of the embryos had no influence on these results. Treatment of cultures with both steroids decreased DOPA formation in a dose-dependent manner without altering the sex difference. These results suggest that cultured diencephalic dopaminergic neurons develop sex differences in the activity of tyrosine hydroxylase. This sexual dimorphism is initiated independently of the action of gonadal steroid hormones. Sex hormones exert an additional modulatory influence on the activity of the enzyme but do not abolish or reverse sex differences. Therefore, the concept of a purely epigenetic mode of sexual differentiation of the mammalian brain needs to be broadened to incorporate other mechanisms, such as the cell-autonomous fulfillment of a sex-specific genetic program.

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References

  • Abreu P, Hernandez G, Calzadilla CH, Alonso R (1988) Reproductive hormones control striatal tyrosine hydroxylase activity in the male rat. Neurosci Lett 95:213–217

    Google Scholar 

  • Ahnert-Hilger G, Engele J, Reisert I, Pilgrim Ch (1986) Different developmental schedules of dopaminergic and noradrenergic neurons in dissociation culture of fetal rat midbrain and hind-brain. Neuroscience 17:157–165

    Google Scholar 

  • Arai Y, Gorski RA (1968) Critical exposure time for androgenization of the rat hypothalamus determined by antiandrogen injection. Proc Soc Exp Biol Med 127:590–593

    Google Scholar 

  • Arita J, Kimura F (1987) Direct inhibitory effect of long term estradiol treatment on dopamine synthesis in tuberoinfundibular dopaminergic neurons: in vitro studies using hypothalamic slices. Endocrinology 121:692–698

    Google Scholar 

  • Arnold AP, Gorski RA (1984) Gonadal steroid induction of structural sex differences in the central nervous system. Annu Rev Neurosci 7:413–442

    Google Scholar 

  • Baum MJ, Woutersen PJA, Slob AK (1991) Sex difference in whole-body androgen content in rats on fetal days 18 and 19 without evidence that androgen passes from males to females. Biol Reprod 44:747–751

    Google Scholar 

  • Berthois Y, Katzenellenbogen JA, Katzenellenbogen BS (1986) Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc Natl Acad Sci USA 83:2496–2500

    Google Scholar 

  • Beyer C, Pilgrim Ch, Reisert I (1991) Dopamine content and metabolism in mesencephalic and diencephalic cell cultures: sex differences and effects of sex steroids. J Neurosc 11:1325–1333

    Google Scholar 

  • Blum M, McEwen BS, Roberts JL (1987) Transcriptional analysis of tyrosine hydroxylase gene expression in the tuberoinfundibular dopaminergic neurons of the rat arcuate nucleus after estrogen treatment. J Biol Chem 262:817–821

    Google Scholar 

  • Brawer J, Bertley J, Beaudet A (1986) Testosterone inhibition of tyrosine hydroxylase expression in the hypothalamic arcuate nucleus. Neurosci Lett 67:313–318

    Google Scholar 

  • Carlsson A, Davis JN, Kehr W, Lindquist M, Atack CV (1972) Simultaneous measurement of tyrosine and tryptophan hydroxylase activities in brain in vivo using an inhibitor of the aromatic amino acid decarboxylase. Naunyn Schmiedebergs Arch Pharmacol 275:153–168

    Google Scholar 

  • Crowley WR, O'Donohue TL, Jacobowitz DM (1978) Sex differences in catecholamine content in discrete brain nuclei of the rat: effects of neonatal castration or testosterone treatment. Acta Endocrinol (Copenh) 89:20–28

    Google Scholar 

  • Demarest KT, Moore KE (1980) Accumulation of L-dopa in the median eminence: an index of tuberoinfundibular dopaminergic nerve activity. Endocrinology 106:463–468

    Google Scholar 

  • Demarest KT, McKay DW, Riegle GD, Moore KE (1981) Sexual differences in tuberoinfundibular dopamine nerve activity induced by neonatal androgen exposure. Neuroendocrinology 32:108–113

    Google Scholar 

  • Döhler KD, Srivastava SS, Shryne JE, Jarzab B, Sipos A, Gorski RA (1984) Differentiation of the sexually dimorphic nucleus in the preoptic area of the rat brain is inhibited by postnatal treatment with an estrogen antagonist. Neuroendocrinology 38:297–301

    Google Scholar 

  • Döhler KD, Coquelin A, Davis F, Hines M, Shryne JE, Sickmöller PM, Jarzab B, Gorski RA (1986) Fre- and postnatal influence of an estrogen antagonist and an androgen antagonist on differentiation of the sexually dimorphic nucleus of the preoptic area in male and female rats. Neuroendocrinology 42:443–448

    Google Scholar 

  • Engel J, Ahlenius S, Almgren O, Carlsson A (1979) Effects of gonadectomy and hormone replacement on brain monoamine synthesis in male rats. Pharmacol Biochem Behav 10:149–154

    Google Scholar 

  • Engele J, Pilgrim C, Kirsch M, Reisert I (1989a) Different developmental potentials of diencephalic and mesencephalic dopaminergic neurons in vitro. Brain Res 483:98–109

    Google Scholar 

  • Engele J, Pilgrim C, Reisert I (1989b) Sexual differentiation of mesencephalic neurons in vitro: effects of sex and gonadal hormones. Int J Dev Neurosci 7:603–611

    Google Scholar 

  • Eusterschulte B, Reisert I, Pilgrim C (1992) Absence of sex differences in size of the genital ducts of the rat prior to embryonic day 15.5–16.0. Tissue Cell 24:483–489

    Google Scholar 

  • Fabre-Nys C, Adkins-Regan E (1987) Antisteroid action in brain and changes in animal behavior. In: Agarwal MK (ed) Receptor mediated antisteroid action. de Gruyter, Berlin, pp 435–468

    Google Scholar 

  • Friedman WJ, Dreyfus CF, McEwen BS, Black IB (1989) Developmental regulation of tyrosine hydroxylase in the mediobasal hypothalamus. Dev Brain Res 48:177–185

    Google Scholar 

  • Furr BJA, Jordan VC (1984) The pharmacology and clinical uses of tamoxifen. Pharmacol Ther 25:127–205

    Google Scholar 

  • Gudelsky GA, Porter JC (1981) Sex-related difference in the release of dopamine into hypophysial portal blood. Endocrinology 109:1394–1398

    Google Scholar 

  • Herdon HJ, Wilson CA (1985) Changes in hypothalamic dopamine D-2 receptors during sexual maturation in male and female rats.Brain Res 343:151–153

    Google Scholar 

  • Kalra SP, Kalra PS (1983) Neural regulation of luteinizing hormone secretion in the rat. Endocrinol Rev 4:311–351

    Google Scholar 

  • Kolbinger W, Trepel M, Beyer C, Pilgrim C, Reisert I (1991) The influence of genetic sex on sexual differentiation of diencephalic dopaminergic neurons in vitro and in vivo. Brain Res 544:349–352

    Google Scholar 

  • Kordon C, Glowinski J (1972) Role of hypothalamic monoaminergic neurones in the gonadotrophin release-regulation mechanisms. Neuropharmacology 11:153–162

    Google Scholar 

  • Lacau de Mengido I, Becú-Villalobos D, Libertun C (1987) Sexual differences in the dopaminergic control of luteinizing hormone secretion in the developing rat. Dev Brain Res 35:91–95

    Google Scholar 

  • Leret ML, González MI, Tranque P, Fraile A (1987) Influence of sexual differentiation on striatal and limbic catecholamines. Comp Biochem Physiol 86C:299–303

    Google Scholar 

  • Lloyd T, Weisz J (1978) Direct inhibition of tyrosine hydroxylase activity by catechol estrogens. J Biol Chem 253:4841–4843

    Google Scholar 

  • Luine VN, McEwen BS, Black IB (1977) Effect of 17β-estradiol on hypothalamic tyrosine hydroxylase activity. Brain Res 120:188–192

    Google Scholar 

  • MacLusky NJ, Naftolin F (1981) Sexual differentiation of the central nervous system. Science 211:1294–1303

    Google Scholar 

  • Mainwaring WIP (1977) Modes of action of antiandrogens: a survey. In: Martini L, Motta M (eds) Androgens and antiandrogens. Raven Press, New York, pp 151–161

    Google Scholar 

  • Moore KE (1987) Hypothalamic dopaminergic neuronal systems. In: Meltzer HY (ed) Psychopharmacology. The third generation of progress. Raven Press, New York, pp 127–139

    Google Scholar 

  • Morrell JI, Rosenthal MF, McCabe JT, Harrington CA, Chikaraishi DM, Pfaff DW (1989) Tyrosine hydroxylase mRNA in the neurons of the tuberoinfundibular region and zona incerta examined after gonadal steroid hormone treatment. Mol Endocrinol 3:1426–1433

    Google Scholar 

  • Reisert I, Pilgrim C (1991) Sexual differentiation of monoaminergic neurons-genetic or epigenetic? Trends Neurosci 14:468–473

    Google Scholar 

  • Reisert I, Engele J, Pilgrim C (1989) Early sexual differentiation of diencephalic dopaminergic neurons of the rat in vitro. Cell Tissue Res 255:411–417

    Google Scholar 

  • Simerly RB (1989) Hormonal control of the development and regulation of tyrosine hydroxylase expression within a sexually dimorphic population of dopaminergic cells in the hypothalamus. Mol Brain Res 6:297–310

    Google Scholar 

  • Simerly RB, Swanson LW, Gorski RA (1985a) The distribution of monoaminergic cells and fibers in a periventricular preoptic nucleus involved in the control of gonadotropin release: Immunocytochemical evidence for a dopaminergic sexual dimorphism. Brain Res 330:55–64

    Google Scholar 

  • Simerly RB, Swanson LW, Handa RJ, Gorski RA (1985b) Influence of perinatal androgen on the sexually dimorphic distribution of tyrosine hydroxylase-immunoreactive cells and fibers in the anteroventral periventricular nucleus of the rat. Neuroendocrinology 40:501–510

    Google Scholar 

  • Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Anal Biochem 150:76–85

    Google Scholar 

  • Toran-Allerand CD, Gerlach JL, McEwen BS (1980) Autoradiographic localization of [3H]estradiol related to steroid responsiveness in cultures of the newborn mouse hypothalamus and preoptic area. Brain Res 184:517–522

    Google Scholar 

  • Vaccari A, Brotman S, Cimino J, Timiras PS (1977) Sex differentiation of neurotransmitter enzymes in central and peripheral nervous systems. Brain Res 132:176–185

    Google Scholar 

  • Weisz J, Ward IL (1980) Plasma testosterone and progesterone titers of pregnant rats, their male and female fetuses, and neonatal offspring. Endocrinology 106:306–316

    Google Scholar 

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  1. Abteilung Anatomie und Zellbiologie der Universität, Albert-Einstein-Allee 11, W-7900, Ulm, Federal Republic of Germany

    Cordian Beyer, Beate Eusterschulte, Christof Pilgrim & Ingrid Reisert

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  1. Cordian Beyer
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  2. Beate Eusterschulte
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  3. Christof Pilgrim
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  4. Ingrid Reisert
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Beyer, C., Eusterschulte, B., Pilgrim, C. et al. Sex steroids do not alter sex differences in tyrosine hydroxylase activity of dopaminergic neurons in vitro. Cell Tissue Res. 270, 547–552 (1992). https://doi.org/10.1007/BF00645057

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  • DOI: https://doi.org/10.1007/BF00645057

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