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Use of Complimentary In Vitro and In Vivo Methods for Assessing Neuroendocrine Disruptors

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In Vitro Neurotoxicology

Part of the book series: Methods in Pharmacology and Toxicology ((MIPT))

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Abstract

It is often necessary to use a variety of techniques to more completely assess a specific area of study. In recent years, this has been increasingly true for the field of neuroendocrinology. Anatomical and molecular methodologies are often used in conjunction with in vitro and in vivo physiological approaches to gain meaningful information regarding factors controlling or altering neuroendocrine functions. When used together in a given study, in vitro and in vivo methods can be complimentary to one another and, thus, provide vital information. The focus of this chapter will be to demonstrate how both in vitro and in vivo techniques can be used in this complimentary fashion to provide new and important insights into basic neuroendocrine events and their mechanisms of action. Furthermore, we will demonstrate how these techniques can be used to help better understand the sites of action and the effects of specific toxic substances that alter neuroendocrine function.

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References

  1. Dees, W. L. and Skelley, C. W. (1990) Effects of ethanol during the onset of female puberty. Neuroendocrinology 51, 64–69

    Article  PubMed  CAS  Google Scholar 

  2. Tannenbaum, G. S., Guyda, H. J., and Posner, B. I. (1983) Insulin-like growth factors: a role in growth hormone negative feedback and body weight regulation via brain. Science 220, 77–79

    Article  PubMed  CAS  Google Scholar 

  3. Schoenle, E., Zapf, J., Hauri, C., Steiner, T., and Froesch, E. R. (1985) Comparison of in vivo effects of insulin-like growth factor I and II and of growth hormone in hypophysectomized rats. Acta Endocrinol. 108, 167–174

    PubMed  CAS  Google Scholar 

  4. Philipps, A. F., Persson, B., Hall, K., et al. (1988) The effects of biosynthetic insulin-like growth factor 1. Supplementation of somatic growth, maturation, and erthropoiesis on neonatal rat. Pediatr. Res. 23, 298–305

    PubMed  CAS  Google Scholar 

  5. Hizuka, N., Takano, K., Shizume, K., et al. (1986) Insulin-like growth factor I stimulates growth in normal growing rats. Eur. J. Pharmacol. 125, 143–146

    Article  PubMed  CAS  Google Scholar 

  6. Behringer, R. R., Lewin, T. L., Quaife, C. J., Palmiter, R. D., Brinster, R. L., and D’Ercole, A. J. (1990) Expression of insulin-like growth factor 1 stimulates normal somatic growth in growth hormone deficient transgenic mice. Endocrinology 127, 1033–1040

    Article  PubMed  CAS  Google Scholar 

  7. Bohannon, N. J., Figlewicz, D. P., Corp, E. S., Wilcox, J., Porte, C. D., Jr., and Baskin, D G. (1986) Identification of binding sites for an insulin-like growth factor (IGF-1) in the median eminence of the rat brain by quantitative autoradiography. Endocrinology 119, 943–945

    Article  PubMed  CAS  Google Scholar 

  8. Marks, J. L., Porte, D., Jr., and Baskin, D. G. (1991) Localization of type 1 insulin-like growth factor receptor messenger RNA in the adult rat brain by in situ hybridization. Mol. Endocrinol. 5, 1158–1168

    Article  PubMed  CAS  Google Scholar 

  9. Negro-Vilar, A., Ojeda, S. R., and McCann, S. M. (1979) Catecholaminergic modulation of luteinizing hormone-releasing hormone release by median eminence terminals in vitro. Endocrinology 128, 1749–1757.

    Article  Google Scholar 

  10. Hiney, J. K., Ojeda, S. R., and Dees, W. L. (1991) Insulin-like growth factor-1: a possible metabolic signal involved in the regulation of female puberty. Neuroendocrinology 54, 420–423

    Article  PubMed  CAS  Google Scholar 

  11. Duenas, M., Luquin, S., Chowen, J. A., Torres-Aleman, I., Naftolin, F., and Garcia-Segura, L. M. (1994) Gonadal hormone regulation of insulin-like growth factor-1-like immunoreactivity in hypothalamic astroglia of developing and adult rats. Neuroendocrinology 59, 528–538.

    Article  PubMed  CAS  Google Scholar 

  12. Antunes-Rodrigues, J. and McCann, S. M. (1970) Water, sodium chloride, and food intake induced by injections of cholinergic and adrenergic drugs into the third ventricle of the rat brain. Proc. Soc. Exp. Biol. Med. 133, 1464–1469.

    PubMed  CAS  Google Scholar 

  13. Harms, P. G. and Ojeda, S. R. (1974) Methods for cannulation of the rat jugular vein. J. Appl. Physiol. 309, 261–263.

    Google Scholar 

  14. Hiney, J. K., Srivastava, V., Nyberg, C. L., Ojeda, S. R., and Dees, W. L. (1996) Insulin-like growth factor-1 of peripheral origin acts centrally to accelerate the initiation of female puberty. Endocrinology 137, 3717–3727.

    Article  PubMed  CAS  Google Scholar 

  15. Andrews, W. W. and Ojeda, S. R. (1981) A detailed analysis of the serum LH secretory profile in conscious, free-moving female rats during the time of puberty. Endocrinology 109, 2032–2039.

    Article  PubMed  CAS  Google Scholar 

  16. Terasawa, E., Bridson, W. E., Nass, T. E., Noonan, J. J., and Dierschke, D. J. (1984) Developmental changes in the luteinizing hormone secretory pattern in peripubertal female rhesus monkeys: comparisons between gonadally intact and ovariectomized animals. Endocrinology 115, 2233–2240.

    Article  PubMed  CAS  Google Scholar 

  17. Urbanski, H. F. and Ojeda, S. R. (1985) The juvenile peripubertal transition period in the female rat: Establishment of a diurnal pattern of pulsatile LH secretion. Endocrinology 117, 644–649.

    Article  PubMed  CAS  Google Scholar 

  18. Danilovich, N., Wernsing, D., Coschigano, J. J., Kopchick, J. J., and Bartke, A. (1998) Deficits in female reproductive function in GH-R-KO mice; role of IGF-1. Endocrinology 140, 2637–2640.

    Article  Google Scholar 

  19. Wilson, M. E. (1998) Premature elevation in serum insulin-like growth factor-1 advances first ovulation in rhesus monkeys. J. Endocr. 158, 247–257.

    Article  PubMed  CAS  Google Scholar 

  20. Xiaowei, X., Ingram, R. L., and Sonntag, W. E. (1995) Ethanol suppresses growth hormone-mediated cellular responses in liver slices. Alcoholism: Clin. Exp. Res. 19, 1246–1251.

    Article  Google Scholar 

  21. De, A., Boyadjieva, N. I., and Sarkar, D. K. (1999) Effects of ethanol on alphadrenergic and beta-adrenergic agonist-stimulated beta endorphin release and cAMP production in hypothalamic cells in primary cultures. Alcoholism: Clin. Exper Res. 23, 46–51.

    CAS  Google Scholar 

  22. Frasor, J. M., Anderson, R. A., and Russell, L. D. (1992) Ethanol impairs secretory activity of Sertoli celss in conventional and bicameral culture. Biol. Reprod. 46(Suppl. 1), 171.

    Google Scholar 

  23. Hiney, J. K. and Dees, W. L. (1991) Ethanol inhibits luteinizing hormone-releasing hormone from the median eminence of prepubertal female rats in vitro: Investigation of its actions on norepinephrine and prostaglandin-E2. Endocrinology 128, 1404–1408.

    Article  PubMed  CAS  Google Scholar 

  24. Nyberg, C. L., Hiney, J. K., Minks, J. E., and Dees, W. L. (1993) Ethanol alters N-methyl-dl-aspartic acid-induced LH secretion of luteinizing hormone releasing hormone and the onset of puberty in the female rat. Neuroendocrinology 57, 863–868.

    Article  PubMed  CAS  Google Scholar 

  25. Rettori, V., Kamat, A., and McCann, S. M. (1994) Nitric oxide mediates the stimulation of luteinizing-hormone releasing hormone release induced by glutamic acid in vitro. Brain Res. Bull. 33, 501–503.

    Article  PubMed  CAS  Google Scholar 

  26. Canteros, G., Rettori, V., Franchi, A., et al. (1995) Ethanol inhibits luteinizing hormone-releasing hormone (LHRH) secretion by blocking the response of LHRH neuronal terminals to nitric oxide. PNAS 92, 3416–3420.

    Article  PubMed  CAS  Google Scholar 

  27. Hiney, J. K., Srivastava, V., Lara, T., and Dees, W. L. (1998) Ethanol blocks the central action of IGF-1 to induce luteinizing hormone secretion in the prepubertal female rat. Life Sci. 62, 301–308.

    Article  PubMed  CAS  Google Scholar 

  28. Dees, W. L., Skelley, C. W., Hiney, J. K., and Johnston, C. A. (1990) Actions of ethanol on hypothalamic and pituitary hormones in prepubertal female rats. Alcohol 7, 21–25

    Article  PubMed  CAS  Google Scholar 

  29. Soszynski, P. A. and Frohman, L. A. (1992) Inhibitory effects of ethanol on the growth hormone (GH)-releasing hormone-GH-Insulin-like growth factor-I axis in the rat. Endocrinology 131, 2603–2608.

    Article  PubMed  CAS  Google Scholar 

  30. Dees, W. L., Rettori, V., Kozlowski, G. P., and McCann, S. M. (1985) Ethanol and the pulsatile release of luteinizing hormone, follicle stimulating hormone and prolactin in ovariectomized rats. Alcohol 2, 641–646.

    Article  PubMed  CAS  Google Scholar 

  31. Dees, W. L., Skelley, C. W., Rettori, V., Kentroti, M. S., and McCann, S. M. (1988) Influence of ethanol on growth hormone secretion in adult and prepubertal female rats. Neuroendocrinology 48, 495–499.

    Article  PubMed  CAS  Google Scholar 

  32. Srivastava, V., Hiney, J. K., Nyberg, C. L., and Dees, W. L. (1995) Effect of ethanol on the synthesis of insulin-like growth factor 1 (IGF-1) and the IGF-1 receptor in late prepubertal female rats: a correlation with serum IGF-1. Alcoholism: Clin. Exp. Res. 19, 1467–1473.

    Article  CAS  Google Scholar 

  33. Dees, W. L., Dissen, G. A., Hiney, J. K., Lara, F., and Ojeda, S. R. (2000) Alcohol ingestion inhibits the increased secretion of puberty-related hormones in the developing female Rhesus monkey. Endocrinology 141, 1325–1331.

    Article  PubMed  CAS  Google Scholar 

  34. Urbanski, H. F. and Ojeda, S. R. (1987) Activation of luteinizing hormone-releasing hormone release advances the onset of female puberty. Neuroendocrinology 46, 273–276.

    Article  PubMed  CAS  Google Scholar 

  35. Gay, V. L. and Plant, T. M. (1987) N-Methyl-d,l-aspartate elicits hypothalamic gonadotropin-releasing hormone release in prepubertal male rhesus monkeys (Macaca mulatta). Endocrinology 120, 2289–2296.

    Article  PubMed  CAS  Google Scholar 

  36. Claypool, L. E. and Terasawa, E. (1989) N-Methyl-dl-aspartate (NMDA) induces LHRH release as measured by in vivo push-pull perfusion in the stalk-median eminence of pre and peripubertal female rhesus monkeys. Biol. Reprod. 40(Suppl.), 83.

    Google Scholar 

  37. Price, M. T., Olney, J. W., and Cicero, T. J. (1978) Acute elevations of serum luteinizing hormone induced by kainic acid, N-methyl-aspartic acid or homosysteic acid. Neuroendocrinology 26, 352–358.

    Article  PubMed  CAS  Google Scholar 

  38. Ondo, J. G., Wheeler, D. D., and Dom, R. M. (1988) Hypothalamic site of action for N-methyl-d-aspartate (NMDA) on LH secretion. Life Sci. 43, 2283–2286.

    Article  PubMed  CAS  Google Scholar 

  39. Wilson, R. C. and Knobil, E. (1982) Acute effects of N-methyl-dl-aspartate on the release of pituitary gonadotropins and prolactin in the adult female rhesus monkey. Brain Res. 248, 177–179.

    Article  PubMed  CAS  Google Scholar 

  40. Tal, J., Price, M T., and Olney, J. W. (1983) Neuroactive amino acids influence gonadotropin output by a suprapituitary mechanism in either rodents or primates. Brain Res. 273, 170–182.

    Article  Google Scholar 

  41. Bourgiuignon, J. P., Gerard, A., and Franchimont, P. (1989) direct activation of gonadotropin-releasing hormone secretion through different receptors to neuroexcitatory amino acids. Neuroendocrinology 49, 402–408

    Article  Google Scholar 

  42. Donoso, A. O., Lopez, F. J., and Negro-Vilar, A. (1990) Glutamate receptors of the non-N-methyl-d-aspartic acid type mediate the increase in luteinizing hormone-releasing hormone release by excitatory amino acids in vitro. Endocrinology 126, 414–420.

    Article  PubMed  CAS  Google Scholar 

  43. Lopez, F. J., Donoso, A. O., and Negro-Vilar, A. (1990) Endogenous excitatory amino acid neurotransmission regulates the estradiol-induced LH surge in ovariectomized rats Endocrinology 126, 1771–1773.

    Article  PubMed  CAS  Google Scholar 

  44. Lovinger, D. M., White, G., and Weight, F. F. (1989) Ethanol inhibits NMDA-activated ion current in hippocampal neurons. Science 128, 1541–1547.

    Google Scholar 

  45. Leslie, S. W., Brwon, L. M., Dildy, J. E., and Sims, J. S. (1990) Ethanol and neuronal calcium channels. Alcohol 7, 233–236.

    Article  PubMed  CAS  Google Scholar 

  46. Simson, P. E., Criswell, H. E., Johnson, K. B., Hicks, R. E., and Breese, G. R. (1991) Ethanol inhibits NMDA-evoked electrophysiological activity in vivo. J. Pharmacol. Exp. Ther. 257, 225–231.

    PubMed  CAS  Google Scholar 

  47. Nyberg, C. L., Srivastava, V., Hiney, J. K., Lara, F., and Dees, W. L. (1995) N-Methyl-aspartic acid receptor messenger ribonucleic acid levels and luteinizing hormone release in immature female rats. Effects of stage of pubertal development and exposure to ethanol. Endocrinology 136, 2874–2880.

    Article  PubMed  CAS  Google Scholar 

  48. Swerdloff, R., Batt, R., and Bray, G. (1976) Reproductive hormonal function in the genetically obese (ob/ob) mouse. Endocrinology 103, 542–547.

    Article  Google Scholar 

  49. Swerdloff, R. S., Peterson, M., Vera, A., Batt, R. A. L., Heber, D., and Bray, G. (1978) The hypothalamic-pituitary axis in genetically obese (ob/ob) mice: Response to luteinizing hormone-releasing hormone. Endocrinology 103, 542–547.

    Article  PubMed  CAS  Google Scholar 

  50. Johnson, L. M. and Sidnam, R. L. (1979) A reproductive endocrine profile in the diabetes (db) mutant mouse. Biol. Reprod. 20, 552–559.

    Article  PubMed  CAS  Google Scholar 

  51. Batt, R., Everard, D., Gillies, G., Wilkinson, M., Wilson, C., and Yeo, T. (1982) Investigation into the hypogonadism of the obese mouse (genotype ob/ob). J. Reprod. Fertil. 64, 363–371.

    Article  PubMed  CAS  Google Scholar 

  52. Chehab, F. F., Lim, M. E., and Ronghua, L. (1996) Correction of the sterility defect in homozygous obese female mice. Nature Genet. 12, 318–320.

    Article  PubMed  CAS  Google Scholar 

  53. Barash, I. A., Cheung, C. C., Weigle, D. S., et al. (1996) Leptin is a metabolic signal to the reproductive system. Endocrinology 137, 3144–3147.

    Article  PubMed  CAS  Google Scholar 

  54. Mounzith, K., Ronghua, L., and Chehab, F. F. (1997) Leptin treatment rescues the sterility of genetically obese ob/ob males. Endocrinology 138, 1190–1193.

    Article  Google Scholar 

  55. Chehab, F. F., Mounzih, K., Ronghua, L., and Lim, M. E. (1997) Early onset of reproductive function in normal female mice treated with leptin. Science 275, 88–90.

    Article  PubMed  CAS  Google Scholar 

  56. Ahima, R. S., Dushay, J., Flier, S. N., Prabakaran, D., and Flier, J. S. (1997) Leptin accelerates the onset of puberty in normal female mice. J. Clin. Invest. 99, 391–395.

    Article  PubMed  CAS  Google Scholar 

  57. Yu, W. H., Kimura, M., Walczewska, A., Karanth, S., and McCann, S. M. (1997) Role of leptin in hypothalamic-pituitary function. Proc. Natl. Acad. Sci. USA 94, 1023–1028.

    Article  PubMed  CAS  Google Scholar 

  58. Carro, E., Pinilla, L., Seoane, L. M., et al. (1997) Influence of endogenous leptin tone on the estrous cycle and luteinizing hormone pulsatility in female rats. Neuroendocrinology 66, 375–377.

    Article  PubMed  CAS  Google Scholar 

  59. Cheung, C. C., Thornton, J. E., Kuijper, J. L., Weigle, D. S., Clifton, D. K., and Steiner, R. A. (1997) Leptin is a metabolic gate for the onset of puberty in the female. Endocrinology 138, 855–858.

    Article  PubMed  CAS  Google Scholar 

  60. Dearth, R. K., Hiney, J. K., and Dees, W. L. (2000) Leptin acts centrally to induce the prepubertal secretion of luteinizing hormone in the female rat. Peptides 21, 387–392.

    Article  PubMed  CAS  Google Scholar 

  61. Hiney, J. K., Dearth, R. K., Lara, F., Wood, S., Srivastava, V., and Dees, W. L. (1999) Effects of ethanol on leptin secretion and the leptin-induced luteinizing hormone (LH) release from the late juvenile female rats. Alcoholism: Clin. Exp. Res. 23, 1785–1792.

    CAS  Google Scholar 

  62. Rettori, V., Gimeno, M., Lyson, K., and McCann, S. M. (1992) Nitric oxid mediates norepinephrine-induced prostaglandin E2 release from the hypothalamus. Proc. Natl. Acad. Sci. USA 89, 11,543–11,546.

    Article  PubMed  CAS  Google Scholar 

  63. Lomniczi, A., Mastronardi, C. A., Faletti, A. G., et al. (2000) Inhibitory pathways and the inhibition of luteinizing hormone-releasing hormone release by alcohol. Proc. Natl. Acad. Sci. USA 97, 2337–2342.

    Article  PubMed  CAS  Google Scholar 

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© 2004 Humana Press Inc., Totowa, NJ

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Les Dees, W., Hiney, J.K., Dearth, R.K., Srivastava, V.K. (2004). Use of Complimentary In Vitro and In Vivo Methods for Assessing Neuroendocrine Disruptors. In: Tiffany-Castiglioni, E., Hollinger, M.A. (eds) In Vitro Neurotoxicology. Methods in Pharmacology and Toxicology. Humana Press. https://doi.org/10.1385/1-59259-651-7:267

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  • DOI: https://doi.org/10.1385/1-59259-651-7:267

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-047-2

  • Online ISBN: 978-1-59259-651-5

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