Development of an animal model of hypothyroxinemia during pregnancy in Wistar rats

Abstract

Background: Hypothyroxinemia is a subclinical thyroid hormone deficiency in which the mother has inadequate levels of T₄ during pregnancy. The fetus relies entirely on the mother’s T₄ hormone level for early neurodevelopment. Isolated maternal hypothyroxinemia (IMH) in the first trimester of pregnancy can lead to lower intelligence, lower motor scores, and a higher risk of mental illness in descendants. Here, we focus on the autism-like behavior of IMH offspring.

Methods: The animals were administered 1 ppm of propylthiouracil (PTU) for 9 weeks. Then, the concentrations of T₃, T₄, and thyroid-stimulating hormone (TSH) were detected using enzyme-linked immunosorbent assay (ELISA) to verify the developed animal model of IMH. We performed four behavioral experiments, including the marble burying test, open-field test, three-chamber sociability test, and Morris water maze, to explore the autistic-like behavior of 40-day-old offspring rats.

Results: The ELISA test showed that the serum T₃ and TSH concentrations in the model group were normal compared with the negative control group, whereas the T₄ concentration decreased. In the behavioral experiments, the number of hidden marbles in the offspring of IMH increased significantly, the frequency of entering the central compartment decreased, and the social ratio decreased significantly.

Conclusion: The animal model of IMH was developed by the administration of 1 ppm of PTU for 9 weeks, and there were autistic-like behavior changes such as anxiety, weakened social ability, and repeated stereotyping in the IMH offspring by 40 days.

Keywords

autism / hypothyroxinemia during pregnancy / IMH animal model / offspring development / T ₄

References

[1] Lazarus J, Brown RS, Daumerie C, Hubalewska-Dydejczyk A, Negro R, Vaidya B. 2014 European thyroid association guidelines for the management of subclinical hypothyroidism in pregnancy and in children. Eur Thyroid J. 2014;3(2):76-94.

[2] Alexander EK, Pearce EN, Brent GA, et al. 2017 guidelines of the American Thyroid Association for the diagnosis and Management of Thyroid Disease during Pregnancy and the postpartum. Thyroid. 2017;27(3):315-389.

[3] Man EB, Brown JF, Serunian SA. Maternal hypothyroxinemia: psychoneurological deficits of progeny. Ann Clin Lab Sci. 1991;21(4):227-239.

[4] Pop VJ, Brouwers EP, Vader HL, Vulsma T, van Baar AL, de Vijlder JJ. Maternal hypothyroxinaemia during early pregnancy and subsequent child development: a 3-year follow-up study. Clin Endocrinol. 2003;59(3):282-288.

[5] Kampouri M, Margetaki K, Koutra K, et al. Maternal mild thyroid dysfunction and offspring cognitive and motor development from infancy to childhood: the Rhea mother-child cohort study in Crete. Greece J Epidemiol Community Health. 2021;75(1):29-35.

[6] Dosiou C, Medici M. Management of endocrine disease: isolated maternal hypothyroxinemia during pregnancy: knowns and unknowns. Eur J Endocrinol. 2017;176(1):R21-R38.

[7] Levie D, Korevaar TIM, Bath SC, et al. Thyroid function in early pregnancy, child IQ, and autistic traits: a meta-analysis of individual participant data. J Clin Endocrinol Metab. 2018;103(8):2967-2979.

[8] Vulsma T, Gons MH, de Vijlder JJ. Maternal-fetal transfer of thyroxine in congenital hypothyroidism due to a total organification defect or thyroid agenesis. N Engl J Med. 1989;321(1):13-16.

[9] Haddow JE, Palomaki GE, Allan WC, et al. Maternal thyroid deficiency during pregnancy and subsequent neuropsychological development of the child. N Engl J Med. 1999;341(8):549-555.

[10] Wang Y, Wang Y, Dong J, et al. Developmental hypothyroxinaemia and hypothyroidism limit dendritic growth of cerebellar Purkinje cells in rat offspring: involvement of microtubule-associated protein 2 (MAP2) and stathmin. Neuropathol Appl Neurobiol. 2014;40(4):398-415.

[11] Han Y, Gao X, Wang X, et al. A systematic review and meta-analysis examining the risk of adverse pregnancy and neonatal outcomes in women with isolated hypothyroxinemia in pregnancy. Thyroid. 2023;33(5):603-614.

[12] Axelstad M, Hansen PR, Boberg J, et al. Developmental neurotoxicity of propylthiouracil (PTU) in rats: relationship between transient hypothyroxinemia during development and long-lasting behavioural and functional changes. Toxicol Appl Pharmacol. 2008;232(1):1-13.

[13] Opazo MC, Gianini A, Pancetti F, et al. Maternal hypothyroxinemia impairs spatial learning and synaptic nature and function in the offspring. Endocrinology. 2008;149(10):5097-5106.

[14] Gilbert ME. Impact of low-level thyroid hormone disruption induced by propylthiouracil on brain development and function. Toxicol Sci. 2011;124(2):432-445.

[15] Casey BM, Thom EA, Peaceman AM, et al. Treatment of subclinical hypothyroidism or hypothyroxinemia in pregnancy. N Engl J Med. 2017;376(9):815-825.

[16] Cooper DS, Pearce EN. Subclinical hypothyroidism and hypothyroxinemia in pregnancy—still no answers. N Engl J Med. 2017;376(9):876-877.

[17] Kawahori K, Hashimoto K, Yuan X, et al. Mild maternal hypothyroxinemia during pregnancy induces persistent DNA hypermethylation in the hippocampal brain-derived neurotrophic factor gene in mouse offspring. Thyroid. 2018;28(3):395-406.

[18] Román GC, Ghassabian A, Bongers-Schokking JJ, et al. Association of gestational maternal hypothyroxinemia and increased autism risk. Ann Neurol. 2013;74(5):733–742.

[19] Royland JE, Parker JS, Gilbert ME. A genomic analysis of subclinical hypothyroidism in hippocampus and neocortex of the developing rat brain. J Neuroendocrinol. 2008;20(12):1319-1338.

[20] Galton VA. Thyroxine metabolism in the rat: effect of varying doses of exogenous thyroxine. Acta Endocrinol. 1975;78(4):714-722.

[21] Frumess RD, Larsen PR. Correlation of serum triiodothyronine (T3) and thyroxine (T4) with biologic effects of thyroid hormone replacement in propylthiouracil-treated rats. Metabolism. 1975;24(4):547-554.

[22] Goodman JH, Gilbert ME. Modest thyroid hormone insufficiency during development induces a cellular malformation in the corpus callosum: a model of cortical dysplasia. Endocrinology. 2007;148(6):2593-2597.

[23] Morris JC, Galton VA. The isolation of thyroxine (T4), the discovery of 3,5,3′-triiodothyronine (T3), and the identification of the deiodinases that generate T3 from T4: an historical review. Endocrine. 2019;66(1):3-9.

[24] Cisternas P, Louveau A, Bueno SM, Kalergis AM, Boudin H, Riedel CA. Gestational hypothyroxinemia affects glutamatergic synaptic protein distribution and neuronal plasticity through neuron-astrocyte interplay. Mol Neurobiol. 2016;53(10):7158-7169.

[25] Wei W, Wang Y, Dong J, et al. Hypothyroxinemia induced by maternal mild iodine deficiency impairs hippocampal myelinated growth in lactational rats. Environ Toxicol. 2015;30(11):1264-1274.

[26] Lavado-Autric R, Ausó E, García-Velasco JV, et al. Early maternal hypothyroxinemia alters histogenesis and cerebral cortex cytoarchitecture of the progeny. J Clin Invest. 2003;111(7):1073-1082.

[27] Glinoer D. What happens to the normal thyroid during pregnancy? Thyroid. 1999;9(7):631-635.

[28] Glinoer D, de Nayer P, Bourdoux P, et al. Regulation of maternal thyroid during pregnancy. J Clin Endocrinol Metab. 1990;71(2):276-287.

[29] Morreale de Escobar G, Obregon MJ, Escobar del Rey F. Role of thyroid hormone during early brain development. Eur J Endocrinol. 2004;151(Suppl 3):U25-U37.

[30] Contempré B, Jauniaux E, Calvo R, Jurkovic D, Campbell S, de Escobar GM. Detection of thyroid hormones in human embryonic cavities during the first trimester of pregnancy. J Clin Endocrinol Metab. 1993;77(6):1719-1722.

[31] Margarity M, Matsokis N, Valcana T. characterization of nuclear triiodothvronine (T,) and tetraiodothyronine (T4) binding in developing brain tissue.

[32] Escobar GMD, Obregón MJ, Rey FED. Maternal thyroid hormones early in pregnancy and fetal brain development. Best Pract Res Clin Endocrinol Metab. 2004;18(2):225-248.

[33] Li G, Liu Y, Su X, Huang S, Liu X, Du Q. Effect of levothyroxine on pregnancy outcomes in pregnant women with hypothyroxinemia: an interventional study. Front Endocrinol (Lausanne). 2022;13:874975.

[34] Medici M, van der Deure WM, Verbiest M, et al. A large-scale association analysis of 68 thyroid hormone pathway genes with serum TSH and FT4 levels. Eur J Endocrinol. 2011;164(5):781-788.

[35] Li P, Cui J, Li L, et al. Association between isolated maternal hypothyroxinemia during the first trimester and adverse pregnancy outcomes in southern Chinese women: a retrospective study of 7051 cases. BMC Pregnancy Childbirth. 2022;22(1):866.

[36] Du J, Ji L, Zhang X, Yuan N, Sun J, Zhao D. Maternal isolated hypothyroxinemia in the first trimester is not associated with adverse pregnancy outcomes, except for macrosomia: a prospective cohort study in China. Front Endocrinol. 2023;14:14.

[37] Toloza FJK, Derakhshan A, Männistö T, et al. Association between maternal thyroid function and risk of gestational hypertension and pre-eclampsia: a systematic review and individual-participant data meta-analysis. Lancet Diabetes Endocrinol. 2022;10(4):243-252.

[38] Finken MJJ, van Eijsden M, Loomans EM, Vrijkotte TGM, Rotteveel J. Maternal hypothyroxinemia in early pregnancy predicts reduced performance in reaction time tests in 5-to 6-year-old offspring. J Clin Endocrinol Metab. 2013;98(4):1417-1426.

[39] Leviton A, Paneth N, Reuss ML, et al. Hypothyroxinemia of prematurity and the risk of cerebral white matter damage. J Pediatr. 1999;134(6):706-711.

RIGHTS & PERMISSIONS

2024 The Author(s). Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.