A deficiency of maternal thyroid hormones (THs) during pregnancy may have severe impacts on fetal brain development. However, the cellular targets of THs and their underlying mechanisms are still unclear. In this study, we found that maternal hypothyroidism during pregnancy in mice inhibited neurogenesis in the embryonic telencephalon and caused learning and memory impairment in the offspring. To explore the underlying mechanisms, we treated cultured mouse embryonic neural stem cells (eNSCs) with a physiological level of 3, 5, 3′-triiodo-L-thyronine (T3). We found that T3 promoted the neuronal differentiation of eNSCs, while inhibiting astrocytic differentiation. In addition, the proliferation and maintenance of eNSCs were inhibited by T3. Furthermore, the TH receptor alpha 1 (TRα1) was detected in the eNSCs both in vivo and in vitro. Silencing TRα1 protein expression with specific siRNA eliminated the effects of T3 on eNSCs. We also found that T3 decreased STAT3 phosphorylation and STAT3-DNA binding activity through TRα1. The over expression of STAT3 attenuated the promotive effects of T3 on neuronal differentiation of eNSCs. Taken together, these results suggest that T3 promotes the neuronal differentiation of eNSCs by inhibiting STAT3 signaling activity through TRα1 and contributes to early neurogenesis in the embryonic telencephalon. Our studies reveal the physiological effects of TH in regulating eNSCs differentiation and suggest that eNSCs are one of the major cellular targets in the central nervous system by which TH influences early brain development. These findings also provide new insights into the mechanisms of neurological deficits caused by TH deficiency during embryogenesis.
HornS, HeuerH. 2009. Thyroid hormone action during brain development: more questions than answers. Mol Cell Endocrinol, 315:19–26.
10.
ContempreB, JauniauxE, CalvoR, JurkovicD, CampbellS, de EscobarGM. 1993. Detection of thyroid hormones in human embryonic cavities during the first trimester of pregnancy. J Clin Endocrinol Metab, 77:1719–1722.
11.
SantiniF, ChiovatoL, GhirriP, LapiP, MammoliC, MontanelliL, ScartabelliG, CeccariniG, CoccoliLet al.1999. Serum iodothyronines in the human fetus and the newborn: evidence for an important role of placenta in fetal thyroid hormone homeostasis. J Clin Endocrinol Metab, 84:493–498.
12.
GaltonVA. 2005. The roles of the iodothyronine deiodinases in mammalian development. Thyroid, 15:823–834.
13.
de EscobarGM, ObregonMJ, del ReyFE. 2004. Maternal thyroid hormones early in pregnancy and fetal brain development. Best Pract Res Clin Endocrinol Metab, 18:225–248.
14.
TrentinAG. 2006. Thyroid hormone and astrocyte morphogenesis. J Endocrinol, 189:189–197.
FernandezM, GiulianiA, PirondiS, D'IntinoG, GiardinoL, AloeL, Levi-MontalciniR, CalzaL. 2004. Thyroid hormone administration enhances remyelination in chronic demyelinating inflammatory disease. Proc Natl Acad Sci U S A, 101:16363–16368.
17.
CalzaL, FernandezM, GiulianiA, AloeL, GiardinoL. 2002. Thyroid hormone activates oligodendrocyte precursors and increases a myelin-forming protein and NGF content in the spinal cord during experimental allergic encephalomyelitis. Proc Natl Acad Sci U S A, 99:3258–3263.
ZoellerRT, RovetJ. 2004. Timing of thyroid hormone action in the developing brain: clinical observations and experimental findings. J Neuroendocrinol, 16:809–818.
20.
ContiL, CattaneoE. 2010. Neural stem cell systems: physiological players or in vitro entities?Nat Rev Neurosci, 11:176–187.
21.
WegnerM, StoltCC. 2005. From stem cells to neurons and glia: a Soxist's view of neural development. Trends Neurosci, 28:583–588.
SunY, HuJ, ZhouL, PollardSM, SmithA. 2011. Interplay between FGF2 and BMP controls the self-renewal, dormancy and differentiation of rat neural stem cells. J Cell Sci, 124:1867–1877.
24.
BonniA, SunY, Nadal-VicensM, BhattA, FrankDA, RozovskyI, StahlN, YancopoulosGD, GreenbergME. 1997. Regulation of gliogenesis in the central nervous system by the JAK-STAT signaling pathway. Science, 278:477–483.
25.
GauthierAS, FurstossO, ArakiT, ChanR, NeelBG, KaplanDR, MillerFD. 2007. Control of CNS cell-fate decisions by SHP-2 and its dysregulation in Noonan syndrome. Neuron, 54:245–262.
NakashimaK, YanagisawaM, ArakawaH, KimuraN, HisatsuneT, KawabataM, MiyazonoK, TagaT. 1999. Synergistic signaling in fetal brain by STAT3-Smad1 complex bridged by p300. Science, 284:479–482.
28.
KimYH, ChungJI, WooHG, JungYS, LeeSH, MoonCH, Suh-KimH, BaikEJ. 2010. Differential regulation of proliferation and differentiation in neural precursor cells by the Jak pathway. Stem Cells, 28:1816–1828.
29.
MenardC, HeinP, PaquinA, SavelsonA, YangXM, LederfeinD, Barnabe-HeiderF, MirAA, SterneckEet al.2002. An essential role for a MEK-C/EBP pathway during growth factor-regulated cortical neurogenesis. Neuron, 36:597–610.
30.
OrtegaJA, AlcantaraS. 2010. BDNF/MAPK/ERK-induced BMP7 expression in the developing cerebral cortex induces premature radial glia differentiation and impairs neuronal migration. Cereb Cortex, 20:2132–2144.
31.
Le BelleJE, OrozcoNM, PaucarAA, SaxeJP, MottahedehJ, PyleAD, WuH, KornblumHI. 2011. Proliferative neural stem cells have high endogenous ROS levels that regulate self-renewal and neurogenesis in a PI3K/Akt-dependant manner. Cell Stem Cell, 8:59–71.
32.
OtaegiG, Yusta-BoyoMJ, Vergano-VeraE, Mendez-GomezHR, CarreraAC, AbadJL, GonzalezM, de la RosaEJ, Vicario-AbejonC, de PabloF. 2006. Modulation of the PI 3-kinase-Akt signalling pathway by IGF-I and PTEN regulates the differentiation of neural stem/precursor cells. J Cell Sci, 119:2739–2748.
33.
IskarosJ, PickardM, EvansI, SinhaA, HardimanP, EkinsR. 2000. Thyroid hormone receptor gene expression in first trimester human fetal brain. J Clin Endocrinol Metab, 85:2620–2623.
34.
WallisK, DudazyS, van HogerlindenM, NordstromK, MittagJ, VennstromB. 2010. The thyroid hormone receptor alpha1 protein is expressed in embryonic postmitotic neurons and persists in most adult neurons. Mol Endocrinol, 24:1904–1916.
35.
MartinezR, EllerC, VianaNB, GomesFC. 2011. Thyroid hormone induces cerebellar neuronal migration and Bergmann glia differentiation through epidermal growth factor/mitogen-activated protein kinase pathway. Eur J Neurosci, 33:26–35.
36.
DavisPJ, LeonardJL, DavisFB. 2008. Mechanisms of nongenomic actions of thyroid hormone. Front Neuroendocrinol, 29:211–218.
37.
LinHY, ShihA, DavisFB, DavisPJ. 1999. Thyroid hormone promotes the phosphorylation of STAT3 and potentiates the action of epidermal growth factor in cultured cells. Biochem J, 338,Pt 2:427–432.
38.
FernandezM, PirondiS, ManservigiM, GiardinoL, CalzaL. 2004. Thyroid hormone participates in the regulation of neural stem cells and oligodendrocyte precursor cells in the central nervous system of adult rat. Eur J Neurosci, 20:2059–2070.
39.
DesouzaLA, LadiwalaU, DanielSM, AgasheS, VaidyaRA, VaidyaVA. 2005. Thyroid hormone regulates hippocampal neurogenesis in the adult rat brain. Mol Cell Neurosci, 29:414–426.
40.
KeeN, TeixeiraCM, WangAH, FranklandPW. 2007. Preferential incorporation of adult-generated granule cells into spatial memory networks in the dentate gyrus. Nat Neurosci, 10:355–362.
GrandbarbeL, BouissacJ, RandM, Hrabe de AngelisM, Artavanis-TsakonasS, MohierE. 2003. Delta-Notch signaling controls the generation of neurons/glia from neural stem cells in a stepwise process. Development, 130:1391–1402.
43.
YoshimatsuT, KawaguchiD, OishiK, TakedaK, AkiraS, MasuyamaN, GotohY. 2006. Non-cell-autonomous action of STAT3 in maintenance of neural precursor cells in the mouse neocortex. Development, 133:2553–2563.
44.
MarteauL, PacaryE, ValableS, BernaudinM, GuillemotF, PetitE. 2010. Angiopoietin-2 regulates cortical neurogenesis in the developing telencephalon. Cereb Cortex, 21:1695–1702.
45.
NguyenL, BessonA, HengJI, SchuurmansC, TeboulL, ParrasC, PhilpottA, RobertsJM, GuillemotF. 2006. p27kip1 independently promotes neuronal differentiation and migration in the cerebral cortex. Genes Dev, 20:1511–1524.
AhmedS. 2009. The culture of neural stem cells. J Cell Biochem, 106:1–6.
48.
CalvoRM, JauniauxE, GulbisB, AsuncionM, GervyC, ContempreB, Morreale de EscobarG. 2002. Fetal tissues are exposed to biologically relevant free thyroxine concentrations during early phases of development. J Clin Endocrinol Metab, 87:1768–1777.
49.
BastianTW, ProhaskaJR, GeorgieffMK, AndersonGW. 2010. Perinatal iron and copper deficiencies alter neonatal rat circulating and brain thyroid hormone concentrations. Endocrinology, 151:4055–4065.
50.
ZhangL, BlomgrenK, KuhnHG, Cooper-KuhnCM. 2009. Effects of postnatal thyroid hormone deficiency on neurogenesis in the juvenile and adult rat. Neurobiol Dis, 34:366–374.
51.
HeF, GeW, MartinowichK, Becker-CataniaS, CoskunV, ZhuW, WuH, CastroD, GuillemotF, FanG, de VellisJ, SunYE. 2005. A positive autoregulatory loop of Jak-STAT signaling controls the onset of astrogliogenesis. Nat Neurosci, 8:616–625.
52.
PeltierJ, O'NeillA, SchafferDV. 2007. PI3K/Akt and CREB regulate adult neural hippocampal progenitor proliferation and differentiation. Dev Neurobiol, 67:1348–1361.
53.
FreitasBC, GerebenB, CastilloM, KalloI, ZeoldA, EgriP, LipositsZ, ZavackiAM, MacielRMet al.2010. Paracrine signaling by glial cell-derived triiodothyronine activates neuronal gene expression in the rodent brain and human cells. J Clin Invest, 120:2206–2217.
54.
CalvoR, ObregonMJ, Ruiz de OnaC, Escobar del ReyF, Morreale de EscobarG. 1990. Congenital hypothyroidism, as studied in rats. Crucial role of maternal thyroxine but not of 3,5,3′-triiodothyronine in the protection of the fetal brain. J Clin Invest, 86:889–899.
55.
AusoE, Lavado-AutricR, CuevasE, Del ReyFE, Morreale De EscobarG, BerbelP. 2004. A moderate and transient deficiency of maternal thyroid function at the beginning of fetal neocorticogenesis alters neuronal migration. Endocrinology, 145:4037–4047.
56.
YamadaT, KunimatsuT, MiyataK, YabushitaS, SukataT, KawamuraS, SekiT, OkunoY, MikamiN. 2004. Enhanced rat Hershberger assay appears reliable for detection of not only (anti-)androgenic chemicals but also thyroid hormone modulators. Toxicol Sci, 79:64–74.
57.
AziziF, AmouzegarA. 2011. Management of hyperthyroidism during pregnancy and lactation. Eur J Endocrinol, 164:871–876.
ShibataM, NakaoH, KiyonariH, AbeT, AizawaS. 2011. MicroRNA-9 regulates neurogenesis in mouse telencephalon by targeting multiple transcription factors. J Neurosci, 31:3407–3422.
60.
KangSM, ChoMS, SeoH, YoonCJ, OhSK, ChoiYM, KimDW. 2007. Efficient induction of oligodendrocytes from human embryonic stem cells. Stem Cells, 25:419–424.
61.
Montero-PedrazuelaA, VeneroC, Lavado-AutricR, Fernandez-LamoI, Garcia-VerdugoJM, BernalJ, Guadano-FerrazA. 2006. Modulation of adult hippocampal neurogenesis by thyroid hormones: implications in depressive-like behavior. Mol Psychiatry, 11:361–371.
62.
LemkineGF, RajA, AlfamaG, TurqueN, HassaniZ, Alegria-PrevotO, SamarutJ, LeviG, DemeneixBA. 2005. Adult neural stem cell cycling in vivo requires thyroid hormone and its alpha receptor. FASEB J, 19:863–865.
63.
FernandezM, ParadisiM, Del VecchioG, GiardinoL, CalzaL. 2009. Thyroid hormone induces glial lineage of primary neurospheres derived from non-pathological and pathological rat brain: implications for remyelination-enhancing therapies. Int J Dev Neurosci, 27:769–778.
64.
Hadj-SahraouiN, SeugnetI, GhorbelMT, DemeneixB. 2000. Hypothyroidism prolongs mitotic activity in the post-natal mouse brain. Neurosci Lett, 280:79–82.
65.
CrisantiP, OmriB, HughesE, MeduriG, HeryC, ClauserE, JacqueminC, SaunierB. 2001. The expression of thyrotropin receptor in the brain. Endocrinology, 142:812–822.
66.
NichaneM, RenX, BellefroidEJ. 2010. Self-regulation of Stat3 activity coordinates cell-cycle progression and neural crest specification. EMBO J, 29:55–67.
67.
IslamO, LooTX, HeeseK. 2009. Brain-derived neurotrophic factor (BDNF) has proliferative effects on neural stem cells through the truncated TRK-B receptor, MAP kinase, AKT, and STAT-3 signaling pathways. Curr Neurovasc Res, 6:42–53.
68.
HirabayashiY, GotohY. 2010. Epigenetic control of neural precursor cell fate during development. Nat Rev Neurosci, 11:377–388.
69.
CaoF, HataR, ZhuP, NakashiroK, SakanakaM. 2010. Conditional deletion of Stat3 promotes neurogenesis and inhibits astrogliogenesis in neural stem cells. Biochem Biophys Res Commun, 394:843–847.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
