Skip to main content
SpringerLink
Log in

The impact of thyroid hormone in seasonal breeding has a restricted transcriptional signature

  • Original Article
  • Published:
Cellular and Molecular Life Sciences Aims and scope Submit manuscript

Abstract

Thyroid hormone (TH) directs seasonal breeding through reciprocal regulation of TH deiodinase (Dio2/Dio3) gene expression in tanycytes in the ependymal zone of the medio-basal hypothalamus (MBH). Thyrotropin secretion by the pars tuberalis (PT) is a major photoperiod-dependent upstream regulator of Dio2/Dio3 gene expression. Long days enhance thyrotropin production, which increases Dio2 expression and suppresses Dio3 expression, thereby heightening TH signaling in the MBH. Short days appear to exert the converse effect. Here, we combined endocrine profiling and transcriptomics to understand how photoperiod and TH control the ovine reproductive status through effects on hypothalamic function. Almost 3000 genes showed altered hypothalamic expression between the breeding- and non-breeding seasons, showing gene ontology enrichment for cell signaling, epigenetics and neural plasticity. In contrast, acute switching from a short (SP) to a long photoperiod (LP) affected the expression of a much smaller core of 134 LP-responsive genes, including a canonical group previously linked to photoperiodic synchronization. Reproductive switch-off at the end of the winter breeding season was completely blocked by thyroidectomy (THX), despite a very modest effect on the hypothalamic transcriptome. Only 49 genes displayed altered expression between intact and THX ewes, including less than 10% of the LP-induced gene set. Neuroanatomical mapping showed that many LP-induced genes were expressed in the PT, independently of the TH status. In contrast, TH-sensitive seasonal genes were principally expressed in the ependymal zone. These data highlight the distinctions between seasonal remodeling effects, which appear to be largely independent of TH, and TH-dependent localised effects which are permissive for transition to the non-breeding state.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

DEG:

Differentially expressed genes

FSH:

Folliculo-stimulating hormone

GPCR:

G-protein-coupled receptor

ISH:

In situ hybridization

LH:

Luteinizing hormone

LP:

Long photoperiod

MBH:

Medio-basal hypothalamus

OVX:

Ovariectomized

PD:

Pars distalis of the pituitary

PRL:

Prolactin

PT:

Pars tuberalis of the pituitary

RIA:

Radioimmunoassay

SP:

Short photoperiod

TH:

Thyroid hormone

T3:

Triiodothyronine

T4:

Thyroxine

TRH:

Thyrotropin-releasing hormone

TSH:

Thyrotropin

THX:

Thyroidectomy/thyroidectomized

ZT:

Zeitgeber time

References

  1. Dardente H, Hazlerigg DG, Ebling FJ (2014) Thyroid hormone and seasonal rhythmicity. Front Endocrinol (Lausanne) 5:19

    Google Scholar 

  2. Shinomiya A, Shimmura T, Nishiwaki-Ohkawa T, Yoshimura T (2014) Regulation of seasonal reproduction by hypothalamic activation of thyroid hormone. Front Endocrinol (Lausanne) 5:12

    Google Scholar 

  3. Wood S, Loudon A (2014) Clocks for all seasons: unwinding the roles and mechanisms of circadian and interval timers in the hypothalamus and pituitary. J Endocrinol 222:R39–R59

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Lincoln GA, Clarke IJ, Hut RA, Hazlerigg DG (2006) Characterizing a mammalian circannual pacemaker. Science 314:1941–1944

    Article  CAS  PubMed  Google Scholar 

  5. Herwig A, de Vries EM, Bolborea M, Wilson D, Mercer JG, Ebling FJ, Morgan PJ, Barrett P (2013) Hypothalamic ventricular ependymal thyroid hormone deiodinases are an important element of circannual timing in the Siberian hamster (Phodopus sungorus). PLoS One 8:e62003

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Saenz de Miera C, Hanon EA, Dardente H, Birnie M, Simonneaux V, Lincoln GA, Hazlerigg DG (2013) Circannual variation in thyroid hormone deiodinases in a short-day breeder. J Neuroendocrinol 25:412–421

    Article  CAS  PubMed  Google Scholar 

  7. Saenz de Miera C, Monecke S, Bartzen-Sprauer J, Laran-Chich MP, Pevet P, Hazlerigg DG, Simonneaux V (2014) A circannual clock drives expression of genes central for seasonal reproduction. Curr Biol 24:1500–1506

    Article  CAS  PubMed  Google Scholar 

  8. Wood SH, Christian HC, Miedzinska K, Saer BR, Johnson M, Paton B, Yu L, McNeilly J, Davis JR, McNeilly AS, Burt DW, Loudon AS (2015) Binary switching of calendar cells in the pituitary defines the phase of the circannual cycle in mammals. Curr Biol 25:2651–2662

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Dupre SM, Miedzinska K, Duval CV, Yu L, Goodman RL, Lincoln GA, Davis JR, McNeilly AS, Burt DW, Loudon AS (2010) Identification of Eya3 and TAC1 as long-day signals in the sheep pituitary. Curr Biol 20:829–835

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Dardente H, Wyse CA, Birnie MJ, Dupre SM, Loudon AS, Lincoln GA, Hazlerigg DG (2010) A molecular switch for photoperiod responsiveness in mammals. Curr Biol 20:2193–2198

    Article  CAS  PubMed  Google Scholar 

  11. Masumoto KH, Ukai-Tadenuma M, Kasukawa T, Nagano M, Uno KD, Tsujino K, Horikawa K, Shigeyoshi Y, Ueda HR (2010) Acute induction of Eya3 by late-night light stimulation triggers TSHbeta expression in photoperiodism. Curr Biol 20:2199–2206

    Article  CAS  PubMed  Google Scholar 

  12. Lechan RM, Fekete C (2005) Role of thyroid hormone deiodination in the hypothalamus. Thyroid 15:883–897

    Article  CAS  PubMed  Google Scholar 

  13. Hanon EA, Lincoln GA, Fustin JM, Dardente H, Masson-Pevet M, Morgan PJ, Hazlerigg DG (2008) Ancestral TSH mechanism signals summer in a photoperiodic mammal. Curr Biol 18:1147–1152

    Article  CAS  PubMed  Google Scholar 

  14. Nicholls TJ, Follett BK, Goldsmith AR, Pearson H (1988) Possible homologies between photorefractoriness in sheep and birds: the effect of thyroidectomy on the length of the ewe’s breeding season. Reprod Nutr Dev 28:375–385

    Article  CAS  PubMed  Google Scholar 

  15. Bechtold DA, Loudon AS (2007) Hypothalamic thyroid hormones: mediators of seasonal physiology. Endocrinology 148:3605–3607

    Article  CAS  PubMed  Google Scholar 

  16. Karsch FJ, Bittman EL, Foster DL, Goodman RL, Legan SJ, Robinson JE (1984) Neuroendocrine basis of seasonal reproduction. Recent Prog Horm Res 40:185–232

    CAS  PubMed  Google Scholar 

  17. Dardente H (2012) Melatonin-dependent timing of seasonal reproduction by the pars tuberalis: pivotal roles for long day lengths and thyroid hormones. J Neuroendocrinol 24:249–266

    Article  CAS  PubMed  Google Scholar 

  18. Moenter SM, Woodfill CJ, Karsch FJ (1991) Role of the thyroid gland in seasonal reproduction: thyroidectomy blocks seasonal suppression of reproductive neuroendocrine activity in ewes. Endocrinology 128:1337–1344

    Article  CAS  PubMed  Google Scholar 

  19. Billings HJ, Viguie C, Karsch FJ, Goodman RL, Connors JM, Anderson GM (2002) Temporal requirements of thyroid hormones for seasonal changes in LH secretion. Endocrinology 143:2618–2625

    Article  CAS  PubMed  Google Scholar 

  20. Nakao N, Ono H, Yamamura T, Anraku T, Takagi T, Higashi K, Yasuo S, Katou Y, Kageyama S, Uno Y, Kasukawa T, Iigo M, Sharp PJ, Iwasawa A, Suzuki Y, Sugano S, Niimi T, Mizutani M, Namikawa T, Ebihara S, Ueda HR, Yoshimura T (2008) Thyrotrophin in the pars tuberalis triggers photoperiodic response. Nature 452:317–322

    Article  CAS  PubMed  Google Scholar 

  21. Dupre SM, Burt DW, Talbot R, Downing A, Mouzaki D, Waddington D, Malpaux B, Davis JR, Lincoln GA, Loudon AS (2008) Identification of melatonin-regulated genes in the ovine pituitary pars tuberalis, a target site for seasonal hormone control. Endocrinology 149:5527–5539

    Article  CAS  PubMed  Google Scholar 

  22. Klosen P, Bienvenu C, Demarteau O, Dardente H, Guerrero H, Pevet P, Masson-Pevet M (2002) The mt1 melatonin receptor and RORbeta receptor are co-localized in specific TSH-immunoreactive cells in the pars tuberalis of the rat pituitary. J Histochem Cytochem 50:1647–1657

    Article  CAS  PubMed  Google Scholar 

  23. Henningsen JB, Gauer F, Simonneaux V (2016) RFRP neurons—the doorway to understanding seasonal reproduction in mammals. Front Endocrinol (Lausanne) 7:36

    Google Scholar 

  24. Chatonnet F, Flamant F, Morte B (2015) A temporary compendium of thyroid hormone target genes in brain. Biochim Biophys Acta 1849:122–129

    Article  CAS  PubMed  Google Scholar 

  25. Visser WE, Friesema EC, Visser TJ (2011) Minireview: thyroid hormone transporters: the knowns and the unknowns. Mol Endocrinol 25:1–14

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Rodriguez EM, Blazquez JL, Pastor FE, Pelaez B, Pena P, Peruzzo B, Amat P (2005) Hypothalamic tanycytes: a key component of brain-endocrine interaction. Int Rev Cytol 247:89–164

    Article  CAS  PubMed  Google Scholar 

  27. Parkinson TJ, Follett BK (1994) Effect of thyroidectomy upon seasonality in rams. J Reprod Fertil 101:51–58

    Article  CAS  PubMed  Google Scholar 

  28. Gereben B, Zeold A, Dentice M, Salvatore D, Bianco AC (2008) Activation and inactivation of thyroid hormone by deiodinases: local action with general consequences. Cell Mol Life Sci 65:570–590

    Article  CAS  PubMed  Google Scholar 

  29. Webster JR, Moenter SM, Woodfill CJ, Karsch FJ (1991) Role of the thyroid gland in seasonal reproduction. II. thyroxine allows a season-specific suppression of gonadotropin secretion in sheep. Endocrinology 129:176–183

    Article  CAS  PubMed  Google Scholar 

  30. Mullur R, Liu YY, Brent GA (2014) Thyroid hormone regulation of metabolism. Physiol Rev 94:355–382

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Pijoan PJ, Williams HL (1984) Effects of shearing on the reproductive activity and plasma prolactin levels of Dorset ewes. Br Vet J 140:444–449

    Article  CAS  PubMed  Google Scholar 

  32. Ravault JP (1976) Prolactin in the ram: seasonal variations in the concentration of blood plasma from birth until three years old. Acta Endocrinol (Copenh) 83:720–725

    CAS  Google Scholar 

  33. Curlewis JD (1992) Seasonal prolactin secretion and its role in seasonal reproduction: a review. Reprod Fertil Dev 4:1–23

    Article  CAS  PubMed  Google Scholar 

  34. Ben-Jonathan N, Hnasko R (2001) Dopamine as a prolactin (PRL) inhibitor. Endocr Rev 22:724–763

    Article  CAS  PubMed  Google Scholar 

  35. Barrett P, Ebling FJ, Schuhler S, Wilson D, Ross AW, Warner A, Jethwa P, Boelen A, Visser TJ, Ozanne DM, Archer ZA, Mercer JG, Morgan PJ (2007) Hypothalamic thyroid hormone catabolism acts as a gatekeeper for the seasonal control of body weight and reproduction. Endocrinology 148:3608–3617

    Article  CAS  PubMed  Google Scholar 

  36. Murphy M, Jethwa PH, Warner A, Barrett P, Nilaweera KN, Brameld JM, Ebling FJ (2012) Effects of manipulating hypothalamic triiodothyronine concentrations on seasonal body weight and torpor cycles in Siberian hamsters. Endocrinology 153:101–112

    Article  CAS  PubMed  Google Scholar 

  37. Dardente H, Klosen P, Pevet P, Masson-Pevet M (2003) MT1 melatonin receptor mRNA expressing cells in the pars tuberalis of the European hamster: effect of photoperiod. J Neuroendocrinol 15:778–786

    Article  CAS  PubMed  Google Scholar 

  38. Bockmann J, Bo Ckers TM, Winter C, Wittkowski W, Winterhoff H, Deufel T, Kreutz MR (1997) Thyrotropin expression in hypophyseal pars tuberalis-specific cells is 3,5,3′-triiodothyronine, thyrotropin-releasing hormone, and Pit-1 independent. Endocrinology 138:1019–1028

    Article  CAS  PubMed  Google Scholar 

  39. Ikegami K, Liao XH, Hoshino Y, Ono H, Ota W, Ito Y, Nishiwaki-Ohkawa T, Sato C, Kitajima K, Iigo M, Shigeyoshi Y, Yamada M, Murata Y, Refetoff S, Yoshimura T (2014) Tissue-specific posttranslational modification allows functional targeting of thyrotropin. Cell Rep 9:801–810

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Davis PJ, Goglia F, Leonard JL (2016) Nongenomic actions of thyroid hormone. Nat Rev Endocrinol 12:111–121

    Article  CAS  PubMed  Google Scholar 

  41. Reshetnyak AV, Murray PB, Shi X, Mo ES, Mohanty J, Tome F, Bai H, Gunel M, Lax I, Schlessinger J (2015) Augmentor alpha and beta (FAM150) are ligands of the receptor tyrosine kinases ALK and LTK: hierarchy and specificity of ligand-receptor interactions. Proc Natl Acad Sci USA 112:15862–15867

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Guan J, Umapathy G, Yamazaki Y, Wolfstetter G, Mendoza P, Pfeifer K, Mohammed A, Hugosson F, Zhang H, Hsu AW, Halenbeck R, Hallberg B, Palmer RH (2015) FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. Elife 4:e09811

    Article  PubMed  PubMed Central  Google Scholar 

  43. Labas V, Grasseau I, Cahier K, Gargaros A, Harichaux G, Teixeira-Gomes AP, Alves S, Bourin M, Gerard N, Blesbois E (2015) Qualitative and quantitative peptidomic and proteomic approaches to phenotyping chicken semen. J Proteomics 112:313–335

    Article  CAS  PubMed  Google Scholar 

  44. Chen Z, Shamsi FA, Li K, Huang Q, Al-Rajhi AA, Chaudhry IA, Wu K (2011) Comparison of camel tear proteins between summer and winter. Mol Vis 17:323–331

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Berasain C, Avila MA (2014) Amphiregulin. Semin Cell Dev Biol 28:31–41

    Article  CAS  PubMed  Google Scholar 

  46. Dardente H (2007) Does a melatonin-dependent circadian oscillator in the pars tuberalis drive prolactin seasonal rhythmicity? J Neuroendocrinol 19:657–666

    Article  CAS  PubMed  Google Scholar 

  47. Clark SD, Duangdao DM, Schulz S, Zhang L, Liu X, Xu YL, Reinscheid RK (2011) Anatomical characterization of the neuropeptide S system in the mouse brain by in situ hybridization and immunohistochemistry. J Comp Neurol 519:1867–1893

    Article  CAS  PubMed  Google Scholar 

  48. Pulkkinen V, Ezer S, Sundman L, Hagstrom J, Remes S, Soderhall C, Greco D, Haglund C, Kere J, Arola J (2014) Neuropeptide S receptor 1 (NPSR1) activates cancer-related pathways and is widely expressed in neuroendocrine tumors. Virchows Arch 465:173–183

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Ingham PW, McMahon AP (2001) Hedgehog signaling in animal development: paradigms and principles. Genes Dev 15:3059–3087

    Article  CAS  PubMed  Google Scholar 

  50. Bolborea M, Dale N (2013) Hypothalamic tanycytes: potential roles in the control of feeding and energy balance. Trends Neurosci 36:91–100

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Goodman T, Hajihosseini MK (2015) Hypothalamic tanycytes-masters and servants of metabolic, neuroendocrine, and neurogenic functions. Front Neurosci 9:387

    Article  PubMed  PubMed Central  Google Scholar 

  52. Alvarez-Buylla A, Ihrie RA (2014) Sonic hedgehog signaling in the postnatal brain. Semin Cell Dev Biol 33:105–111

    Article  CAS  PubMed  Google Scholar 

  53. Hazlerigg DG, Lincoln GA (2011) Hypothesis: cyclical histogenesis is the basis of circannual timing. J Biol Rhythms 26:471–485

    Article  PubMed  Google Scholar 

  54. Hazlerigg DG, Wyse CA, Dardente H, Hanon EA, Lincoln GA (2013) Photoperiodic variation in CD45-positive cells and cell proliferation in the mediobasal hypothalamus of the Soay sheep. Chronobiol Int 30:548–558

    Article  CAS  PubMed  Google Scholar 

  55. Batailler M, Derouet L, Butruille L, Migaud M (2016) Sensitivity to the photoperiod and potential migratory features of neuroblasts in the adult sheep hypothalamus. Brain Struct Funct 221:3301–3314

    Article  CAS  PubMed  Google Scholar 

  56. Sousa-Ferreira L, de Almeida LP, Cavadas C (2014) Role of hypothalamic neurogenesis in feeding regulation. Trends Endocrinol Metab 25:80–88

    Article  CAS  PubMed  Google Scholar 

  57. Remaud S, Gothie JD, Morvan-Dubois G, Demeneix BA (2014) Thyroid hormone signaling and adult neurogenesis in mammals. Front Endocrinol (Lausanne) 5:62

    Google Scholar 

  58. Dentice M, Luongo C, Huang S, Ambrosio R, Elefante A, Mirebeau-Prunier D, Zavacki AM, Fenzi G, Grachtchouk M, Hutchin M, Dlugosz AA, Bianco AC, Missero C, Larsen PR, Salvatore D (2007) Sonic hedgehog-induced type 3 deiodinase blocks thyroid hormone action enhancing proliferation of normal and malignant keratinocytes. Proc Natl Acad Sci USA 104:14466–14471

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Wilkens S (2015) Structure and mechanism of ABC transporters. F1000Prime Rep 7:14 (eCollection 2015)

    Article  PubMed  PubMed Central  Google Scholar 

  60. Bellia F, Vecchio G, Rizzarelli E (2014) Carnosinases, their substrates and diseases. Molecules 19:2299–2329

    Article  PubMed  Google Scholar 

  61. Bonfanti L, Peretto P, De Marchis S, Fasolo A (1999) Carnosine-related dipeptides in the mammalian brain. Prog Neurobiol 59:333–353

    Article  CAS  PubMed  Google Scholar 

  62. Amparan D, Avram D, Thomas CG, Lindahl MG, Yang J, Bajaj G, Ishmael JE (2005) Direct interaction of myosin regulatory light chain with the NMDA receptor. J Neurochem 92:349–361

    Article  CAS  PubMed  Google Scholar 

  63. Yamamura T, Yasuo S, Hirunagi K, Ebihara S, Yoshimura T (2006) T(3) implantation mimics photoperiodically reduced encasement of nerve terminals by glial processes in the median eminence of Japanese quail. Cell Tissue Res 324:175–179

    Article  CAS  PubMed  Google Scholar 

  64. Klosen P, Sebert ME, Rasri K, Laran-Chich MP, Simonneaux V (2013) TSH restores a summer phenotype in photoinhibited mammals via the RF-amides RFRP3 and kisspeptin. FASEB J 27:2677–2686

    Article  CAS  PubMed  Google Scholar 

  65. Dardente H, Birnie M, Lincoln GA, Hazlerigg DG (2008) RFamide-related peptide and its cognate receptor in the sheep: cDNA cloning, mRNA distribution in the hypothalamus and the effect of photoperiod. J Neuroendocrinol 20:1252–1259

    Article  CAS  PubMed  Google Scholar 

  66. Revel FG, Saboureau M, Pevet P, Simonneaux V, Mikkelsen JD (2008) RFamide-related peptide gene is a melatonin-driven photoperiodic gene. Endocrinology 149:902–912

    Article  CAS  PubMed  Google Scholar 

  67. Smith JT (2012) The role of kisspeptin and gonadotropin inhibitory hormone in the seasonal regulation of reproduction in sheep. Domest Anim Endocrinol 43:75–84

    Article  CAS  PubMed  Google Scholar 

  68. Decourt C, Anger K, Robert V, Lomet D, Bartzen-Sprauer J, Caraty A, Dufourny L, Anderson G, Beltramo M (2016) No evidence that RFamide-related peptide 3 directly modulates LH secretion in the Ewe. Endocrinology 157:1566–1575

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank the staff of the CIRE platform for assistance with surgical procedures, Damien Capo, Olivier Lasserre and Didier Dubreuil from the Unité Expérimentale PAO no. 1297 (EU0028) for taking care of the animals and blood sampling, staff of the IGF sequencing platform (Montpellier, France) and Shona Wood (Manchester University, UK) for help with bioinformatics.

Funding

This work was supported by a Career Integration Grant (No. 320553) from the FP7-Marie-Curie actions (H.D.).

Author information

Authors and Affiliations

  1. PRC, INRA, CNRS, IFCE, Université de Tours, 37380, Nouzilly, France

    Didier Lomet, Juliette Cognié, Didier Chesneau & Hugues Dardente

  2. MGX-Montpellier GenomiX, Institut de Génomique Fonctionnelle, 34094, Montpellier, France

    Emeric Dubois

  3. Department of Arctic and Marine Biology, University of Tromsø, 9037, Tromsø, Norway

    David Hazlerigg

Authors
  1. Didier Lomet
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Juliette Cognié
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Didier Chesneau
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Emeric Dubois
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. David Hazlerigg
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hugues Dardente
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

DL, DC & HD carried out the molecular lab work; JC carried out surgeries; ED performed the RNAseq experiments; HD designed the study, performed data analysis and drafted the manuscript; DH contributed intellectual support and helped draft the manuscript. All authors gave final approval for publication.

Corresponding author

Correspondence to Hugues Dardente.

Ethics declarations

Conflict of interest

The authors have no conflict of interest.

Ethical approval

All experimental procedures were performed in accordance with international (directive 2010/63/UE) and national legislation (décret no. 2013–118) governing the ethical use of animals in research (authorization no. E37–175-2 and no. A38 801). All procedures used in this work were evaluated by a local ethics committee (Comité d’Ethique en Expérimentation Animale Val de Loire) and approved by the Ministry of Higher Education and Research (Project No. 00710.02).

Electronic supplementary material

Below is the link to the electronic supplementary material.

18_2017_2667_MOESM1_ESM.pdf

Suppl. Fig.S1(associated to Fig. 1): Transcriptomics of the non-breeding to breeding transition (Experiment 1). (A) Procedure for dissecting the MBH block. The two pictures on the left are ventral views of the ovine brain; the two pictures on the right are coronal slices. Details and abbreviations are provided in the panel. (B) Procedure for the RNAseq analysis. First, RNA were individually extracted from each MBH block resulting in 18 samples: 6 RNA samples for each of the 3 groups (May, Aug & Nov). Then, for each of the 3 groups, RNA were randomly pooled in 2 equimolar mixes from 3 animals. This resulted in 6 cDNA libraries, which were sequenced and submitted to alignment and statistical analysis as indicated. (C) Normalized RNAseq counts for genes analyzed by qPCR in Fig. 1D. Note the broad correlation between the two datasets. Slc7a8 was absent from this genome assembly (Oar3.1). Suppl. Fig.S2 (associated to Fig. 2): Impact of TH on transcriptomics of the breeding to non-breeding transition (Experiment 2). (A) Validation of the THX procedure using RIA for TSH and T4 in intact ewes (pilot Experiment for Experiment 2). The upper panel shows temporal profiles for both hormones 1.5 month prior to surgery and 2.5 months after surgery. The red dotted line corresponds to the limit of the T4 assay sensitivity (4.7 ng/ml). The histogram (lower panel) shows mean ± sem for the two periods. (B) Characterization of the hypothyroid state by qRT-PCR analysis of select genes within the pars distalis (PD) of the pituitary. The picture on the left indicates that the caudalmost part of the PD was used. Note the > 175-fold increase in Tshb, the 50-70% decreases in Dio2 and Hr. Also note that relative levels of Lhb and Fshb are not impacted by photoperiod or THX. (C) Hormonal profiling of the response in plasma levels for PRL. Neither photoperiod nor treatment had any statistically significant impact (data not shown). The orange line indicates photoperiod. (D) Procedure for the RNAseq analysis. First, RNA were individually extracted from each MBH block resulting in 24 samples: 8 RNA samples for each of the 3 groups (SP, LP & LP-THX). Then, for each of the 3 groups, RNA were randomly pooled in 2 equimolar mixes from 4 animals. This results in 6 cDNA libraries, which are sequenced and submitted to alignment and statistical analysis as indicated. (E) qRT-PCR analysis of Hr, Slc16a2, Kiss1 and Npvf in the MBH. Suppl Fig.S3 (associated to Fig. 3): Investigating the time-specific role of TH on the photoperiodic transcriptional network of the MBH (Experiment 3). Non-breeding to breeding transition: (A) Hormonal profiling of the seasonal response in plasma levels for PRL (left) and TSH (right) following THX. The orange line indicates photoperiod. Breeding to non-breeding transition: (B) Analysis of individual TSH mean levels during the 7 weeks before (i.e. 14 samples) and 7 weeks after (i.e. 14 samples) shearing in Sham-operated and THX ewes (left and right panels, respectively). TSH levels were significantly decreased after shearing in THX animals while they were significantly - albeit modestly, check the y-axis - increased in Sham ewes (pairwise t-test comparison). (C) Mean weight of ewes in the experimental group sham-operated or THX before the transition to breeding. Two-way ANOVA reveals a trend towards lower weight in THX animals at the end of experiment. (D) Hormonal profiling of the seasonal response in plasma levels for PRL following THX before the transition to non-breeding. The dotted line indicates time of shearing; the orange line indicates photoperiod. (E) Characterization of the hypothyroid state by qRT-PCR analysis of select genes within the pars distalis (PD) of the pituitary. The picture on the left indicates that the caudalmost part of the PD was used. Note the > 40-fold increase in Tshb, the 50-70% decreases in Dio2 and Hr. Also note that relative levels of Lhb and Fshb are not impacted by THX (PDF 231 kb)

18_2017_2667_MOESM2_ESM.docx

Suppl.Table S1: This folder includes 3 sheets. (i) List of qRT-PCR primers and size of the PCR amplicons (Fig. 1D, Fig. 2B, Fig. 2D & Fig. 2E). (ii) List of probes used for ISH (Fig. 3D & Fig.S4). (iii) Results of the two-way ANOVA for ISH data of Fig. 3D (DOCX 24 kb)

18_2017_2667_MOESM3_ESM.xlsx

Suppl.Table S2: RNAseq data and pathway analysis for the non-breeding to breeding transition (associated with Fig. 1). Folder includes 12 sheets: (i) M vs A: DEG using EdgeR. (ii) M vs A: DEG using DESeq2. (iii) A vs N: DEG using EdgeR. (iv) A vs N: DEG using DESeq2. (v) M vs N: DEG using EdgeR. (vi) M vs N: DEG using DESeq2. (vii) up-regulated pathways and GO terms (Cpdb analysis): M/A comparison. (viii) down-regulated pathways and GO terms (Cpdb analysis): M/Acomparison. (ix) up-regulated pathways and GO terms (Cpdb analysis): A/N comparison. (x) down-regulated pathways and GO terms (Cpdb analysis): A/N comparison. (xi) up-regulated pathways and GO terms (Cpdb analysis): M/N comparison. (xii) down-regulated pathways and GO terms (Cpdb analysis): M/N comparison (XLSX 1014 kb)

18_2017_2667_MOESM4_ESM.xlsx

Suppl.Table S3: RNAseq data and pathway analysis for the breeding to non-breeding transition / long day exposure (associated with Fig. 2). Folder includes 4 sheets: (i) LP vs SP: list of DEG using EdgeR. (ii) LP vs SP: list of DEG using DESeq2, p value < 0.05. (iii) LP vs LP-THX: list of DEG using EdgeR. (iv) LP vs LP-THX: list of DEG using DESeq2, pVal < 0.05 (XLSX 6282 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lomet, D., Cognié, J., Chesneau, D. et al. The impact of thyroid hormone in seasonal breeding has a restricted transcriptional signature. Cell. Mol. Life Sci. 75, 905–919 (2018). https://doi.org/10.1007/s00018-017-2667-x

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00018-017-2667-x

Keywords

Search

Navigation