RESUMO
Men with nonalcoholic fatty liver disease (NAFLD) are more exposed to nonalcoholic steatohepatitis (NASH) and liver fibrosis than women. However, the underlying molecular mechanisms of NALFD sex dimorphism are unclear. We combined gene expression, histological and lipidomic analyses to systematically compare male and female liver steatosis. We characterized hepatosteatosis in three independent mouse models of NAFLD, ob/ob and lipodystrophic fat-specific (PpargFΔ/Δ) and whole-body PPARγ-null (PpargΔ/Δ) mice. We identified a clear sex dimorphism occurring only in PpargΔ/Δ mice, with females showing macro- and microvesicular hepatosteatosis throughout their entire life, while males had fewer lipid droplets starting from 20 weeks. This sex dimorphism in hepatosteatosis was lost in gonadectomized PpargΔ/Δ mice. Lipidomics revealed hepatic accumulation of short and highly saturated TGs in females, while TGs were enriched in long and unsaturated hydrocarbon chains in males. Strikingly, sex-biased genes were particularly perturbed in both sexes, affecting lipid metabolism, drug metabolism, inflammatory and cellular stress response pathways. Most importantly, we found that the expression of key sex-biased genes was severely affected in all the NAFLD models we tested. Thus, hepatosteatosis strongly affects hepatic sex-biased gene expression. With NAFLD increasing in prevalence, this emphasizes the urgent need to specifically address the consequences of this deregulation in humans.
Assuntos
Hepatopatia Gordurosa não Alcoólica/patologia , PPAR gama/deficiência , Caracteres Sexuais , Animais , Modelos Animais de Doenças , Ácidos Graxos/metabolismo , Feminino , Regulação da Expressão Gênica , Hormônios Esteroides Gonadais/metabolismo , Inflamação/patologia , Gotículas Lipídicas/metabolismo , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Hepatopatia Gordurosa não Alcoólica/genética , PPAR gama/metabolismo , Fenótipo , Transdução de Sinais , Triglicerídeos/metabolismoRESUMO
Thyroid hormone deficiency is known to deeply affect cerebellum post-natal development. We present here a detailed analysis of the phenotype of a recently generated mouse model, expressing a dominant-negative TRα1 mutation. Although hormonal level is not affected, the cerebellum of these mice displays profound alterations in neuronal and glial differentiation, which are reminiscent of congenital hypothyroidism, indicating a predominant function of this receptor isoform in normal cerebellum development. Some of the observed effects might result from the cell autonomous action of the mutation, while others are more likely to result from a reduction in neurotrophic factor production.
Assuntos
Cerebelo/crescimento & desenvolvimento , Mutação , Receptores alfa dos Hormônios Tireóideos/fisiologia , Animais , Diferenciação Celular , Movimento Celular , Proliferação de Células , Hipotireoidismo Congênito/etiologia , Camundongos , Camundongos Transgênicos , Bainha de Mielina/fisiologia , Neuroglia/fisiologia , Oligodendroglia/citologia , Fator de Transcrição PAX2/análise , Isoformas de Proteínas/genética , Isoformas de Proteínas/fisiologia , Células de Purkinje/citologia , Sinapses/fisiologia , Receptores alfa dos Hormônios Tireóideos/genética , Ácido gama-Aminobutírico/fisiologiaRESUMO
Previous clinical observations and data from mouse models with defects in lipid metabolism suggested that epineurial adipocytes may play a role in peripheral nervous system myelination. We have used adipocyte-specific Lpin1 knockout mice to characterize the consequences of the presence of impaired epineurial adipocytes on the myelinating peripheral nerve. Our data revealed that the capacity of Schwann cells to establish myelin, and the functional properties of peripheral nerves, were not affected by compromised epineurial adipocytes in adipocyte-specific Lpin1 knockout mice. To evaluate the possibility that Lpin1-negative adipocytes are still able to support endoneurial Schwann cells, we also characterized sciatic nerves from mice carrying epiblast-specific deletion of peroxisome proliferator-activated receptor gamma, which develop general lipoatrophy. Interestingly, even the complete loss of adipocytes in the epineurium of peroxisome proliferator-activated receptor gamma knockout mice did not lead to detectable defects in Schwann cell myelination. However, probably as a consequence of their hyperglycemia, these mice have reduced nerve conduction velocity, thus mimicking the phenotype observed under diabetic condition. Together, our data indicate that while adipocytes, as regulators of lipid and glucose homeostasis, play a role in nerve function, their presence in epineurium is not essential for establishment or maintenance of proper myelin.
Assuntos
Adipócitos/metabolismo , Bainha de Mielina/metabolismo , Nervos Periféricos/citologia , Células de Schwann/metabolismo , Adipócitos/citologia , Animais , Camundongos , Camundongos Knockout , Proteínas Nucleares/deficiência , Proteínas Nucleares/genética , Fosfatidato Fosfatase/deficiência , Fosfatidato Fosfatase/genética , Reação em Cadeia da Polimerase em Tempo Real , Células de Schwann/citologia , Nervo Isquiático/citologia , Nervo Isquiático/metabolismoRESUMO
3,5,3'-Triido-L: -thyronine (T3) exerts pleiotropic actions on development and homeostasis mostly via its nuclear receptors, TRalpha1, TRbeta1, and TRbeta2, encoded by the THRA and THRB genes. Muouse genetics data outline the contrasting functions of THRA and THRB, and suggest that these are dictated by both the respective abundance of the receptor isoforms in a given cell type and the differences in the intrinsic properties of the receptors. The diversity of consequences of either hypothyroidism or THRA/THRB mutation is astonishing, suggesting that TR controls a large number of genes and that the repertoire of target gene differs from one tissue to another. In order to distinguish between the direct and indirect actions of TH in vivo, we use the CRE/LoxP recombination system to control the expression of a mutant TRalpha1 receptor with dominant negative properties. Ubiquitous expression of this mutation in heterozygous mice recapitulates many consequences of TH deficiency, except in tissues where TRbeta is highly expressed.
Assuntos
Mutação , Receptores alfa dos Hormônios Tireóideos/genética , Receptores alfa dos Hormônios Tireóideos/metabolismo , Receptores beta dos Hormônios Tireóideos/metabolismo , Tri-Iodotironina/metabolismo , Animais , Integrases/genética , Camundongos , Camundongos Transgênicos , Modelos Animais , Recombinação Genética , Receptores beta dos Hormônios Tireóideos/genéticaRESUMO
BACKGROUND: The peroxisome proliferator-activated receptor γ (PPARγ) is a ligand-dependent transcription factor involved in many aspects of metabolism, immune response and development. Numerous studies relying on tissue-specific invalidation of the Pparg gene have shown distinct facets of its activity, whereas the effects of its systemic inactivation remain unexplored due to embryonic lethality. By maintaining PPARγ expression in the placenta, we recently generated a mouse model carrying Pparg full body deletion (PpargΔ/Δ), which in contrast to a previously published model is totally deprived of any form of adipose tissue. Herein, we propose an in-depth study of the metabolic alterations observed in this new model. METHODS: Young adult mice, both males and females analyzed separately, were first phenotyped for their gross anatomical alterations. Systemic metabolic parameters were analyzed in the blood, in static and in dynamic conditions. A full exploration of energy metabolism was performed in calorimetric cages as well as in metabolic cages. Our study was completed by expression analyses of a set of specific genes. MAIN FINDINGS: PpargΔ/Δ mice show a striking complete absence of any form of adipose tissue, which triggers a complex metabolic phenotype including increased lean mass with organomegaly, hypermetabolism, urinary energy loss, hyperphagia, and increased amino acid metabolism. PpargΔ/Δ mice develop severe type 2 diabetes, characterized by hyperglycemia, hyperinsulinemia, polyuria and polydispsia. They show a remarkable metabolic inflexibility, as indicated by the inability to shift substrate oxidation between glucose and lipids, in both ad libitum fed state and fed/fasted/refed transitions. Moreover, upon fasting PpargΔ/Δ mice enter a severe hypometabolic state. CONCLUSIONS: Our data comprehensively describe the impact of lipoatrophy on metabolic homeostasis. As such, the presented data on PpargΔ/Δ mice gives new clues on what and how to explore severe lipodystrophy and its subsequent metabolic complications in human.
Assuntos
Diabetes Mellitus Tipo 2/genética , Transtornos do Metabolismo dos Lipídeos/genética , Tamanho do Órgão/genética , PPAR gama/genética , Tecido Adiposo/anatomia & histologia , Animais , Diabetes Mellitus Tipo 2/metabolismo , Metabolismo Energético/genética , Feminino , Deleção de Genes , Glucose/metabolismo , Transtornos do Metabolismo dos Lipídeos/metabolismo , Lipodistrofia/genética , Lipodistrofia/metabolismo , Camundongos , GravidezRESUMO
Thyroid hormones act directly on transcription by binding to TRalpha1, TRbeta1, and TRbeta2 nuclear receptors, regulating many aspects of postnatal development and homeostasis. To analyze precisely the implication of the widely expressed TRalpha1 isoform in this pleiotropic action, we have generated transgenic mice with a point mutation in the TRalpha1 coding sequence, which is expressed only after CRE/loxP-mediated DNA recombination. The amino acid change prevents interaction between TRalpha1 and histone acetyltransferase coactivators and the release of corepressors. Early expression of this dominant-negative receptor deeply affects postnatal development and adult homeostasis, recapitulating many aspects of congenital and adult hypothyroidism, except in tissues and cells where TRbeta1 and TRbeta2 are predominantly expressed. Both respective abundance and intrinsic properties of TRalpha1 and TRbeta1/2 seem to govern specificity of action.
Assuntos
Genes Dominantes , Hipotireoidismo/genética , Receptores alfa dos Hormônios Tireóideos/agonistas , Alelos , Substituição de Aminoácidos , Animais , Arginina/química , Arginina/genética , Osso e Ossos/anormalidades , Integrases/genética , Leucina/química , Leucina/genética , Camundongos , Camundongos Transgênicos , Mutação Puntual , Recombinação Genética , Tamoxifeno/farmacologia , Receptores alfa dos Hormônios Tireóideos/genética , Receptores beta dos Hormônios Tireóideos/metabolismoRESUMO
PPARγ regulates multiple aspects of skin physiology, including sebocyte differentiation, keratinocyte proliferation, epithelial stem cell survival, adipocyte biology, and inflammatory skin responses. However, the effects of its global deletion, namely of nonredundant key functions of PPARγ signaling in mammalian skin, are yet unknown because of embryonic lethality. Here, we describe the skin and hair phenotype of a whole-body PPARγ-null mouse (PpargΔ/Δ), obtained by preserving PPARγ expression in the placenta. PpargΔ/Δ mice exhibited total lipoatrophy and complete absence of sebaceous glands. Right after birth, hair follicle (HF) morphogenesis was transiently delayed, along with reduced expression of HF differentiation markers and of transcriptional regulators necessary for HF development. Later, adult PpargΔ/Δ mice developed scarring alopecia and severe perifollicular inflammation. Skin analyses in other models of lipodystrophy, AZIPtg/+ and Adipoq-Cretg/+Ppargfl/fl mice, coupled with skin graft experiments, showed that the early defects observed in hair morphogenesis were caused by the absence of adipose tissue. In contrast, the late alteration of HF cycle and appearance of inflammation were observed only in PpargΔ/Δ mice and likely were due to the lack sebaceous glands. Our findings underscore the increasing appreciation for the importance of adipose tissue-mediated signals in HF development and function.
Assuntos
Folículo Piloso/crescimento & desenvolvimento , Lipodistrofia/patologia , Morfogênese , PPAR gama/fisiologia , Animais , Diferenciação Celular , Modelos Animais de Doenças , Homeostase , Camundongos , Camundongos Knockout , PPAR gama/genéticaRESUMO
Peroxisome proliferator-activated receptor γ (PPARγ) is a nuclear receptor involved in diverse biological processes including adipocyte differentiation, glucose homeostasis, and inflammatory responses. Analyses of PPARγ knockout animals have been so far preempted by the early embryonic death of PPARγ-/- embryos as a consequence of the severe alteration of their placental vasculature. Using Sox2Cre/PPARγL2/L2 mice, we obtained fully viable PPARγ-null mice through specific and total epiblastic gene deletion, thereby demonstrating that the placental defect is the unique cause of PPARγ-/- embryonic lethality. The vasculature defects observed in PPARγ-/- placentas at embryonic d 9.5 correlated with an unsettled balance of pro- and antiangiogenic factors as demonstrated by increased levels of proliferin (Prl2c2, PLF) and decreased levels of proliferin-related protein (Prl7d1, PRP), respectively. To analyze the role of PPARγ in the later stage of placental development, when its expression peaks, we treated pregnant wild-type mice with the PPARγ agonist rosiglitazone. This treatment resulted in a disorganization of the placental layers and an altered placental microvasculature, accompanied by the decreased expression of proangiogenic genes such as Prl2c2, vascular endothelial growth factor, and Pecam1. Together our data demonstrate that PPARγ plays a pivotal role in controlling placental vascular proliferation and contributes to its termination in late pregnancy.
Assuntos
Neovascularização Fisiológica/genética , PPAR gama/fisiologia , Placenta/irrigação sanguínea , Animais , Perda do Embrião/genética , Perda do Embrião/metabolismo , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/genética , Camadas Germinativas/metabolismo , Idade Gestacional , Glicoproteínas/genética , Glicoproteínas/metabolismo , Hipoglicemiantes/farmacologia , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Especificidade de Órgãos/genética , PPAR gama/agonistas , PPAR gama/genética , PPAR gama/metabolismo , Placenta/metabolismo , Gravidez , Prolactina , Rosiglitazona , Tiazolidinedionas/farmacologiaRESUMO
The type 3 deiodinase (D3) is an enzyme that inactivates thyroid hormones (TH) and is highly expressed during development and in the central nervous system. D3-deficient (D3KO) mice develop markedly elevated serum T(3) level in the perinatal period. In adulthood, circulating T(4) and T(3) levels are reduced due to functional deficits in the thyroid axis and peripheral tissues (i.e. liver) show evidence of decreased TH action. Given the importance of TH for brain development, we aimed to assess TH action in the brain of D3KO mice at different developmental stages and determine to what extent it correlates with serum TH parameters. We used a transgenic mouse model (FINDT3) that expresses the reporter gene ß-galactosidase (ß-gal) in the central nervous system as a readout of local TH availability. Together with experiments determining expression levels of TH-regulated genes, our results show that after a state of thyrotoxicosis in early development, most regions of the D3KO brain show evidence of decreased TH action at weaning age. However, later in adulthood and in old age, the brain again manifests a thyrotoxic state, despite reduced serum TH levels. These region-specific changes in brain TH status during the life span of the animal provide novel insight into the important role of the D3 in the developing and adult brain. Our results suggest that, even if serum concentrations of TH are normal or low, impaired D3 activity may result in excessive TH action in multiple brain regions, with potential consequences of altered neural function that may be of clinical relevance to neurological and neuroendocrine disorders.
Assuntos
Encéfalo/metabolismo , Iodeto Peroxidase/deficiência , Tiroxina/metabolismo , Tri-Iodotironina/metabolismo , Análise de Variância , Animais , Northern Blotting , Feminino , Hibridização In Situ , Camundongos , Camundongos Knockout , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Glândula Tireoide/metabolismoRESUMO
In vertebrates, the active form of thyroid hormone (T3) acts directly on transcription by changing the conformation of the TR nuclear receptors (TR alpha 1, TR beta 1 et TR beta 2) which are encoded by the THRA and THRB genes. These receptors are bound to DNA at specific response elements in a ligand independent manner. Mouse genetics have clarified the respective function of each receptor isoform, mainly reflecting THRA and THRB expression patterns. It also revealed the importance of negative regulation by unliganded receptors not only in pathological but also in physiological situations.
Assuntos
Camundongos/genética , Receptores dos Hormônios Tireóideos/genética , Tri-Iodotironina/farmacologia , Animais , Núcleo Celular/química , DNA/metabolismo , Expressão Gênica/efeitos dos fármacos , Camundongos Knockout , Mutação , Conformação Proteica/efeitos dos fármacos , Receptores dos Hormônios Tireóideos/química , Receptores dos Hormônios Tireóideos/fisiologia , Receptores alfa dos Hormônios Tireóideos/química , Receptores alfa dos Hormônios Tireóideos/genética , Receptores alfa dos Hormônios Tireóideos/fisiologia , Receptores beta dos Hormônios Tireóideos/química , Receptores beta dos Hormônios Tireóideos/genética , Receptores beta dos Hormônios Tireóideos/fisiologiaRESUMO
peroxisome proliferator-activated receptors (PPARs) are nuclear receptors acting as lipid sensors. Besides its metabolic activity in peripheral organs, the PPAR beta/delta isotype is highly expressed in the brain and its deletion in mice induces a brain developmental defect. Nevertheless, exploration of PPARbeta action in the central nervous system remains sketchy. The lipid content alteration observed in PPARbeta null brains and the positive action of PPARbeta agonists on oligodendrocyte differentiation, a process characterized by lipid accumulation, suggest that PPARbeta acts on the fatty acids and/or cholesterol metabolisms in the brain. PPARbeta could also regulate central inflammation and antioxidant mechanisms in the damaged brain. Even if not fully understood, the neuroprotective effect of PPARbeta agonists highlights their potential benefit to treat various acute or chronic neurological disorders. In this perspective, we need to better understand the basic function of PPARbeta in the brain. This review proposes different leads for future researches.
RESUMO
Thyroid hormones act directly on gene transcription in the post-natal developing cerebellum, controlling neuronal, and glial cell differentiation. We have combined three experimental approaches to identify the target genes that are underlying this phenomenon: 1) a microarray analysis of gene expression to identify hormone responsive genes in the cerebellum of Pax8-/- mice, a transgenic mouse model of congenital hypothyroidism; 2) a similar microarray analysis on primary culture of cerebellum neurons; and 3) a bioinformatics screen of conserved putative-binding sites in the mouse genome. This identifies surprisingly a small set of target genes, which, for some of them, might be key regulators of cerebellum development and neuronal differentiation.
Assuntos
Envelhecimento/fisiologia , Cerebelo/fisiologia , Regulação da Expressão Gênica no Desenvolvimento , Neurônios/fisiologia , Receptores dos Hormônios Tireóideos/genética , Tiroxina/genética , Tri-Iodotironina/genética , Animais , Animais Recém-Nascidos , Cerebelo/crescimento & desenvolvimento , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Camundongos Knockout , Análise de Sequência com Séries de Oligonucleotídeos , Fator de Transcrição PAX8 , Fatores de Transcrição Box Pareados/deficiência , Fatores de Transcrição Box Pareados/genética , RNA/genética , RNA/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Thyroid hormones are involved in the regulation of many physiological processes and regulate gene transcription by binding to their nuclear receptors TRalpha and TRbeta. In the absence of triiodothyronine (T3), the unliganded receptors (aporeceptors) do bind DNA and repress the transcription of target genes. The role of thyroid hormone aporeceptors as repressors was observed in hypothyroid adult mice, but its physiological relevance in nonpathological hypothyroid conditions remained to be determined. Here we show that, in the normal mouse fetus, TRalpha aporeceptors repress heart rate as well as the expression of TRbeta and several genes encoding ion channels involved in cardiac contractile activity. Right after birth, when T3 concentration sharply increases, liganded TRalpha (holoreceptors) turn on the expression of some of these same genes concomitantly with heart rate increase. These data describe a physiological situation under which conversion of TRalpha from apo-receptors into holo-receptors, upon changes in T3 availability, plays a determinant role in a developmental process.