RESUMO
Acute nociception is essential for survival by warning organisms against potential dangers, whereas tissue injury results in a nociceptive hypersensitivity state that is closely associated with debilitating disease conditions, such as chronic pain. Transient receptor potential (Trp) ion channels expressed in nociceptors detect noxious thermal and chemical stimuli to initiate acute nociception. The existing hypersensitivity model suggests that under tissue injury and inflammation, the same Trp channels in nociceptors are sensitized through transcriptional and posttranslational modulation, leading to nociceptive hypersensitivity. Unexpectedly and different from this model, we find that in Drosophila larvae, acute heat nociception and tissue injury-induced hypersensitivity involve distinct cellular and molecular mechanisms. Specifically, TrpA1-D in peripheral sensory neurons mediates acute heat nociception, whereas TrpA1-C in a cluster of larval brain neurons transduces the heat stimulus under the allodynia state. As a result, interfering with synaptic transmission of these brain neurons or genetic targeting of TrpA1-C blocks heat allodynia but not acute heat nociception. TrpA1-C and TrpA1-D are two splicing variants of TrpA1 channels and are coexpressed in these brain neurons. We further show that Gq-phospholipase C signaling, downstream of the proalgesic neuropeptide Tachykinin, differentially modulates these two TrpA1 isoforms in the brain neurons by selectively sensitizing heat responses of TrpA1-C but not TrpA1-D. Together, our studies provide evidence that nociception and noncaptive sensitization could be mediated by distinct sensory neurons and molecular sensors.
Assuntos
Nociceptividade , Canais de Potencial de Receptor Transitório , Animais , Drosophila/fisiologia , Neurônios , Nociceptividade/fisiologia , Nociceptores/fisiologia , Transdutores , Canais de Potencial de Receptor Transitório/genéticaRESUMO
Sleep is a fundamental, evolutionarily conserved, plastic behavior that is regulated by circadian and homeostatic mechanisms as well as genetic factors and environmental factors, such as light, humidity, and temperature. Among environmental cues, temperature plays an important role in the regulation of sleep. This review presents an overview of thermoreception in animals and the neural circuits that link this process to sleep. Understanding the influence of temperature on sleep can provide insight into basic physiologic processes that are required for survival and guide strategies to manage sleep disorders.
Assuntos
Ritmo Circadiano , Sono , Animais , Ritmo Circadiano/fisiologia , Temperatura , Sono/fisiologia , Homeostase/fisiologia , PlásticosRESUMO
The transient receptor potential A1 (TRPA1) channel is an evolutionarily conserved detector of temperature and irritant chemicals. Here, we show that two specific isoforms of TRPA1 in Drosophila are H2O2 sensitive and that they can detect strong UV light via sensing light-induced production of H2O2. We found that ectopic expression of these H2O2-sensitive Drosophila TRPA1 (dTRPA1) isoforms conferred UV sensitivity to light-insensitive HEK293 cells and Drosophila neurons, whereas expressing the H2O2-insensitive isoform did not. Curiously, when expressed in one specific group of motor neurons in adult flies, the H2O2-sensitive dTRPA1 isoforms were as competent as the blue light-gated channelrhodopsin-2 in triggering motor output in response to light. We found that the corpus cardiacum (CC) cells, a group of neuroendocrine cells that produce the adipokinetic hormone (AKH) in the larval ring gland endogenously express these H2O2-sensitive dTRPA1 isoforms and that they are UV sensitive. Sensitivity of CC cells required dTRPA1 and H2O2 production but not conventional phototransduction molecules. Our results suggest that specific isoforms of dTRPA1 can sense UV light via photochemical production of H2O2. We speculate that UV sensitivity conferred by these isoforms in CC cells may allow young larvae to activate stress response--a function of CC cells--when they encounter strong UV, an aversive stimulus for young larvae.
Assuntos
Proteínas de Drosophila/fisiologia , Proteínas de Choque Térmico HSP90/fisiologia , Peróxido de Hidrogênio/metabolismo , Isoformas de Proteínas/fisiologia , Raios Ultravioleta , Animais , Drosophila , Células HEK293 , Humanos , Processos FotoquímicosRESUMO
The metallic conductive filament (CF) model, which serves as an important conduction mechanism for realizing synaptic functions in electronic devices, has gained recognition and is the subject of extensive research. However, the formation of CFs within the active layer is plagued by issues such as uncontrolled and random growth, which severely impacts the stability of the devices. Therefore, controlling the growth of CFs and improving the performance of the devices have become the focus of that research. Herein, a synaptic device based on polyvinylpyrrolidone (PVP)/graphene oxide quantum dot (GO QD) nanocomposites is proposed. Doping GO QDs in the PVP provides a large number of active centers for the reduction of silver ions, which allows, to a certain extent, the growth of CFs to be controlled. Because of this, the proposed device can simulate a variety of synaptic functions, including the transition from long-term potentiation to long-term depression, paired-pulse facilitation, post-tetanic potentiation, transition from short-term memory to long-term memory, and the behavior of the "learning experience". Furthermore, after being bent repeatedly, the devices were still able to simulate multiple synaptic functions accurately. Finally, the devices achieved a high recognition accuracy rate of 89.39% in the learning and inference tests, producing clear digit classification results.
RESUMO
Controllable contraception in male animals was demonstrated through the utilization of gold nanorods' photothermal effect to accomplish mild testicular hyperthermia. However, the challenges arising from testicular administration and the non-biodegradability of nanoparticles hinder further clinical implementation. Therefore, a straightforward, non-invasive, and enhanced contraception approach is required. This study explores the utilization of human heavy chain ferritin (HFn) nanocarriers loaded with aggregation-induced emission luminogens (AIEgens) for noninvasive, controllable male contraception guided by Near-Infrared-II (NIR-II) fluorescence imaging. The HFn-caged AIEgens (HFn@BBT) are delivered via intravenous injection and activated by near-infrared irradiation. Lower hyperthermia treatment induces partial damage to the testes and seminiferous tubules, reducing fertility indices by approximately 100% on the 7th day, which gradually recovers to 80% on the 60th day. Conversely, implementation of elevated hyperthermia therapy causes total destruction of both testes and seminiferous tubules, leading to a complete loss of fertility on the 60th day. Additionally, the use of AIEgens in NIR-II imaging offers improved fluorescence efficiency and penetration depth. The findings of this study hold significant promise for the advancement of safe and effective male contraceptive methods, addressing the need for noninvasive and controllable approaches to reproductive health and population control.
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Primary cicatricial alopecias (PCAs) are a group of permanent hair loss disorders, of which the pathogenesis is still poorly understood. The alopecia and excoriation (AE) mouse strain is a dominant mutant generated from ethyl nitrosourea mutagenesis. AE mice exhibit a progressive alopecia phenotype similar to that seen in PCAs, resulting from a point mutation in the gasdermin A3 gene. Mutant mice begin to show alopecia on the head from postnatal day 22 and experience complete hair loss by the age of 6 months, along with hyperkeratosis and catagen delay. The results of a histological examination showed that bulge stem cells in AE skin are gradually depleted, as indicated by decreased keratin 15 and CD34 expression, and reduced bromodeoxyuridine label-retaining cells in the AE bulge. In addition, AE mice display an inflammatory condition in the skin from postnatal day 7, including elevated tumor necrosis factor-α and monocyte chemotactic protein-1 mRNA levels and significantly increased macrophages and dendritic cell number. Immune privilege in the bulge was also compromised in AE skin. Consistently, after treatment with the immunosuppressive agent, cyclosporine A, immune privilege collapse, stem cell destruction, and alopecia phenotype of AE mice were all rescued. Collectively, our data demonstrate that immune-mediated destruction of bulge stem cells plays a crucial role in the pathogenesis of alopecia in AE mice, and this strain might be an interesting model for PCAs, especially for lichen planopilaris.
Assuntos
Alopecia/genética , Dermatite/genética , Mutação Puntual/genética , Proteínas/genética , Células-Tronco/patologia , Alopecia/imunologia , Alopecia/patologia , Animais , Ciclosporina/farmacologia , Dermatite/imunologia , Dermatite/patologia , Fármacos Dermatológicos/farmacologia , Genótipo , Folículo Piloso/patologia , Camundongos , Camundongos Endogâmicos , Células-Tronco/imunologiaRESUMO
Smart windows with light management and indoor solar heating modulation capacities are of paramount importance for building energy conservation. Thermochromic poly(N-isopropylacrylamide) (PNIPAm) hydrogel smart windows exhibit advantages of the relatively suitable transition temperature of 32 °C, high cost-effective and automatic passive sunlight regulation, but sustain slow response rate and unsatisfactory solar modulation efficiency. Herein, a strategy of one-step copolymerization of NIPAm and different olefine acids (OA) using reverse atom transfer radical polymerization method is developed to fabricate various chain/microparticle hybrids (CMH) for liquid energy-saving windows. Synergetic mechanisms of thermal-induced dissolution and aggregation of linear polymer chains integrated with water capture and release of microgel particles contribute to tunable light-scattering behaviors and adaptive solar modulation. Without any post-treatment, the as-prepared poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAm-co-AA))-based CMH suspension is injected into sandwich glass to construct energy-saving windows, which exhibits appreciated near-room-temperature transition (26.7 °C), rapid response (5 s), extraordinary luminous transmittance (91.5%), and solar modulation efficiency (85.8%), resulting in a substantial decline of indoor temperature of 24.5 °C in simulation experiment. Combining the versatile strategy with flexible adjustment on transition temperature, multifarious P(NIPAm-co-OA)-based CMH windows with eminent light management capacity are obtained. This work will powerfully promote the development and renovation of energy-efficient windows.
RESUMO
Adult stem cells are essential for tissue maintenance and repair. Although genetic pathways for controlling adult stem cells are extensively investigated in various tissues, much less is known about how mechanosensing could regulate adult stem cells and tissue growth. Here, we demonstrate that shear stress sensing regulates intestine stem cell proliferation and epithelial cell number in adult Drosophila. Ca2+ imaging in ex vivo midguts shows that shear stress, but not other mechanical forces, specifically activates enteroendocrine cells among all epithelial cell types. This activation is mediated by transient receptor potential A1 (TrpA1), a Ca2+-permeable channel expressed in enteroendocrine cells. Furthermore, specific disruption of shear stress, but not chemical, sensitivity of TrpA1 markedly reduces proliferation of intestinal stem cells and midgut cell number. Therefore, we propose that shear stress may act as a natural mechanical stimulation to activate TrpA1 in enteroendocrine cells, which, in turn, regulates intestine stem cell behavior.
Assuntos
Células-Tronco Adultas , Proteínas de Drosophila , Drosophila , Canais Iônicos , Animais , Proliferação de Células , Intestinos/citologia , Estresse Mecânico , Canais Iônicos/metabolismo , Proteínas de Drosophila/metabolismoRESUMO
Protein phosphatase 2A (PP2A) is one of the most abundant serine/threonine phosphatases, with a critical role in embryonic development and human disease. There are two isoforms of the catalytic subunit of PP2A, Ppp2ca and Ppp2cb. Null mutation of Ppp2ca leads to early embryonic lethality at E6.5, hindering functional study of PP2A beyond this stage. We generated conditional null alleles of Ppp2ca and Ppp2cb by flanking with loxP sites exons 3 to 5 of Ppp2ca and exon 3 of Ppp2cb. Ppp2ca(fl/fl) mice did not display any visible phenotype. Homozygous mutants in which Cre-mediated excision resulted in global deletion of Ppp2ca displayed embryonic lethality and developmental defects similar to those previously reported. Ppp2cb(Δ/Δ) mice generated by the same strategy did not display any obvious morphological or physiological defects. These mouse strains can serve as important genetic tools to study the roles of PP2A during development and disease in a spatial- or temporal-specific manner.
Assuntos
Alelos , Isoenzimas/genética , Proteína Fosfatase 2/genética , Animais , Sequência de Bases , Southern Blotting , Western Blotting , Primers do DNA , Camundongos , Camundongos Knockout , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Calcium (Ca(2+)) influx through Ca(v)1.2 L-type Ca(2+) channels is an important event for cardiac excitation-contraction (E-C) coupling. The functional regulation of Ca(v)1.2 is controlled by multiple kinases and phosphatases. It has been well documented that phosphorylation of Ca(v)1.2 by PKA or other kinases is sufficient for the upregulation of channel activity. However, little is known about the role of protein phosphatases in counterbalancing the phosphorylation of Ca(v)1.2, especially the degree to which protein phosphatase 2A (PP2A)-mediated dephosphorylation is involved in the regulation of Ca(v)1.2 in the mouse heart. Here, we report a physical interaction between PP2A and the C-terminus of Ca(v)1.2 in mouse heart extracts as revealed by coimmunoprecipitation. This interaction was further confirmed by the observation that PP2A and Ca(v)1.2 are colocalized in isolated mouse cardiomyocytes. Specifically, PP2A was bound at serine 1866 in the C-terminus of Ca(v)1.2, and PP2A-induced Ca(v)1.2 dephosphorylation at serine 1866 was observed in mouse cardiomyocytes. Importantly, the density of L-type calcium current increased in line with the increase in the phosphorylation at serine 1866 of Ca(v)1.2 in cardiac-specific PP2A Cα knockout mice. These phenomena were reproduced by treatment with okadaic acid, a PP2A inhibitor, in H9c2 cells. In summary, our data reveal the functional role of PP2A in cardiac Ca(v)1.2 regulation.
Assuntos
Canais de Cálcio Tipo L/metabolismo , Miócitos Cardíacos/metabolismo , Proteína Fosfatase 2/metabolismo , Serina/metabolismo , Animais , Cálcio/metabolismo , Canais de Cálcio Tipo L/genética , Linhagem Celular , Camundongos , Camundongos Knockout , Fosforilação , Proteína Fosfatase 2/genética , Serina/genéticaRESUMO
Suppression of programmed cell death is critical for the final maturation of red blood cells and depends largely on the anti-apoptotic effects of EpoR-STAT5-Bcl-x(L) signaling. As the major eukaryotic serine/threonine phosphatase, protein phosphatase 2A (PP2A) regulates multiple cellular processes, including apoptosis. However, whether PP2A plays a role in preventing erythroid cells from undergoing apoptosis remains to be elucidated. We conditionally inactivated the catalytic subunit α of PP2A (PP2Acα), which is the predominant form of PP2Ac, during early embryonic hematopoiesis. Loss of PP2Acα in hematopoietic cells perturbed definitive erythropoiesis characterized by fetal liver atrophy, reduced Ter119(+) cell number, abnormal expression patterns of molecular markers, less colony formation, and a reduction in definitive globin expression. Levels of erythropoiesis-promoting cytokines and initial seeding with hematopoietic progenitors remained unchanged in PP2Acα(TKO) fetal livers. We noted impaired expansion of the fetal erythroid compartment, which was associated with increased apoptosis of committed erythroid cells. Mechanistically, PP2Acα depletion markedly reduced Tyr(694) phosphorylation of STAT5 and expression of Bcl-x(L). Unexpectedly, PP2Acα-deficient embryos did not manifest any early embryonic vascular defects. Collectively, these data provide direct loss-of-function evidence demonstrating the importance of PP2Acα for the survival of committed erythroid cells during fetal liver erythropoiesis.
Assuntos
Células Eritroides/citologia , Eritropoese/fisiologia , Fígado/embriologia , Fosfoproteínas Fosfatases/metabolismo , Fator de Transcrição STAT5/metabolismo , Transdução de Sinais/fisiologia , Animais , Western Blotting , Separação Celular , Embrião de Mamíferos , Células Eritroides/metabolismo , Citometria de Fluxo , Imuno-Histoquímica , Fígado/metabolismo , Camundongos , Proteína Fosfatase 2C , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Mechanical nociception is essential for animal survival. However, the forces involved in nociceptor activation and the underlying mechanotransduction mechanisms remain elusive. Here, we address these problems by investigating nocifensive behavior in Drosophila larvae. We show that strong poking stimulates nociceptors with a mixture of forces including shear stress and stretch. Unexpectedly, nociceptors are selectively activated by shear stress, but not stretch. Both the shear stress responses of nociceptors and nocifensive behavior require transient receptor potential A1 (TrpA1), which is specifically expressed in nociceptors. We further demonstrate that expression of mammalian or Drosophila TrpA1 in heterologous cells confers responses to shear stress but not stretch. Finally, shear stress activates TrpA1 in a membrane-delimited manner, through modulation of membrane fluidity. Together, our study reveals TrpA1 as an evolutionarily conserved mechanosensitive channel specifically activated by shear stress and suggests a critical role of shear stress in activating nociceptors to drive mechanical nociception.
Assuntos
Nociceptores , Canais de Potencial de Receptor Transitório , Animais , Nociceptores/metabolismo , Drosophila/metabolismo , Nociceptividade/fisiologia , Mecanotransdução Celular , Canal de Cátion TRPA1/metabolismo , Canais de Potencial de Receptor Transitório/metabolismo , Mamíferos/metabolismoRESUMO
Sleep behavior is characterized by long-term quiescence and increased arousal threshold, and it is homeostatically regulated. The sleep rebound after deprivation is utilized to verify the abilities to maintain homeostasis. This protocol shows how to build a programmed mechanic oscillation system and detailed procedures to conduct sleep deprivation in Drosophila. This deprivation system is featured by its programming flexibility. The knowledge of electronic circuits and a certain level of programming are both required to fulfill this protocol. For complete details on the use and execution of this protocol, please refer to Jin et al. (2021).
Assuntos
Comportamento Animal/fisiologia , Pesquisa Comportamental , Modelos Animais de Doenças , Drosophila/fisiologia , Privação do Sono/fisiopatologia , Animais , Pesquisa Comportamental/instrumentação , Pesquisa Comportamental/métodos , Desenho de Equipamento , Feminino , Homeostase/fisiologia , MasculinoRESUMO
Sleep is an essential and evolutionarily conserved behavior that is modulated by many environmental factors. Ambient temperature shifting usually occurs during climatic or seasonal change or travel from high-latitude area to low-latitude area that affects animal physiology. Increasing ambient temperature modulates sleep in both humans and Drosophila. Although several thermosensory molecules and neurons have been identified, the neural mechanisms that integrate temperature sensation into the sleep neural circuit remain poorly understood. Here, we reveal that prolonged increasing of ambient temperature induces a reversible sleep reduction and impaired sleep consolidation in Drosophila via activating the internal thermosensory anterior cells (ACs). ACs form synaptic contacts with a subset of posterior dorsal neuron 1 (DN1p) neurons and release acetylcholine to promote wakefulness. Furthermore, we identify that this subset of DN1ps promotes wakefulness by releasing CNMamide (CNMa) neuropeptides to inhibit the Dh44-positive pars intercerebralis (PI) neurons through CNMa receptors. Our study demonstrates that the AC-DN1p-PI neural circuit is responsible for integrating thermosensory inputs into the sleep neural circuit. Moreover, we identify the CNMa signaling pathway as a newly recognized wakefulness-promoting DN1 pathway.
Assuntos
Drosophila melanogaster/citologia , Drosophila melanogaster/fisiologia , Neurônios/metabolismo , Sono/fisiologia , Sensação Térmica/fisiologia , Vigília/fisiologia , Animais , Proteínas de Drosophila/metabolismo , Feminino , Masculino , Vias Neurais , Neuropeptídeos/metabolismo , Transdução de SinaisRESUMO
Transcripts of noxious stimulus-detecting TrpA1 channels are alternatively spliced. Despite the importance of nociception for survival, the in vivo significance of expressing different TrpA1 isoforms is largely unknown. Here, we develop a novel genetic approach to generate Drosophila knockin strains expressing single TrpA1 isoforms. Drosophila TrpA1 mediates heat and UVC-triggered nociception. We show that TrpA1-C and TrpA1-D, two alternative isoforms, are co-expressed in nociceptors. When examined in heterologous cells, both TrpA1-C and TrpA1-D are activated by heat and UVC. By contrast, analysis of knockin flies reveals the striking functional specificity; TrpA1-C mediates UVC-nociception, whereas TrpA1-D mediates heat-nociception. Therefore, in vivo functions of TrpA1-C and TrpA1-D are different from each other and are different from their in vitro properties. Our results indicate that a given sensory stimulus preferentially activates a single TrpA1 isoform in vivo and that polymodal nociception requires co-expression of TrpA1 isoforms, providing novel insights of how alternative splicing regulates nociception.
Assuntos
Processamento Alternativo , Proteínas de Drosophila/genética , Drosophila melanogaster/fisiologia , Canais Iônicos/genética , Nociceptividade , Animais , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/genética , Canais Iônicos/metabolismo , Isoformas de Proteínas/genética , Análise de Célula ÚnicaRESUMO
The evolutionarily conserved TRPA1 channel can sense various stimuli including temperatures and chemical irritants. Recent results have suggested that specific isoforms of Drosophila TRPA1 (dTRPA1) are UV-sensitive and that their UV sensitivity is due to H2O2 sensitivity. However, whether such UV sensitivity served any physiological purposes in animal behavior was unclear. Here, we demonstrate that H2O2-sensitive dTRPA1 isoforms promote avoidance of UV when adult Drosophila females are selecting sites for egg-laying. First, we show that blind/visionless females are still capable of sensing and avoiding UV during egg-laying when intensity of UV is high yet within the range of natural sunlight. Second, we show that such vision-independent UV avoidance is mediated by a group of bitter-sensing neurons on the proboscis that express H2O2-sensitive dTRPA1 isoforms. We show that these bitter-sensing neurons exhibit dTRPA1-dependent UV sensitivity. Importantly, inhibiting activities of these bitter-sensing neurons, reducing their dTRPA1 expression, or reducing their H2O2-sensitivity all significantly reduced blind females' UV avoidance, whereas selectively restoring a H2O2-sensitive isoform of dTRPA1 in these neurons restored UV avoidance. Lastly, we show that specifically expressing the red-shifted channelrhodopsin CsChrimson in these bitter-sensing neurons promotes egg-laying avoidance of red light, an otherwise neutral cue for egg-laying females. Together, these results demonstrate a physiological role of the UV-sensitive dTRPA1 isoforms, reveal that adult Drosophila possess at least two sensory systems for detecting UV, and uncover an unexpected role of bitter-sensing taste neurons in UV sensing.
Assuntos
Células Quimiorreceptoras/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Oviposição/genética , Canais de Cátion TRPC/genética , Raios Ultravioleta , Animais , Células Quimiorreceptoras/efeitos dos fármacos , Células Quimiorreceptoras/efeitos da radiação , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiologia , Feminino , Peróxido de Hidrogênio/farmacologia , Canais Iônicos , Locomoção , Oviposição/efeitos da radiação , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Canal de Cátion TRPA1 , Canais de Cátion TRPC/metabolismo , PaladarRESUMO
Studies using in vitro cultured oocytes have indicated that the protein phosphatase 2A (PP2A), a major serine/threonine protein phosphatase, participates in multiple steps of meiosis. Details of oocyte maturation regulation by PP2A remain unclear and an in vivo model can provide more convincing information. Here, we inactivated PP2A by mutating genes encoding for its catalytic subunits (PP2Acs) in mouse oocytes. We found that eliminating both PP2Acs caused female infertility. Oocytes lacking PP2Acs failed to complete 1(st) meiotic division due to chromosome misalignment and abnormal spindle assembly. In mitosis, PP2A counteracts Aurora kinase B/C (AurkB/C) to facilitate correct kinetochore-microtubule (KT-MT) attachment. In meiosis I in oocyte, we found that PP2Ac deficiency destabilized KT-MT attachments. Chemical inhibition of AurkB/C in PP2Ac-null oocytes partly restored the formation of lateral/merotelic KT-MT attachments but not correct KT-MT attachments. Taken together, our findings demonstrate that PP2Acs are essential for chromosome alignments and regulate the formation of correct KT-MT attachments in meiosis I in oocytes.
Assuntos
Cinetocoros/metabolismo , Meiose , Microtúbulos/metabolismo , Oócitos/metabolismo , Proteína Fosfatase 2/genética , Subunidades Proteicas/genética , Sequência de Aminoácidos , Animais , Aurora Quinase B/genética , Aurora Quinase B/metabolismo , Aurora Quinase C/genética , Aurora Quinase C/metabolismo , Cromossomos de Mamíferos/metabolismo , Cromossomos de Mamíferos/ultraestrutura , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Infertilidade Feminina/genética , Infertilidade Feminina/metabolismo , Cinetocoros/ultraestrutura , Camundongos , Camundongos Transgênicos , Microtúbulos/ultraestrutura , Mitose , Oócitos/ultraestrutura , Cultura Primária de Células , Proteína Fosfatase 2/metabolismo , Subunidades Proteicas/metabolismo , Transdução de Sinais , Fuso Acromático/metabolismo , Fuso Acromático/ultraestruturaRESUMO
Our understanding of how metabolic switches occur in the failing heart is still limited. Here, we report the emblematic pattern of metabolic alternations in two different mouse models. PP2Acα deficient hearts exhibited a dramatic decrease in the levels of mRNA encoding for transporters and enzymes involved in glucose utilization, which compensated by higher expression levels of genes controlling fatty acid utilization. These features were partly reproduced in cultured PP2Acα KD cardiomyocytes. Equivalently, a decrease in the expression of most of the transporters and enzymes controlling both glucose and fatty acid metabolism were observed in TAC model.
Assuntos
Coração/fisiopatologia , Miocárdio/metabolismo , Pressão/efeitos adversos , Proteína Fosfatase 2/deficiência , Adaptação Fisiológica , Animais , Cardiomiopatias/genética , Cardiomiopatias/metabolismo , Cardiomiopatias/patologia , Cardiomiopatias/fisiopatologia , Metabolismo Energético , Hipertrofia/genética , Hipertrofia/metabolismo , Hipertrofia/patologia , Hipertrofia/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Miocárdio/patologia , Especificidade de Órgãos , Estresse Fisiológico , Fatores de TempoRESUMO
Colonic immune homeostasis is essential for normal gastrointestinal tract functioning. In this study, we report that specific gene targeting of phosphatase and tensin homolog (PTEN) in smooth muscle cells caused age-related colonic lymphoid hyperplasia followed by global immune activation in mice. Beginning at 5 weeks of age, these mutant mice displayed massive neutrophil infiltration in the colonic lamina propria. The gene expression levels of pro-inflammatory cytokines and chemokines, including those code for monocyte chemotactic protein-1 (Mcp-1), stromal cell-derived factor 1α (Sdf-1α), and chemokine (C-C motif) ligand 28 (Ccl-28), were upregulated in the colon. Accordingly, permeability and proliferation of the colonic epithelium was compromised. These abnormalities were alleviated to a great extent when the mutants were crossed with Akt1-null mice, indicating that the pathogenesis was mediated by Akt1 signaling. Our results suggest that in smooth muscle cells, PTEN is crucial for maintaining colonic immune homeostasis.
Assuntos
Neoplasias do Colo/genética , Inflamação/genética , PTEN Fosfo-Hidrolase/genética , Pseudolinfoma/genética , Animais , Quimiocina CCL2/biossíntese , Quimiocina CXCL12/biossíntese , Neoplasias do Colo/imunologia , Neoplasias do Colo/patologia , Regulação Neoplásica da Expressão Gênica , Inflamação/imunologia , Inflamação/patologia , Camundongos , Miócitos de Músculo Liso/imunologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Neutrófilos/imunologia , Neutrófilos/patologia , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Pseudolinfoma/patologia , Transdução de Sinais/genéticaRESUMO
N-ethyl-N-nitrosourea (ENU) mutagenesis is a phenotype-driven approach with potential to assign function to every locus in the mouse genome. In this article, we describe a new mutation, Pug, as a mouse model for X-linked hypophosphatemic rickets (XLH) in human. Mice carrying the Pug mutation exhibit abnormal phenotypes including growth retardation, hypophosphatemia and decreased bone mineral density (BMD). The new mutation was mapped to X-chromosome between 65.4 cM and 66.6 cM, where Phex gene resides. Sequence analysis revealed a unique T-to-C transition mutation resulting in Phe-to-Ser substitution at amino acid 80 of PHEX protein. In vitro studies of Pug mutation demonstrated that PHEX(pug) was incompletely glycosylated and sequestrated in the endoplasmic reticulum region of cell, whereas wild-type PHEX could be fully glycosylated and transported to the plasma membrane to exert its function as an endopeptidase. Taken together, the Pug mutant directly confirms the role of Phex in phosphate homeostasis and normal skeletal development and may serves as a new disease model of human hypophosphatemic rickets.