RESUMO
Heterochromatin is a key architectural feature of eukaryotic chromosomes critical for cell type-specific gene expression and genome stability. In the mammalian nucleus, heterochromatin segregates from transcriptionally active genomic regions and exists in large, condensed, and inactive nuclear compartments. However, the mechanisms underlying the spatial organization of heterochromatin need to be better understood. Histone H3 lysine 9 trimethylation (H3K9me3) and lysine 27 trimethylation (H3K27me3) are two major epigenetic modifications that enrich constitutive and facultative heterochromatin, respectively. Mammals have at least five H3K9 methyltransferases (SUV39H1, SUV39H2, SETDB1, G9a and GLP) and two H3K27 methyltransferases (EZH1 and EZH2). In this study, we addressed the role of H3K9 and H3K27 methylation in heterochromatin organization using a combination of mutant cells for five H3K9 methyltransferases and an EZH1/2 dual inhibitor, DS3201. We showed that H3K27me3, which is normally segregated from H3K9me3, was redistributed to regions targeted by H3K9me3 after the loss of H3K9 methylation and that the loss of both H3K9 and H3K27 methylation resulted in impaired condensation and spatial organization of heterochromatin. Our data demonstrate that the H3K27me3 pathway safeguards heterochromatin organization after the loss of H3K9 methylation in mammalian cells.
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Epigênese Genética , Heterocromatina , Animais , Heterocromatina/genética , Histonas/metabolismo , Lisina/metabolismo , Mamíferos/genética , Metilação , Histona Metiltransferases/metabolismoRESUMO
The calyx of Held, a large glutamatergic presynaptic terminal in the auditory brainstem undergoes developmental changes to support the high action-potential firing rates required for auditory information encoding. In addition, calyx terminals are morphologically diverse, which impacts vesicle release properties and synaptic plasticity. Mitochondria influence synaptic plasticity through calcium buffering and are crucial for providing the energy required for synaptic transmission. Therefore, it has been postulated that mitochondrial levels increase during development and contribute to the morphological-functional diversity in the mature calyx. However, the developmental profile of mitochondrial volumes and subsynaptic distribution at the calyx of Held remains unclear. To provide insight on this, we developed a helper-dependent adenoviral vector that expresses the genetically encoded peroxidase marker for mitochondria, mito-APEX2, at the mouse calyx of Held. We developed protocols to detect labeled mitochondria for use with serial block face scanning electron microscopy to carry out semiautomated segmentation of mitochondria, high-throughput whole-terminal reconstruction, and presynaptic ultrastructure in mice of either sex. Subsequently, we measured mitochondrial volumes and subsynaptic distributions at the immature postnatal day (P)7 and the mature (P21) calyx. We found an increase of mitochondria volumes in terminals and axons from P7 to P21 but did not observe differences between stalk and swelling subcompartments in the mature calyx. Based on these findings, we propose that mitochondrial volumes and synaptic localization developmentally increase to support high firing rates required in the initial stages of auditory information processing.SIGNIFICANCE STATEMENT Elucidating the developmental processes of auditory brainstem presynaptic terminals is critical to understanding auditory information encoding. Additionally, morphological-functional diversity at these terminals is proposed to enhance coding capacity. Mitochondria provide energy for synaptic transmission and can buffer calcium, impacting synaptic plasticity; however, their developmental profile to ultimately support the energetic demands of synapses following the onset of hearing remains unknown. Therefore, we created a helper-dependent adenoviral vector with the mitochondria-targeting peroxidase mito-APEX2 and expressed it at the mouse calyx of Held. Volumetric reconstructions of serial block face electron microscopy data of immature and mature labeled calyces reveal that mitochondrial volumes are increased to support high firing rates upon maturity.
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Mitocôndrias/fisiologia , Tamanho Mitocondrial/fisiologia , Terminações Pré-Sinápticas/fisiologia , Sinapses/fisiologia , Potenciais de Ação , Animais , Axônios/metabolismo , Axônios/ultraestrutura , Tronco Encefálico/crescimento & desenvolvimento , Tronco Encefálico/ultraestrutura , Cálcio/fisiologia , Fenômenos Eletrofisiológicos/fisiologia , Metabolismo Energético/fisiologia , Feminino , Vetores Genéticos , Processamento de Imagem Assistida por Computador , Masculino , Camundongos , Mitocôndrias/ultraestrutura , Plasticidade Neuronal , Terminações Pré-Sinápticas/ultraestruturaRESUMO
BACKGROUND AND OBJECTIVE: The periodontal ligament (PDL) is an essential tissue for tooth function. However, the 3-dimensional ultrastructure of these PDL collagen bundles on a mesoscale is not clear. We investigated the 3-dimensional ultrastructure of these collagen bundles and quantitatively analyzed their histomorphometry using focused ion beam/scanning electron microscope (FIB/SEM) tomography. MATERIAL AND METHODS: The PDLs of the first mandibular molar of male C57BL/6 mice were analyzed using FIB/SEM tomography. The serial images of the collagen bundles so obtained were reconstructed. The collagen bundles were analyzed quantitatively using 3-dimensional histomorphometry. RESULTS: Collagen bundles of the PDL demonstrated multiple branched structures, rather than a single rope-like structure, and were wrapped in cytoplasm sheets. The structure of the horizontal fiber of the collagen bundle was an extensive meshwork. In contrast, the oblique and apical fibers of the collagen bundle showed a chain-like structure. The area and the minor and major axis lengths of cross-sections of the horizontal fiber, as determined from 3-dimensional images, were significantly different from those of the oblique and apical fibers. CONCLUSION: These findings indicate that collagen bundles in horizontal fiber areas have high strength and that the tooth is firmly anchored to the alveolar bone by the horizontal fibers, but is not secured evenly to the alveolar bone. The tooth is firmly anchored around the cervical area, creating a "slingshot-like structure." This study has provided further insights into the structure of the PDL and forms the basis for the development of more effective therapies for periodontal tissue regeneration.
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Colágeno/ultraestrutura , Ligamento Periodontal/ultraestrutura , Dente , Animais , Tomografia com Microscopia Eletrônica , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
In mammals, definitive erythropoiesis first occurs in fetal liver (FL), although little is known about how the process is regulated. FL consists of hepatoblasts, sinusoid endothelial cells and hematopoietic cells. To determine niche cells for fetal liver erythropoiesis, we isolated each FL component by flow cytometry. mRNA analysis suggested that Dlk-1-expressing hepatoblasts primarily expressed EPO and SCF, genes encoding erythropoietic cytokines. EPO protein was detected predominantly in hepatoblasts, as assessed by ELISA and immunohistochemistry, and was not detected in sinusoid endothelial cells and hematopoietic cells. To characterize hepatoblast function in FL, we analyzed Map2k4(-/-) mouse embryos, which lack hepatoblasts, and observed down-regulation of EPO and SCF expression in FL relative to wild-type mice. Our observations demonstrate that hepatoblasts comprise a niche for erythropoiesis through cytokine secretion.
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Citocinas/biossíntese , Eritropoese , Feto/fisiologia , Células-Tronco Hematopoéticas/fisiologia , Fígado/embriologia , Animais , Regulação para Baixo , Citometria de Fluxo , MAP Quinase Quinase 4/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICRRESUMO
Motile cilia of multiciliated epithelial cells undergo synchronized beating to produce fluid flow along the luminal surface of various organs. Each motile cilium consists of an axoneme and a basal body (BB), which are linked by a "transition zone" (TZ). The axoneme exhibits a characteristic 9+2 microtubule arrangement important for ciliary motion, but how this microtubule system is generated is not yet fully understood. Here we show that calmodulin-regulated spectrin-associated protein 3 (CAMSAP3), a protein that can stabilize the minus-end of a microtubule, concentrates at multiple sites of the cilium-BB complex, including the upper region of the TZ or the axonemal basal plate (BP) where the central pair of microtubules (CP) initiates. CAMSAP3 dysfunction resulted in loss of the CP and partial distortion of the BP, as well as the failure of multicilia to undergo synchronized beating. These findings suggest that CAMSAP3 plays pivotal roles in the formation or stabilization of the CP by localizing at the basal region of the axoneme and thereby supports the coordinated motion of multicilia in airway epithelial cells.
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Cílios/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Microtúbulos/metabolismo , Citoesqueleto de Actina/metabolismo , Animais , Axonema/fisiologia , Corpos Basais/fisiologia , Células Epiteliais/metabolismo , Feminino , Masculino , Camundongos , Camundongos Endogâmicos ICR , Camundongos Transgênicos , Movimento/fisiologia , Traqueia/fisiologiaRESUMO
Targeted temperature management (TTM), or therapeutic hypothermia, is one of the most potent neuroprotective approaches after ischemic and traumatic brain injuries. TTM has been applied clinically with various methods, but effective achievement and maintenance of the target temperature remain challenging. Furthermore, timing of cooling and target body and brain temperature to optimize effectiveness for neuroprotection and to minimize side effects are yet to be standardized. Focal brain cooling is a potential strategy to minimize adverse effects of systemic TTM. In this study, we report on a focal brain cooling device for animals and its effectiveness of focal cooling in several animal models of ischemic cerebral stroke. A focal brain cooling device was constructed using a Peltier's element, a thermoelectric heat pump. The device was validated for its cooling ability, and optimal settings to induce an effective intracranial temperature were determined using male Sprague-Dawley rats. Transient and permanent middle cerebral artery occlusions were experimentally induced, and focal brain cooling was applied using the device varying the timing and duration of cooling. The stroke-induced infarct and edema volumes were evaluated from Nissl-stained cryosections. The focal brain cooling device was able to decrease and subsequently maintained cerebral hypothermia in free-moving rats without altering the core temperature. The device with validated intracranial temperatures produced neuroprotective effects in the acute phase of ischemic neural death, reperfusion injury, progressing damage to the penumbra, and edema formation. In conclusion, our validated focal cooling device enabled rapid and accurate cerebral TTM in rats. Using this device, we were able to test the neuroprotective effect of focal TTM in several pathological stages of cerebral ischemia, which warrants further studies to develop clinically feasible TTM procedures for patients with cerebral stroke.
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Hipotermia Induzida/métodos , Infarto da Artéria Cerebral Média/terapia , Animais , Encéfalo/patologia , Infarto da Artéria Cerebral Média/patologia , Masculino , Ratos Sprague-DawleyRESUMO
Rett syndrome (RTT) is a neurodevelopmental disorder mainly caused by mutations in the gene encoding the transcriptional regulator Methyl-CpG-binding protein 2 (MeCP2), located on the X chromosome. Many RTT patients have breathing abnormalities, such as apnea and breathing irregularity, and respiratory infection is the most common cause of death in these individuals. Previous studies showed that MeCP2 is highly expressed in the lung, but its role in pulmonary function remains unknown. In this study, we found that MeCP2 deficiency affects pulmonary gene expression and structures. We also found that Mecp2-null mice, which also have breathing problems, often exhibit inflammatory lung injury. These injuries occurred in specific sites in the lung lobes. In addition, polarizable foreign materials were identified in the injured lungs of Mecp2-null mice. These results indicated that aspiration might be a cause of inflammatory lung injury in Mecp2-null mice. On the other hand, MeCP2 deficiency affected the expression of several neuromodulator genes in the lower brainstem. Among them, neuropeptide substance P (SP) immunostaining was reduced in Mecp2-null brainstem. These findings suggest that alteration of SP expression in brainstem may be involved in autonomic dysregulation, and may be one of the causes of aspiration in Mecp2-null mice.
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Proteína 2 de Ligação a Metil-CpG/genética , Pneumonia/genética , Transtornos Respiratórios/genética , Síndrome de Rett/genética , Animais , Tronco Encefálico/metabolismo , Feminino , Expressão Gênica , Humanos , Pulmão/metabolismo , Pulmão/patologia , Pulmão/fisiopatologia , Masculino , Proteína 2 de Ligação a Metil-CpG/deficiência , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pneumonia/mortalidade , Pneumonia/fisiopatologia , Respiração , Transtornos Respiratórios/mortalidade , Transtornos Respiratórios/fisiopatologia , Síndrome de Rett/metabolismo , Síndrome de Rett/fisiopatologia , Substância P/genética , Substância P/metabolismo , Taxa de SobrevidaRESUMO
The accurate comprehension of normal tissue provides essential data to analyse abnormalities such as disease and regenerative processes. In addition, understanding the proper structure of the target tissue and its microenvironment may facilitate successful novel treatment strategies. Many studies have examined the nature and structure of periodontal ligaments (PDLs); however, the three-dimensional (3D) structure of cells in normal PDLs remains poorly understood. In this study, we used focused ion beam/scanning electron microscope tomography to investigate the whole 3D ultrastructure of PDL cells along with quantitatively analysing their structural properties and ascertaining their orientation to the direction of the collagen fibre. PDL cells were shown to be in contact with each other, forming a widespread mesh-like network between the cementum and the alveolar bone. The volume of the cells in the horizontal fibre area was significantly larger than in other areas, whereas the anisotropy of these cells was lower than in other areas. Furthermore, the orientation of cells to the PDL fibres was not parallel to the PDL fibres in each area. As similar evaluations are recognized as being challenging using conventional two-dimensional methods, these novel 3D findings may contribute necessary knowledge for the comprehensive understanding and analysis of PDLs.
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Ligamento Periodontal/citologia , Animais , Osso e Ossos/citologia , Osso e Ossos/metabolismo , Microambiente Celular/fisiologia , Colágeno/metabolismo , Cemento Dentário/metabolismo , Tomografia com Microscopia Eletrônica/métodos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ligamento Periodontal/metabolismoRESUMO
Endocrine and endothelial cells of the anterior pituitary gland frequently make close appositions or contacts, and the secretory granules of each endocrine cell tend to accumulate at the perivascular regions, which is generally considered to facilitate secretory functions of these cells. However, three-dimensional relationships between the localization pattern of secretory granules and blood vessels are not fully understood. To define and characterize these spatial relationships, we used scanning electron microscopy (SEM) three-dimensional reconstruction method based on focused ion-beam slicing and scanning electron microscopy (FIB/SEM). Full three-dimensional cellular architectures of the anterior pituitary tissue at ultrastructural resolution revealed that about 70% of endocrine cells were in apposition to the endothelial cells, while almost 30% of endocrine cells were entirely isolated from perivascular space in the tissue. Our three-dimensional analyses also visualized the distribution pattern of secretory granules in individual endocrine cells, showing an accumulation of secretory granules in regions in close apposition to the blood vessels in many cases. However, secretory granules in cells isolated from the perivascular region tended to distribute uniformly in the cytoplasm of these cells. These data suggest that the cellular interactions between the endocrine and endothelial cells promote an uneven cytoplasmic distribution of the secretory granules.
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Grânulos Citoplasmáticos/metabolismo , Células Endócrinas/ultraestrutura , Células Endoteliais/ultraestrutura , Adeno-Hipófise/ultraestrutura , Animais , Simulação por Computador , Células Endócrinas/classificação , Células Endócrinas/patologia , Células Endoteliais/classificação , Células Endoteliais/patologia , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Varredura , Adeno-Hipófise/anatomia & histologia , Ratos , Ratos WistarRESUMO
An important consideration in regeneration therapy is the fact that the tissue surrounding an organ supports its function. Understanding the structure of the periosteum can contribute to more effective bone regeneration therapy. As a cellular source, the periosteum also assists bone growth and fracture healing; this further necessitates its direct contact with the bone. However, its anchoring strength appears to be inexplicably stronger than expected. In this study, we used focused ion beam/scanning electron microscope tomography to investigate ultrathin serial sections as well as the three dimensional ultrastructure of the periosteum to clarify the architecture of its anchoring strength, as such assessments are challenging using conventional methods. We discovered perforating fibres that arise from the bone surface at 30 degree angles. Additionally, the fibres across the osteoblast layer were frequently interconnected to form a net-like structure. Fibroblast processes were observed extending into the perforating fibres; their morphologies were distinct from those of typical fibroblasts. Thus, our study revealed novel ultrastructures of the periosteum that support anchorage and serve as a cellular source as well as a mechanical stress transmitter.
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Fibroblastos/ultraestrutura , Periósteo/ultraestrutura , Crânio/ultraestrutura , Animais , Fibroblastos/metabolismo , Masculino , Microscopia Eletroquímica de Varredura , Periósteo/metabolismo , Ratos , Ratos Sprague-Dawley , Crânio/metabolismoRESUMO
Methyl-CpG-binding protein 2 (MeCP2) is an epigenetic regulator of gene expression that is essential for normal brain development. Mutations in MeCP2 lead to disrupted neuronal function and can cause Rett syndrome (RTT), a neurodevelopmental disorder. Previous studies reported cardiac dysfunction, including arrhythmias in both RTT patients and animal models of RTT. In addition, recent studies indicate that MeCP2 may be involved in cardiac development and dysfunction, but its role in the developing and adult heart remains unknown. In this study, we found that Mecp2-null ESCs could differentiate into cardiomyocytes, but the development and further differentiation of cardiovascular progenitors were significantly affected in MeCP2 deficiency. In addition, we revealed that loss of MeCP2 led to dysregulation of endogenous cardiac genes and myocardial structural alterations, although Mecp2-null mice did not exhibit obvious cardiac functional abnormalities. Furthermore, we detected methylation of the CpG islands in the Tbx5 locus, and showed that MeCP2 could target these sequences. Taken together, these results suggest that MeCP2 is an important regulator of the gene-expression program responsible for maintaining normal cardiac development and cardiomyocyte structure.
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Arritmias Cardíacas/genética , Epigênese Genética , Predisposição Genética para Doença , Proteína 2 de Ligação a Metil-CpG/genética , Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Animais , Arritmias Cardíacas/metabolismo , Arritmias Cardíacas/patologia , Diferenciação Celular , Ilhas de CpG , Metilação de DNA , Modelos Animais de Doenças , Células-Tronco Embrionárias/metabolismo , Células-Tronco Embrionárias/patologia , Feminino , Perfilação da Expressão Gênica , Humanos , Proteína 2 de Ligação a Metil-CpG/deficiência , Camundongos , Camundongos Knockout , Miocárdio/patologia , Miócitos Cardíacos/patologia , Miosinas/genética , Miosinas/metabolismo , Transdução de Sinais , Proteínas com Domínio T/genética , Proteínas com Domínio T/metabolismoRESUMO
The endoplasmic reticulum (ER) and mitochondria associate at multiple contact sites to form specific domains known as mitochondria-ER associated membranes (MAMs) that play a role in the regulation of various cellular processes such as Ca2+ transfer, autophagy, and inflammation. Recently, it has been suggested that MAMs are also involved in mitochondrial dynamics, especially fission events. Cytological analysis showed that ER tubules were frequently located close to each other in mitochondrial fission sites that accumulate fission-related proteins. Three-dimensional (3D) imaging of ER-mitochondrial contacts in yeast mitochondria by using cryo-electron tomography also showed that ER tubules were attached near the constriction site, which is considered to be a fission site1). MAMs have been suggested to play a role in the initiation of mitochondrial fission, although the molecular relationships between MAMs and the mitochondrial fission process have not been established. Although an ER-mitochondrial membrane association has also been observed at the fission site in mammalian mitochondria, the detailed organization of MAMs around mammalian mitochondria remains to be established. To visualize the 3D distribution of the ER-mitochondrial contacts around the mitochondria, especially around the constriction site in mammalian cells, we attempted 3D structural analysis of the mammalian cytoplasm using high-resolution focused ion-beam scanning electron microscopy (FIB-SEM) tomography, and observed the distribution pattern of ER contacts around the mammalian mitochondrial constriction site.Rat hepatocytes and HeLa cells were used. Liver tissue was obtained from male rats (Wistar, 6W) fixed by transcardial perfusion of 2% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4) under deep anesthesia. HeLa cells were fixed with the same fixative. The specimens were then stained en bloc to enhance membrane contrast and embedded in epoxy resin2). The surface of the specimens was freshly exposed using an ultramicrotome and examined by FIB/SEM (Quanta 3D FEG, FEI, USA). Ion-beam milling and image acquisition cycles were performed under the following conditions. The milling was performed with a gallium ion beam at 30 kV with a current of 100 pA, with a milling pitch of 10 nm/step. Material contrast images using backscattered electrons (BSE) were acquired at a landing energy of 2 keV with a bias voltage of 1.5-2.5 kV using a vCD detector. The remaining acquisition parameters were as follows: beam current = 11 pA, dwell time = 6-30 µs/pixel, image size = 1024 × 883 pixel (5.9 × 5.1 µm), pixel size = 5.8 nm/pixel. The resultant image stack was processed using Avizo 6.3 and Amira 5.4(FEI, USA).Reconstructed volume showed the existence of several constriction sites on mitochondria in both chemically fixed normal hepatocytes and HeLa cells. Each material contrast image of specimen surfaces showed two types of membrane associations between the ER and mitochondria. The first was an osmiophilic bridge-like structure; these bridges were approximately 50 nm in length, and they connected the ER membrane and the mitochondrial outer membrane (OMM). The second was a close apposition (< 20 nm) of the ER membrane and the OMM. Membrane segmentation revealed the 3D distribution of the membrane contacts; 10 to 20% of the mitochondrial surface was occupied by ER contacts. No fundamental difference was observed between hepatocytes and HeLa cells in the distribution pattern of the contacts. Although ER-contacts and bridge-like structures were occasionally found to accumulate around the mitochondrial constriction area, we did not observe any ring-like ER tubules around the mammalian mitochondrial constriction site, as in yeast. These results suggest that the role of ER-membrane associations in the mitochondrial fission process may differ between mammals and yeast.
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IntroductionMitochondrial fission and fusion events are fundamental mechanisms for quality control of mitochondrial functions. Mitochondrial DNA (mtDNA) usually divides in offspring mitochondria after fission and mtDNA dynamics are thought to be coordinated with mitochondrial turnover. Recently, several candidate mechanisms for the relationship between mtDNA division and mitochondrial fission have been suggested ([1], 2012). The dynamics of mtDNA or nucleoids can be observed using fluorescent microscopy, but the ultrastructural aspects of their coordination remain unclear. Although visualization of mtDNA at the electron microscopic level is an important step in understanding how mtDNA division and mitochondrial division are coordinated, it is quite difficult to observe using conventional electron microscopic methods. In the present study, we attempted to establish correlative light and electron microscopy (CLEM) observation to visualize the three-dimensional localization of the mtDNA /nucleoid within mitochondria at electron microscopic resolution using a combination of immuno-electron and focused-ion beam scanning electron microscopy (FIB/SEM) tomography methods. Materials & methodsHeLa cells were fixed using 4% paraformaldehyde and 0.05% glutaraldehyde in 0.1 M phosphate buffer, and then immunohistochemically labeled with anti-TFAM IgG antibody (Abnova, USA) or anti-DNA IgM antibody (Progen, Germany). The cells were then reacted with biotin-labeled secondary antibodies. The immunoreactivities were visualized using two methods: the ABC method and streptavidin Fluoro-Nanogold. Immunohistochemically labeled specimens were then observed using light microscopy. These specimens were then developed using a Gold Enhancement kit (Nanoprobe, USA) for 150 s. Specimens for electron microscopy were stained using the ROTO method, embedded in resin, and subjected to FIB/SEM tomography (Quanta 3D FEG, FEI). 3D reconstruction was performed using the software Amira (FEI). Results & discussionWe were not able to identify a nucleoid-like structure within mitochondria, even in a complete 3D reconstruction using FIB/SEM with conventional staining. In CLEM observations, immunoreaction (IR) products were correlatively observed under LM and EM. Pre-embedding immuno-electron microscopy showed DAB and gold IR in the matrix of some mitochondria. Interestingly, IR products were observed in the globular region of the mitochondrial matrix (approximately 0.4 µm in diameter), frequently localizing in the peripheral end of the mitochondrial matrix, adjacent to the inner membrane. Using the post-embedding immunogold method, gold labels were also observed in a portion of the matrix adjacent to the mitochondrial inner membrane. These immunocytochemical results were concordant with our fluorescent microscopic observations.
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Induced pluripotent stem cells (iPSCs) are a promising tool for regenerative medicine. Use of iPSC lines for future hematotherapy will require examination of their hematopoietic potential. Adult skin fibroblast somatic cells constitute a source of iPSCs that can be accessed clinically without ethical issues. Here, we used different methods to compare mesodermal and hematopoietic potential by embryoid body formation of five iPSC lines established from adult mouse tail-tip fibroblasts (TTFs). We observed variation in proliferation and in expression of genes (Brachyury, Tbx1, Gata1, Klf1, Csf1r) and proteins (Flk1, Ter119 and CD45) among TTF-derived lines. 256H18 iPSCs showed highest proliferation and most efficient differentiation into mesodermal and hematopoietic cells, while expression levels of the pluripotency genes Oct3/4, Sox2, Klf4 and Nanog were lowest among lines analyzed. By contrast, the 212B2 line, transduced with c-Myc, showed lowest proliferation and differentiation potential, although expression levels of Oct3/4, Sox2 and Klf4 were highest. Overall, we find that mesodermal and hematopoietic potential varies among iPSCs from an identical tissue source and that c-Myc expression likely underlies these differences.