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1.
Neuroscience ; 489: 84-97, 2022 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-35218884

RESUMO

Microtubules deliver essential resources to and from synapses. Three-dimensional reconstructions in rat hippocampus reveal a sampling bias regarding spine density that needs to be controlled for dendrite caliber and resource delivery based on microtubule number. The strength of this relationship varies across dendritic arbors, as illustrated for area CA1 and dentate gyrus. In both regions, proximal dendrites had more microtubules than distal dendrites. For CA1 pyramidal cells, spine density was greater on thicker than thinner dendrites in stratum radiatum, or on the more uniformly thin terminal dendrites in stratum lacunosum moleculare. In contrast, spine density was constant across the cone shaped arbor of tapering dendrites from dentate granule cells. These differences suggest that thicker dendrites supply microtubules to subsequent dendritic branches and local dendritic spines, whereas microtubules in thinner dendrites need only provide resources to local spines. Most microtubules ran parallel to dendrite length and associated with long, presumably stable mitochondria, which occasionally branched into lateral dendritic branches. Short, presumably mobile, mitochondria were tethered to microtubules that bent and appeared to direct them into a thin lateral branch. Prior work showed that dendritic segments with the same number of microtubules had elevated resources in subregions of their dendritic shafts where spine synapses had enlarged, and spine clusters had formed. Thus, additional microtubules were not required for redistribution of resources locally to growing spines or synapses. These results provide new understanding about the potential for microtubules to regulate resource delivery to and from dendritic branches and locally among dendritic spines.


Assuntos
Dendritos , Espinhas Dendríticas , Animais , Dendritos/fisiologia , Hipocampo , Microtúbulos , Células Piramidais/fisiologia , Ratos , Sinapses/fisiologia
2.
Sci Data ; 2: 150046, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26347348

RESUMO

Resurgent interest in synaptic circuitry and plasticity has emphasized the importance of 3D reconstruction from serial section electron microscopy (3DEM). Three volumes of hippocampal CA1 neuropil from adult rat were imaged at X-Y resolution of ~2 nm on serial sections of ~50-60 nm thickness. These are the first densely reconstructed hippocampal volumes. All axons, dendrites, glia, and synapses were reconstructed in a cube (~10 µm(3)) surrounding a large dendritic spine, a cylinder (~43 µm(3)) surrounding an oblique dendritic segment (3.4 µm long), and a parallelepiped (~178 µm(3)) surrounding an apical dendritic segment (4.9 µm long). The data provide standards for identifying ultrastructural objects in 3DEM, realistic reconstructions for modeling biophysical properties of synaptic transmission, and a test bed for enhancing reconstruction tools. Representative synapses are quantified from varying section planes, and microtubules, polyribosomes, smooth endoplasmic reticulum, and endosomes are identified and reconstructed in a subset of dendrites. The original images, traces, and Reconstruct software and files are freely available and visualized at the Open Connectome Project (Data Citation 1).


Assuntos
Hipocampo/anatomia & histologia , Neurópilo , Animais , Processamento de Imagem Assistida por Computador , Microscopia Eletrônica , Ratos , Software
3.
Cardiovasc Pathol ; 23(5): 267-71, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24985899

RESUMO

Thrombus aspiration in the setting of primary percutaneous coronary intervention is a recently recommended technique that facilitates thrombus removal from the culprit lesions in acute myocardial infarction (AMI) patients. Thrombectomy specimens from 50 patients with symptoms of AMI lasting usually not more than 12 h were examined by methods of routine histology, immunohistochemistry (IHC), and electron microscopy (ELMI). In 36 patients, there were fresh thrombi, in 10 older thrombi (8 of them with simultaneous presence of a fresh thrombi) and in 3 atheroma material only (in additional 7 patients atheroma material was admixed to the thrombi), and in one patient, there was carcinoma embolus. To help to distinguish between fresh and older thrombi, we recommend IHC (presence of macrophages and endothelia) and ELMI (loss of density of the erythrocyte matrix and presence of macrophages). On the other hand, changes of neutrophils (IHC degranulation/lysis) and of platelets (ELMI degranulation) appear early and thus contribute little to distinguishing between fresh and older thrombi. It could be concluded that, in a substantial proportion of patients with AMI, there is a discrepancy between duration of the symptoms and microscopic picture of the coronary thrombus. The thrombus may apparently be symptomless for a period of days or even weeks.


Assuntos
Trombose Coronária/patologia , Infarto do Miocárdio/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Trombose Coronária/cirurgia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , Infarto do Miocárdio/cirurgia , Intervenção Coronária Percutânea , Trombectomia
5.
J Comp Neurol ; 521(2): 448-64, 2013 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-22740128

RESUMO

Although the extracellular space in the neuropil of the brain is an important channel for volume communication between cells and has other important functions, its morphology on the micron scale has not been analyzed quantitatively owing to experimental limitations. We used manual and computational techniques to reconstruct the 3D geometry of 180 µm(3) of rat CA1 hippocampal neuropil from serial electron microscopy and corrected for tissue shrinkage to reflect the in vivo state. The reconstruction revealed an interconnected network of 40-80 nm diameter tunnels, formed at the junction of three or more cellular processes, spanned by sheets between pairs of cell surfaces with 10-40 nm width. The tunnels tended to occur around synapses and axons, and the sheets were enriched around astrocytes. Monte Carlo simulations of diffusion within the reconstructed neuropil demonstrate that the rate of diffusion of neurotransmitter and other small molecules was slower in sheets than in tunnels. Thus, the non-uniformity found in the extracellular space may have specialized functions for signaling (sheets) and volume transmission (tunnels).


Assuntos
Espaço Extracelular/fisiologia , Ácido Glutâmico/metabolismo , Hipocampo/metabolismo , Neurópilo/metabolismo , Algoritmos , Animais , Membrana Celular/ultraestrutura , Difusão , Processamento de Imagem Assistida por Computador , Microscopia Eletrônica , Método de Monte Carlo , Rede Nervosa/fisiologia , Rede Nervosa/ultraestrutura , Neurotransmissores/metabolismo , Ratos , Software , Frações Subcelulares/metabolismo
6.
Neuron ; 67(6): 1009-20, 2010 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-20869597

RESUMO

Complete reconstructions of vertebrate neuronal circuits on the synaptic level require new approaches. Here, serial section transmission electron microscopy was automated to densely reconstruct four volumes, totaling 670 µm(3), from the rat hippocampus as proving grounds to determine when axo-dendritic proximities predict synapses. First, in contrast with Peters' rule, the density of axons within reach of dendritic spines did not predict synaptic density along dendrites because the fraction of axons making synapses was variable. Second, an axo-dendritic touch did not predict a synapse; nevertheless, the density of synapses along a hippocampal dendrite appeared to be a universal fraction, 0.2, of the density of touches. Finally, the largest touch between an axonal bouton and spine indicated the site of actual synapses with about 80% precision but would miss about half of all synapses. Thus, it will be difficult to predict synaptic connectivity using data sets missing ultrastructural details that distinguish between axo-dendritic touches and bona fide synapses.


Assuntos
Hipocampo/ultraestrutura , Neurópilo/ultraestrutura , Animais , Animais Recém-Nascidos , Axônios/ultraestrutura , Dendritos/ultraestrutura , Processamento Eletrônico de Dados/métodos , Técnicas In Vitro , Masculino , Microscopia Eletrônica de Transmissão/métodos , Ratos , Ratos Long-Evans , Sinapses/ultraestrutura
8.
Pathol Res Pract ; 203(10): 753-8, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17656038

RESUMO

Carcinosarcomas (CSs) are rare biphasic neoplasms of the female genital tract, with the presence of malignant epithelial and mesenchymal components. The occurrence of non-epithelial or non-mesenchymal differentiations in CSs is extremely rare. A case of a 54-year-old woman with a history of treatment for breast carcinoma is reported. The patient presented with a uterine tumor showing histological, immunohistochemical, and ultrastructural features of an extremely rare CS, with signs of melanocytic differentiation. The lesion was a 7.0 x 5.6 cm polypoid tumor mass arising from the endometrium and metastasizing in the abdominal cavity as a malignant melanoma - the most aggressive component of CS. Despite adjuvant chemotherapy, she died 8 months after the initial diagnosis due to the metastatic spread of CS. In line with current knowledge, we assess the melanocytic differentiation as one of a hypothetic differential line with the ability of multidirectional stem cell differentiation. We discuss a possible pathogenesis of this lesion and the role of tamoxifen in the tumor development as well. To the best of our knowledge, only one case of uterine CS with melanocytic differentiation has been reported in the English literature.


Assuntos
Neoplasias Abdominais/secundário , Carcinossarcoma/patologia , Diferenciação Celular , Melanócitos/ultraestrutura , Melanoma/secundário , Tumor Mulleriano Misto/patologia , Neoplasias Uterinas/patologia , Neoplasias Abdominais/tratamento farmacológico , Neoplasias Abdominais/cirurgia , Carcinossarcoma/tratamento farmacológico , Carcinossarcoma/cirurgia , Quimioterapia Adjuvante , Evolução Fatal , Feminino , Humanos , Imuno-Histoquímica , Melanoma/tratamento farmacológico , Melanoma/cirurgia , Microscopia Eletrônica , Pessoa de Meia-Idade , Tumor Mulleriano Misto/tratamento farmacológico , Tumor Mulleriano Misto/cirurgia , Neoplasias Uterinas/tratamento farmacológico , Neoplasias Uterinas/cirurgia
9.
J Neurosci ; 24(17): 4233-41, 2004 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-15115819

RESUMO

Locations of a distinctive mode of trans-endocytosis involving dendrites, axons, and glia were quantified through serial section electron microscopy. Short vesicular or long vermiform evaginations emerged from dendrites and axons and were engulfed by presynaptic or neighboring axons, astrocytes, and, surprisingly, a growth cone to form double-membrane structures called spinules. In total, 254 spinules were evaluated in 326 microm(3) of stratum radiatum in area CA1 of mature rat hippocampus. Spinules emerged from spine heads (62%), necks (24%), axons (13%), dendritic shafts (1%), or nonsynaptic protrusions (<1%) and invaginated into axons (approximately 90%), astrocytic processes (approximately 8%), or a growth cone (approximately 1%). Coated pits occurred on the engulfing membrane at the tips of most spinules (69%), and double-membrane structures occurred freely in axonal and astrocytic cytoplasm, suggesting trans-endocytosis. Spinule locations differed among mushroom and thin spines. For mushroom spines, most (84%) of the spinules were engulfed by presynaptic axons, 16% by neighboring axons, and none by astrocytic processes. At thin spines, only 17% of the spinules were engulfed by presynaptic axons, whereas 67% were engulfed by neighboring axons and 14% by astrocytic processes. Spinules engulfed by astrocytic processes support the growing evidence that perisynaptic glia interact directly with synapses at least on thin spines. Spinules with neighboring axons may provide a mechanism for synaptic competition in the mature brain. Trans-endocytosis of spinules by presynaptic axons suggest retrograde signaling or coordinated remodeling of presynaptic and postsynaptic membranes to remove transient perforations and assemble the postsynaptic density of large synapses on mushroom spines.


Assuntos
Invaginações Revestidas da Membrana Celular/ultraestrutura , Endocitose/fisiologia , Hipocampo/fisiologia , Hipocampo/ultraestrutura , Animais , Astrócitos/ultraestrutura , Axônios/fisiologia , Axônios/ultraestrutura , Vesículas Citoplasmáticas/ultraestrutura , Dendritos/fisiologia , Dendritos/ultraestrutura , Retículo Endoplasmático Liso/ultraestrutura , Cones de Crescimento/ultraestrutura , Masculino , Neurópilo/ultraestrutura , Ratos , Ratos Long-Evans
10.
Brain Res Brain Res Rev ; 39(1): 29-54, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12086707

RESUMO

Altered dendritic spines are characteristic of traumatized or diseased brain. Two general categories of spine pathology can be distinguished: pathologies of distribution and pathologies of ultrastructure. Pathologies of spine distribution affect many spines along the dendrites of a neuron and include altered spine numbers, distorted spine shapes, and abnormal loci of spine origin on the neuron. Pathologies of spine ultrastructure involve distortion of subcellular organelles within dendritic spines. Spine distributions are altered on mature neurons following traumatic lesions, and in progressive neurodegeneration involving substantial neuronal loss such as in Alzheimer's disease and in Creutzfeldt-Jakob disease. Similarly, spine distributions are altered in the developing brain following malnutrition, alcohol or toxin exposure, infection, and in a large number of genetic disorders that result in mental retardation, such as Down's and fragile-X syndromes. An important question is whether altered dendritic spines are the intrinsic cause of the accompanying neurological disturbances. The data suggest that many categories of spine pathology may result not from intrinsic pathologies of the spiny neurons, but from a compensatory response of these neurons to the loss of excitatory input to dendritic spines. More detailed studies are needed to determine the cause of spine pathology in most disorders and relationship between spine pathology and cognitive deficits.


Assuntos
Dendritos/patologia , Doenças do Sistema Nervoso/etiologia , Doenças do Sistema Nervoso/patologia , Animais , Dendritos/diagnóstico por imagem , Humanos , Ultrassonografia
11.
Proteomics ; 2(1): 85-93, 2002 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11788995

RESUMO

The phagocytosis of pathogens by macrophages classically initiates maturation of the phagosome that involves a dynamic exchange of phagosomal components with intracellular compartments of the endocytic pathway. The intracellular microorganisms have developed sophisticated mechanisms to sense environmental conditions and respond to them by phenotypic alterations that ensure their adaptation, survival and proliferation inside the cell. They have learned also to utilise host cellular components to ensure own survival. Recent results suggest that the Bcg locus/Nramp1 gene (natural resistance-associated macrophage protein 1) controls natural resistance to infection by Francisella tularensis LVS (live vaccine strain) and its effect is opposite to that observed for other Bcg/Nramp1-controlled pathogens such as several mycobacterial species, Leischmania donovani, and Salmonella typhimurium. In the case of F. tularensis LVS infection, the mutant allele of the Bcg locus (Bcg(s)/Nramp1(s)) is associated with natural resistance and, inversely, the wild type allele (Bcg(r)/Nramp1(r)) confers susceptibility. To determine whether differential allelic expression of the Bcg locus/Nramp1 gene modifies the composition of F. tularensis LVS-containing phagosomes (FCP), we have utilised an approach where we isolated FCP from infected Bcg congenic B10R (Bcg(r)/Nramp1(r)) and B10S (Bcg(s)/Nramp1(s)) macrophages of susceptible and resistant phenotype, respectively. Comparative proteomic analysis of the two phagosomal compartments with subsequent mass spectrometric analysis allowed identification of several proteins typical for FCP from B10R macrophages. They include a bacterial hypothetical 23 kDa protein, 60 kDa chaperonin GroEL, and host putative proteins that appeared to be mitochondrial ATP synthase beta-chain and NADH-ubiquinone oxidoreductase based on high cross-species homology. High abundance of the hypothetical 23 kDa protein correlates with the susceptible phenotype and, possibly, pathogenicity of F. tularensis LVS. The results demonstrate that F. tularensis LVS can exploit ion transport function of Bcg/Nramp1 to its own advantage.


Assuntos
Alelos , Vacinas Bacterianas , Proteínas de Transporte de Cátions/genética , Suscetibilidade a Doenças/imunologia , Francisella tularensis/imunologia , Fagossomos/metabolismo , Proteoma , Eletroforese em Gel Bidimensional , Citometria de Fluxo , Francisella tularensis/genética , Francisella tularensis/metabolismo , Microscopia Eletrônica , Fagossomos/ultraestrutura , Fenótipo
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