Localization of Actin Filaments in the Central Nervous System Using Phalloidin and Correlative Light and Electron Microscopy

1999 ◽  
Vol 5 (S2) ◽  
pp. 498-499
Author(s):  
Francisco Capani ◽  
Maryann E. Martone ◽  
Thomas J. Deerinck ◽  
Mark H. Ellisman

Determination of the fine organization of actin networks is important for understanding different functional aspects of the central nervous system (CNS) such as plastic events. Early studies investigating the localization of actin filaments at the ultrastructural level relied on structural methods or antibodies raised against actin isoforms. More recently, many investigators have employed the actin-binding peptide phalloidin conjugated to various fluorescent molecules. By conjugating phalloidin to the fluorophore eosin, we have been able to localize f-actin at the electron microscopic level using photooxidation of diaminobenzidine (DAB) by eosin. The goal of this study is to provide a better description of the cellular and subcellular localization of actin filaments at the light and electron microscopy level based on the high binding affinity of phalloidin for actin and utilizing this novel method.Tissues were obtained from male Sprague Dawley rats that were perfused transcardially under deep anesthesia with normal rat Ringer's solution at 35°C followed by fixative. The fixative contained 4 % paraformaldehyde and different concentrations of glutaraldehyde, ranging from 0.1- 2.5%. Sections of cerebellum, striatum and hippocampus which were cut at a thickness of 50-80 μm with a Vibratome were incubated with phalloidin-eosin in a solution of 0.05% in 0.5% cold water fish gelatin/50mM glycine-PBS (“working buffer”) for 2 hrs. As a control, phalloidin-eosin was omitted for one set of tissues.

BIOCELL ◽  
2008 ◽  
Vol 32 (1) ◽  
pp. 1-8 ◽  
Author(s):  
FRANCISCO CAPANI ◽  
EZEQUIEL SARACENO ◽  
VALERIA ROMINA BOTI ◽  
LAURA AON-BERTOLINO ◽  
JUAN CARLOS FERN罭DEZ ◽  
...  

2000 ◽  
Vol 853 (2) ◽  
pp. 245-268 ◽  
Author(s):  
Julia Serrano ◽  
L.Otto Uttenthal ◽  
Alfredo Martı́nez ◽  
A.Patricia Fernández ◽  
Javier Martı́nez de Velasco ◽  
...  

1995 ◽  
Vol 37 (2) ◽  
pp. 137-143
Author(s):  
C.L.P. Lancellotti ◽  
C.E.P. Corbett ◽  
M.I.S. Duarte

Histopathological and ultrastructural studies of 23 patients who died with clinical diagnosis of measles were carried out. In 12 cases viral nucleocapsids were searched by electron microscopy and detected in 100% of the cases in the lungs and in 50% of the cases in the central nervous system. They were mostly intranuclear. Histopathological changes associated to neurological alterations and the detection of virion are discussed in relation to acute and delayed clinical manifestations.


Author(s):  
Minkyo Jung ◽  
Doory Kim ◽  
Ji Young Mun

Actin networks and actin-binding proteins (ABPs) are most abundant in the cytoskeleton of neurons. The function of ABPs in neurons is nucleation of actin polymerization, polymerization or depolymerization regulation, bundling of actin through crosslinking or stabilization, cargo movement along actin filaments, and anchoring of actin to other cellular components. In axons, ABP–actin interaction forms a dynamic, deep actin network, which regulates axon extension, guidance, axon branches, and synaptic structures. In dendrites, actin and ABPs are related to filopodia attenuation, spine formation, and synapse plasticity. ABP phosphorylation or mutation changes ABP–actin binding, which regulates axon or dendritic plasticity. In addition, hyperactive ABPs might also be expressed as aggregates of abnormal proteins in neurodegeneration. Those changes cause many neurological disorders. Here, we will review direct visualization of ABP and actin using various electron microscopy (EM) techniques, super resolution microscopy (SRM), and correlative light and electron microscopy (CLEM) with discussion of important ABPs in neuron.


2016 ◽  
Vol 36 (8) ◽  
pp. 753-760 ◽  
Author(s):  
Fernanda Menezes de Oliveira e Silva ◽  
Dayane Alcantara ◽  
Rafael Cardoso Carvalho ◽  
Phelipe Oliveira Favaron ◽  
Amilton Cesar dos Santos ◽  
...  

Abstract: This study describes the development of the central nervous system in guinea pigs from 12th day post conception (dpc) until birth. Totally, 41 embryos and fetuses were analyzed macroscopically and by means of light and electron microscopy. The neural tube closure was observed at day 14 and the development of the spinal cord and differentiation of the primitive central nervous system vesicles was on 20th dpc. Histologically, undifferentiated brain tissue was observed as a mass of mesenchymal tissue between 18th and 20th dpc, and at 25th dpc the tissue within the medullary canal had higher density. On day 30 the brain tissue was differentiated on day 30 and the spinal cord filling throughout the spinal canal, period from which it was possible to observe cerebral and cerebellar stratums. At day 45 intumescences were visualized and cerebral hemispheres were divided, with a clear division between white and gray matter in brain and cerebellum. Median sulcus of the dorsal spinal cord and the cauda equina were only evident on day 50. There were no significant structural differences in fetuses of 50 and 60 dpc, and animals at term were all lissencephalic. In conclusion, morphological studies of the nervous system in guinea pig can provide important information for clinical studies in humans, due to its high degree of neurological maturity in relation to its short gestation period, what can provide a good tool for neurological studies.


2017 ◽  
Vol 5 (1) ◽  
pp. 74-78
Author(s):  
V. Tsymbaliuk ◽  
V. Semenova ◽  
L. Pichkur ◽  
O. Velychko ◽  
D. Egorova

The review summarizes the current concepts of cell-tissue and molecular features of development of demyelinating processes in the central nervous system related to multiple sclerosis and its animal model – allergic encephalomyelitis. An analysis of recently published studies of this pathology, carried out with light and electron microscopy and immunohistochemical and molecular genetic methods, is given. New methodological approaches to the study of the pathomorhological aspects of demyelinating disorders allowed receiving in-depth understanding of the etiology and mechanisms of demyelination processes in the brain and spinal cord tissues at the cellular level and identifying the ways to develop effective modern methods of pathogenetic treatment of these diseases using cell therapy.


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