Molecular organization of perineuronal net of the oculomotor neurons in the frog

Parvalbumin-positive (PV+) interneurons play a pivotal role in orchestrating windows of experience-dependent brain plasticity during development. Critical period closure is marked by the condensation of a perineuronal net (PNN) tightly enwrapping subsets of PV+ neurons, both acting as a molecular brake on plasticity and maintaining mature PV+ cell signaling. As much of the molecular organization of PNNs exists at length scales near or below the diffraction limit of light microscopy, we developed a superresolution imaging and analysis platform to visualize the structural organization of PNNs and the synaptic inputs perforating them in primary visual cortex. We identified a structural trajectory of PNN maturation featuring a range of net structures, which was accompanied by an increase in Synaptotagmin-2 (Syt2) signals on PV+ cells suggestive of increased inhibitory input between PV+ neurons. The same structural trajectory was followed by PNNs both during normal development and under conditions of critical period delay by total sensory deprivation or critical period acceleration by deletion ofMeCP2, the causative gene for Rett syndrome, despite shifted maturation levels under these perturbations. Notably, superresolution imaging further revealed a decrease in Syt2 signals alongside an increase in vesicular glutamate transporter-2 signals on PV+ cells inMeCP2-deficient animals, suggesting weaker recurrent inhibitory input between PV+ neurons and stronger thalamocortical excitatory inputs onto PV+ cells. These results imply a latent imbalanced circuit signature that might promote cortical silencing in Rett syndrome before the functional regression of vision.

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Bimolecular and Monomolecular Lipid Films: Selected Area Electron Diffraction

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100063603 ◽

1969 ◽

Vol 27 ◽

pp. 338-339

Author(s):

Robert M. Glaeser ◽

David W. Deamer

Keyword(s):

Electron Diffraction ◽

Membrane Structure ◽

Molecular Organization ◽

Membrane Material ◽

X Ray Diffraction ◽

Membrane Systems ◽

X Ray ◽

Selected Area Electron Diffraction ◽

Lipid Films ◽

Ultimate Objective

In the investigation of the molecular organization of cell membranes it is often supposed that lipid molecules are arranged in a bimolecular film. X-ray diffraction data obtained in a direction perpendicular to the plane of suitably layered membrane systems have generally been interpreted in accord with such a model of the membrane structure. The present studies were begun in order to determine whether selected area electron diffraction would provide a tool of sufficient sensitivity to permit investigation of the degree of intermolecular order within lipid films. The ultimate objective would then be to apply the method to single fragments of cell membrane material in order to obtain data complementary to the transverse data obtainable by x-ray diffraction.

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Evaluation of Freezing Methods by Low Temperature X-Ray Diffraction

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100079176 ◽

1977 ◽

Vol 35 ◽

pp. 330-333

Author(s):

T. Gulik-Krzywicki ◽

M.J. Costello

Keyword(s):

Biological Materials ◽

Molecular Organization ◽

X Ray Diffraction ◽

Glass Capillary ◽

X Ray ◽

Rate Dependent ◽

Before And After ◽

Freeze Etching ◽

Diffraction Patterns ◽

Set Up

Freeze-etching electron microscopy is currently one of the best methods for studying molecular organization of biological materials. Its application, however, is still limited by our imprecise knowledge about the perturbations of the original organization which may occur during quenching and fracturing of the samples and during the replication of fractured surfaces. Although it is well known that the preservation of the molecular organization of biological materials is critically dependent on the rate of freezing of the samples, little information is presently available concerning the nature and the extent of freezing-rate dependent perturbations of the original organizations. In order to obtain this information, we have developed a method based on the comparison of x-ray diffraction patterns of samples before and after freezing, prior to fracturing and replication.Our experimental set-up is shown in Fig. 1. The sample to be quenched is placed on its holder which is then mounted on a small metal holder (O) fixed on a glass capillary (p), whose position is controlled by a micromanipulator.

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On the structural organization of biological pumps and channels

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100110878 ◽

1984 ◽

Vol 42 ◽

pp. 138-141

Author(s):

G. Zampighi ◽

M. Kreman

Keyword(s):

Active Transport ◽

Transport System ◽

Plasma Membranes ◽

Transport Proteins ◽

Molecular Organization ◽

Passive Transport ◽

Concentration Gradients ◽

Cell Functions ◽

Natural Concentration ◽

Active Transport System

The plasma membranes of most animal cells contain transport proteins which function to provide passageways for the transported species across essentially impermeable lipid bilayers. The channel is a passive transport system which allows the movement of ions and low molecular weight molecules along their concentration gradients. The pump is an active transport system and can translocate cations against their natural concentration gradients. The actions and interplay of these two kinds of transport proteins control crucial cell functions such as active transport, excitability and cell communication. In this paper, we will describe and compare several features of the molecular organization of pumps and channels. As an example of an active transport system, we will discuss the structure of the sodium and potassium ion-activated triphosphatase [(Na+ +K+)-ATPase] and as an example of a passive transport system, the communicating channel of gap junctions and lens junctions.

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High-resolution measurement of birefringent fine structure in living cells using a new polarized light microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100136647 ◽

1995 ◽

Vol 53 ◽

pp. 54-55

Author(s):

Rudolf Oldenbourg

Keyword(s):

Light Microscope ◽

Fluorescent Dyes ◽

Polarized Light ◽

Processing System ◽

Video Camera ◽

Molecular Organization ◽

Computer Assisted ◽

Image Recording ◽

Birefringent Crystal ◽

Technological Advances

The recent renaissance of the light microsope is fueled in part by technological advances in components on the periphery of the microscope, such as the laser as illumination source, electronic image recording (video), computer assisted image analysis and the biochemistry of fluorescent dyes for labeling specimens. After great progress in these peripheral parts, it seems timely to examine the optics itself and ask how progress in the periphery facilitates the use of new optical components and of new optical designs inside the microscope. Some results of this fruitful reflection are presented in this symposium.We have considered the polarized light microscope, and developed a design that replaces the traditional compensator, typically a birefringent crystal plate, with a precision universal compensator made of two liquid crystal variable retarders. A video camera and digital image processing system provide fast measurements of specimen anisotropy (retardance magnitude and azimuth) at ALL POINTS of the image forming the field of view. The images document fine structural and molecular organization within a thin optical section of the specimen.

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Faculty Opinions recommendation of CD151 accelerates breast cancer by regulating alpha 6 integrin function, signaling, and molecular organization.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1115403.571429 ◽

2008 ◽

Author(s):

C Michael DiPersio

Keyword(s):

Breast Cancer ◽

Molecular Organization

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Faculty Opinions recommendation of Perineuronal net digestion with chondroitinase restores memory in mice with tau pathology.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.725268556.793504202 ◽

2015 ◽

Author(s):

Robert Petersen

Keyword(s):

Tau Pathology ◽

Perineuronal Net

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Hybrid Biopolymer and Lipid Nanoparticles with Improved Transfection Efficacy for mRNA

Cells ◽

10.3390/cells9092034 ◽

2020 ◽

Vol 9 (9) ◽

pp. 2034 ◽

Cited By ~ 5

Author(s):

Christian D. Siewert ◽

Heinrich Haas ◽

Vera Cornet ◽

Sara S. Nogueira ◽

Thomas Nawroth ◽

...

Keyword(s):

Small Angle ◽

Messenger Rna ◽

Physicochemical Characterization ◽

Single Step ◽

Model Systems ◽

Molecular Organization ◽

Hybrid Nanoparticles ◽

Polymer Mixture ◽

Internal Organization ◽

Transfection Efficacy

Hybrid nanoparticles from lipidic and polymeric components were assembled to serve as vehicles for the transfection of messenger RNA (mRNA) using different portions of the cationic lipid DOTAP (1,2-Dioleoyl-3-trimethylammonium-propane) and the cationic biopolymer protamine as model systems. Two different sequential assembly approaches in comparison with a direct single-step protocol were applied, and molecular organization in correlation with biological activity of the resulting nanoparticle systems was investigated. Differences in the structure of the nanoparticles were revealed by thorough physicochemical characterization including small angle neutron scattering (SANS), small angle X-ray scattering (SAXS), and cryogenic transmission electron microscopy (cryo-TEM). All hybrid systems, combining lipid and polymer, displayed significantly increased transfection in comparison to lipid/mRNA and polymer/mRNA particles alone. For the hybrid nanoparticles, characteristic differences regarding the internal organization, release characteristics, and activity were determined depending on the assembly route. The systems with the highest transfection efficacy were characterized by a heterogenous internal organization, accompanied by facilitated release. Such a system could be best obtained by the single step protocol, starting with a lipid and polymer mixture for nanoparticle formation.

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