Reconstruction and Morphometric Analysis of Hippocampal Neurons from Mice Expressing Fluorescent Proteins

2014 ◽  
pp. 251-262 ◽  
Author(s):  
Nataliya Golovyashkina ◽  
Frederik Sündermann ◽  
Roland Brandt ◽  
Lidia Bakota
Keyword(s):  
Morphometric Analysis ◽  
Hippocampal Neurons ◽  
Fluorescent Proteins

scholarly journals Voltage Imaging with a NIR-Absorbing Phosphine Oxide Rhodamine Voltage Reporter

2019 ◽  
Author(s):  
Monica Gonzalez ◽  
Alison Walker ◽  
Kevin Cao ◽  
Julia Lazzari-Dean ◽  
Nick Settineri ◽  
...  
Keyword(s):  
Phosphine Oxide ◽  
Optical Sensors ◽  
Hippocampal Neurons ◽  
Near Infrared ◽  
Fluorescent Dyes ◽  
Fluorescent Proteins ◽  
Ex Vivo ◽  
Physiological Activity ◽  
Voltage Sensitivity ◽  
Voltage Imaging

Near infrared (NIR) fluorophores may hold the key for non-invasive optical imaging of deep structures in intact organisms with high spatial and temporal resolution. Yet, developing fluorescent dyes that emit and absorb light at wavelengths greater than 700 nm and that respond to biochemical and biophysical events in living systems remains an outstanding challenge. Here, we report the design, synthesis, and application of NIR-absorbing and -emitting, sulfonated, phosphine-oxide (po) rhodamines for voltage imaging in thick tissue from the central nervous system. We find po-rhodamine based voltage reporters, or poRhoVRs, display NIR excitation and emission profiles at greater than 700 nm, show best-in class voltage sensitivity (up to 43% ΔF/F per 100 mV in HEK cells), and can be combined with existing optical sensors, like Ca<sup>2+</sup>-sensitive fluorescent proteins (GCaMP), and actuators, like light-activated opsins ChannelRhodopsin-2 (ChR2). Simultaneous voltage and Ca<sup>2+</sup> imaging reveals differences in activity dynamics in rat hippocampal neurons, and pairing poRhoVR with blue-light based ChR2 affords all-optical electrophysiology. In <i>ex vivo</i> retinas isolated from a mouse model of retinal degeneration, poRhoVR, together with GCaMP-based Ca<sup>2+</sup> imaging and traditional multi-electrode array (MEA) recording, can provide a comprehensive physiological activity profile of neuronal activity. Taken together, these experiments establish that poRhoVR will open new horizons in optical interrogation of cellular and neuronal physiology in intact systems.

scholarly journals Voltage Imaging with a NIR-Absorbing Phosphine Oxide Rhodamine Voltage Reporter

2019 ◽  
Author(s):  
Monica Gonzalez ◽  
Alison Walker ◽  
Kevin Cao ◽  
Julia Lazzari-Dean ◽  
Nick Settineri ◽  
...  
Keyword(s):  
Phosphine Oxide ◽  
Optical Sensors ◽  
Hippocampal Neurons ◽  
Near Infrared ◽  
Fluorescent Dyes ◽  
Fluorescent Proteins ◽  
Ex Vivo ◽  
Physiological Activity ◽  
Voltage Sensitivity ◽  
Voltage Imaging

Near infrared (NIR) fluorophores may hold the key for non-invasive optical imaging of deep structures in intact organisms with high spatial and temporal resolution. Yet, developing fluorescent dyes that emit and absorb light at wavelengths greater than 700 nm and that respond to biochemical and biophysical events in living systems remains an outstanding challenge. Here, we report the design, synthesis, and application of NIR-absorbing and -emitting, sulfonated, phosphine-oxide (po) rhodamines for voltage imaging in thick tissue from the central nervous system. We find po-rhodamine based voltage reporters, or poRhoVRs, display NIR excitation and emission profiles at greater than 700 nm, show best-in class voltage sensitivity (up to 43% ΔF/F per 100 mV in HEK cells), and can be combined with existing optical sensors, like Ca<sup>2+</sup>-sensitive fluorescent proteins (GCaMP), and actuators, like light-activated opsins ChannelRhodopsin-2 (ChR2). Simultaneous voltage and Ca<sup>2+</sup> imaging reveals differences in activity dynamics in rat hippocampal neurons, and pairing poRhoVR with blue-light based ChR2 affords all-optical electrophysiology. In <i>ex vivo</i> retinas isolated from a mouse model of retinal degeneration, poRhoVR, together with GCaMP-based Ca<sup>2+</sup> imaging and traditional multi-electrode array (MEA) recording, can provide a comprehensive physiological activity profile of neuronal activity. Taken together, these experiments establish that poRhoVR will open new horizons in optical interrogation of cellular and neuronal physiology in intact systems.

scholarly journals Morphometry of ventral hippocampal neurons of CA1-CA4 fields in domestic cattle

2016 ◽  
Vol 72 (12) ◽  
pp. 773-776
Author(s):  
Iwona Łuszczewska-Sierakowska ◽  
Agata Wawrzyniak ◽  
Anna Charuta ◽  
Marcin R. Tatara ◽  
Mariusz Klepacki ◽  
...  
Keyword(s):  
Surface Area ◽  
Cell Nucleus ◽  
Morphometric Analysis ◽  
Hippocampal Neurons ◽  
Bos Taurus ◽  
Average Diameter ◽  
Novel Approach ◽  
Pyramidal Layer ◽  
The Central Nervous System ◽  
The Individual

The aim of the study was a morphometric analysis of the ventral hippocampal neurons of the individual CA1-CA4 fields in domestic cattle (Bos Taurus; N = 6). The hippocampus in cattle is formed by a sizable arched invagination of the medial wall of the lateral ventricle of the brain. The brains were removed and analyzed conventionally with a light microscope. The samples were stained by Nissl’s method. The morphometric analysis of the neurons of the hippocampal CA1-CA4 fields included the following parameters: the area of nervous cells and the area of the cell nucleus in μm2; the nucleus-to-cell ratio in %; the average diameter and perimeter of the nervous cell in μm. The morphometric investigations indicate that the cells of the pyramidal layer in the CA1-CA4 fields of the hippocampus in adult domestic cattle differ in terms of their size, shape, and surface area, as well as the surface area of the cell nucleus. The size of cells in CA1 was the largest, fluctuating around 22 μm, whereas in CA4 it amounted to about 19 μm. Cells in CA1 and CA2 had the largest diameter of about 24 µm, whereas cells in the CA4 field had the smallest diameter of about 20 µm. The results obtained suggest a novel approach to studying the morphometric properties of the hippocampus in domestic cattle. Morphometric studies of the central nervous system (CNS) are regarded as a valuable source of data on the function of environmental and pharmacological factors and their effects on several structures of the CNS.

Morphometric Analysis of the Hepatocytes from Fish Exposed to Arsenic

1976 ◽  
Vol 34 ◽  
pp. 282-283
Author(s):  
Elsie M. B. Sorensen
Keyword(s):  
Morphometric Analysis ◽  
Arsenic Concentration ◽  
Ultrastructural Changes ◽  
Sodium Arsenate ◽  
Lepomis Cyanellus ◽  
Mammalian Counterpart ◽  
Intracellular Changes ◽  
Green Sunfish ◽  
Organic Pesticides

The detoxification capacity of the liver is well documented for a variety of substances including ethanol, organic pesticides, drugs, and metals. The piscean liver, although less enzymatically active than the mammalian counterpart (1), contains endoplasmic reticulum with an impressive repertoire of oxidizing, reducing, and conjugating abilities (2). Histopathologic changes are kncwn to occur in fish hepatocytes following in vivo exposure to arsenic (3); however, ultrastructural changes have not been reported. This study involved the morphometric analysis of intracellular changes in fish parynchymal hepatocytes and correlation with arsenic concentration in the liver.Green sunfish (Lepomis cyanellus, R.) were exposed to 0, 30, or 60 ppm arsenic (as sodium arsenate) at 20°C for 1, 2, or 3 week intervals before removal of livers for quantification of the arsenic burden (using neutron activation analysis) and morphometric analysis of ultrastructural alterations. Livers were cut into 1 mm cubes for fixation, dehydration, and embedding.

ATRA-induced increase in intracellular Ca2+concentration in primary hippocampal neurons

2010 ◽  
Vol 34 (8) ◽  
pp. S74-S74
Author(s):  
Tingyu Li ◽  
Xiaojuan Zhang ◽  
Xuan Zhang ◽  
Jian Hea ◽  
Yang Bi Youxue Liu ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Primary Hippocampal Neurons

Fluorescent proteins for in vivo imaging, where's the biliverdin?

2020 ◽  
Vol 48 (6) ◽  
pp. 2657-2667
Author(s):  
Felipe Montecinos-Franjola ◽  
John Y. Lin ◽  
Erik A. Rodriguez
Keyword(s):  
Hydrogen Peroxide ◽  
In Vivo Imaging ◽  
Near Infrared ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Fluorescent Imaging ◽  
Infrared Light ◽  
Red Fluorescent Protein ◽  
Color Palette

Noninvasive fluorescent imaging requires far-red and near-infrared fluorescent proteins for deeper imaging. Near-infrared light penetrates biological tissue with blood vessels due to low absorbance, scattering, and reflection of light and has a greater signal-to-noise due to less autofluorescence. Far-red and near-infrared fluorescent proteins absorb light &gt;600 nm to expand the color palette for imaging multiple biosensors and noninvasive in vivo imaging. The ideal fluorescent proteins are bright, photobleach minimally, express well in the desired cells, do not oligomerize, and generate or incorporate exogenous fluorophores efficiently. Coral-derived red fluorescent proteins require oxygen for fluorophore formation and release two hydrogen peroxide molecules. New fluorescent proteins based on phytochrome and phycobiliproteins use biliverdin IXα as fluorophores, do not require oxygen for maturation to image anaerobic organisms and tumor core, and do not generate hydrogen peroxide. The small Ultra-Red Fluorescent Protein (smURFP) was evolved from a cyanobacterial phycobiliprotein to covalently attach biliverdin as an exogenous fluorophore. The small Ultra-Red Fluorescent Protein is biophysically as bright as the enhanced green fluorescent protein, is exceptionally photostable, used for biosensor development, and visible in living mice. Novel applications of smURFP include in vitro protein diagnostics with attomolar (10−18 M) sensitivity, encapsulation in viral particles, and fluorescent protein nanoparticles. However, the availability of biliverdin limits the fluorescence of biliverdin-attaching fluorescent proteins; hence, extra biliverdin is needed to enhance brightness. New methods for improved biliverdin bioavailability are necessary to develop improved bright far-red and near-infrared fluorescent proteins for noninvasive imaging in vivo.

Immunohistochemical localization of lysosomal cysteine protease cathepsins B and L in monkey hippocampal neurons after transient ischemia

1999 ◽  
Vol 19 (3) ◽  
pp. 302-310
Author(s):  
Yukihiko Kohda ◽  
Katsuhiro Tsuchiya ◽  
Junkoh Yamashita ◽  
Masaki Yoshida ◽  
Takashi Ueno ◽  
...  
Keyword(s):  
Cysteine Protease ◽  
Hippocampal Neurons ◽  
Immunohistochemical Localization ◽  
Transient Ischemia ◽  
Lysosomal Cysteine Protease

1019: Visualization of the Neurovascular Bundles and Major Pelvic Ganglion with Fluorescent Proteins after Penile Injection

2006 ◽  
Vol 175 (4S) ◽  
pp. 328-328 ◽  
Author(s):  
Hugo H. Davila ◽  
Maggie Mamcarz ◽  
Irving Nadelhaft ◽  
Raoul Salup ◽  
Jorge Lockhart ◽  
...  
Keyword(s):  
Fluorescent Proteins ◽  
Pelvic Ganglion ◽  
Major Pelvic Ganglion ◽  
Neurovascular Bundles

Stimulation with EphB-Fc induces spine maturation in cultured rat hippocampal neurons

2007 ◽  
Author(s):  
Clara L. Essmann ◽  
Inmaculada Segura ◽  
Stefan Weinges ◽  
Amparo Acker-Palmer
Keyword(s):  
Hippocampal Neurons
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