Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Relationships between the Firing of Identified Striatal Interneurons and Spontaneous and Driven Cortical Activities In Vivo

doi: 10.1523/JNEUROSCI.2440-12.2012

Figure Lengend Snippet: Spontaneous activity of identified striatal cholinergic interneurons. Ai–Ci , Juxtacellularly labeled cholinergic interneurons, identified by their colocalization of fluorescent labeling for neurobiotin (NB) and choline acetyltransferase (ChAT),

Article Snippet: Extracellular recordings of the action potentials (“spikes”) of individual neurons (i.e., single-unit activities) in the dorsal striatum were made using glass electrodes (10–30 MΩ in situ ; tip diameter ~1.2 μ m) containing 0.5 m NaCl solution and neurobiotin (1.5% w/v; Vector Laboratories).

Techniques: Activity Assay, Labeling

Journal: The Journal of Physiology

Article Title: Cell-type specific expression of ATP-sensitive potassium channels in the rat hippocampus

doi: 10.1111/j.1469-7793.1999.315ae.x

Figure Lengend Snippet: Morphological and electrophysiological characterization of CA1 pyramidal cells, interneurones and dentate granule cells Top row, hippocampal slice stained with DAPI. The white frames enclose the regions in which the somata of CA1 pyramidal cells ( A) , interneurones from stratum radiatum CA1 ( B ) and dentate granule cells ( C ) were located. Second row, morphology of a pyramidal cell ( A ), an interneurone ( B ) and a granule cell ( C ). The cells were filled with neurobiotin, coupled to avidin-fluorescein, and imaged using a two-photon microscope. Axons are marked by white arrows. Note that the interneurone ( B ) is shown at a higher magnification and that the distal dendrites of this cell were cut during slicing. Third row, spontaneous action potentials recorded in the three cell types. Fourth row, trains of action potentials evoked by 400 ms current pulses of 50 pA, as indicated by the bars.

Article Snippet: To label the cells for morphological studies, 1 % neurobiotin tracer (Vector Labs, Burlingame, CA, USA) was added to the internal solution.

Techniques: Staining, Avidin-Biotin Assay, Microscopy, Mass Spectrometry

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Chemical synaptic activity modulates nearby electrical synapses

doi: 10.1073/pnas.0734299100

Figure Lengend Snippet: Electrical synapses at neighboring terminals differ in junctional conductance. ( A ) Unitary electrical EPSP amplitude was not correlated with difference in time (Δ t ) between the peaks of the presynaptic spike and the unitary electrical EPSP ( n = 95). ( B ) Intraterminal recordings (club ending) obtained after an initial recording of the antidromic spike in the M-cell lateral dendrite (M-cell, Left ) showed that the amplitude of the antidromic coupling potential also varied in amplitude (1–4, Right ). The amplitude varied from 0.9 to 7 mV, averaging 4.7 ± 0.28 mV (SEM, n = 21). ( C – F ) Transfer of Neurobiotin at neighboring club endings differs dramatically, indicating that they differ in permeability. ( C ) View of the bifurcation of the lateral dendrite (LD, dark branches) obtained with Nomarski optics and revealing differences of labeling between two neighboring club endings. Labeled (arrowhead) and unlabeled (arrow) fibers terminate in the same area of lateral dendrite. ( D ) Tetanic stimulation that causes LTP of the electrical EPSPs increases the number of labeled terminals. The plot summarizes data from five experiments (empty circles) in which Neurobiotin was injected into both M-cells, but only one eighth nerve was tetanized [TET (abscissa), tetanized side; non-TET (ordinate), control side; red circle and lines show means and SDs]. Both M-cells were injected sequentially within an interval of only 5–10 min using the same injection parameters. ( E and F ) Images of the distal portions of the lateral dendrites of the left ( E ) and right ( F ) M-cells from one such experiment where the left eighth nerve was tetanized. No terminals were detected on the nontetanized side in the same 50-μm section (image is in pseudocolor to improve contrast).

Article Snippet: For anatomical identification or dye coupling evaluation, auditory afferents and M-cells were intracellularly injected with the tracer Neurobiotin (cation M r : 286; Vector Laboratories).

Techniques: Permeability, Labeling, Injection

Journal: Cell and tissue research

Article Title: Cell proliferation and cytoarchitectural remodeling during spinal cord reconnection in the fresh-water turtle Trachemys dorbignyi

doi: 10.1007/s00441-011-1173-y

Figure Lengend Snippet: Retrograde labeling with neurobiotin reveals neurons and neural processes in the rostral stump. a One neuron appears to be Golgi-like stained, whereas another one only shows small fluorescent granules distributed in the perikaryon and main process ( arrow

Article Snippet: A neurobiotin solution (10 mM in saline; Vector Laboratories, Burlingame, Calif., USA) was pressure injected (Picospritzer III, Parker Instrumentation) with a glass micropipette placed within the caudal stumps of the spinal cord 20–30 days following injury ( ).

Techniques: Labeling, Staining

Journal: Frontiers in Cellular Neuroscience

Article Title: Anisotropic Panglial Coupling Reflects Tonotopic Organization in the Inferior Colliculus

doi: 10.3389/fncel.2018.00431

Figure Lengend Snippet: Astrocyte coupling. (A) Scheme depicting the central part of the inferior colliculus. The orientation of “length” and “width” is indicated. A schematic network is represented by asterisks. The patch-clamped cell is marked with a filled asterisk. The tonotopic organization is indicated by dotted lines. (B) The tracer neurobiotin diffused from a patch-clamped astrocyte (AF568) to neighboring cells. (C) Properties of IC networks. The network size depended on the duration of patch-clamping and concomitant injection of neurobiotin into an astrocyte (C 1 ) . The increase of network size was slowed during prolonged tracer filling. Note: In two experiments, astrocytes lacked any coupling (red squares). The dotted line represents a logarithmic fit ( R 2 = 0.2342). IC networks exhibited a high cell density (C 2 ) . The vast majority of networks had a non-spheroidal extension (D 1 ) exhibiting an oval shape oriented orthogonally to the tonotopic axis (D 2 ) . In (D 1 ) one coordinate pair is located outside the dimensions of the diagram (width/length: 553/565). Length: orthogonal to tonotopic axis. Width: longitudinal to tonotopic axis [see (A) ]. n represents the number of experiments and is provided within the diagrams. Shown are mean values ± SEM.

Article Snippet: In most experiments, intracellular solution contained a cocktail of the gap junction-permeable tracer neurobiotin (1%, Vector Laboratories, Inc.) and the gap junction-impermeable dye alexa fluor 568 (100 μM, Invitrogen).

Techniques: Injection

Journal: PLoS ONE

Article Title: Shaking-B misexpression increases the formation of gap junctions but not chemical synapses between auditory sensory neurons and the giant fiber of Drosophila melanogaster

doi: 10.1371/journal.pone.0198710

Figure Lengend Snippet: Quantification of GF synaptic inputs and branching. (A) Diagrammatic view of the GF dendrite (cyan) and JON axons (red spots), similar to that shown in the box in Fig 1D . The JO-A group of axons (magenta) is NB-coupled to the GF. The GCI neurons are also NB-coupled to the GF. The posterodorsally-projecting GF neurite (n) is cropped out of subsequent images. Anterior (A) is towards the top, lateral (L) to the right. (B-C). Confocal views of a typical preparation. The right GF axon was injected with a mixture of Neurobiotin (NB: blue) and Alexa Fluor 488 –coupled dextran (DA488: green), making it cyan in color. JO15-GAL4 was used to drive expression of Strawberry-tagged Brp-short (Brp: red) in JO-A and JO-B subgroups of JON axons, labeling putative active zones at chemical synapses. Blue NB passes retrogradely into a subgroup of the JO-A axons, so their red active zones appear magenta, or white when apposed to the GF. (B) Dorsal 3D view of the complete image stack of frontal slices, with anterior (A) towards the top, lateral (L) to the right. The subsequent panels represent single frontal slices taken at the antero-posterior positions indicated by the arrows: 1, at the anterior end of the GF dendrite, 2, midway along the dendrite in the region of medial branches, and 3, at the posterior end of the dendrite, where it bends dorsally and extends a ventral branch. In these panels, dorsal is to the top and lateral to the right. (C) Single confocal slices taken at the indicated regions, with dorsal (D) towards the top, lateral (L) to the right. Regions of overlap of Brp signal with the cyan GF dendrites (ie. putative synaptic contacts) appear white. (D) Thresholded signal. (E) NB signal removed to show only putative active zones (AZ: pink) on the GF. The total AZ volume over the length of the GF dendrite was subsequently quantified. (F) Single pixel outline of the GF dendrites, used as an approximation for surface area. Dendrites projecting medially more than 5 um were masked (pale green) and quantified separately. Scale bars: 20 μm in A, 10 μm in B-E.

Article Snippet: One of the GF axons was impaled with a sharp glass microelectrode, the tip of which was filled with a mixture of 3% Neurobiotin Tracer (NB) (Vector Labs, SP-1120) and 3% Dextran Alexa Fluor 488 (DA488) (10,000 MW, Bioanalytical Instruments, D22910) diluted in distilled water.

Techniques: Injection, Expressing, Labeling

Journal: The Journal of comparative neurology

Article Title: Vocal-Motor and Auditory Connectivity of the Midbrain Periaqueductal Gray in a Teleost Fish

doi: 10.1002/cne.23202

Figure Lengend Snippet: Charting of label resulting from a neurobiotin injection to the medial PAG. Labeled cells are indicated as large dots, fibers as thin lines, and putative terminals as small dots. All sections are coronal, with planes of section as indicated in the inset

Article Snippet: To characterize the connectivity, both antero- and retrograde, of neurons in the PAG, we made focal iontophoretic injections of the neuronal tracer Neurobiotin (Vector Labs, Burlingame, CA).

Techniques: Injection, Labeling

Journal: The Journal of comparative neurology

Article Title: Vocal-Motor and Auditory Connectivity of the Midbrain Periaqueductal Gray in a Teleost Fish

doi: 10.1002/cne.23202

Figure Lengend Snippet: Summary of the major afferent and efferent connections of the PAG, as revealed by neurobiotin injections. Auditory, vocal-motor and other structures are shown separately, as indicated in the key. Relative density of each connection is indicated by the

Article Snippet: To characterize the connectivity, both antero- and retrograde, of neurons in the PAG, we made focal iontophoretic injections of the neuronal tracer Neurobiotin (Vector Labs, Burlingame, CA).

Techniques:

Journal: The Journal of comparative neurology

Article Title: Vocal-Motor and Auditory Connectivity of the Midbrain Periaqueductal Gray in a Teleost Fish

doi: 10.1002/cne.23202

Figure Lengend Snippet: Charting of label resulting from a neurobiotin injection to the lateral PAG. Labeled cells are indicated as large dots, fibers as thin lines, and putative terminals as small dots. All sections are coronal, with planes of section as indicated in the inset

Article Snippet: To characterize the connectivity, both antero- and retrograde, of neurons in the PAG, we made focal iontophoretic injections of the neuronal tracer Neurobiotin (Vector Labs, Burlingame, CA).

Techniques: Injection, Labeling

Journal: The Journal of comparative neurology

Article Title: Vocal-Motor and Auditory Connectivity of the Midbrain Periaqueductal Gray in a Teleost Fish

doi: 10.1002/cne.23202

Figure Lengend Snippet: Photomicrographs of label resulting from a neurobiotin injection to the lateral PAG. All sections are coronal, oriented as indicated in A , and have been counterstained with cresyl violet. A , low power image showing the injection site in the lateral PAG.

Article Snippet: To characterize the connectivity, both antero- and retrograde, of neurons in the PAG, we made focal iontophoretic injections of the neuronal tracer Neurobiotin (Vector Labs, Burlingame, CA).

Techniques: Injection

Journal: The Journal of comparative neurology

Article Title: Vocal-Motor and Auditory Connectivity of the Midbrain Periaqueductal Gray in a Teleost Fish

doi: 10.1002/cne.23202

Figure Lengend Snippet: Photomicrographs of label resulting from a neurobiotin injection to the medial PAG. All sections are coronal, oriented as indicated in A , and have been counterstained with cresyl violet. Examples of putative terminal boutons are indicated with arrowheads.

Article Snippet: To characterize the connectivity, both antero- and retrograde, of neurons in the PAG, we made focal iontophoretic injections of the neuronal tracer Neurobiotin (Vector Labs, Burlingame, CA).

Techniques: Injection

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Activity of Neurochemically Heterogeneous Dopaminergic Neurons in the Substantia Nigra during Spontaneous and Driven Changes in Brain State

doi: 10.1523/JNEUROSCI.4423-08.2009

Figure Lengend Snippet: Spontaneous activity of identified dopaminergic neurons in the substantia nigra. A , Recording of spontaneous unit activity (DA Unit) of a typical calbindin-negative dopaminergic neuron during robust slow-wave activity recorded in the ECoG. Cortical activity is dominated by a ~1 Hz oscillation, as shown in the ECoG power spectrum (pECoG). Unit activity is irregular, as shown by a relatively flat autocorrelogram (AC; left in 50 ms bins, right in 5 ms bins), and the neuron did not fire preferentially during either the active (Act) or inactive (Inact) component of the cortical slow oscillation (activity histogram, AH). Average action potential waveform (filter settings 300–5000 Hz) of the recorded neuron is shown in the inset. After recording, this neuron was revealed to be a calbindin-negative dopaminergic neuron, as shown in the digital micrographs of Neurobiotin, tyrosine hydroxylase (TH), and calbindin reactivity (arrow points to neuron). Neurobiotin was later visualized with diaminobenzidine (DAB) to form a permanent reaction product for light microscopy. B , A typical calbindin-positive dopaminergic neuron. Note the similarity in electrophysiological characteristics between units in A and B . C , An identified calbindin-negative dopaminergic neuron that showed oscillatory activity at ~1Hz, as shown by the peaks in the ACs. This neuron preferentially fired in time with the inactive component of the cortical slow oscillation, as shown in the AH. D , Schematic of sagittal sections of the substantia nigra pars compacta (SNc) and substantia nigra pars reticulata (SNr) showing approximate positions of all identified calbindin-negative (green, n = 40) and calbindin-positive (blue, n = 16) dopaminergic neurons, and whether their responses to a pinch stimulus (circles) or pinch and electrical stimuli (triangles) were tested or not (squares). Note that the calbindin-positive neurons were located mainly within the dorsal tier of SNc. Calibration of ECoG and DA unit in A also applies to B and C . ML (mediolateral) numbers in C denote positions with respect to midline. R, Rostral; C, caudal; D, dorsal; V, ventral.

Article Snippet: Fluorescent conjugates were used to visualize Neurobiotin (streptavidin-CY3; 1:1000, 4°C, overnight; Zymed), TH immunoreactivity (AlexaFluor 488-conjugated donkey anti-mouse; 1:1000, 4°C, overnight; Invitrogen) and CB immunoreactivity (CY5-conjugated donkey anti-rabbit or CY5-conjugated donkey anti-goat; 1:500, 4°C, overnight; Jackson Immunoresearch Laboratories).

Techniques: Activity Assay, Mass Spectrometry, Activated Clotting Time Assay, Light Microscopy

Journal: Frontiers in Neural Circuits

Article Title: Striatal Distribution and Cytoarchitecture of Dopamine Receptor Subtype 1 and 2: Evidence from Double-Labeling Transgenic Mice

doi: 10.3389/fncir.2017.00057

Figure Lengend Snippet: The receptors’ expression, neural firing pattern and dendritic branches of D1-tdTomato and D2-enhanced fluorescent protein (eGFP) positive neurons in the striatum. (A) D1 receptor and D1-tdTomato signal were co-expressed in the striatal medium-sized spiny neurons (MSNs). (B) D2 receptor and D2-eGFP signal were co-expressed in the MSNs. Scale bar = 10 μm. (C) A D1-tdTomato positive neuron was recorded by patch clamp pipette in differential interference contrast (DIC) image mode. Scale bar = 20 μm. (D) A D2-eGFP positive neuron was recorded by patch clamp pipette in DIC image mode. Scale bar = 20 μm. (E) The same neuron from (C) showed typical delay firing pattern in current clamp mode. (F) The same neuron from (D) also showed typical delay firing pattern and smaller rheobase comparing with the cell in (E) in current clamp mode. (G) Cell filling with neurobiotin 350 and 3D reconstruction outlined the dendritic branches of a D1-tdTomato positive neuron. Scale bar = 40 μm. (H) Cell filling with neurobiotin 350 and 3D reconstruction outlined the dendritic branches of a D2-eGFP positive neuron. Scale bar = 40 μm.

Article Snippet: Cell Filling for Electrophysiologically Characterized Neurons Some MSNs were labeled by adding 0.5% neurobiotin 350 (SP-1155, Vector laboratories, Burlingame, CA, USA) to the internal solution.

Techniques: Expressing, Patch Clamp, Transferring

Journal: JCI Insight

Article Title: Satiety induced by bile acids is mediated via vagal afferent pathways

doi: 10.1172/jci.insight.132400

Figure Lengend Snippet: Cocaine and amphetamine regulated transcript and glutamate are expressed by NG neurons. A representative NG neuron is identified after ( A ) Neurobiotin-350 tracer injection following recording. ( B ) Cocaine and amphetamine regulated transcript (CART) staining; ( C ) merged image of neurobiotin and CART immunostaining; ( D ) glutamate staining; ( E ) merged image of neurobiotin and glutamate; and ( F ) triple merged image of neurobiotin, CART, and glutamate staining. Eight neurons from 3 rats were traced and stained.

Article Snippet: Neurobiotin-350 tracer (SP-1151) was obtained from Vector Laboratories.

Techniques: Injection, Staining, Immunostaining

Journal: bioRxiv

Article Title: Group II metabotropic glutamate receptors modulate sound evoked and spontaneous activity in the mouse inferior colliculus

doi: 10.1101/2020.06.03.130724

Figure Lengend Snippet: Example recording sites labelled with Fluoro-Gold ( A ), Neurobiotin 350 ( B ), and Neurobiotin 488 ( C ). Dashed lines indicate the boundaries of IC and its subregions: parts of the central IC nucleus - area 1 and area 2, as well as dorsal (d) and lateral (lc) cortex. Scale = 500 μm.

Article Snippet: Recording site labelingAfter neuronal activity was assessed, recording sites were labeled with either 1% Neurobiotin 350 in 1 M NaCl (Vector laboratories, Burlingame, CA; product SP-1155), 1% Neurobiotin 488 in 1 M NaCl (Vector laboratories, product SP-1125-2), or 0.1 % Fluoro-Gold in saline (Fluorochrome, Denver, CO).

Techniques:

Journal: Journal of Cell Science

Article Title: AII amacrine cells discriminate between heterocellular and homocellular locations when assembling connexin36-containing gap junctions

doi: 10.1242/jcs.133066

Figure Lengend Snippet: Homocellular AII–AII coupling is disrupted in the KO-Cx36-EGFP retina. Example results of AII cell tracer-coupling experiments (asterisks represent Alexa-Fluor-488-injected AII cells; the tracer neurobiotin is shown in magenta). (A,D,G,J) Rotated projections showing the injected AII (green), Cx36–EGFP clusters (green), and neurobiotin-containing coupled cells (magenta). (B,E,H,K) and (C,F,I,L) collapsed confocal stacks derived from the proximal and distal INL, respectively. AII cells in the KO-Cx36-EGFP retina have functional heterocellular gap junctions between AII and ON cone bipolar cells but lack homocellular AII–AII gap junctions (H,I). Dye-injected AII cells in KO-Cx36-EGFP (G) and KO mice (J) contain higher amounts of neurobiotin as they lack homocellular (KO and KO-Cx36-EGFP) and heterocellular gap junctions (KO). This makes the outline of these cells appear in magenta as opposed to green (due to the co-injected Alexa Fluor 488) for injected AII cells in the WT and HET-Cx36-EGFP genotypes. (M,N) Graph indicating the number of AII cells (M) and ON cone bipolar (CB) cells (N) coupled to the injected AII cell. Homocellular AII–AII gap junctions are completely lacking in the KO-Cx36-EGFP retina. The reduction in the number of coupled bipolar cells in this genotype could be attributed to the absence of homocellular coupling (see Fig. 5 ). A t -test was used to compare means with the mean in the WT: *** P

Article Snippet: Microelectrodes (120–180 MΩ resistance) were filled with 5 mM Alexa Fluor 488 and 4% (w/v) neurobiotin (Vector Laboratories, Burlingame, CA) or 7.5 mM Alexa Fluor 555 or 594.

Techniques: Injection, Derivative Assay, Functional Assay, Mouse Assay

Journal: Scientific Reports

Article Title: Depolarizing GABA/glycine synaptic events switch from excitation to inhibition during frequency increases

doi: 10.1038/srep21753

Figure Lengend Snippet: Morphology of E13.5 and E17.5 MNs and their canonical counterparts that were used in the simulations. ( A 1 ) Photograph of a neurobiotin-filled E13.5 MN as revealed with fluorescent dye. ( A 2 ) Morphology of the canonical E13.5 MN that was used in the simulation. ( B 1-2 ) The same disposition as in ( A 1-2 ) but for E17.5 MNs.

Article Snippet: Immunohistochemistry Four E13.5 MNs and four E17.5 MNs were stained during the whole-cell recordings with pipettes containing neurobiotin (0.4%, CliniSciences, Montrouge, France) diluted in the intracellular medium.

Techniques: