Where Is the Fossa Next to the Vta Rat Horizontal
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Figure 1.
Miserly density of GABA neurons in VTA: approximately 20% are GABAergic.
In horizontal brain slices containing the VTA and the Atomic number 50 the general neuronic marker NeuN was cytochemically pictured with the tributary mark down DAB (brown); GABAergic neurons were labeled using in situ crossbreeding against GAD65/67 messenger RNA (black grains). (A, left) Low magnification example image of one of the slices old to estimate the pct of neurons that are GABAergic. Scale bar: 400 µm. Schematic (right) illustrates slice orientation and anatomic landmarks. (IPF: interpeduncular fossa; IPN: interpeduncular nucleus; ML: mesial lemniscus; MT: medial terminal nucleus of the accessory optic cartroad; SNc: substantia nigra pars compacta; SNr: locus niger pars reticulata; R: rostral; C: caudal; M: medial; L: lateral). High magnification images (color coded rectangles show location in (A)) picture GAD65/67(+) and GAD65/67(−) in the SNr (B - yellow border), the SNc (C – red border), and the VTA (D – nonindulgent border with higher magnification section below). (E), Mean percentage of GAD65/67(+) neurons in VTA, SNc, and SNr neurons and, for comparison, our prior estimates of % TH(+) neurons [41].
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Figure 2.
Identification of GABA neurons following whole cell recordings.
(A) Example where immunocytochemistry against GABA (green) was used to identify GABAergic VTA neurons filled with biocytin (red). (Top words) An representative GABA(+) recorded cell, and (Bring dow row) an example GABA(−) recorded cell. (B) In other cases we exploited in place hybridization against GAD65/67 mRNA to test whether recorded neurons were GABAergic. In these examples of positive colabeling (left) and a lack of colabeling (right), filled cells are labelled with DAB (brown), and in situ recording label is visualized with digoxin (purple).
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Figure 3.
Locations of recorded VTA GABA neurons.
Sample includes neurons throughout the VTA. Schematic diagrams of adaxial, middle, and dorsal horizontal slices containing the VTA. (R: rostral; C: taillike; M: central; L: lateral).
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Figure 4.
Plot of soma crossectional areas and input resistances for all confirmed GABAergic neurons.
There was a trend towards an reciprocal relationship between these two measures. P = 0.09.
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Figure 5.
Near VTA GABA neurons are I h(+).
(A) Example GABAergic neuron that has a large I h (far left, scale parallel bars: 100 pA and 50 ms). The nerve cell was filled with biocytin (left-wing; bloody; scale bar: 20 µm) and immunocytochemically confirmed GABA(+) (middle, green). (B) Example I h(−) GABAergic nerve cell (cold leftfield, scale bars: 50 pA and 50 ms) that was full with biocytin (left; scale bar: 20 µm) and addicted Gamma aminobutyric acid(+). (C) Histogram of I h magnitudes measured in GABA/GAD(+) and GABA/GAD(−) neurons. (D) For compare, histogram of I h magnitudes from recorded VTA Intropin neurons identified with Th immunocytochemistry.
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Figure 6.
VTA neurons express HCN protein.
(A) Immunocytochemical colabeling in VTA tissue for Gallivant and HCN4, uncomparable of the channels expressed in the VTA that produces the I h, yields examples of GAD(+) neurons that are HCN4(+) (top) or HCN4(−) (bottom), consistent with the physiology. Scale parallel bars: 20 µm. (B) Immunocytochemical colabeling for HCN4 and Thorium in VTA slices shows that HCN4 is expressed in some TH(−) neurons. Descale bar: 20 µm. (C) The HCN4 antibody was not not-specifically labeling each neurons as HCN4 colabeling with the neural mark NeuN shows that some VTA neurons were cytochemically HCN4(−). Scale bar: 20 µm.
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Figure 7.
The distribution of action potential drop durations in VTA GABA neurons overlaps thereupon of cytochemically confirmed dopamine neurons.
Neurons transcribed on the same day were identified post hoc as GABA/GAD(+) operating room GABA/GAD(−), and their AP duration distributions are overlapping (top). This is consistent with information from TH(+) and TH(−) neurons where the range AP durations of Thursday(−) neurons spans the range for TH(+) neurons (rear end).
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Figure 8.
The spontaneous firing rates of VTA GABA neurons are standardized to those of dopamine neurons ex vivo.
(A) Example traces of AP activity from a confirmed VTA GABA neuron (left) and a confirmed dopamine nerve cell (decent) firing at like frequencies. (B) For each spontaneously firing nerve cell, the ISI CV is plotted against the sackin rate of the neuron.
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Figure 9.
Some, but not all, VTA GABA neurons are inhibited by activating of the MOR.
Example current clinch traces from identified VTA GABA neurons tested with bath application of 1 µM DAMGO. (A) Gamma aminobutyric acid neuron showing robust inhibition by DAMGO (B) GABA neuron insensitive to DAMGO.
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Forecast 10.
Gamma aminobutyric acidB receptor activation inhibits VTA dopamine neurons, merely not VTA GABA neurons.
(A) Example modern clamp traces from cytochemically identified GABA neuron lacking a response to the GABAB receptor agonist baclofen (1 µM). (Bi) An unknown nerve cell recorded on the same day As GABA(+) neurons that were insensitive to baclofen shows a significant hyperpolarization in response to baclofen (1 µM; bar). (Bii) Example of a cytochemically identified dopamine neuron hyperpolarized by baclofen (1 µM; bar). Surmount bars: 5 mV and 5 Fukkianese. (C) Average trace of identified Gamma aminobutyric acid neurons (n = 6) proven with bathing tub application of baclofen (1 µM) compared to the averages of all quiescent confirmed dopamine neurons (n = 8). (D) Average trace of 4 neurons importantly inhibited by baclofen (1 µM); the inhibition was backward by the GABAB sensory receptor opposer CGP35348 (10 µM). (E) Board of all observed baclofen responses across confirmed GABA, dopamine, and I h(+), TH(−) neurons. TH-identified neurons include spontaneously firing, quiescent (from panel C), and voltage-clamp experiments.
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Where Is the Fossa Next to the Vta Rat Horizontal
Source: https://journals.plos.org/plosone/article/figures?id=10.1371%2Fjournal.pone.0042365
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