polyclonal anti β1 adrenergic receptor  (Alomone Labs)

Journal: iScience

Article Title: Adipocyte-released adipomes in Chagas cardiomyopathy: Impact on cardiac metabolic and immune regulation

doi: 10.1016/j.isci.2024.109672

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Article Snippet: Rabbit Polyclonal Anti-β1-Adrenergic Receptor , Cell Signaling Technology , Cat#12271; RRID: AB_2797865.

Techniques: Infection, Clinical Proteomics, Recombinant, Electron Microscopy, SYBR Green Assay, Lysis, Protease Inhibitor, Plasmid Preparation, XF Assay, Isolation, Quantitation Assay, Bicinchoninic Acid Protein Assay, Software

Journal: Neurobiology of Stress

Article Title: β1-adrenoceptor expression on GABAergic interneurons in primate dorsolateral prefrontal cortex: potential role in stress-induced cognitive dysfunction

doi: 10.1016/j.ynstr.2024.100628

Figure Lengend Snippet: β1-AR expression across inhibitory neuron types in layer III dlPFC Maximum projections of confocal microscopy photomicrographs depicting multiple label immunofluorescence (MLIF) in deep layer III dlPFC in a young macaque (8 years), depicting β1-AR with parvalbumin (PV) (A) , calbindin (CB) (B) , or calretinin (CR) (C) . Yellow arrow, neuron double labeled for β1-AR (green), and PV, CB or CR (red). White arrowhead, neuron positive for PV, CB, or CR only. White double-headed arrow, pyramidal-like neuron positive for β1-AR and negative for PV, CB, or CR. White single-headed arrow, pyramidal-like neuron positive for β1-AR and lightly labeled by CB. (D) Schematic depicting exemplar sampling site (blue) in the mid to posterior principal sulcus (ps) of the dlPFC. Top, lateral surface of a macaque brain. Left, example coronal slice with sampling region along the principal sulcus (blue). Right, schematic of a cortical column depicting layer III, the region sampled for quantitative analysis. (E) Percent of PV, CB, or CR neurons that are β1-AR negative (light green, expression lower than sampled “neuropil” regions), β1-AR intermediate (intermediate green, expression above sampled “neuropil” regions, but less than expression observed in pyramidal-like neurons), or β1-AR strong (expression greater than β1-AR pyramidal-like neurons), sampled from two young monkeys (M1, 8 years, left; M2, 10 years, right). (F) Total β1-AR + neurons (β1-AR strong + β1-AR intermediate), averaged across M1 and M2, shown with standard deviation. In monkey 1 (M1), we analyzed 90 CB cells, 251 CR cells, and 250 PV cells. In monkey 2 (M2), we analyzed 136 CB cells, 211 CR cells, and 250 PV cells. Scale bars, 50 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To label β1-AR, we used the Alomone polyclonal rabbit anti-β1-AR at 1:100 (Alomone, cat# AAR-023, RRID: AB_2340886 ), which has been previously used in macaque PFC ( ).

Techniques: Expressing, Confocal Microscopy, Immunofluorescence, Labeling, Sampling, Standard Deviation

Journal: Neurobiology of Stress

Article Title: β1-adrenoceptor expression on GABAergic interneurons in primate dorsolateral prefrontal cortex: potential role in stress-induced cognitive dysfunction

doi: 10.1016/j.ynstr.2024.100628

Figure Lengend Snippet: Preliminary data showing that β1-AR stimulation increases the firing of a small subset of FS neurons and reduces the firing of RS neurons in dlPFC (A) The ODR working memory task, (B) The iontophoretic electrode: a carbon fiber and surrounding glass micropipettes for drug delivery, with a tip diameter of around 40 μm. (C) The dlPFC recording site (PS = principal sulcus; AS = arcuate sulcus). The insert shows waveforms of regular spiking (RS) and fast spiking (FS) neurons. (D) An example of a regular spiking Delay cell, with sustained delay-related firing for its preferred direction only. (E-G) show that xamoterol decreased the firing of regular spiking neurons (n = 11), reducing firing rate ( E , F , delay firing: R-two-way ANOVA, F directionxdrug (1, 10) = 13.448, p = 0.0043; Sidak's multiple comparisons: preferred direction, p < 0.0389 and non-preferred direction, p = 0.7599) and spatial tuning ( G , paired t -test, n = 11, P = 0.000591). The dark grey area, Cue epoch; light grey area, Delay epoch. (H-J) show that xamoterol also reduced the firing rate of most fast spiking neurons, reducing both firing rate ( H , I delay firing: R-two-way ANOVA, F directionxdrug (1, 4) = 27.696, p = 0.0062; Sidak's multiple comparisons: preferred direction, p < 0.0342 and non-preferred direction, p = 0.33) and spatial tuning ( J , paired t -test, P = 0.0363). (K-M) A single cell example of a fast spiking neuron that showed increased firing with xamoterol, showing that both its delay-related firing ( K , L , control vs. xamoterol@40 nA condition: two-way ANOVA, F directionxdrug (1,13) = 1.58, p = 0.2555; F drug (1,13) = 8.499, p = 0.0268; F direction (1,13) = 17.39, p = 0.0059; Sidak's multiple comparisons: preferred direction, p = 0.0163 and non-preferred direction, p = 0.73), and spatial tuning ( M ) were increased by xamoterol. The neuron did not return to control levels of firing after xamoterol was washed out, suggesting potential second messenger actions.

Article Snippet: To label β1-AR, we used the Alomone polyclonal rabbit anti-β1-AR at 1:100 (Alomone, cat# AAR-023, RRID: AB_2340886 ), which has been previously used in macaque PFC ( ).

Techniques:

Journal: Neurobiology of Stress

Article Title: β1-adrenoceptor expression on GABAergic interneurons in primate dorsolateral prefrontal cortex: potential role in stress-induced cognitive dysfunction

doi: 10.1016/j.ynstr.2024.100628

Figure Lengend Snippet: β1-AR expression in high-likelihood inhibitory dendrites of layer III dlPFC (A-G) Electron microscopy photomicrographs depicting β1-AR expression in high-likelihood inhibitory dendrites (pseudocolored pink) of young macaque layer III dlPFC (8-10y). Dendrites were classified as high-likelihood inhibitory based on multiple criteria, including multiple asymmetric synapses (presumed excitatory, black arrows) on the dendritic shaft, and no apparent spines in plane or in nearby sections. β1-AR immunogold [or immunoperoxidase nickel DAB (C)] labeling (arrowheads) indicate the presence of β1-AR expression in the cytoplasm (grey arrowheads), in extrasynaptic segments of the membrane (orange arrowheads), or in or near the post-synaptic density (red arrowheads). Label that appeared very close to a membrane but not clearly adhered was color coded with hatching to capture that they may be undergoing trafficking to a specific membrane compartment. Red/grey hatching captures possible trafficking to the synapse, and orange/grey hatching captures possible trafficking to an extrasynaptic location. ax, axon (pseudocolored blue); mt, mitochondria. Scale bars, 200 nm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To label β1-AR, we used the Alomone polyclonal rabbit anti-β1-AR at 1:100 (Alomone, cat# AAR-023, RRID: AB_2340886 ), which has been previously used in macaque PFC ( ).

Techniques: Expressing, Electron Microscopy, Labeling, Membrane

Journal: Neurobiology of Stress

Article Title: β1-adrenoceptor expression on GABAergic interneurons in primate dorsolateral prefrontal cortex: potential role in stress-induced cognitive dysfunction

doi: 10.1016/j.ynstr.2024.100628

Figure Lengend Snippet: β1-AR expression in PV neurons of layer III dlPFC (A) Confocal microscopy photomicrographs of deep layer III dlPFC parvalbumin (PV) neurons (red) with robust β1-AR expression (green) in the cytoplasm of the somata. Scale bars, 15 μm (B) Electron microscopy photomicrograph of a PV soma and proximal dendritic segment (pseudocolored pink), labeled with the immunoperoxidase DAB (dark precipitate, black arrowheads). Cytoplasmic (grey arrowheads) and extrasynaptic β1-AR (orange arrowhead) are labeled with immunogold (dark quantal structures). The soma and proximal segment receive asymmetric (presumably excitatory) synapses (black arrows) from axons (pseudocolored blue). Scale bar, 500 nm. (C) PV dendrite expressing β1-AR (labeled by DAB, black arrowheads), receiving multiple asymmetric synapses on the dendritic shaft, featuring synaptic β1-AR expression (red arrowhead). (D) PV dendrite expressing cytoplasmic β1-AR (grey arrowhead). ax, axon; mt, mitochondria. Scale bars for C-D, 200 nm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To label β1-AR, we used the Alomone polyclonal rabbit anti-β1-AR at 1:100 (Alomone, cat# AAR-023, RRID: AB_2340886 ), which has been previously used in macaque PFC ( ).

Techniques: Expressing, Confocal Microscopy, Electron Microscopy, Labeling

Journal: Neurobiology of Stress

Article Title: β1-adrenoceptor expression on GABAergic interneurons in primate dorsolateral prefrontal cortex: potential role in stress-induced cognitive dysfunction

doi: 10.1016/j.ynstr.2024.100628

Figure Lengend Snippet: Systemic administration of the β1-AR antagonist, nebivolol, protects working memory performance from stress exposure. (A) Nebivolol on its own had no effect compared to vehicle with 0.01 mg/kg or 0.1 mg/kg, but significantly improved performance following 1.0 mg/kg. The range of scores for vehicle was 60–73% correct; for 0.01 mg/kg, 63–77% correct, for 0.1 mg/kg, 57–73% correct, and for 1.0 mg/kg, 70–80% correct. (B) Pretreatment with 0.01 mg/kg nebivolol, which had no effect on its own, prevented the impairment on delayed response performance caused by the pharmacological stressor, FG7142. The range of scores for vehicle + vehicle was 63–70% correct; for FG7142+vehicle, 40–57% correct, for 0.01 mg/kg nebivolol + vehicle, 63–77% correct, and for 0.01 nebivolol mg/kg + FG7142, 57–77% correct. (C) Pretreatment with 0.1 mg/kg nebivolol, which had no effect on its own, prevented the impairment on delayed response performance caused by the pharmacological stressor, FG7142. The range of scores for vehicle + vehicle was 63–73% correct; for FG7142+vehicle, 40–53% correct, for 0.1 mg/kg nebivolol + vehicle, 70–73% correct, and for 0.1 nebivolol mg/kg + FG7142, 67–80% correct. ** signifies different from vehicle control; ⌘ signifies different from FG7142+vehicle.

Article Snippet: To label β1-AR, we used the Alomone polyclonal rabbit anti-β1-AR at 1:100 (Alomone, cat# AAR-023, RRID: AB_2340886 ), which has been previously used in macaque PFC ( ).

Techniques:

Journal: Neurobiology of Stress

Article Title: β1-adrenoceptor expression on GABAergic interneurons in primate dorsolateral prefrontal cortex: potential role in stress-induced cognitive dysfunction

doi: 10.1016/j.ynstr.2024.100628

Figure Lengend Snippet: Working model of β1-AR mechanisms in layer III dlPFC We hypothesize that high levels of NE release during stress engage low affinity β1-ARs, which take dlPFC “offline” to switch control of behavior to more primitive circuits, e.g. mediated by the amygdala. The data indicate that β1-AR can reduce the firing of dlPFC Delay cells by at least two mechanisms: (A) The current study shows that β1-AR are expressed on GABAergic interneurons, including PV-expressing interneurons, where β1-AR drive GABAergic inhibition of pyramidal cell firing. (B) Our previous work has shown that β1-ARs are also highly concentrated on the dendritic spines of pyramidal cells, that weaken recurrent excitation by increasing cAMP-calcium opening of nearby K + channels, reducing the recurrent excitatory connections needed for working memory. As interneurons are activated by pyramidal cells as well as by β1-AR, the loss of pyramidal cell firing would ultimately reduce PV interneuron firing as well, where the entire microcircuit may have low levels of firing. Note that the roles of calretinin (CR)- and calbindin (CB)-expressing interneurons are still to be determined, but β1-AR excitation of CB-expressing interneurons would theoretically add to pyramidal cell inhibition.

Article Snippet: To label β1-AR, we used the Alomone polyclonal rabbit anti-β1-AR at 1:100 (Alomone, cat# AAR-023, RRID: AB_2340886 ), which has been previously used in macaque PFC ( ).

Techniques: Expressing, Inhibition