human m1 achr  (Alomone Labs)

Journal: Brain and Behavior

Article Title: Muscarinic acetylcholine receptors are expressed by most parvalbumin-immunoreactive neurons in area MT of the macaque

doi: 10.1002/brb3.225

Figure Lengend Snippet: Most m1 AChR-immunoreactive neurons (panel B, magenta) in the middle temporal visual area (MT) are not immunoreactive for parvalbumin (PV; panel A, green). This image was captured in layer 5 of area MT using a 40× objective. There are seven PV neurons in the imaged field (+, A). Note that the cell body marked with a ∼ in panel A would not have been counted in this study as it does not have a clearly defined boundary within the imaging plane, although it is a PV neuron. Five of these seven PV neurons are also immunoreactive for the m1 AChR (+ in B and C). Two are m1-immunonegative (arrows in B). A striking feature of this image is the number of neurons that are immunoreactive for m1 AChRs (B, C), but not PV (A). Although these singly labeled m1-ir neurons occasionally appear pyramidal (*), it is generally not possible to determine cell morphology (or, therefore, cell class) from the immunoreactivity for m1 AChRs. The strongly fluorescent regions circled in B and C would not have been counted as they do not appear somatic in shape. These regions appear in only one channel and can thus be distinguished from the lipofuscin autofluorescence (∧ in A, B, and C) which is equally strong but appears in all channels. Scale bar = 100 μ m.

Article Snippet: We detected m1 muscarinic ACh receptors (m1 AChRs) using a polyclonal antibody raised in rabbit against amino acids 227–353 of the intracellular loop i3 of the human m1 AChR, obtained from Alomone Labs (Jerusalem, Israel, catalog #AMR-001, lot # AN-05).

Techniques: Imaging, Labeling

Journal: Brain and Behavior

Article Title: Muscarinic acetylcholine receptors are expressed by most parvalbumin-immunoreactive neurons in area MT of the macaque

doi: 10.1002/brb3.225

Figure Lengend Snippet: Quantification of m1 AChR expression by parvalbumin neurons. The graphs show the percentage of parvalbumin (PV) neurons encountered, by cortical layer, that were also immunoreactive for m1 AChRs in areas V1 (A) and the middle temporal visual area (MT) (B). Error bars represent SEM. Number of PV neurons in each graph is the same: V1 = 293 PV neurons, MT = 293 PV neurons.

Article Snippet: We detected m1 muscarinic ACh receptors (m1 AChRs) using a polyclonal antibody raised in rabbit against amino acids 227–353 of the intracellular loop i3 of the human m1 AChR, obtained from Alomone Labs (Jerusalem, Israel, catalog #AMR-001, lot # AN-05).

Techniques: Expressing

Journal: Brain and Behavior

Article Title: Muscarinic acetylcholine receptors are expressed by most parvalbumin-immunoreactive neurons in area MT of the macaque

doi: 10.1002/brb3.225

Figure Lengend Snippet: Quantification of the parvalbumin-immunoreactive population as a percentage of m1 AChR-expressing neurons. The graphs show the percentage of m1 AChR-expressing neurons encountered, by cortical layer, that were also immunoreactive for parvalbumin in areas V1 (A) and the middle temporal visual area (MT) (B). Error bars represent SEM. Number of m1 AChR-ir neurons: V1 = 520, MT = 1081.

Article Snippet: We detected m1 muscarinic ACh receptors (m1 AChRs) using a polyclonal antibody raised in rabbit against amino acids 227–353 of the intracellular loop i3 of the human m1 AChR, obtained from Alomone Labs (Jerusalem, Israel, catalog #AMR-001, lot # AN-05).

Techniques: Expressing

Journal: Brain and Behavior

Article Title: Muscarinic acetylcholine receptors are expressed by most parvalbumin-immunoreactive neurons in area MT of the macaque

doi: 10.1002/brb3.225

Figure Lengend Snippet: The distributions of soma sizes are similar between the populations of singly and dually labeled neurons in V1 (A) or the middle temporal visual area (MT) (B). In these histograms of soma sizes, measured along the long axis, it can be seen that in V1 (A) the distributions for singly-labeled parvalbumin (PV)- and m1 AChR-ir neurons, and for dually immunoreactive neurons are unimodal and centered around 12–14 microns. In MT these distributions are shifted to the right and somewhat skewed, with a long tail at the end representing soma sizes greater than 20 μ m. This tail of the distribution has a very small N , but all three groups (single PV-ir, single m1AChR-ir and dual PV/m1-ir) are represented in the population of neurons with large somata, offering no evidence for a bi-modal distribution of soma size in either cortical area. N for V1: m1 single = 52, PV single = 11, dual = 50 neurons. N for MT: m1 single = 52, PV single = 12, dual = 45 neurons.

Article Snippet: We detected m1 muscarinic ACh receptors (m1 AChRs) using a polyclonal antibody raised in rabbit against amino acids 227–353 of the intracellular loop i3 of the human m1 AChR, obtained from Alomone Labs (Jerusalem, Israel, catalog #AMR-001, lot # AN-05).

Techniques: Labeling

Journal: Brain and Behavior

Article Title: Muscarinic acetylcholine receptors are expressed by most parvalbumin-immunoreactive neurons in area MT of the macaque

doi: 10.1002/brb3.225

Figure Lengend Snippet: Qualitative detail of single-label immunoperoxidase reactivity for m1 ACh receptors, panels A and B) and parvalbumin (C and D) in visual areas V1 (A and C) and the middle temporal visual area (MT) (B and D). The micrograph in panel A shows m1 AChR-immunoreactivity in layer 5 of area V1. Panel B shows m1 AChR-immunoreactivity in layer 3 of MT. In both panels the characteristic stained cytoplasmic ring around an immunonegative nuclear region is evident (asterisks). When immunoreactive dendrites are evident, they are usually (arrowhead in A), but not always (arrowhead in B) visibly attached to a cell body. There was no axonal staining evident in either cortical area. Parvalbumin immunoreactivity (C and D) on the other hand, is evident in cell bodies, dendrites (arrowheads in C and D) and axons (arrows in D). Panel C is a micrograph captured in layer 2 of area V1. The parvalbumin immunoreactivity shown in Panel D is of layer 5 in MT. Scale bar = 20 μ m.

Article Snippet: We detected m1 muscarinic ACh receptors (m1 AChRs) using a polyclonal antibody raised in rabbit against amino acids 227–353 of the intracellular loop i3 of the human m1 AChR, obtained from Alomone Labs (Jerusalem, Israel, catalog #AMR-001, lot # AN-05).

Techniques: Staining

Journal: bioRxiv

Article Title: Uncovering a neural circuit controlling adult quiescent neural stem cell activation in the subventricular zone

doi: 10.1101/2021.12.27.474262

Figure Lengend Snippet: Muscarinic 1 receptor (M1) activation regulates the activity of qNSCs in SVZ niche. A . Immunofluorescence staining for VCAM 1 (Purple) and ChRM1 (green) in the SVZ niche from P30 C57BL/6J mice. B . Immunofluorescence staining for VCAM 1 (Purple) and ChRM1 (green) from the SVZ NSCs culture. C . Western blot detection of pEGFR (PhosphoY1068) in treated samples with carbachol (15μM) and carbachol (15μM)/pirenzepine (20 μM) of SVZ NSCs cultures in proliferation media collected after 12 hours. D . Analysis of normalized quantification of pEGFR(PhosphoY1068)/β-actin protein intensity in treated samples with carbachol (15μM) and carbachol (15μM)/pirenzepine (20 μM) of SVZ NSCs cultures from panel C. *P = 0.0107, n=4, Unpaired t-test. E . Immunofluorescence staining for EdU (purple) and Ki67 (green) in treated samples with carbachol (15μM) and carbachol (15μM)/pirenzepine (20μM) of SVZ NSCs cultures in proliferation media collected after one day. F . Analysis of normalized quantification of Ki67 + cells per well in treated samples with carbachol (15μM) and carbachol (15μM)/pirenzepine (20 μM) of SVZ NSCs cultures from panel E. P = n.s., n=5, Unpaired t-test. G . Analysis of normalized quantification of EdU + /Ki67 + cells ratio per well in control and treated samples with carbachol (15μM) of SVZ NSCs cultures from panel E. **P = 0.0012, n=6, Unpaired t-test. H . Western blot detection of pEGFR(PhosphoY1068)/β-actin in treated samples with carbachol (15μM) and carbachol (15μM)/pirenzepine (20μM) of SVZ NSCs cultures in differentiation media collected every 12 hours for 36 hours. I . Analysis of normalized quantification of pEGFR(PhosphoY1068)/β-actin protein intensity in treated samples with carbachol (15μM) and carbachol (15μM)/pirenzepine (20μM) of SVZ NSCs cultures from panel H. P

Article Snippet: We used primary antibodies to GFP (#GFP-1020, 1:400, AVES lab), Choline Acetyltransferase Antibody (#AB144P, 1:250, Millipore sigma), tdTomato [16D7] (#EST203, 1:200, Kerafast), RFP (#600-401-379, 1:250, Rockland), Calretinin (#ab702, 1:200, Abcam), calretinin (#6B3, 1:250, Swant), Calretinin (#MCA-3G9, 1:250, EnCor Biotechnology), RFP (#ab62341, 1:200, Abcam), Doublecortin (#AB2253, 1:250, Millipore), Doublecortin (#4604S, 1:200, Cell Signaling Technology), Ki67 (ab15580, 1:250: Abcam), Ki67 (#CPCA-Ki67, 1:200, EnCor Biotechnology), beta Actin HRP (#MA515739HRP (BA3R), 1:5000, Thermo Fisher Scientific), HRP-conjugated Beta Actin (#HRP-60008, 1:5000, Proteintech), EGFR (phospho Y1068) (#ab5644, 1:250, Abcam), EGFR (phospho Y1068) [Y38] (#ab32430, 1:250, Abcam), CD106 (#550547, 1:100, BD Diagnostic Systems), GFAP (#GFAP, 1:500, Aves Labs), M1 Muscarinic Receptor (#AMR-001, Alomone Labs).

Techniques: Activation Assay, Activity Assay, Immunofluorescence, Staining, Mouse Assay, Western Blot