Journal: Frontiers in Neuroanatomy

Article Title: Differences in Noradrenaline Receptor Expression Across Different Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fnana.2020.574130

Figure Lengend Snippet: Antibody information.

Article Snippet: Membranes were blocked with Intercept Blocking Buffer (LiCor 927-70001) and then incubated overnight with anti-α1A adrenergic receptor (Alomone Labs AAR-015), anti-α2A adrenergic receptor (Alomone Labs AAR-020), anti-β1 adrenergic receptor (Alomone Labs AAR-023), or anti-β2 adrenergic receptor (Alomone Labs AAR-016) pre-incubated with peptide plus 1% BSA, or pre-incubated with 1% BSA alone.

Techniques:

Journal: Frontiers in Neuroanatomy

Article Title: Differences in Noradrenaline Receptor Expression Across Different Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fnana.2020.574130

Figure Lengend Snippet: Density of adrenergic receptors across different layers of the FEF. The number of neurons per mm 2 that express a given receptor across FEF layers. α2A adrenergic receptors (α2ARs) and β2 adrenergic receptors (β2Rs) are more abundant than either α1A adrenergic receptors (α1ARs) or β1 adrenergic receptors (β1Rs) across layers II through V. There are no obvious differences in expression across layers other than the predictably low expression of all receptor classes in layer I where there are few neurons.

Article Snippet: Membranes were blocked with Intercept Blocking Buffer (LiCor 927-70001) and then incubated overnight with anti-α1A adrenergic receptor (Alomone Labs AAR-015), anti-α2A adrenergic receptor (Alomone Labs AAR-020), anti-β1 adrenergic receptor (Alomone Labs AAR-023), or anti-β2 adrenergic receptor (Alomone Labs AAR-016) pre-incubated with peptide plus 1% BSA, or pre-incubated with 1% BSA alone.

Techniques: Expressing

Journal: Frontiers in Neuroanatomy

Article Title: Differences in Noradrenaline Receptor Expression Across Different Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fnana.2020.574130

Figure Lengend Snippet: Expression of adrenergic receptors across cell types and layers. (A) The number of different classes of cell types per mm 2 across FEF layers. General classes of pyramidal neurons (neurogranin and RP) are more abundant than any other class of neuron across layers II through VI. There is low expression of all receptor classes in layer I where there are few neurons. (B) For each of the four adrenergic receptors (pale to dark: α1A, α2A, β1, β2) we quantified the proportion of each cell type that expressed that receptor across all cortical layers in the FEF. We found that expression was very consistent for any given receptor/cell type pair.

Article Snippet: Membranes were blocked with Intercept Blocking Buffer (LiCor 927-70001) and then incubated overnight with anti-α1A adrenergic receptor (Alomone Labs AAR-015), anti-α2A adrenergic receptor (Alomone Labs AAR-020), anti-β1 adrenergic receptor (Alomone Labs AAR-023), or anti-β2 adrenergic receptor (Alomone Labs AAR-016) pre-incubated with peptide plus 1% BSA, or pre-incubated with 1% BSA alone.

Techniques: Expressing

Journal: Frontiers in Neuroanatomy

Article Title: Differences in Noradrenaline Receptor Expression Across Different Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fnana.2020.574130

Figure Lengend Snippet: Expression of adrenergic receptors in FEF. From left to right: expression of α1A, α2A, β1, and β2 adrenergic receptors (α1AR, α2AR, β1R, and β2R, respectively) in macaque FEF. Images show a cross-section of all layers of cortex and are oriented with the pial surface at the top and white matter at the bottom. The α2A and β2 adrenergic receptors had strong, punctate staining of cell bodies, with little to no background labeling of processes. While the α1A and β1 adrenergic receptors also had strong, punctate staining of cell bodies, there was also staining of the surrounding processes (dendrites and axons), which resulted in a higher amount of background signal. Scale bar = 100 μm for all panels.

Article Snippet: Membranes were blocked with Intercept Blocking Buffer (LiCor 927-70001) and then incubated overnight with anti-α1A adrenergic receptor (Alomone Labs AAR-015), anti-α2A adrenergic receptor (Alomone Labs AAR-020), anti-β1 adrenergic receptor (Alomone Labs AAR-023), or anti-β2 adrenergic receptor (Alomone Labs AAR-016) pre-incubated with peptide plus 1% BSA, or pre-incubated with 1% BSA alone.

Techniques: Expressing, Staining, Labeling

Journal: Frontiers in Neuroanatomy

Article Title: Differences in Noradrenaline Receptor Expression Across Different Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fnana.2020.574130

Figure Lengend Snippet: Expression of adrenergic receptors on pyramidal neurons. (A) Panels show expression of α1A, α2A, β1, and β2 adrenergic receptors (α1AR, α2AR, β1R, and β2R, respectively) from top to bottom with pyramidal neuron markers (RP, neurogranin, and SMI-32) from left to right. RP and neurogranin are both putative general markers of pyramidal neurons and SMI-32 is a marker for putative long-range projecting pyramidal neurons. All adrenergic receptors are labeled in green, and all pyramidal neurons are labeled in magenta. (B) Quantification of the proportion of each neuron class that expressed each receptor class. Chi-squared tests were performed using pooled neuron counts across all animals. All four adrenergic receptors were expressed significantly more highly on long-range projecting pyramidal neurons than either class of general pyramidal neuron. Significance levels are noted as *** p < 0.001; ** p < 0.01; and * p < 0.05. Scale bar = 100 μm for all panels.

Article Snippet: Membranes were blocked with Intercept Blocking Buffer (LiCor 927-70001) and then incubated overnight with anti-α1A adrenergic receptor (Alomone Labs AAR-015), anti-α2A adrenergic receptor (Alomone Labs AAR-020), anti-β1 adrenergic receptor (Alomone Labs AAR-023), or anti-β2 adrenergic receptor (Alomone Labs AAR-016) pre-incubated with peptide plus 1% BSA, or pre-incubated with 1% BSA alone.

Techniques: Expressing, Marker, Labeling

Journal: Frontiers in Neuroanatomy

Article Title: Differences in Noradrenaline Receptor Expression Across Different Neuronal Subtypes in Macaque Frontal Eye Field

doi: 10.3389/fnana.2020.574130

Figure Lengend Snippet: Expression of adrenergic receptors on inhibitory interneurons. (A) Panels show expression of α1A, α2A, β1, and β2 adrenergic receptors (α1AR, α2AR, β1R, and β2R, respectively) from top to bottom with inhibitory interneuron markers (parvalbumin, calbindin and calretinin) from left to right. All adrenergic receptors are labeled in green, and all inhibitory interneurons are labeled in magenta. (B) Quantification of the proportion of each neuron class that expressed each receptor class. Chi-squared tests were performed using pooled neuron counts across all animals. Lines above the bars show the significance of different comparisons. Black lines indicate significant differences between the expression of different receptors within a neuron class; gray lines indicate significant differences of expression of a specific receptor across different neuron classes. The shade of gray indicates which receptor class is being compared and matches the shading of the bars: from light to dark—α1AR, α2AR, β1R, and β2R. Significance levels are noted as *** p < 0.001; ** p < 0.01; and * p < 0.05. Scale bar = 100 μm for all panels.

Article Snippet: Membranes were blocked with Intercept Blocking Buffer (LiCor 927-70001) and then incubated overnight with anti-α1A adrenergic receptor (Alomone Labs AAR-015), anti-α2A adrenergic receptor (Alomone Labs AAR-020), anti-β1 adrenergic receptor (Alomone Labs AAR-023), or anti-β2 adrenergic receptor (Alomone Labs AAR-016) pre-incubated with peptide plus 1% BSA, or pre-incubated with 1% BSA alone.

Techniques: Expressing, Labeling