Journal: Molecular Pain

Article Title: Systemic administration of a β2-adrenergic receptor agonist reduces mechanical allodynia and suppresses the immune response to surgery in a rat model of persistent post-incisional hypersensitivity

doi: 10.1177/1744806921997206

Figure Lengend Snippet: Effects of β2 adrenergic receptor (AR) agonist on enhanced spinal microglia activation in a rat model of persistent postoperative pain. Sections of spinal cord were collected from rats 8 days following plantar incision and following treatment with DβH-saporin to deplete spinal noradrenergic terminals or control IgG-saporin. Rats were chronically administered clenbuterol (0.5 mg/kg, 2×/day, i.p.) or saline vehicle 6 days prior to and for 8 days after plantar incision. Depletion of spinal noradrenergic fibers was verified immunohistochemically with an antibody against dopamine β hydroxylase (DβH, (a)–(c)). Representative confocal images of IBA1-IR (blue, (d)–(f)) and phospho-p38 MAPK-IR (purple, (g)–(i)) in the ipsilateral spinal cord of incision rats. Localization of p38 MAPK in microglia was confirmed by colocalization with an antibody against the cell surface antigen CD11b (green, inset in (h)). Quantification of IBA1-IR in ipsilateral and contralateral spinal cord of rats with incision (j). Data represent mean ± SEM, n = 3 rats per group. Two way ANOVA indicated effect of group: p < 0.001 but not side p = 0.184 or interaction: p = 0.59 with SNK pairwise comparisons *p < 0.001 versus Incision+ DβH-saporin+ vehicle. Quantification of phospho-p38 MAPK microglial in the ipsilateral and contralateral spinal cord of rats with incision (k). Data represent mean ± SEM, n = 3 rats per group. Two way ANOVA indicate effect of group: p = 0.002 but not side p = 0.398 or interaction: p = 0.67 with SNK pairwise comparisons * p < 0.005 versus Incision+ DβH-saporin+ vehicle.

Article Snippet: We used a previously characterized antibody against β2 adrenergic receptors (AAR-016, β2-AR, 1:1000, rabbit anti-mouse, Alomone Labs; Jerusalem, Israel).

Techniques: Activation Assay

Journal: Molecular Pain

Article Title: Systemic administration of a β2-adrenergic receptor agonist reduces mechanical allodynia and suppresses the immune response to surgery in a rat model of persistent post-incisional hypersensitivity

doi: 10.1177/1744806921997206

Figure Lengend Snippet: Beta 2-adrenergic receptor immunoreactivity in the spinal cord of rats under naïve conditions and two days following plantar incision. Transverse section of L4 spinal cord of rat reacted with antibody against β2 adrenergic receptor ((a), β2-AR, green). There is a high density of immunoreactivity in cellular profiles throughout dorsal and ventral horn. There is also dense immunoreactivity in axon terminals within the lateral portion of the superficial laminae (arrow) and ependymal cells in the vicinity of the central canal (Arrowhead). Note lack of staining for β2-AR in motor neurons within the ventral horn (asterisk). Higher magnification confocal images show β2-AR-IR ((c), green) is present in a subpopulation of neurons ((d), NeuN, purple) in the dorsal spinal cord. Most β2-AR-IR cellular profiles colocalized with NeuN with the exception of a few non-neuronal profiles with morphology typical of microglia (arrows, (c)–(f)). β2-AR-IR non-neuronal cellular profiles in the spinal cord colocalized with the microglial marker IBA1 (red, (e) and (f)). Arrows in F indicate IBA1 negative neuronal cellular profiles. Representative images of β2 mRNA and DAPI in the dorsal spinal cord (g) with high power image showing colocalization with a subset of nuclei (h).

Article Snippet: We used a previously characterized antibody against β2 adrenergic receptors (AAR-016, β2-AR, 1:1000, rabbit anti-mouse, Alomone Labs; Jerusalem, Israel).

Techniques: Staining, Marker

Journal: Molecular Pain

Article Title: Systemic administration of a β2-adrenergic receptor agonist reduces mechanical allodynia and suppresses the immune response to surgery in a rat model of persistent post-incisional hypersensitivity

doi: 10.1177/1744806921997206

Figure Lengend Snippet: β2-adrenergic receptor immunoreactivity (β2AR-IR) in hindpaw of rats under naïve conditions and following plantar incision. Skin sections were obtained from the hind paw of naïve rats and incision rats two days following surgery. Sixteen-μm-thick sections were stained with antibodies against β2AR-IR (green, (a)–(c)), IBA1 (red, (d)–(f)) to label all monocytes/and macrophage, CD68 (blue, (g)–(i)) for activated M1 macrophage) and DAPI ((j) and (k)) to label all nuclei. β2-AR IR was present in keratinocytes of both naïve and incision rats. Two days following plantar incision there were increased β2-AR IR cellular profiles in predominantly the dermal layers of the skin. Higher magnification confocal images ((c), (f), (i), and (l)) indicate colocalization of β2-AR in IBA1 + cells and a subset of which express CD68-IR. Note in naïve skin IBA1-IR was primarily present at the epidermal/dermal interface and had reduced dermal cellularity (DAPI + cells) compared to skin adjacent to the wound in incision rats.

Article Snippet: We used a previously characterized antibody against β2 adrenergic receptors (AAR-016, β2-AR, 1:1000, rabbit anti-mouse, Alomone Labs; Jerusalem, Israel).

Techniques: Staining

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Propranolol enhanced adipogenesis instead of induction of apoptosis of hemangiomas stem cells

doi:

Figure Lengend Snippet: Figure 2

Article Snippet: The primary antibodies included rabbit anti-human β1 and β2 AR (Novus Biologicals), rabbit anti human Caspase 3 and Caspase 9, rabbit anti human Bcl-2 (Cell Signaling Technology), and rabbit anti human VEGF (EMD Millopore).

Techniques:

Journal: PLoS ONE

Article Title: MicroRNA-185 and 342 Inhibit Tumorigenicity and Induce Apoptosis through Blockade of the SREBP Metabolic Pathway in Prostate Cancer Cells

doi: 10.1371/journal.pone.0070987

Figure Lengend Snippet: A, Both miR-185 and 342 inhibited mRNA expression of SREBP-1, SREBP-2, FASN, HMGCR and AR in LNCaP and C4-2B prostate cancer cells determined by qRT-PCR. -: non-transfected; NC: miR-negative control. The relative mRNA expression (fold) was assigned as 1.0 in non-transfected cells. Data were normalized to 18S rRNA and represent the mean ± SD of three independent duplicate experiments. **, P < 0.005 significant differences from NC. B, MiR-185 and 342 inhibited precursor (125 kDa) and mature (68 kDa) forms of SREBP-1 and SREBP-2, FASN, HMGCR and AR expression in LNCaP and C4-2B cells assayed by Western blot. β2-microglobulin (β2M) was used as a loading control. C, MiR-185 and 342 inhibitors (antisense oligonucleotides against miR-185 and 342) increased SREBP-1, SREBP-2, FASN, HMGCR and AR expression in LNCaP and C4-2B cells determined by qRT-PCR. The relative mRNA expression (fold) was assigned as 1.0 in non-transfected cells. Data were normalized to 18S rRNA and represent the mean ± SD of three independent duplicate experiments. **, P < 0.005 significant differences from NC. D, Expression of intrinsic miR-185 and 342 in RWPE-1, LNCaP and C4-2B cells. qRT-PCR results showed that relative expression of miR-185 and 342 was significantly decreased in prostate cancer cells compared with normal/non-cancerous RWPE-1. Lower expression of both miRNAs was observed in aggressive C4-2B compared with LNCaP cells. The relative miRNA expression (fold) was assigned as 1.0 in RWPE-1 cells. **, P < 0.005 significant differences from RWPE-1. Data were normalized to RNU6B control and represent the mean ± SD of three independent experiments performed in quadruplicate.

Article Snippet: Primary antibodies anti-SREBP-1, SREBP-2, FASN, HMGCR, AR and β2-microglobulin (β2M) (Santa Cruz Biotechnology, Santa Cruz, CA), and secondary antibodies which were conjugated with horseradish peroxidase (GE Healthcare, Piscataway, NJ) were used.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Negative Control, Western Blot