Journal: Neuroscience

Article Title: Palatable food affects HPA axis responsivity and forebrain neurocircuitry in an estrous cycle-specific manner in female rats

doi: 10.1016/j.neuroscience.2018.05.030

Figure Lengend Snippet: A history of limited sucrose intake (LSI) increases FosB/deltaFosB-immunolabeling in basolateral and central amygdala specifically in proestrus/estrus. The impact of prior sucrose (vs. water) on the density of FosB/deltaFosB-positive cells in the BLA (A,B), CeA (C,D), PVN (E,F), and MeA (G,H). Representative images shown in B, D, F, and H were taken at 50× magnification from a water D1/D2 rat; scale bar = 100 um. *p < 0.05 vs. water, #p < 0.05 vs. D1/D2. n = 7–17/group. Abbreviations: BLA, basolateral amygdala; CeA, central amygdala; ec, external capsule (denoted by dashed line); ic, internal capsule; PVN, paraventricular hypothalamic nucleus; opt, optic tract; MeA, medial amygdala; III, third ventricle.

Article Snippet: The following day, sections were rinsed in KPBS and incubated in biotinylated goat anti-rabbit secondary antibody (1:500 for pCREB, 1:250 for FosB/DeltaFosB, product #BA1000, Vector Laboratories, Burlingame, CA) for 1 h. Following rinsing with KPBS, sections were incubated in avidin-biotin-peroxidase (Vectastain ABC solution, Vector Laboratories, Burlingame, CA) for 1 h, rinsed again in KPBS, and reacted with 3,3’-diaminobenzidine (Sigma-Aldrich, Perth, WA).

Techniques: Immunolabeling

Journal: Neuroscience

Article Title: Palatable food affects HPA axis responsivity and forebrain neurocircuitry in an estrous cycle-specific manner in female rats

doi: 10.1016/j.neuroscience.2018.05.030

Figure Lengend Snippet: The impact of LSI and/or estrous cycle stage on the density of FosB/deltaFosB-positive cells (number of cells/mm 2 ) in multiple stress- and reward-regulatory brain regions.

Article Snippet: The following day, sections were rinsed in KPBS and incubated in biotinylated goat anti-rabbit secondary antibody (1:500 for pCREB, 1:250 for FosB/DeltaFosB, product #BA1000, Vector Laboratories, Burlingame, CA) for 1 h. Following rinsing with KPBS, sections were incubated in avidin-biotin-peroxidase (Vectastain ABC solution, Vector Laboratories, Burlingame, CA) for 1 h, rinsed again in KPBS, and reacted with 3,3’-diaminobenzidine (Sigma-Aldrich, Perth, WA).

Techniques:

Journal: Neuroscience

Article Title: Palatable food affects HPA axis responsivity and forebrain neurocircuitry in an estrous cycle-specific manner in female rats

doi: 10.1016/j.neuroscience.2018.05.030

Figure Lengend Snippet: Exploratory Bayesian network considered the contribution of both FosB/deltaFosB and pCREB in all the brain regions at once. Predicted significant relationships between proteins within and across brain regions are shown in blue for water control rats, in red for sucrose-fed rats, and in yellow when predicted in both water control and sucrose-fed. Separate Bayesian networks were created for D1/D2 (A) and P/E (B), and a Pearson’s Chi-squared test for homogeneity indicated that the D1/D2 and P/E networks were significantly different (p < 0.01). As an example, for the predicted network during D1/D2, MeA pCREB expression is related to PVN pCREB expression for rats drinking water, but not sucrose (denoted by the circled blue box in panel A). In contrast, for the predicted network during P/E, MeA pCREB is related to PVN pCREB for rats drinking sucrose, but not water (denoted by the circled red box in panel B). Abbreviations: PVN, paraventricular hypothalamic nucleus; PFC, prefrontal cortex; NAc, nucleus accumbens; MeA, medial amygdala; CeA, central amygdala; BSTpr, bed nucleus of the stria terminalis, principal subdivision; BSTad, bed nucleus of the stria terminalis, anterodorsal subdivision; BLA, basolateral amygdala; FosB, FosB/deltaFosB

Article Snippet: The following day, sections were rinsed in KPBS and incubated in biotinylated goat anti-rabbit secondary antibody (1:500 for pCREB, 1:250 for FosB/DeltaFosB, product #BA1000, Vector Laboratories, Burlingame, CA) for 1 h. Following rinsing with KPBS, sections were incubated in avidin-biotin-peroxidase (Vectastain ABC solution, Vector Laboratories, Burlingame, CA) for 1 h, rinsed again in KPBS, and reacted with 3,3’-diaminobenzidine (Sigma-Aldrich, Perth, WA).

Techniques: Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Transcriptional regulation of the chemokine co-receptor CCR5 by the cAMP/PKA/CREB pathway

doi: 10.1016/j.biopha.2011.03.009

Figure Lengend Snippet: Forskolin stimulation leads to increased nuclear accumulation of pCREB-1α,Δ and pCREM-α/β in TF-1 cells. (A) Analysis of the kinetics of CREB-1 phosphorylation reveals negligible pCREB-1 in the untreated sample. Following forskolin stimulation, maximum pCREB-1 is found at 1 h. Heightened pCREB-1 levels persist up to 6 h followed by dephosphorylation at 9 h. β-actin serves as a normalization control. (B) Analysis of the kinetics of pCREM accumulation reveals a bell-shaped curve. Following stimulation, an exponential increase occurs in nuclear pCREM levels. Maximal accumulation is observed at 1 h. Thereafter, levels drop rapidly up to 6 h. β-actin serves as a loading control.

Article Snippet: An equal amount of protein for all samples was run on a 10% sodium dodecyl sulfate–polyacrylamide gel and transferred to a 0.45-μm Immobilon-P polyvinylidene difluoride membrane followed by probing with a rabbit polyclonal pCREB-1 antibody (Upstate Chemicon, Temecula, CA).

Techniques: De-Phosphorylation Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Transcriptional regulation of the chemokine co-receptor CCR5 by the cAMP/PKA/CREB pathway

doi: 10.1016/j.biopha.2011.03.009

Figure Lengend Snippet: Inhibition of PKA abrogates nuclear accumulation of pCREB-1α,Δ. Nuclear pCREB-1 levels increase in response to forskolin stimulation for 1 h. This response is partly abrogated by pretreatment with H-89 (2.34-fold). pCREB-1 levels are not affected by pretreatment with other kinase inhibitors. Samples treated with inhibitors alone exhibit no significant change in comparison to untreated cells (data not shown).

Article Snippet: An equal amount of protein for all samples was run on a 10% sodium dodecyl sulfate–polyacrylamide gel and transferred to a 0.45-μm Immobilon-P polyvinylidene difluoride membrane followed by probing with a rabbit polyclonal pCREB-1 antibody (Upstate Chemicon, Temecula, CA).

Techniques: Inhibition