Journal: ImmunoHorizons

Article Title: Pituitary Adenylate Cyclase-Activating Polypeptide Prevents Mortality Caused by Septic Peritonitis in Mice.

doi: 10.4049/immunohorizons.1900044

Figure Lengend Snippet: FIGURE 5. Effects of PACAP on plasma levels of ACTH and corticosterone after CLP.

Article Snippet: Measurement of plasma corticosterone and ACTH levels Measurements of plasma corticosterone and ACTH were performed using ELISA kits (corticosterone; Abcam, Cambridge, MA and ACTH; OriGene Technologies, Rockville, MD).

Techniques: Clinical Proteomics

Journal: International Journal of Molecular Sciences

Article Title: DHCR24, a Key Enzyme of Cholesterol Synthesis, Serves as a Marker Gene of the Mouse Adrenal Gland Inner Cortex

doi: 10.3390/ijms24020933

Figure Lengend Snippet: Phenotypic analyses of Dhcr24 cKO mice. ( A ) Oil Red O staining showing lipid droplets in the adrenal glands of euthyroid P35 mice. ( B ) Plasma levels of corticosterone, ACTH, and the ACTH/corticosterone ratio in euthyroid P35 mice. Each dot (WT) and circle (cKO) represents data from one mouse. Data are shown with the mean. Scale bars, 210 µm. WT: wild-type mice, cKO: Dhcr24 conditional knockout mice. ns, no significant difference.

Article Snippet: Hormones were measured using the corticosterone EIA kit (K014-H1, Arbor Assays, Ann Arbor, MI, USA) and the mouse ACTH ELISA kit (NBP3-14759, Novus Biologicals, Centennial, CO, USA) according to manufacturers’ instructions.

Techniques: Staining, Clinical Proteomics, Knock-Out

Journal: Archives of Histology and Cytology

Article Title: Localization of ATP-sensitive K⁺ channel subunits in rat pituitary gland

doi: 10.1679/aohc.76.53

Figure Lengend Snippet: Fig. 6. Immunofluorescence double staining shows the expression of ACTH as red (Alexa 594, A, D, G, J) and Kir6.1, Kir6.2, SUR2A, and SUR2B as green (Alexa 488, B, E, H, K) in the anterior lobe of the pituitary gland. The merged images (right column, C, F, I, L) indicate that Kir6.1 (C), Kir6.2 (F), SUR2A (I), and/or SUR2B (L) colocalize with ACTH. Scale bar: 50 μm.

Article Snippet: To determine whether the KATP channel subunit molecules colocalize with ACTH cells, PRL cells, FSH cells, and GH cells, sections were incubated with a mixture of each KATP channel subunit’s antibodies with mouse anti-human ACTH (Bio-Rad AbD Serotec.

Techniques: Immunofluorescence, Double Staining, Expressing

Journal: Archives of Histology and Cytology

Article Title: Localization of ATP-sensitive K⁺ channel subunits in rat pituitary gland

doi: 10.1679/aohc.76.53

Figure Lengend Snippet: Fig. 9. Immunofluorescence triple staining shows the expression of SUR2A (A, E) and SUR2B (I, M) as blue (Alexa 350), ACTH as green (Alexa 488, B, F, J, N), and Kir6.1 (C, K) and Kir6.2 (G, O) as red (Alexa 555) in the anterior lobe of the pituitary gland. The merged images (fourth column) indicate that Kir6.1/SUR2A (D), Kir6.2/SUR2A (H), Kir6.1/SUR2B (L), and Kir6.2/SUR2B (P) colocalize with ACTH in corticotrophs (arrows). Scale bar: 40 μm.

Article Snippet: To determine whether the KATP channel subunit molecules colocalize with ACTH cells, PRL cells, FSH cells, and GH cells, sections were incubated with a mixture of each KATP channel subunit’s antibodies with mouse anti-human ACTH (Bio-Rad AbD Serotec.

Techniques: Immunofluorescence, Staining, Expressing

Journal: Biosensors & bioelectronics

Article Title: Bioelectronic sensor mimicking the human neuroendocrine system for the detection of hypothalamic-pituitary-adrenal axis hormones in human blood.

doi: 10.1016/j.bios.2020.112071

Figure Lengend Snippet: Fig. 1. Schematic of the HPA axis of the human neuroendocrine system and bio electronic sensor system. In the human HPA axis, the hypothalamus, pituitary, and adre nal cortex communicate with interactions between CRHR1 or MC2R and CRH or ACTH, respectively. The bioelectronic sensor system mimics this system, in which CRHR1- and MC2R-embedded nanodiscs (CRHR1NDs and MC2RNDs) are immobilized on the floating electrode-based CNT-FET, and applied to detect CRH and ACTH, respectively.

Article Snippet: CRHR1 and MC2R cDNA were purchased from Origene (USA), and the MSP1E3D1 bacterial expression vector (pMSP1E3D1) was purchased from Addgene (USA).

Techniques:

Journal: Biosensors & bioelectronics

Article Title: Bioelectronic sensor mimicking the human neuroendocrine system for the detection of hypothalamic-pituitary-adrenal axis hormones in human blood.

doi: 10.1016/j.bios.2020.112071

Figure Lengend Snippet: Fig. 2. Characterization of CRHR1 and MC2R expressed in HEK-293 cells and E. coli (a) Dose- dependent responses of CRHR1 expressed in HEK- 293 cells to CRH (n ¼ 3, *p < 0.05, **p < 0.01, ***p < 0.001). (b) Selectivity of CRHR1 expressed in HEK- 293 cells after the addition of various hormones and neurotransmitters (n ¼ 3, CRH: corticotropin- releasing hormone, ACTH: adrenocorticotropic hor mone, DA: dopamine, EP: epinephrine, SE: seroto nin). (c) Dose-dependent responses of MC2R expressed in HEK-293 cells to ACTH (n ¼ 3, *p < 0.05, **p < 0.01, ***p < 0.001). (d) Selectivity of MC2R expressed in HEK-293 cells to various hor mones and neurotransmitters (n ¼ 3). (e) Western blot analysis of CRHR1 and MC2R expressed in E. coli using V5 Ab. The band indicates the molecular weight of CRHR1 (left image). The band corresponds to the molecular weight of MC2R (right image). (f) FE-SEM image of CRHR1NDs. The NDs had a dis coidal shape, and their average size was approxi mately 20 nm.

Article Snippet: CRHR1 and MC2R cDNA were purchased from Origene (USA), and the MSP1E3D1 bacterial expression vector (pMSP1E3D1) was purchased from Addgene (USA).

Techniques: Western Blot, Molecular Weight

Journal: Antioxidants & redox signaling

Article Title: Pterostilbene Decreases the Antioxidant Defenses of Aggressive Cancer Cells In Vivo: A Physiological Glucocorticoids- and Nrf2-Dependent Mechanism.

doi: 10.1089/ars.2015.6437

Figure Lengend Snippet: FIG. 3. Pter bioavailability in brain and the pituitary gland and its effect on ACTH synthesis and POMC ex- pression in AtT-20 cells. (A) Whole brain and pituitary gland levels of Pter after its i.v. administration (30 mg/kg) (n = 6–7 mice; *Significantly different p < 0.01, comparing hypophysis vs. whole brain levels). Data in (A) correspond to tissues obtained from nontumor-bearing mice (results obtained in A2058 or MeWo melanoma-bearing mice were not significantly different, not shown). (B) Effect of Pter on ACTH production by AtT-20 cells in a 24 h-period (mea- sured between 24 and 48 h or between 48 and 72 h). (C) Effect of Pter on POMC expression and (D) POMC protein levels (72 h of culture time) in AtT-20 cells (n = 6 for B, C, and D; *Significantly different p < 0.01, comparing +Pter vs. controls). For the in vitro experiments (B, C, and D), Pter (15 lM, see under the Results section) and CRH (100 nM) were added at time 0 and every 24 h through the culture time and were present, after each addition, for only 30 min. After the 30-min period, culture flasks were washed out (thrice with PBS) and the medium was renewed (controls received identical treatment). POMC, proopiomelanocortin.

Article Snippet: For Western blotting (see also above), POMC mouse monoclonal antibody (clone 2B2) from Origen was used.

Techniques: Expressing, In Vitro