erβ selective antagonist phtpp  (Tocris)

Journal: Brain research

Article Title: Estrogen receptor α regulates the IKKs/NF-kB activity involved in the development of mechanical allodynia induced by REM sleep deprivation in rats.

doi: 10.1016/j.brainres.2024.149269

Figure Lengend Snippet: Fig. 3. ERα but not ERβ regulates BMS-345541-prevented REMSD-induced allodynia in female rats. A) Effect of intrathecal injection of BMS-345541 pretreatment (preTx BMS 10 µg) in the establishment of mechanical allodynia induced by REM sleep deprivation (REMSD) for two days in intact female rats (F), female rats with vehicle (Veh), rats treated with ICI (ICI+preTx BMS), rats treated with a selective ERα antagonist (MPP+preTx BMS) or rats treated with a selective ERβ antagonist (PHTPP+preTx BMS). B) Area under the curve (AUC) obtained from time course in A. C) Effect of intrathecal injection of BMS-345541 pretreatment (preTx BMS 10 µg) in the establishment of mechanical allodynia induced by REMSD in female rats with vehicle (Veh), non-ovariectomized female rats (F), ovariectomized rats (OVX) and ovariectomized rats in the presence of ERα agonist (OVX+PPT). D) Area under the curve obtained from time course in C. Data are expressed as the mean ± SEM (n = 6). *** p < 0.001 versus Veh group; # # # p < 0.001 versus intact female rats with pretreatment (F) by one-way ANOVA followed by the Tukey test.

Article Snippet: PHTPP [4-[2-fenil-5,7-bis(trifluorometil) pirazolo[1,5-a]-pirimidin-3-il]fenol], a selective ERβ antagonist (Cat. No SC-204191, Santa Cruz Biotecnology, Dallas, TX) and PPT(1,3,5-Tris (4-hydroxyphenyl)-4-propyl-1H-pyrazole), a selective ERα agonist (Cat. No. SC-297946, Santa Cruz Biotechnology, Dallas, TX) were dissolved in 1 % DMSO in saline.

Techniques: Injection

Journal: Neurobiology of Stress

Article Title: Estrogen receptor beta in the central amygdala regulates the deleterious behavioral and neuronal consequences of repeated social stress in female rats

doi: 10.1016/j.ynstr.2023.100531

Figure Lengend Snippet: Experimental Timelines. Two separate experiments were completed to determine the role of CeA ERβ signaling in the development of hypervigilant behavioral responses to repeated social stress in females. A) Rats in Experiment #1 were first assessed in a marble burying test (MB) to establish baseline levels of anxiety-like behavior, then were exposed to five consecutive days of WS or CON (15 min/day). Sessions were recorded and behaviors were assessed on days #1 (D1) and #5 (D5). WS/CON rats underwent a post-WS/CON MB test to determine how exposure to repeated WS affects measures of hypervigilant and anxiety-like behaviors. Six days after the final WS/CON exposure, rats were re-exposed to the context in which they originally experienced either WS (but in the absence of the resident) or CON for 15 min. Brains were collected 30 min after the start of WS/CON context exposure. B) For Experiment #2, all rats were implanted with indwelling bilateral cannulas into the CeA. Following surgical recovery, a pre-stress testing regimen commenced to assess baseline sucrose preference (SPT) and acoustic startle response (ASR). Subsequently, rats were exposed to five consecutive days of WS/CON (15 min/day) with local microinjection of either vehicle or the ERβ antagonist, PHTPP, into the CeA 1 h prior to each WS/CON session. Post-WS/CON behavioral testing consisted of the elevated plus maze (EPM), SPT, ASR, and MB, each separated by 24 h, to determine if ERβ blockade, through PHTPP, prevented the development of hypervigilant, anxiety-like, and reward motivated behaviors. Tissue was collected at rest two days following the final behavioral test. CeA: Central Amygdala; ERβ: estrogen receptor beta.

Article Snippet: One hour prior to each WS/CON exposure, females in Experiment #2 received either vehicle (10% dimethyl sulfoxide [DMSO], Fisher Scientific, Pittsburgh, PA) in saline, or the potent and selective ERβ antagonist, 4-[2-Phenyl-5,7- bis (trifluoromethyl)pyrazolol[1,5- a ]pyrimidin-3-yl]phenol (PHTPP, 10 μM dissolved in 10% DMSO, TOCRIS, Minneapolis, MN), microinjected bilaterally into the CeA (0.5 μL over 1 min with a 1-min wait post-infusion) ( ).

Techniques: Microinjection

Journal: Neurobiology of Stress

Article Title: Estrogen receptor beta in the central amygdala regulates the deleterious behavioral and neuronal consequences of repeated social stress in female rats

doi: 10.1016/j.ynstr.2023.100531

Figure Lengend Snippet: Blocking ERβ in the CeA reduces the sensitization to increased spontaneous WS-evoked burying during repeated WS exposure . Spontaneous burying behavior was increased among WS rats compared to CON during day #1 (D1) exposure to WS/CON, and this acute response was unaffected by pre-treatment with the ERβ antagonist PHTPP ( A ). Burying also presented significantly quicker in WS rats with a longer latency to begin burying observed in the WS rats treated with PHTPP ( B ). There were no group differences observed in rearing behavior ( C ). However, on the fifth day of WS exposure, PHTPP attenuates the WS-evoked burying response ( D ), along with an overall stress effect on latency to begin burying ( E ) and no significant differences between the groups with regards to rearing behavior ( F ). When directly comparing the burying behavior between D1 and D5, there was a interaction between treatment and time, with WS + VEH rats increasing burying from D1 to D5 which this behavioral response was prevented with PHTPP treatment ( G, H ) (n = 12–13/group with the exception of statistical outliers, ʄ p < 0.05 WS + VEH vs. WS + PHTPP, * p < 0.05, **** p < 0.0001). ERβ: estrogen receptor beta; CeA: central amygdala; WS: witness stress; D1: day 1; CON: control.

Article Snippet: One hour prior to each WS/CON exposure, females in Experiment #2 received either vehicle (10% dimethyl sulfoxide [DMSO], Fisher Scientific, Pittsburgh, PA) in saline, or the potent and selective ERβ antagonist, 4-[2-Phenyl-5,7- bis (trifluoromethyl)pyrazolol[1,5- a ]pyrimidin-3-yl]phenol (PHTPP, 10 μM dissolved in 10% DMSO, TOCRIS, Minneapolis, MN), microinjected bilaterally into the CeA (0.5 μL over 1 min with a 1-min wait post-infusion) ( ).

Techniques: Blocking Assay, Control

Journal: Neurobiology of Stress

Article Title: Estrogen receptor beta in the central amygdala regulates the deleterious behavioral and neuronal consequences of repeated social stress in female rats

doi: 10.1016/j.ynstr.2023.100531

Figure Lengend Snippet: A history of WS and ERβ antagonism in the CeA differentially affect hypervigilant, anxiety- and depressive-like behaviors. There were no significant effects of WS on behavior in the EPM, with no differences in the percentage of time spent in the open arms or ( A ) in general motor behaviors as evidenced by overall distance traveled ( B ). Further, behavior on the EPM was not regulated by pharmacological blockade of ERβ during WS exposure, as evidenced by a lack of effect induced by PHTPP. Repeated WS produced a persistent increases in anxiety-like behavior evidenced by the hypervigilant phenotype indicated by an enhanced acoustic startle response. Startle velocity was increased in WS rats and blocking ERβ with PHTPP during each WS session prevented the development of heightened ASR following stress ( C ). Further, a history of repeated WS exposure resulted in a significant decrease in the percent of sucrose preferred in the post-stress test but this reduction in reward seeking/depressive-like behaviors was prevented by PHTPP ( D ). Behaviors were not altered in the single post-stress marble burying task, with no group differences in the number of marbles buried ( E ) or the duration of burying ( F ). Brains were collected under resting conditions 48 h after the final behavioral task and CeA tissue was prepared for Western blot analysis. PHTPP caused a a significant long-term reduction in resting levels of CRF (G) with no effects on resting levels of proCRF (H). There was also a compensatory increase in ERβ at rest in both control and WS rats ( I ) that were treated with PHTPP. (n = 12–13/group with the exception of statistical outliers, + main effect of drug, * p < 0.05, ** p < 0.005, *** p < 0.0005, **** p < 0.0001). WS: witness stress; ERβ: estrogen receptor beta; CeA: central amygdala; EPM: elevated plus maze; ASR: acoustic startle response; MB: marble burying; CRF: corticotropin releasing factor.

Article Snippet: One hour prior to each WS/CON exposure, females in Experiment #2 received either vehicle (10% dimethyl sulfoxide [DMSO], Fisher Scientific, Pittsburgh, PA) in saline, or the potent and selective ERβ antagonist, 4-[2-Phenyl-5,7- bis (trifluoromethyl)pyrazolol[1,5- a ]pyrimidin-3-yl]phenol (PHTPP, 10 μM dissolved in 10% DMSO, TOCRIS, Minneapolis, MN), microinjected bilaterally into the CeA (0.5 μL over 1 min with a 1-min wait post-infusion) ( ).

Techniques: Produced, Blocking Assay, Western Blot, Control