immobility measurement device id-tech  (BIOSEB Inc)

Journal: iScience

Article Title: ER stress in mouse serotonin neurons triggers a depressive phenotype alleviated by ketamine targeting eIF2α signaling

doi: 10.1016/j.isci.2024.109787

Figure Lengend Snippet: Activation of ERS in DR in mouse models based on lipopolysaccharide (LPS)-induced neuroinflammation or chronic exposure to corticosterone stress (CORT) (A) Treatment schedule. Different groups of mice were treated with i) a single dose of LPS (0.83 mg/kg, ip) and examined 12 h post-administration, or ii) chronic CORT exposure in the drinking water for 28 days and examined on day 36. (B) In the tail suspension test (TST), the immobility time was compared between Veh ( n = 6) and LPS ( n = 6) or Veh ( n = 10) and CORT ( n = 9) mice. (C) BDNF protein levels in the DR were analyzed by Western blot (WB) and compared between Veh ( n = 5) and LPS ( n = 9) or Veh ( n = 5) and CORT ( n = 8) mice. (D) BiP protein levels in the DR were analyzed by WB and compared between Veh ( n = 5) and LPS ( n = 9) or Veh ( n = 5) and CORT ( n = 9) mice. (E) GRP94 protein levels in the DR were analyzed by WB and compared between Veh ( n = 5) and LPS ( n = 9) or Veh ( n = 5) and CORT ( n = 9) mice. (F) CHOP protein levels in the DR were analyzed by WB and compared between Veh ( n = 5) and LPS ( n = 9) or Veh ( n = 5) and CORT ( n = 8) mice. (G) eIF2α and p -eIF2α protein levels in the DR were analyzed by WB and compared between Veh ( n = 5) and LPS ( n = 9) or Veh ( n = 5) and CORT ( n = 9) mice. (H) eEF2 and p -eEF2 protein levels in the DR were analyzed by WB and compared between Veh ( n = 5) and LPS ( n = 9) or Veh ( n = 5) and CORT ( n = 9) mice. Data are expressed as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 by two-tailed t-test, see .

Article Snippet: Sessions were videotaped for 6 min and the immobility time was measured (Smart, Panlab, Cornella, Spain).

Techniques: Activation Assay, Suspension, Western Blot, Two Tailed Test

Journal: iScience

Article Title: ER stress in mouse serotonin neurons triggers a depressive phenotype alleviated by ketamine targeting eIF2α signaling

doi: 10.1016/j.isci.2024.109787

Figure Lengend Snippet: Activating ERS in DR induces a depressive-like phenotype in mice (A) Schematic time course diagram. Mice were injected with 1μL tunicamycin (Tm, 200 ƞg/μl) or vehicle (Veh, 4% DMSO in artificial cerebrospinal fluid-aCSF) into the dorsal raphe nucleus (DR) and examined after 1, 3, and 7 days. (B) In the tail suspension test (TST), the immobility time of Veh ( n = 11) and Tm ( n = 6) mice was compared. (C) In the dark-light box (DLB), latency to enter and the number of entries into the light chamber were compared between Veh ( n = 11) and Tm ( n = 6) mice. (D) During the 15 min observation period, the total distance and mean speed were recorded for Veh ( n = 10) and Tm ( n = 6) mice in the open field test (OFT). (E) BiP protein levels in the DR were analyzed by Western blot (WB) and compared between Veh ( n = 11) and Tm ( n = 6) mice. (F) GRP94 protein levels in the DR were analyzed by WB and compared between Veh ( n = 8) and Tm ( n = 5) mice. (G) eIF2α and p -eIF2α protein levels in the DR were analyzed by WB and compared between Veh ( n = 8) and Tm ( n = 6) mice. (H) eEF2 and p -eEF2 protein levels in the DR were analyzed by WB and compared between Veh ( n = 8) and Tm ( n = 6) mice. (I) Photomicrographs showing TPH-positive or GABA-positive cells expressing Egr1 mRNA ( 33 P-oligonucleotide silver grains) in the DR of Veh and Tm mice. Scale bar: 20 μm. White arrowheads indicate TPH-positive or GABA-positive cells co-localizing with Egr1 mRNA. (J) The total number of TPH-positive or GABA-positive cells and the percentage of TPH- or GABA-positive cells expressing Egr1 mRNA were analyzed in Veh ( n = 5) and Tm ( n = 4) mice. Data are expressed as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 by one-way ANOVA, see . See also Figures S1‒S3 .

Article Snippet: Sessions were videotaped for 6 min and the immobility time was measured (Smart, Panlab, Cornella, Spain).

Techniques: Injection, Suspension, Western Blot, Expressing

Journal: iScience

Article Title: ER stress in mouse serotonin neurons triggers a depressive phenotype alleviated by ketamine targeting eIF2α signaling

doi: 10.1016/j.isci.2024.109787

Figure Lengend Snippet: Ketamine alleviates depressive-like phenotype by reducing ERS in raphe 5-HT neurons (A) Treatment schedule. Mice were injected with 1 μL Tm (200 ƞg/μl) or Veh (aCSF+DMSO 4%) into the DR, 24 h later received Ketamine (Ket, 10 mg/kg, ip) or Veh (saline solution, ip), and examined 30 min or 48 h post-Ket. (B) In the TST, the immobility time was recorded in Ket-treated Veh and Tm mice ( n = 5 to 8 per group). (C) BDNF protein levels in the DR were analyzed by WB. They were compared between Veh and Tm mice treated with Ket ( n = 5 to 8 per group). (D) Representative coronal midbrain sections showing TPH-positive cells expressing different NMDAR subunits including GluN1, GluN2B, and GluN2D mRNA in the DR of wild-type mice. The bottom row shows high magnification photomicrographs of the frames in the top row. The + symbol indicates TPH- and NMDA subunits-positive cells. Scale bars: low = 100 μm, high = 20 μm. (E) BiP protein levels in the DR were analyzed by WB and compared between Veh and Tm mice treated with Ket ( n = 4 to 5 per group). (F) GRP94 protein levels in the DR were analyzed by WB and compared between Ket-treated Veh and Tm mice ( n = 4 to 5 per group). (G) eIF2α and p -eIF2α protein levels in the DR were analyzed by WB and compared between Ket-treated Veh and Tm mice ( n = 4 to 5 per group). (H) eEF2 and p -eEF2 protein levels in the DR were analyzed by WB and compared between Ket-treated Veh and Tm mice ( n = 4 to 5 per group). Data are expressed as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 by one-way ANOVA, see . See also Figures S6 and .

Article Snippet: Sessions were videotaped for 6 min and the immobility time was measured (Smart, Panlab, Cornella, Spain).

Techniques: Injection, Saline, Expressing

Journal: iScience

Article Title: ER stress in mouse serotonin neurons triggers a depressive phenotype alleviated by ketamine targeting eIF2α signaling

doi: 10.1016/j.isci.2024.109787

Figure Lengend Snippet: eIF2α signaling in 5-HT neurons is critical to the antidepressant effects of ketamine (A) Treatment schedule. Mice were injected with 1 μL Tm (200 ƞg/μl) or Veh (aCSF+DMSO 4%) into the DR and received two doses of ISRIB (PERK inhibitor, 0.25 mg/kg, ip) or Salubrinal (SAL, eIF2α-GADD34:PP1 phosphatase inhibitor, 1 mg/kg, ip). Ket (10 mg/kg, ip) was administered 24 h later and mice were examined 30 min after Ket administration. (B) BiP and GRP94 protein levels in mouse DR were analyzed by WB and the effects of ISRIB or SAL on Ket action were compared ( n = 7 per group). (C) eIF2α and p -eIF2α protein levels in mouse DR were analyzed by WB and the effects of ISRIB or SAL on Ket action were compared ( n = 7 per group). (D) eEF2 and p -eEF2 protein levels in mouse DR were analyzed by WB and the effects of ISRIB or SAL on Ket action were compared ( n = 7 per group). (E) In the TST, the immobility time was recorded in mice ( n = 7 to 14 per group). (F) BDNF protein levels in DR were analyzed by WB. They were compared between Veh and Tm mice treated with ISRIB or SAL and Ket ( n = 5 to 7 per group). (G) Representative images of coronal sections of the mPFC showing the mRNA expression of BDNF, TrkB, Neuritin, PSD95, VEGF, and Egr1 in the mouse model of Tm assessed by in situ hybridization. Scale bar: 1 mm. The relative density of mRNA expression of the different transcripts was compared between Veh and Tm mice treated with ISRIB or SAL and Ket ( n = 4 to 5 per group). Data are expressed as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001 by one-way ANOVA, see . See also Figures S8‒S14 .

Article Snippet: Sessions were videotaped for 6 min and the immobility time was measured (Smart, Panlab, Cornella, Spain).

Techniques: Injection, Expressing, In Situ Hybridization

Journal: The Journal of Neuroscience

Article Title: Altered DNA Methylation in the Developing Brains of Rats Genetically Prone to High versus Low Anxiety

doi: 10.1523/JNEUROSCI.1157-15.2019

Figure Lengend Snippet: Perinatal methyl donor depletion improves LR rats' adult anxiety- and depression-like behavior. In an attempt to reduce DNA methylation levels in developing LR pups, LR mothers were fed a diet severely depleted of methyl donors (or standard rat chow) from late pregnancy through weaning. Their adult offspring (pDEP and pCON; males n = 15, females LR-pDEP n = 14, LR-pCON n = 16) were later evaluated in several emotional behavior assays (A–L). A, In male offspring, perinatal methyl donor deprivation led LRs to display greater exploratory behavior in the OFT compared with controls. B, C, The LR-pDEP males also showed less anxiety-like behavior, with a trend to spend more time in the center of the open field and significantly more time in the open arms of the EPM. D, Perinatal methyl donor depletion led male LRs to display more social exploration in the Social Interaction Test. E, F, During FST, male LR-pDEP show reduced immobility compared with LR-pCON, indicated less depression-like behavior, although the groups did not differ in the Sucrose Preference Test. FST was also significantly affected by the weight of the animal. G–L, Perinatal methyl donor depletion elicited similar effects on LR females as it did in males, with female LR-pDEP offspring showing increased novelty-induced locomotion, more time spent in the center of the OF, less FST immobility and greater sucrose preference compared with LR-pCON females. Because the perinatal methyl donor deplete diet led to reduced body weight in exposed offspring, we used linear modeling for statistical analysis and considered diet, sex, and weight as factors. Through all behavioral measures, sex was not found to have a significant effect and weight only have a significant effect on FST. Last, the perinatal methyl donor depletion study was repeated in HR offspring, which had limited effect on their behavioral phenotype (Figure 4-1). #p = 0.0576; *p < 0.05; **p < 0.01; compared with control. ∧p < 0.05, considering effect of weight.

Article Snippet: We focused on the immobility measure because it is classically considered to be an indicator of behavioral despair and depressive-like behavior ( Porsolt et al., 1977 ), and it can be clearly defined from active coping measures, such as swimming and climbing ( Cryan et al., 2005 ).

Techniques: DNA Methylation Assay, Control

Journal: The Journal of Neuroscience

Article Title: Altered DNA Methylation in the Developing Brains of Rats Genetically Prone to High versus Low Anxiety

doi: 10.1523/JNEUROSCI.1157-15.2019

Figure Lengend Snippet: Transiently suppressing Dnmt3b expression in the developing amygdala leads to reduced anxiety-like behavior in adulthood. An siRNA was injected into the early postnatal amygdala to transiently suppress expression of the DNMT, Dnmt3b, to test whether this would lead to less anxiety- and/or depression-like behavior in adulthood (Dnmt3b siRNA: n = 15 males; n = 14 females; control siRNA n = 14 males; n = 15 females; A–L). A, In males, early-life DNMT3b knockdown (KD) leads to greater novelty-induced exploration in adult animals compared with controls. B, C, The siRNA Dnmt3b KD males also showed less anxiety-like behavior, spending more time in the center of the open field and more time in the open arms of the EPM. D–F, The early-life DNMT3b KD did not affect male rats' behavior in the Social Interaction Test, FST, or Sucrose Preference Test. G–L, Female animals were less affected by the transient early-life DNMT3b KD with the early DNMT3b KD females and control females displaying similar behavior across all tests, except increased immobility in FST. *p < 0.05 compared with control.

Article Snippet: We focused on the immobility measure because it is classically considered to be an indicator of behavioral despair and depressive-like behavior ( Porsolt et al., 1977 ), and it can be clearly defined from active coping measures, such as swimming and climbing ( Cryan et al., 2005 ).

Techniques: Expressing, Injection, Control, Knockdown