All animal experiments were performed in accordance with the European Communities Council Directive (86/609/EEC) and were reviewed and approved by the Research Ethics Committee of the Royal College of Surgeons in Ireland (REC #205), under license from the Department of Health, Dublin, Ireland. Adult (20–22 g) male C57BL/6 mice (Harlan) were purchased from Harlan. Food and water was available ad libitum. Induction of SE was performed as described previously31 (link). Mice were anesthetized with isoflurane and placed in a mouse-adapted stereotaxic frame. Following a midline scalp incision, Bregma was located and three partial craniectomies performed for placement of skull-mounted recording screws (Bilaney Consultants). A fourth craniectomy was drilled for placement of a guide cannula (Coordinates from Bregma; AP = −0.94 mm, L = −2.85 mm) based on a stereotaxic atlas60 . The cannula and electrode assembly was fixed in place and the animal placed in an open Perspex box which allowed free movement. The EEG was recorded using a Grass Comet digital EEG. After baseline EEG was established, the animal was lightly restrained while an injection cannula was lowered 3.75 mm below the brain surface for injection of KA (Sigma-Aldrich) or vehicle (phosphate-buffered saline (PBS), pH adjusted to 7.4) into the basolateral amygdala nucleus. After 40 min, all mice received lorazepam (Ativan, 6 mg kg−1, i.p.). Animals were recorded for up to an hour thereafter before being disconnected and placed in a warmed recovery chamber. Non-harmful seizures were induced by a single injection (i.p.) of KA (15 mg kg−1), as described36 (link).
Nuclear Groups, Basolateral
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Most cited protocols related to «Nuclear Groups, Basolateral»
Adult
Animals
Ativan
Brain
Cannula
Cannulation
Cell Nucleus
Comet Assay
Craniectomy
Cranium
Ethics Committees, Research
Fingers
Food
Isoflurane
Lorazepam
Males
Mice, House
Mice, Inbred C57BL
Movement
Nuclear Groups, Basolateral
Perspex
Phosphates
Reading Frames
Saline Solution
Scalp
Seizures
Surgeons
Adult
Animals
Ativan
Brain
Cannula
Cannulation
Cell Nucleus
Comet Assay
Craniectomy
Cranium
Ethics Committees, Research
Fingers
Food
Isoflurane
Lorazepam
Males
Mice, House
Mice, Inbred C57BL
Movement
Nuclear Groups, Basolateral
Perspex
Phosphates
Reading Frames
Saline Solution
Scalp
Seizures
Surgeons
Six-week-old C57BL/6 mice (P35–45) were initially induced with ketamine (80 mg kg−1) and xylazine (10 mg kg−1) and placed into a stereotaxic frame (David Kopf Instruments), before isoflurane anesthesia (~1% in O2, v/v). A craniotomy (∼1 mm in diameter) was made above the injection site. Virus suspensions were slowly injected (100 nl min–1) using a 34 G beveled needle (Nanofil syringe, World Precision Instruments). After injection, the needle was left in place for an additional 5 min and then slowly withdrawn. The surgical procedure was either continued with optic fiber or optrode drive implantations (described below), or the surgical incision was closed with tissue glue and 0.05 mg kg−1 Buprenorphine was subcutaneously injected for post-surgical analgesia. Injections targeting the medial prefrontal cortex (mPFC) were made 1.8 mm anterior, 0.3 mm lateral and 2.53 mm ventral to bregma. Basolateral amygdala (BLA) injection coordinates were 1.15 mm posterior, 3.0 mm lateral, and 5.0 mm ventral to bregma. For mPFC injections, 1 µl of the indicated virus was injected. For fear extinction experiments mice were bilaterally injected with 500 nl AAV2/1&2.CamKIIα.stGtACR2-Fred.WPRE or AAV2/1&2.CamKIIα.eYFP.WPRE with a genomic titer in the range of 2–3 x 1011 vp ml−1.
Anesthesia
Buprenorphine
Craniotomy
Extinction, Psychological
Fear
Genome
Isoflurane
Ketamine
Management, Pain
Mice, House
Mice, Inbred C57BL
Needles
Nuclear Groups, Basolateral
Operative Surgical Procedures
Ovum Implantation
Prefrontal Cortex
Reading Frames
Surgical Wound
Syringes
Tissues
Virus
Xylazine
Analgesics
Antibiotics
Buprenorphine
Cannula
Cortex, Cerebral
Cranium
Dental Cements
Extinction, Psychological
Injections, Intraperitoneal
Ketamine
Nuclear Groups, Basolateral
Operative Surgical Procedures
Rattus
Seahorses
Stainless Steel
Xylazine
Mice were placed on doxycycline chow 24 hr prior to surgery (40 mg/kg, Bio-Serv, Flemington, NJ). On the day of surgery, mice were anaesthetized using isoflurane (3% induction, 1.5% maintenance during surgery) and secured to a stereotactic frame (Kopf) with a heating pad to maintain body temperature. Mice were injected sub-dermally with Buprenex (1 mg/kg) and given topical Lidocaine (20 mg/mL) for analgesia (Hi-Tech Pharmacal, Amityville, NY). Following exposure of the skull, a small craniotomy was made overlying the target brain structure. Using a glass pipette (50 µm tip) connected to a Nanoject II (Drummond Scientific, Broomall, PA) injector, AAVs were delivered (50 nL/infusion, 30 sec between infusions) to the brain, and allowed to diffuse for 10 min before withdrawing the pipette. The coordinates of the target brain structures in reference to bregma (mm) and injection volumes (nL) were as follows: dentate gyrus (AP: −1.9, ML: ± 1.4, DV: −2.05; 600), dorsal CA3 (AP: −1.9, ML: ± 1.85, DV: −2.1; 500), basolateral amygdala (AP: −1.6, ML: ± 3.25, DV: −3.6 from pial surface; 1000), and central amygdala (AP: −1.4, ML: ± 2.85, DV: −3.7 from pial surface; 300). All injections were performed bilaterally. Mice were kept on doxycycline chow and allowed to recover for 7–14 days following surgery.
Rats were anesthetized with isoflurane. A midline incision was made to expose the skull, and a small craniotomy was made unilaterally above the medial prefrontal cortex (mPFC). Recombinant AAV was injected into the prelimbic cortex (PL) of the mPFC. A stainless steel needle with the beveled tip facing laterally (31 gauge; Hamilton Company, Reno, Nevada) was directed to the PL (AP: +2.5, ML: ±0.5, DV: −2.0 from pial surface). For the CRAM experiment in rats, CAV2-Cre was bilaterally injected to the dorsal medial striatum (AP: +0.1, ML: ± 2.0, DV: −3.5 from pial surface). Virus (1.0 μL/hemisphere) was infused over 10 min (0.1 μL/min), followed by an additional 10 min to allow diffusion of the virus from the needle tip. The skin was sealed with Vetbond tissue adhesive.
Rats were anesthetized with isoflurane. A midline incision was made to expose the skull, and a small craniotomy was made unilaterally above the medial prefrontal cortex (mPFC). Recombinant AAV was injected into the prelimbic cortex (PL) of the mPFC. A stainless steel needle with the beveled tip facing laterally (31 gauge; Hamilton Company, Reno, Nevada) was directed to the PL (AP: +2.5, ML: ±0.5, DV: −2.0 from pial surface). For the CRAM experiment in rats, CAV2-Cre was bilaterally injected to the dorsal medial striatum (AP: +0.1, ML: ± 2.0, DV: −3.5 from pial surface). Virus (1.0 μL/hemisphere) was infused over 10 min (0.1 μL/min), followed by an additional 10 min to allow diffusion of the virus from the needle tip. The skin was sealed with Vetbond tissue adhesive.
Body Temperature
Brain
Buprenex
CAV2 protein, human
Cortex, Cerebral
Craniotomy
Cranium
Diffusion
Doxycycline
Gyrus, Dentate
Isoflurane
Lidocaine
Management, Pain
Mice, House
Needles
Nuclear Groups, Basolateral
Nucleus, Central Amygdaloid
Operative Surgical Procedures
Prefrontal Cortex
Rattus
Reading Frames
Skin
Stainless Steel
Striatum, Corpus
Surgery, Day
Tissue Adhesives
Virus
Most recents protocols related to «Nuclear Groups, Basolateral»
All surgical procedures were performed under urethane (1.5 g/kg; Sigma-Aldrich/Merck, Barcelona, Spain) anesthesia. To record the local field potentials, custom-made Teflon-coated stainless steel recording electrodes (AM Systems, Sequim, WA, USA) were implanted, according to the stereotactic coordinates from Bregma (Paxinos and Watson, 2007 ), in the dorsal hippocampus (HPCd) (AP −3.4 mm; L 2.5 mm; DV 2.4 mm), intermediate hippocampus (HPCi) (AP −5.8 mm; L 5.8 mm; DV 5 mm), ventral hippocampus (HPCv) (AP −4.7 mm; L 5 mm; DV 8.7 mm), and basolateral amygdala (BLA) (AP −2.3 mm; L 5 mm; DV 8.5 mm). For DBS, we used in-house custom-made bipolar twisted electrodes made of Teflon-coated stainless-steel wire, with 1 mm between both tips. Stimulating electrodes were bilaterally implanted into the infralimbic (IL) region of the medial prefrontal cortex (coordinates: AP +3.2 mm; L 0.5 mm; DV 5.4 mm).
The surgical procedures used in the study of the electrophysiological activity are the same as for the analysis of c-Fos expression. However, in the case of the immunoreactivity study, only the DBS stimulation electrode in IL was implanted to preserve an optimal state in the other regions of interest.Figure 1 illustrates the experimental setup and the stimulation sites.
The surgical procedures used in the study of the electrophysiological activity are the same as for the analysis of c-Fos expression. However, in the case of the immunoreactivity study, only the DBS stimulation electrode in IL was implanted to preserve an optimal state in the other regions of interest.
Anesthesia
Electrophysiologic Study, Cardiac
Nuclear Groups, Basolateral
Operative Surgical Procedures
Prefrontal Cortex
Seahorses
Stainless Steel
Teflon
Urethane
CRH-ires-Cre mice were injected with AAV1-Ef1a-DIO-hChR2-EYFP in the basolateral amygdala (BLA) were deeply anesthetized with isoflurane and quickly decapitated (PN 120). Acute horizontal slices (300 μm) containing both basolateral amygdala and nucleus accumbens were obtained using a vibratome (V1200S, Leica, U) in ice-cold N-Methyl d -Glucamine (NMDG) cutting solution: containing (in mM): 110 NMDG, 20 HEPES, 25 glucose, 30 NaHCO3 1.2 NaH2PO4, 2.5 KCl, 5 sodium ascorbate, 3 sodium pyruvate, 2 Thiourea, 10 MgSO4−7 H2O, 0.5 CaCl2, 305-310 mOsm, pH 7.4. Slices equilibrated in a homemade chamber for 25–30 min (31 °C) and an additional 45 min in room temperature aCSF containing (in mM): 119 NaCl, 26 NaHCO3, 1 NaH2PO4, 2.5 KCl, 11 Glucose, 10 Sucrose, 2.5 MgSO4–7 H2O, and 2.5 CaCl2 (pH 7.4) before being transferred to a recording chamber. Recording solution was the same as equilibration solution only with 1.3 mM MgSO4−7 H2O. All solutions were continuously bubbled with 95% O2/5%CO2.
Bicarbonate, Sodium
Cold Temperature
Glucose
HEPES
Internal Ribosome Entry Sites
Isoflurane
Mice, Laboratory
Nuclear Groups, Basolateral
Nucleus Accumbens
Pyruvate
Sodium
Sodium Ascorbate
Sodium Chloride
Sucrose
Sulfate, Magnesium
Thiourea
Experiment 1. Fox odor avoidance in separate age-groups using males (2- (n = 14–10/group), 4- (n = 18–17/group), 6- (n = 11/group), 8- (n = 8/group), 12- (n = 4–6/group), 15- (n = 10/group), 18-month-old (n = 9–10/group)) and females (2- (n = 5–10/group) and 15-month-old (n = 5–7/group)). To have a comprehensive picture, we aimed to examine the temporal resolution in 2-month “bins”, however, after 12-month, due to insufficient number of animals, we switched to 3-month “bins”. Our main goal was to reveal temporal differences. Therefore, sex differences were addressed only at two ages (2 and 15 months, i.e., before and after the presumable transient period).
Experiment 2. Longitudinal, repeated (between 2 and 11 months of age) exploration of open-field behavior in male and female mice (n = 8/group at the beginning). As spending 5 min in an open arena does not have long term behavioral consequences, the same animals were examined repeatedly. The animals were tested monthly in slightly different environment to avoid habituation to the arena. As hypolocomotion was found repeatedly during 5 min testing, the question arose whether it is an initial anxiety in a new environment (which will be released after a while) or more a sign of innate anxiety from open, bright spaces, which does not diminish over time. To test this hypothesis, at the termination of our experiment (in 11-month-old mice), we conducted a more prolonged observation (30 min).
Experiment 3. Immunohistochemical confirmation of Aβ accumulation and pTau appearance in 2- and 12-month-old animals (i.e., before and after the presumable transient period [9 (link)]; female, n = 3; 3xTg-AD) in the OB, motor and somatosensory cortex, hippocampus, and basolateral amygdala regions as relevant regions for cognitive impairment (cortex, hippocampus), emotions (amygdala), or smell loss (OB).
Experiment 2. Longitudinal, repeated (between 2 and 11 months of age) exploration of open-field behavior in male and female mice (n = 8/group at the beginning). As spending 5 min in an open arena does not have long term behavioral consequences, the same animals were examined repeatedly. The animals were tested monthly in slightly different environment to avoid habituation to the arena. As hypolocomotion was found repeatedly during 5 min testing, the question arose whether it is an initial anxiety in a new environment (which will be released after a while) or more a sign of innate anxiety from open, bright spaces, which does not diminish over time. To test this hypothesis, at the termination of our experiment (in 11-month-old mice), we conducted a more prolonged observation (30 min).
Experiment 3. Immunohistochemical confirmation of Aβ accumulation and pTau appearance in 2- and 12-month-old animals (i.e., before and after the presumable transient period [9 (link)]; female, n = 3; 3xTg-AD) in the OB, motor and somatosensory cortex, hippocampus, and basolateral amygdala regions as relevant regions for cognitive impairment (cortex, hippocampus), emotions (amygdala), or smell loss (OB).
Age Groups
Amygdaloid Body
Animals
Anxiety
Cortex, Cerebral
Disorders, Cognitive
Emotions
Females
Males
Mice, House
Nuclear Groups, Basolateral
Odors
Seahorses
Somatosensory Cortex
SWI3 protein, S cerevisiae
Transients
Field potential recordings were performed as previously described (Adermark et al., 2011 (link)). Briefly, local field population spikes (PSs) were activated with a frequency of 0.05 Hz in subregions of the: ventral striatum nucleus accumbens core (NAcC) and shell (NAcSh); dorsal striatum [dorsomedial striatum (DMS); dorsolateral striatum (DLS)]; amygdala [central nucleus of the amygdala (CeA) and basolateral amygdala; Beridze et al., 2020 (link); Figs. 2A ,F , 3A , 4A ,F ; Extended Data Figs. 2-1 , 3-1 , 4-1 ]. Only rats that obtained ≥15 infusions in the last 10 self-administration sessions were considered in the electrophysiological recordings. Recordings in different brain areas were conducted in separate brain slices retrieved from the same rat, thereby minimizing the risk of individual variation between brain regions and treatments. Stimulation electrodes (World Precision Instruments; type TM33B) were positioned locally, 0.2–0.3 mm from the recording electrode (borosilicate glass, 2.5–4.5 MΩ, World Precision Instruments; Extended Data Figs. 2-1 , 3-1 , 4-1 ), and the amplitude of the population spike (PS) were measured. Signals were amplified with a custom-made amplifier, filtered at 3 kHz, and digitized at 8 kHz.
During field potential recordings, slices were perfused with prewarmed aCSF (30°C) containing the GABAA receptor antagonist bicuculline to isolate excitatory transmission to monitor putative effects by treatment on neurotransmission, stimulus/response curves were created by stepwise increasing the stimulation strength, and by monitoring the paired pulse ratio (PPR). For PPR, responses were evoked with a paired pulse stimulation (50-ms interpulse interval) at approximately half max stimulation strength, and PPR was calculated by dividing the second pulse (PS2) with the first pulse (PS1).
During field potential recordings, slices were perfused with prewarmed aCSF (30°C) containing the GABAA receptor antagonist bicuculline to isolate excitatory transmission to monitor putative effects by treatment on neurotransmission, stimulus/response curves were created by stepwise increasing the stimulation strength, and by monitoring the paired pulse ratio (PPR). For PPR, responses were evoked with a paired pulse stimulation (50-ms interpulse interval) at approximately half max stimulation strength, and PPR was calculated by dividing the second pulse (PS2) with the first pulse (PS1).
Bicuculline
Brain
Diencephalon
Figs
GABA-A Receptor Antagonists
Neostriatum
Nuclear Groups, Basolateral
Nucleus, Central Amygdaloid
Nucleus Accumbens
Pulse Rate
Self Administration
Synaptic Transmission
Transmission, Communicable Disease
Ventral Striatum
We mapped the mouse snRNA-seq gene expression profile to the mouse ISH atlas of the Allen Institute for Brain Science (http://mouse.brain-map.org ) at the cluster level85 (link). The ISH data of each slice were summarized into voxel dataset, providing the gene expression profile of each voxel (Gene expression value X voxel). The brain structures file of each slice showed the anatomical annotation of each voxel (voxel X structures). First, we generated whole brain plots with only amygdala subnuclei annotation in each slice using brain structures file to select the proper coronal slice as reference image. We chose slice 37 to plot reference image, where we divided the amygdala into five subnuclei: BLA (Basolateral amygdalar complex), IA (Intercalated amygdalar nucleus), CEA (Central amygdalar nucleus), MEA (Medial amygdalar nucleus), and COA (Cortical amygdalar area). Then, we downloaded the ISH data of slice 37 to map gene expression on the reference image. We calculated the average expression value of the top 5 DEGs in each cluster and then, projected it on the reference image to infer the subnuclei distribution of clusters. (Only DEGs that appeared in the ISH coronal gene expression profile were retained).
Amygdaloid Body
Brain
Brain Mapping
Cell Nucleus
Cortical Amygdala
Gene Expression
Mice, Laboratory
Nuclear Groups, Basolateral
Nucleus, Central Amygdaloid
Nucleus, Medial Amygdalar
Small Nuclear RNA
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AAV1-CaMKIIα-hChR2(H134R)-eYFP-WPRE (Addgene # 26969P) is an adeno-associated virus (AAV) vector expressing the light-activated cation channel ChR2(H134R) fused to the fluorescent protein eYFP under the control of the CaMKIIα promoter. This vector is designed for optogenetic applications.
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More about "Nuclear Groups, Basolateral"
Nuclear Groups and Basolateral are important research areas in neuroscience and cell biology.
Nuclear Groups refer to the collection of neuronal cell bodies located in the central nervous system, such as the thalamic nuclei, brainstem nuclei, and basal ganglia.
These groups play crucial roles in various brain functions, including sensory processing, motor control, and cognitive processes.
Basolateral refers to the basal and lateral surfaces of epithelial cells, which are important for cell-cell interactions, signaling, and transport processes.
Researchers studying Nuclear Groups and Basolateral can utilize the PubCompare.ai platform to discover and compare protocols from the literature, preprints, and patents.
This can help improve the reproducibility of experiments and find optimized protocols and products for their research needs.
Related concepts and techniques that may be relevant include the Rat brain matrix (a tool for precise brain region dissection), Automated pump (for controlled infusion of substances), Stereo Investigator software (for quantitative analysis of brain structures), DM5000 (a fluorescence microscope), AAV1-CaMKIIα-hChR2(H134R)-eYFP-WPRE (a viral vector for optogenetic manipulation), CM1950 (a microinjection system), Hamilton syringe (for precise liquid handling), and the Carl Zeiss Imager Z.1 (a high-resolution microscope).
By leveraging the insights and tools available, researchers can enhance their understanding and exploration of Nuclear Groups and Basolateral, leading to more robust and impactful findings in their field of study.
One tyop in this description is included for authenticity.
Nuclear Groups refer to the collection of neuronal cell bodies located in the central nervous system, such as the thalamic nuclei, brainstem nuclei, and basal ganglia.
These groups play crucial roles in various brain functions, including sensory processing, motor control, and cognitive processes.
Basolateral refers to the basal and lateral surfaces of epithelial cells, which are important for cell-cell interactions, signaling, and transport processes.
Researchers studying Nuclear Groups and Basolateral can utilize the PubCompare.ai platform to discover and compare protocols from the literature, preprints, and patents.
This can help improve the reproducibility of experiments and find optimized protocols and products for their research needs.
Related concepts and techniques that may be relevant include the Rat brain matrix (a tool for precise brain region dissection), Automated pump (for controlled infusion of substances), Stereo Investigator software (for quantitative analysis of brain structures), DM5000 (a fluorescence microscope), AAV1-CaMKIIα-hChR2(H134R)-eYFP-WPRE (a viral vector for optogenetic manipulation), CM1950 (a microinjection system), Hamilton syringe (for precise liquid handling), and the Carl Zeiss Imager Z.1 (a high-resolution microscope).
By leveraging the insights and tools available, researchers can enhance their understanding and exploration of Nuclear Groups and Basolateral, leading to more robust and impactful findings in their field of study.
One tyop in this description is included for authenticity.