IHC staining for Trk A, B, and C expression was performed with pan-Trk monoclonal antibody (mAb) clone EPR17341 (Abcam, Cambridge, MA). The antibody is reactive to a homologous region of Trk-A, -B, and –C near the C terminus, and the exact sequence is proprietary. This commercial antibody is not FDA approved for clinical use to our best knowledge. Testis tissue, ganglia of the colonic plexus submucosus and cortical brain tissue were used as positive control tissues. EPR17341 was used at 6ug/ml. All assays were performed on a Leica-Bond-3 (Leica, Buffalo Grove, IL) automated stainer platform using a heat based antigen retrieval method and high pH buffer solution (ER2, Leica). Non-neoplastic lymphocytes, hepatocytes, colorectal epithelium, alveolar epithelium, and renal cortex were used as negative external controls. All available cases with NTRK rearrangements on MSK-IMPACT as well as 20 consecutive tumors without evidence of NTRK fusion on Archer were stained.
Myenteric Plexus
The Myenteric Plexus, also known as the Auerbach's Plexus, is a key component of the enteric nervous system that regulates the motility and function of the gastrointestinal tract.
It is a network of neurones and supporting cells located between the longitudinal and circular muscle layers of the esophagus, stomach, small and large intestines.
The Myenteric Plexus plays a crucial role in coordinating peristaltic movements, secretion of digestive enzymes, and blood flow regulation within the digestive system.
Researching the Myenteric Plexus is essential for understanding the pathophysiology of various gastrointestinal disorders, such as irritable bowle syndrome, Hirschsprung's disease, and gastroparesis.
PubCompare.ai is an invaluable AI-driven platform that can optimize your Myenteric Plexus research by locating relevant protocols from literature, preprints, and patents, and leveraging AI-comparisons to identify the best methods and products.
This powerful tool can enhance the reproductibility and accuracy of your Myenteric Plexus studies.
It is a network of neurones and supporting cells located between the longitudinal and circular muscle layers of the esophagus, stomach, small and large intestines.
The Myenteric Plexus plays a crucial role in coordinating peristaltic movements, secretion of digestive enzymes, and blood flow regulation within the digestive system.
Researching the Myenteric Plexus is essential for understanding the pathophysiology of various gastrointestinal disorders, such as irritable bowle syndrome, Hirschsprung's disease, and gastroparesis.
PubCompare.ai is an invaluable AI-driven platform that can optimize your Myenteric Plexus research by locating relevant protocols from literature, preprints, and patents, and leveraging AI-comparisons to identify the best methods and products.
This powerful tool can enhance the reproductibility and accuracy of your Myenteric Plexus studies.
Most cited protocols related to «Myenteric Plexus»
Antigens
Biological Assay
Brain
Buffaloes
Buffers
Clone Cells
Colon
Epithelium
Ganglia
Gene Rearrangement
Hepatocyte
Immunoglobulins
Kidney Cortex
Lymphocyte
Monoclonal Antibodies
Myenteric Plexus
Neoplasms
Testis
Tissues
We prepared whole-mount preparations of the colonic myenteric plexus by removing the mucosa, submucosa and circular muscle by microdissection. We processed the resulting longitudinal muscle myenteric plexus (LMMP) preparations for immunohistochemistry as described by Gulbransen and Sharkey25 (link) with the antibodies listed in Supplementary Table 1 . We confirmed antibody specificity by pre-absorption with control peptides or in respective knockout animals.
Animals
Antibodies
Antibody Specificity
Colon
Immunohistochemistry
Microdissection
Mucous Membrane
Muscle Tissue
Myenteric Plexus
Peptides
A-factor (Streptomyces)
Biopsy
Blood Vessel
Buffers
Diabetes Mellitus
Duodenum
factor A
Gastric Bypass
Gastroepiploic Artery
Gastrointestinal Diseases
Gastroparesis
Human Body
Malignant Neoplasms
Muscle Tissue
Myenteric Plexus
Obesity
Operative Surgical Procedures
paraform
Patients
Pharmacotherapy
Phosphates
Pylorus
Radiotherapy
Stomach
Tissues
Briefly, rats were decapitated, the whole intestine removed and immediately stored in MEM (Gibco) on ice. The muscle and submucous layer were separated, and the dissected muscle tissue incubated in a collagenase II solution (Worthington) for 2 hours. After vortexing the tissue for 20 seconds the suspension was filtered through a sterile syringe filter (Cell strainer, BD biosciences), pore size 40 µm. The supernatant was centrifuged three times, the obtained myenteric plexus incubated in accutase (PAA Laboratories GmbH) for 20 minutes and dissociated by trituration. In a second centrifugation step, the accutase was removed and replaced with 10 ml culture medium. The cells were kept at standardized densities (1×106 cells/25 cm2 flask) in culture medium adapted for expansion cultures [Neurobasal A (Gibco) supplemented with 2% B27- (without vitamin A, Gibco), 1% albumin (Sigma), 0.25% -2-mercaptoethanol (50 mM, Invitrogen), 0.12% glutamine (200 mM, Sigma), EGF (10 ng/ml, Tebue), bFGF (20 ng/ ml; Tebue), GDNF (10 ng/ml, Tebue), gentamycin and metronidazole (100 µg/ml)]. After two days in vitro, the first floating neurospheres (EnNS’s) were seen, while neurons and radiating glial cells expanded at the bottom.
2-Mercaptoethanol
accutase
Albumins
Cells
Centrifugation
Collagenase
Culture Media
Gentamicin
Glial Cell Line-Derived Neurotrophic Factor
Glutamine
Intestines
Metronidazole
Muscle Tissue
Myenteric Plexus
Neuroglia
Neurons
Rattus
Sterility, Reproductive
Syringes
Tissues
Vision
Vitamin A
All animal care and experimental procedures were in accordance with and approved by the Institutional Animal Care and Use Committee at Virginia Commonwealth University.
All chemicals and reagents were obtained from Sigma Aldrich (St Louis, MO), unless otherwise noted, except cell culture reagents, which were purchased from Gibco (Grand Island, NY). Male Swiss Webster mice (26–30 g, Harlan Sprague Dawley, Inc.) were killed by cervical dislocation. The ileum was immediately dissected and placed in ice-cold Krebs solution (in mM: 118 NaCl, 4.6 KCl, 1.3 NaH2PO4, 1.2 MgSO4 25 NaHCO3, 11 glucose and 2.5 CaCl2) bubbled with carbogen (95% O2/5% CO2). Ileal segments were threaded longitudinally on a plastic rod through the lumen and the longitudinal muscle with the myenteric plexus (LMMP) was gently removed using a cotton-tipped applicator. LMMP strips were rinsed three times in 1 ml Krebs and gathered by centrifugation (350× g, 30 s). LMMP strips were then minced with scissors and digested in 1.3 mg/ml collagenase type II (Worthington) and 0.3 mg/ml bovine serum albumin in bubbled Krebs (37°C) for 1 hour, followed by 0.05% trypsin for 7 min. Following each digestion, cells were triturated and collected by centrifuge (350× g for 8 min). Cells were then plated on laminin (BD Biosciences) and poly-D-lysine coated coverslips in Neurobasal A media containing B-27 supplement, 1% fetal bovine serum, 10 ng/ml glial cell line-derived neurotrophic factor (GDNF, Neuromics, Edina, MN), and antibiotic/antimycotic liquid. Half of the cell media was changed every 2–3 days.
All chemicals and reagents were obtained from Sigma Aldrich (St Louis, MO), unless otherwise noted, except cell culture reagents, which were purchased from Gibco (Grand Island, NY). Male Swiss Webster mice (26–30 g, Harlan Sprague Dawley, Inc.) were killed by cervical dislocation. The ileum was immediately dissected and placed in ice-cold Krebs solution (in mM: 118 NaCl, 4.6 KCl, 1.3 NaH2PO4, 1.2 MgSO4 25 NaHCO3, 11 glucose and 2.5 CaCl2) bubbled with carbogen (95% O2/5% CO2). Ileal segments were threaded longitudinally on a plastic rod through the lumen and the longitudinal muscle with the myenteric plexus (LMMP) was gently removed using a cotton-tipped applicator. LMMP strips were rinsed three times in 1 ml Krebs and gathered by centrifugation (350× g, 30 s). LMMP strips were then minced with scissors and digested in 1.3 mg/ml collagenase type II (Worthington) and 0.3 mg/ml bovine serum albumin in bubbled Krebs (37°C) for 1 hour, followed by 0.05% trypsin for 7 min. Following each digestion, cells were triturated and collected by centrifuge (350× g for 8 min). Cells were then plated on laminin (BD Biosciences) and poly-D-lysine coated coverslips in Neurobasal A media containing B-27 supplement, 1% fetal bovine serum, 10 ng/ml glial cell line-derived neurotrophic factor (GDNF, Neuromics, Edina, MN), and antibiotic/antimycotic liquid. Half of the cell media was changed every 2–3 days.
Animals
Antibiotics
Bicarbonate, Sodium
carbogen
Cell Culture Techniques
Cells
Centrifugation
Common Cold
Dietary Supplements
Digestion
Fetal Bovine Serum
Glial Cell Line-Derived Neurotrophic Factor
Glucose
Gossypium
Ileum
Institutional Animal Care and Use Committees
Joint Dislocations
Krebs-Ringer solution
Laminin
Lysine
Males
Matrix Metalloproteinase 13
Mouse, Swiss
Muscle Tissue
Myenteric Plexus
Neck
Poly A
Serum Albumin, Bovine
Sodium Chloride
Sulfate, Magnesium
Trypsin
Most recents protocols related to «Myenteric Plexus»
Distal colon segments were fixed in 0.1M PBS containing 4% paraformaldehyde at room temperature for 3 h. Whole mounts of longitudinal muscle and myenteric plexus were obtained by microdissection and permeabilized with PBS containing 10% horse serum (HS) and 4% Triton X-100 for 2 h at room temperature. Tissues were then incubated with the following primary antibodies: rabbit anti-ChAT (1:1000, a gift from Professor M. Schemann, Hannover, Germany) (Schemann et al., 1993 (link)), mouse anti-neuronal NOS (nNOS; 610308, 1:500; BD Biosciences), rabbit anti-GR (D8H2, 3660S 1:500, Cell Signaling), human anti-Hu (Bodin et al., 2021 (link)) (gift from the CHU of Nantes; 1:5000) diluted in PBS containing 10% horse serum and 0.5% Triton X-100 for 24 or 48 h at room temperature. After washing, tissues were incubated for 2 h at room temperature with the appropriate secondary antibodies, respectively, anti-rabbit CY5 (1:500), anti-rabbit CY3 (1:500) and anti-human FP488 (1:200), and mounted Glycerol 60% (vol/vol) (Thermo Fisher Scientific). Nuclei were stained with 4′ 6-Diamidino-2-phenylindole dihydrochloride (DAPI D9542; 1:10000; Sigma Aldrich, Paris, France).
Antibodies
Cell Nucleus
Colon
DAPI
Equus caballus
Glycerin
Homo sapiens
Microdissection
Mus
Muscle Tissue
Myenteric Plexus
Neurons
NOS1 protein, human
paraform
Rabbits
Serum
Tissues
Triton X-100
The muscularis externa containing myenteric plexuses was prepared from mouse GITs as previously described with some modification (65 (link), 66 (link)). For HILIC-MS/MS analysis, mouse GIT segments were cut open along the attachment line of the mesentery and then placed onto a cold surface with the muscularis externa facing up. The muscularis externa of the GIT segments was isolated by gently scraping the outer layer with watchmaker tweezers under a binocular stereomicroscope. For whole-mount immunostaining, the mouse duodenum was cut open along the mesentery line, pinned onto a rubber plate, and then fixed with 4% paraformaldehyde overnight at 4 °C. The muscularis layer was then gently separated from the GIT segment using watchmaker tweezers and a cotton swab under a binocular stereomicroscope. For RNA extraction, mouse GITs were immersed in saturated ammonium sulfate solution containing 20 mM EDTA and 25 mM sodium citrate (pH5.2) to inhibit RNA degradation. The muscularis externa of the segments was placed over a glass rod and then peeled away using a cotton swab along the attachment line of the mesentery under a binocular stereomicroscope as described (67 ). Isolated muscularis externa was stored in saturated ammonium sulfate solution and then frozen until further use.
Bladder Detrusor Muscle
Cardiac Arrest
Cold Temperature
Duodenum
Edetic Acid
Freezing
Gossypium
Mesentery
Mice, Laboratory
Myenteric Plexus
paraform
RNA Degradation
Rubber
Sodium Citrate
Sulfate, Ammonium
Tandem Mass Spectrometry
Gastric and intestinal tissue of n = 19 guinea pigs were used. The stomach was cut along the greater and smaller curvature and the small and large intestine were cut along the mesenteric border, contents were removed, and the tissue was carefully washed. Tissue samples were pinned mucosal side up in Sylgard®-coated Petri dishes. Mucosa and submucosa were carefully removed under microscopic control (Olympus SZ30 stereo microscope, Olympus Corporation, Hamburg, Germany). Muscle-myenteric plexus preparations (MPPs) were cut in the direction of visible muscle fibers to examine circular muscle motility, and vertically to visible muscle fibers for investigation of longitudinal muscle motility, dissecting MPP preparations of 2 × 1 cm for both gastric and intestinal tissue. For the stomach, two MPPs were cut from both the oral fundus and the aboral antrum, and four MPPs were cut from the corpus. For the intestine, MPPs were cut from the ileum and from the proximal colon.
Threads were knotted on both ends of each MPP that were then put into organ baths containing 12 mL of oxygenated Krebs solution (in mmol/L: 1.2 MgCl2, 2.5 CaCl2, 1.2 NaH2PO4, 117 NaCl, 20 NaHCO3, 11 glucose, 4.7 KCl) at 37 °C. While one thread tied the MPP to the organ bath, the other one was knotted to an isometric force transducer (Spider). Platinum electrodes connected to an electric stimulator (Grass S88 Dual output pulse stimulator, Grass Instruments, RI, USA) were placed at each side of single MPPs for electrical field stimulations (EFS; 30 V, 10 Hz, 0.5 ms individual pulse duration for 10 s). Stimulation parameters were chosen to exclusively stimulate neuronal-mediated smooth muscle activity. After a 30 min equilibration with a preload of 30 mN, the buffer was changed and tissue vitality and responsiveness were proved by EFS repeated three times with 15 to 20 min in between. Ten minutes after the last EFS and when a stable baseline was reached again, 10 µM cocaine was applied to the bath. The motility was recorded for 50 min and then a further EFS was applied, followed by washout. In order to investigate whether the effect of cocaine application was nerve-mediated, in a set of experiments 1 µM tetrodotoxin (TTX) was applied to the bath 20 min prior to cocaine addition, blocking the voltage-gated neuronal Na+ channels.
Threads were knotted on both ends of each MPP that were then put into organ baths containing 12 mL of oxygenated Krebs solution (in mmol/L: 1.2 MgCl2, 2.5 CaCl2, 1.2 NaH2PO4, 117 NaCl, 20 NaHCO3, 11 glucose, 4.7 KCl) at 37 °C. While one thread tied the MPP to the organ bath, the other one was knotted to an isometric force transducer (Spider). Platinum electrodes connected to an electric stimulator (Grass S88 Dual output pulse stimulator, Grass Instruments, RI, USA) were placed at each side of single MPPs for electrical field stimulations (EFS; 30 V, 10 Hz, 0.5 ms individual pulse duration for 10 s). Stimulation parameters were chosen to exclusively stimulate neuronal-mediated smooth muscle activity. After a 30 min equilibration with a preload of 30 mN, the buffer was changed and tissue vitality and responsiveness were proved by EFS repeated three times with 15 to 20 min in between. Ten minutes after the last EFS and when a stable baseline was reached again, 10 µM cocaine was applied to the bath. The motility was recorded for 50 min and then a further EFS was applied, followed by washout. In order to investigate whether the effect of cocaine application was nerve-mediated, in a set of experiments 1 µM tetrodotoxin (TTX) was applied to the bath 20 min prior to cocaine addition, blocking the voltage-gated neuronal Na+ channels.
Antral
Bath
Bicarbonate, Sodium
Buffers
Cavia
Cocaine
Colon
Electricity
Glucose
Hyperostosis, Diffuse Idiopathic Skeletal
Ileum
Intestines
Krebs-Ringer solution
Large Intestine
Magnesium Chloride
Mesentery
Microscopy
Motility, Cell
Mucous Membrane
Muscle Tissue
Myenteric Plexus
Nervousness
Neurons
Platinum
Poaceae
Pulse Rate
Smooth Muscles
Sodium Chloride
Spiders
Stimulations, Electric
Stomach
Tetrodotoxin
Tissues
Transducers
Small intestinal tissue from n = 14 guinea pigs was used to obtain primary culture of enteric neurons as described elsewhere (Kugler et al., 2015, 2018).
Briefly, after mechanical separation of longitudinal muscle-myenteric plexus preparation and following enzymatic digestion, cell culture dishes (Ibidi) were seeded with 200 µL of myenteric ganglia suspension. This was incubated in medium M199 supplemented with 10% fetal bovine serum (FBS) (Gibco), 50–100 ng mL−1 mouse nerve growth factor 7S (Alomone labs, Jerusalem, Israel), 5 mg mL−1 Glucose, 100 U mL−1 Penicillin, 100 mg mL−1 Streptomycin (Gibco) and 2 mM arabinose-C-furanoside (Sigma–Aldrich, St. Louis, MO, USA). The neurons were cultured in vitro under standard culture conditions (5% CO2; 37 °C; humidity 95%). Medium with additives was changed every second day. The cultures were grown for at least two weeks to obtain interconnected neuronal clusters.
Briefly, after mechanical separation of longitudinal muscle-myenteric plexus preparation and following enzymatic digestion, cell culture dishes (Ibidi) were seeded with 200 µL of myenteric ganglia suspension. This was incubated in medium M199 supplemented with 10% fetal bovine serum (FBS) (Gibco), 50–100 ng mL−1 mouse nerve growth factor 7S (Alomone labs, Jerusalem, Israel), 5 mg mL−1 Glucose, 100 U mL−1 Penicillin, 100 mg mL−1 Streptomycin (Gibco) and 2 mM arabinose-C-furanoside (Sigma–Aldrich, St. Louis, MO, USA). The neurons were cultured in vitro under standard culture conditions (5% CO2; 37 °C; humidity 95%). Medium with additives was changed every second day. The cultures were grown for at least two weeks to obtain interconnected neuronal clusters.
Arabinose
Cavia
Cell Culture Techniques
Digestion
Enzymes
Fetal Bovine Serum
Ganglia
Glucose
Humidity
Hyperostosis, Diffuse Idiopathic Skeletal
Intestines, Small
Mice, House
Muscle Tissue
Myenteric Plexus
Nerve Growth Factors
Neurons
Penicillins
Streptomycin
Tissues
Anesthetized (using sodium pentobarbital, 80 mg/kg, intraperitoneally) adult mice were transcardially perfused with PBS before introducing 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4). Then, the tongue was dissected and post-fixed in 4% paraformaldehyde overnight at 4 °C. The tongues were kept in 30% sucrose solution for 5–6 d. The tissue was mounted in a cryostat embedding medium and snap-frozen in powdered dry ice. The tongues were sectioned using a cryostat (thickness of 30 µm) and kept at 4 °C in PBS for storage until used for immunohistochemistry. For the myenteric plexus, the whole mount of the duodenum was prepared as described in our previous study [19 (link)].
Adult
Buffers
Dry Ice
Duodenum
Freezing
Immunohistochemistry
Mice, House
Myenteric Plexus
paraform
Pentobarbital Sodium
Phosphates
Sucrose
Tissues
Tongue
Top products related to «Myenteric Plexus»
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Collagenase XI is a purified enzyme preparation derived from Clostridium histolyticum. It is used for the dissociation and dispersion of a variety of tissues, including connective tissue.
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Dispase is a highly active neutral protease that can be used to dissociate a variety of cell types from tissue samples or cell culture. It efficiently separates cells while minimizing damage to cell surface antigens, enabling downstream applications such as flow cytometry or cell culture.
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Streptomycin is a broad-spectrum antibiotic used in laboratory settings. It functions as a protein synthesis inhibitor, targeting the 30S subunit of bacterial ribosomes, which plays a crucial role in the translation of genetic information into proteins. Streptomycin is commonly used in microbiological research and applications that require selective inhibition of bacterial growth.
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Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.
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Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It contains a combination of penicillin and streptomycin, which are broad-spectrum antibiotics that inhibit the growth of both Gram-positive and Gram-negative bacteria.
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Penicillin is a type of antibiotic used in laboratory settings. It is a broad-spectrum antimicrobial agent effective against a variety of bacteria. Penicillin functions by disrupting the bacterial cell wall, leading to cell death.
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DMEM (Dulbecco's Modified Eagle's Medium) is a cell culture medium formulated to support the growth and maintenance of a variety of cell types, including mammalian cells. It provides essential nutrients, amino acids, vitamins, and other components necessary for cell proliferation and survival in an in vitro environment.
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More about "Myenteric Plexus"
The Myenteric Plexus, also known as Auerbach's Plexus, is a crucial component of the enteric nervous system that regulates the motility and function of the gastrointestinal tract.
This intricate network of neurons and supporting cells is located between the longitudinal and circular muscle layers of the esophagus, stomach, small and large intestines.
The Myenteric Plexus plays a pivotal role in coordinating peristaltic movements, secretion of digestive enzymes, and blood flow regulation within the digestive system.
Understanding the Myenteric Plexus is essential for unraveling the pathophysiology of various gastrointestinal disorders, such as irritable bowel syndrome, Hirschsprung's disease, and gastroparesis.
Researchers studying the Myenteric Plexus may utilize a variety of tools and techniques, including Collagenase XI, Dispase, Streptomycin, Bovine serum albumin (BSA), Fetal bovine serum (FBS), Penicillin/streptomycin, Penicillin, Nicardipine, Dulbecco's Modified Eagle Medium (DMEM), and DMEM/Nutrient Mixture F-12.
These reagents and media can be invaluable in isolating, culturing, and analyzing Myenteric Plexus cells and tissues.
PubCompare.ai, an AI-driven platform, can optimize Myenteric Plexus research by locating relevant protocols from literature, preprints, and patents, and leveraging AI-comparisons to identify the best methods and products.
This powerful tool can enhance the reproducibility and accuracy of your Myenteric Plexus studies, unlocking new insights into this critical component of the enteric nervous system.
This intricate network of neurons and supporting cells is located between the longitudinal and circular muscle layers of the esophagus, stomach, small and large intestines.
The Myenteric Plexus plays a pivotal role in coordinating peristaltic movements, secretion of digestive enzymes, and blood flow regulation within the digestive system.
Understanding the Myenteric Plexus is essential for unraveling the pathophysiology of various gastrointestinal disorders, such as irritable bowel syndrome, Hirschsprung's disease, and gastroparesis.
Researchers studying the Myenteric Plexus may utilize a variety of tools and techniques, including Collagenase XI, Dispase, Streptomycin, Bovine serum albumin (BSA), Fetal bovine serum (FBS), Penicillin/streptomycin, Penicillin, Nicardipine, Dulbecco's Modified Eagle Medium (DMEM), and DMEM/Nutrient Mixture F-12.
These reagents and media can be invaluable in isolating, culturing, and analyzing Myenteric Plexus cells and tissues.
PubCompare.ai, an AI-driven platform, can optimize Myenteric Plexus research by locating relevant protocols from literature, preprints, and patents, and leveraging AI-comparisons to identify the best methods and products.
This powerful tool can enhance the reproducibility and accuracy of your Myenteric Plexus studies, unlocking new insights into this critical component of the enteric nervous system.