Primary rat neural cells (PRNCs; consisted of 40% neurons and 60% astrocytes) were obtained from BrainBit (E18 rat cortex; Springfield, IL, USA). As described elsewhere15 (link), cells (4 × 104 cells/well) were suspended in 200 μl Neural Medium (NbActive 4, BrainBit) containing 2 mM l- glutamine and 2% B27 in the absence of antibiotics and grown in poly-l-lysine-coated 96-well plates at 37 °C in humidified atmosphere containing 95% O2 and 5% CO2. After 3 days in culture, PRNCs were exposed to 1 μM oxytocin (O4375, Sigma-Aldrich, St. Louis, MI, USA), 1 μM oxytocin + 10 μM atosiban (A3480, Sigma-Aldrich), 10 μM atosiban, and the absence of regents (control) for 3 days at 37 °C. After 6 days in culture (Fig. 1 ), RPNCs were exposed to OGD as described previously15 (link). The cells were initially exposed to OGD medium (glucose-free Dulbecco’s Modified Eagle Medium), then placed in an anaerobic chamber containing 95% N2 and 5% CO2 for 15 min at 37 °C (preincubation), for 90 min at 37 °C (culture medium pH 6.7~6.8; mimicking the acidic environment of ischemic brain in vivo). OGD was terminated by adding 5 mM glucose to medium and cell cultures were re-introduced to the regular CO2 incubator at 37 °C for 2 h. Control cells were incubated in the same buffer containing 5 mM glucose at 37 °C in a regular 95% O2 and 5% CO2 incubator.
Oxytocin
Manufactured by Merck Group
Sourced in United States
Oxytocin is a hormone produced in the hypothalamus and released by the posterior pituitary gland. It plays a crucial role in childbirth and lactation. This synthetic version of the hormone is used for research purposes in laboratories.
Automatically generated - may contain errors
Lab products found in correlation
4 aminopyridine 4 ap, by Merck Group (3 mentions)
Atosiban, by Merck Group (2 mentions)
Isoflurane, by Zoetis (2 mentions)
Nifedipine, by Merck Group (2 mentions)
Capsaicin, by Merck Group (2 mentions)
Acetonitrile, by Merck Group (2 mentions)
Quinpirole, by Merck Group (2 mentions)
Nitrocellulose membrane, by GE Healthcare (2 mentions)
Protein g sepharose 4 fast flow, by GE Healthcare (2 mentions)
Anti rabbit secondary antibody, by Cell Signaling Technology (2 mentions)
Protease inhibitor cocktail, by Cell Signaling Technology (2 mentions)
L 371 257, by Bio-Techne (2 mentions)
Ecl select, by GE Healthcare (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Pgf2α, by Cayman Chemical (2 mentions)
Carbachol, by Merck Group (2 mentions)
Acetylcholine, by Merck Group (2 mentions)
Indomethacin, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
62 protocols using oxytocin
1
Oxytocin and OGD Effects on Primary Rat Neural Cells
2
Oxytocin Modulation of IL-6 Secretion in Ovarian Cancer
3
Milking Mouse Dams for Breast Milk Collection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse milk was collected using a homemade suction apparatus consisting of two tubing segments and a 50 ml conical tube. Suction was created using the first tubing segment where one end was connected to vacuum at negative pressures between −18 to −20 mmHg and the other end of the tubing was inserted into a sealed 50 mL conical tube. The second tubing segment was used as the collection system. One end was inserted into the sealed 50 mL conical tube and the hub end of the tubing was placed around the teat creating a seal during the milking process.
The milking process of the dams was carried out at 10–14 days’ post-parturition. The dams were separated from their pups for 4–5 hours before milking. Isoflurane (Zoetis, Parsippany, NJ) was used for anesthesia. Dams were given 5 IU of oxytocin (Sigma-Aldrich, St. Louis, MO) intraperitoneally to induce milk flow. After 3–5 minutes, gentle finger massage around the mammary glands was used to stimulate milk ejection. The milk was stored at −80°C until further analysis.
The milking process of the dams was carried out at 10–14 days’ post-parturition. The dams were separated from their pups for 4–5 hours before milking. Isoflurane (Zoetis, Parsippany, NJ) was used for anesthesia. Dams were given 5 IU of oxytocin (Sigma-Aldrich, St. Louis, MO) intraperitoneally to induce milk flow. After 3–5 minutes, gentle finger massage around the mammary glands was used to stimulate milk ejection. The milk was stored at −80°C until further analysis.
4
Two-Photon Imaging of Calcium Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bolus load of the calcium indicator Oregon Green 488 BAPTA-1 AM (OGB-1, Invitrogen) was performed as described7 (link). Imaging was performed by using a two-photon microscope (MOM, Sutter, or A1RMP, Nikon) and a mode-locked Ti:Sapphire laser (MaiTai, Spectra Physics or Chamaleon, Coherent, λ = 810 nm). Consecutive xyt-stacks were acquired at a frame rate of 4-7 Hz (pixel size 0.3 - 0.6 μm) through a 40x (0.8 NA, Olympus) or a 16x (0.8 NA, Nikon) water-immersion objective, controlled by ScanImage70 (link) or NIS-Elements AR4.51.00 software (Nikon).
For in vivo experiments, oxytocin (1 μM, Sigma) and the oxytocin receptor antagonist (desGly-NH2,d(CH2)5[D-Tyr2,Thr4]OVT, 250 μM, synthesized and kindly donated by Dr. Maurice Manning, University of Toledo) was diluted in cortex buffer solution7 (link) and applied topically at the craniotomy. The craniotomy was filled with approximately 300 μl of solution and that volume was kept constant until the end of the experiment.
For in vivo experiments, oxytocin (1 μM, Sigma) and the oxytocin receptor antagonist (desGly-NH2,d(CH2)5[D-Tyr2,Thr4]OVT, 250 μM, synthesized and kindly donated by Dr. Maurice Manning, University of Toledo) was diluted in cortex buffer solution7 (link) and applied topically at the craniotomy. The craniotomy was filled with approximately 300 μl of solution and that volume was kept constant until the end of the experiment.
5
Breast Milk Collection and Preservation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Breast milk specimens (whole milk collection) were obtained within four days after birth (colostrum), at 1 month and 6 months, and were immediately frozen (− 80 °C) for long-term storage to avoid lipopolysaccharide (LPS) contamination88 (link). Milk samples were thawed immediately before testing and were centrifuged (10,000× g, 10 min) to obtain surface lipid, interface liquid, and cell-containing pellets. For milk collection from mice, lactating mice were separated from offspring in the same cage with a divider for 3 h prior to milking. Mice were administered with 3 IU of oxytocin (Sigma) via i.p. and within 1 min after oxytocin injection, milk collection was performed using a milking apparatus (KN-591, Natsume Tokyo, Japan) attached to an aspirator.
6
Recombinant Cytokine Stimulation of Cells
7
Oxytocin Enhances Traumatic Memory Reactivation Resilience
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rats received two remodeling sessions beginning with an oxytocin (Sigma-Aldrich, Saint Quentin Fallavier, France) or saline (0.9%) infusion delivered 40 min before the trauma reactivation induced by the exposure to a trauma-associated cue. oxytocin was administered i.c.v. at a dose of 0.1 µg (in 1 µl of saline) chosen on the basis of previous studies17 (link),29 ,30 (link), as well as of preliminary experiments showing that it significantly reduced anxiety in traumatized rats after acute ICV injections.
The infusions was performed via an infusion cannula (25G, extended 2 mm beyond the guide cannula) connected via polyethylene tubing to a 10-µl syringe (Hamilton) during 10 min (1 µl/min) delivered in a new room dimly lighted (15 lux). On the first session (day 29), rats were placed in the white safe box of the inhibitory avoidance apparatus22 (link). On the second session (day 31), they were placed in a novel box and exposed to the SPS-associated tone (metronome, 120 bpm). Upon completion of each session, rats remained for 1 h in the dimly light room before they returned to the colony room.
The infusions was performed via an infusion cannula (25G, extended 2 mm beyond the guide cannula) connected via polyethylene tubing to a 10-µl syringe (Hamilton) during 10 min (1 µl/min) delivered in a new room dimly lighted (15 lux). On the first session (day 29), rats were placed in the white safe box of the inhibitory avoidance apparatus22 (link). On the second session (day 31), they were placed in a novel box and exposed to the SPS-associated tone (metronome, 120 bpm). Upon completion of each session, rats remained for 1 h in the dimly light room before they returned to the colony room.
8
Milk Collection and Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At postnatal day 17, nurses used in the cross-fostering study were anaesthetised with an intraperitoneal injection of 0.1% (w/v) xylazine, 0.5% (w/v) ketaset in 0.9% (w/v) NaCl at 16 µl/g mouse. Milk collection was aided by an intraperitoneal injection of 200 µl 10 IU/ml oxytocin (Sigma Aldrich) in 0.1% PBS. A vacuum pump was used to harvest 100–200 µl milk. Milk fat and protein content were analysed as described [48] (link). After fat removal, protein was diluted 1/5 in 50 mM Tris HCl (pH 8.0), 150 mM NaCl, 1% (v/v) Igepal CA-630, boiled for 10 minutes, and mixed with an equal volume of reducing sample buffer. Samples were run on a 15% Criterion Tris-HCl gel (Bio-Rad Laboratories) alongside pre-stained molecular weight markers. Fixation and drying were performed using standard methods.
After milk collection, nurses were humanely killed by cervical dislocation and pituitary glands harvested. RNA was extracted from pituitaries using TRI Reagent (Sigma Aldrich). Total RNA (1 µg) was DNase-treated with RQ1 RNase-free DNase I (Promega). cDNA was synthesised using random hexamers with Superscript III RNase H− Reverse Transcriptase (Invitrogen). Real-time PCR (qRT-PCR) was used to measure expression of prolactin and growth hormone normalised to β-actin. Reactions were performed in duplicate and analysed as described previously [8] (link).
After milk collection, nurses were humanely killed by cervical dislocation and pituitary glands harvested. RNA was extracted from pituitaries using TRI Reagent (Sigma Aldrich). Total RNA (1 µg) was DNase-treated with RQ1 RNase-free DNase I (Promega). cDNA was synthesised using random hexamers with Superscript III RNase H− Reverse Transcriptase (Invitrogen). Real-time PCR (qRT-PCR) was used to measure expression of prolactin and growth hormone normalised to β-actin. Reactions were performed in duplicate and analysed as described previously [8] (link).
9
Cardiac Progenitor Sphere Generation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For generation of CardioStem spheres,21 0.25 to 0.5×105 mCPCs were seeded on 6‐well dishes with culture medium (35% Iscove's modified Dulbecco's medium, 32.5% DMEM, 32.5% F12, and 3.5% bovine growth serum [HyClone]) with 100 nmol/L oxytocin (Sigma) for 3 days, then trypsinized and cultured on bacterial dishes (P100) for 2 to 3 days to induce CardioStem sphere formation. For rCPCs, 1.0×105 cells were seeded on 6‐well dishes with culture medium (F12 with 10% FCS) with 100 nmol/L oxytocin for 3 days, then trypsinized and cultured on bacterial dishes (P100) for 3 days.
10
Oxytocin Administration in Mice
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!