The GMH injury model and lesion grading system utilized was as we previously described [5 (link)]. Briefly, Clostridium-derived collagenase (Type VII-S collagenase, C2399-1.5 KU, Sigma-Aldrich) was injected into the SVZ of mouse pups at P4 to induce direct spontaneous non-traumatic vessel rupture with intracerebral hemorrhage in the region of the germinal matrix and SVZ. Sham PBS injections were performed to ensure that hemorrhage was a result of collagenase injection and not from mechanical insertion of the needle or dynamics of a fluid injection. The animal-specific injury grading system was used that established a distinction between parenchymal injury, ventricular involvement, and PHH. The grading system has been described [5 (link)], and is as follows: 0 = No lesion or ventricular enlargement; 1 = Lesion volume < 30% of hemispheric cortical tissue ipsilateral to injury site without ventricular involvement; 2 = Lesion volume > 30% of hemispheric cortical tissue ipsilateral to injury site without ventricular involvement; 3 = Lesion extending into the ipsilateral ventricle with no ventricular enlargement; 4 = Lesion extending into the ipsilateral ventricle coupled with unilateral ventriculomegaly; 5 = Lesion extending into both ventricles resulting in bilateral ventriculomegaly (global hydrocephalus).
Collagenase type 7 s
Manufactured by Merck Group
Sourced in United States
Collagenase type VII-S is a laboratory enzyme used for the digestion and dissociation of collagen-rich tissues. It is a mixture of proteolytic enzymes derived from Clostridium histolyticum. The product is suitable for use in cell culture, tissue engineering, and other applications where the breakdown of collagen-based substrates is required.
Automatically generated - may contain errors
Lab products found in correlation
Stereotaxic frame, by Stoelting (2 mentions)
Microm hm550, by Thermo Fisher Scientific (1 mentions)
Stereotaxic frame, by Kopf Instruments (1 mentions)
Cd 1 mice, by Charles River Laboratories (1 mentions)
Stereotactic frame, by Stoelting (1 mentions)
Stereotaxic instrument, by Stoelting (1 mentions)
12 protocols using collagenase type 7 s
1
Clostridium-Induced Germinal Matrix Hemorrhage Model
2
Collagenase-Induced Intracerebral Hemorrhage in Mice
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ICH was induced by injecting collagenase into the left striatum of mice according to a previously described method19 (link)45 (link). Briefly, mice were placed in a stereotaxic frame under anaesthesia with isoflurane. A burr hole was drilled into the skull, and type VII-S collagenase (0.075 U in 0.5 μl saline; Sigma, St. Louis, MO) was infused into the left striatum with a microinfusion pump at a constant rate of 0.1 μl/min. The needle tip was placed for haematoma induction to invade the middle of the striatum without damaging the motor cortex: 0.4 mm anterior and 2.0 mm lateral from the bregma, 3.0 mm below the skull. Sham-operated mice were infused with an equal volume of saline. Body temperature was maintained at 37 ± 0.5 °C during surgery. After surgery, mice were returned to their home cage. Vital conditions of the mice were monitored every day after ICH surgery.
3
Collagenase and Autologous Blood ICH Models
4
Collagenase-Induced Intracerebral Hemorrhage
5
Collagenase-Induced Intracerebral Hemorrhage in Mice
6
Stereotaxic Collagenase Injection and Adropin Treatment
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Stereotaxic Z-axis was modified at 40° from the vertical state to avoid motor cortex damage by needle insertion [20 (link)]. 8-week-old C57BL/6 mice were anesthetized using sevoflurane and put on a stereotaxic frame (DAVID KOPF INSTRUMENTS, CA, USA). After a small incision in the skin, collagenase type VII-S (0.5U, Sigma, St. Louis, MO) was injected (0.2 µl/min, AP + 0.4 mm, ML + 3.8 mm from bregma and DV -3.2 mm from the skull) using 33 G blunt needle. The needle was left for 5 min and removed, then sutured the skin. The mice were allowed to recover. Adropin (Novopro, Shanghai, China) was dissolved in saline and adropin (900 nmol/kg) was administered by intraperitoneal injection at 1 h after collagenase injection.
7
Collagenase-Induced Intracerebral Hemorrhage
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The 8-week-old male CD1 mice (Charles River, Wilmington, USA) weighing 30–32 g were used for the establishment of the ICH model. In the brain tissue surrounding the hematoma in the right basal ganglia region, collagenase was injected into the right basal ganglia to induce ICH. Then, all mice were anesthetized with ketamine (100 mg/kg) and dexmedetomidine (0.5 mg/kg) via intraperitoneal injection. Subsequently, the mice were immobilized on a brain stereotaxic apparatus (RWD, China). A 1-mm crenocloma was drilled, and a 26-gauge needle was inserted stereotactically and 0.075 units of collagenase (type VII-S; Sigma-Aldrich, 9001-12-1, St. Louis, MO, USA) in 0.5 μl sterile phosphate-buffered saline (PBS) was injected into the right basal ganglia. Then the needle was kept at the injection point for 5 min in case of liquid backflow. After the cranial pinhole was blocked with bone wax, and the skin was sutured, 0.4 ml of saline was injected subcutaneously. In sham group, the needles were inserted with equivalent PBS, and the other operations were identical. All animal experiments were approved by the Institutional Animal Care and Use Committee of ShangRao People’s Hospital.
8
Collagenase-Induced Intracerebral Hemorrhage
9
Induction of Intracerebral Hemorrhage in Mice
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ICH was induced by unilateral intrastriatial infusion of collagenase type VII-S (Sigma, St. Louis, MO, USA) as we have described previously [11 (link)]. Briefly, mice were anesthetized with isoflurane (3% induction, 1–1.5% maintenance) and immobilized on a stereotactic frame (Stoelting, Wood Dale, IL, USA). After making a small incision in the skin overlying the skull, a craniotomy was performed 0.5 mm anterior and 2.4 mm right of bregma. Collagenase (0.04 units in 0.2 µl of sterile saline) was infused 3.2 mm ventral from the dura over a 5-min period using a syringe with a 26-gauge blunt tip needle (Hamilton Co., Reno, NV, USA). Rectal temperatures were maintained at 37.0 ± 0.5°C throughout all surgical procedures and mice were allowed to recover in temperature- and humidity-controlled chambers for at least 1 h postoperatively.
10
Intracerebral Collagenase-Induced Hemorrhage
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ICH was induced by means of a stereotactic injection of collagenase type VII-S (Sigma Aldrich, St. Louis, MO). For doing so, mice were anesthetized with isoflurane (1.5–2 %) in a 70 % N2O- and 30 % O2-mixture and placed in a stereotactic frame. A small borehole was drilled in the skull. A 32-Gauge needle (Hamilton 7000 series, Hamilton, Reno, NV) was lowered into the right striatum at the following coordinates from the bregma: 0.0 mm anterior, 2 mm lateral and 3.5 mm in depth. Over a period of 5 min, collagenase (0.1 U in 0.5 µL saline) was injected. The needle was left in situ for 10 min and was then slowly withdrawn over 5 min. After suturing, the animals were allowed to wake up and returned to their cages. Throughout the procedure, a heating lamp was used to maintain body temperature.
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