Zebrafish Brain Dissection: A Technique of Fish Neurobiology
내레이션 대본
– Place a euthanized fish on a dissection bed and use a surgical blade 세스 decapitate the fish at the level of the gills. Hold the head with the ventral side facing up and remove the soft tissues until you see the optic chiasm, a structure made of optic nerves that connects the brain 세스 the eyes.
Cut the optic nerves and remove the eyes. Now, orient the fish with the dorsal side facing up and remove the parts of the skull 세스 isolate the brain. Transfer the brain into a Petri dish containing a dissection medium 세스 maintain a constant pH of the tissue. Observe the parts of the brain.
The olfactory bulbs are a pair of structures at the anterior end connected 세스 the olfactory organs which detect odor signals. The telencephalon includes memory related areas. The habenula relays information from the telencephalon 세스 other parts of the brain.
The optic tectum is a sensory information processing center. The cerebellum plays a 역할 in somatic motor function and balance. And the medulla relays information from the spinal cord 세스 the brain. In the following protocol, we will isolate the brain from an adult zebrafish 세스 collect neural stem cells from different regions.
To begin, prepare a dissection bed by filling a Petri dish with gel packs. Then cover the dish with its corresponding lid and incubate it at minus 20 degrees Celsius. Once the gel is frozen, remove the dish from the freezer and place a clean square of filter paper on top of its lid. Proceed 세스 wrap both the paper and dish with plastic film.
Next, treat all micro-dissection instruments with 70% ethanol. Place these sterilized tools next 세스 a dissecting microscope and position the completed dissection bed under the microscope with optical fiber illumination. Immediately place a previously prepared head specimen on top of the dissection bed and orient it so that its dorsal side is facing down.
Then use scissors 세스 make a longitudinal cut through the soft tissue from the back of the head 세스 the mouth. Afterwards, expose the base of the skull with forceps and remove all of the adjacent tissue. Next, cut one of the lateral walls of the skull starting at the back of the head and moving towards the tectum region of the brain. Repeat this process for the contralateral side. Proceed 세스 cut the optic nerve. And then remove the two lateral most sides of the skull at the level of the tectum
Finally, turn the head ventral side up. And with forceps, peel off the most apical part of the skull 세스 expose the brain. Afterwards, transfer the brain and any remaining parts of the skull 세스 a dish containing dissection medium which is composed of DMM F12 supplemented with penicillin streptomycin. With the plastic handle of a microknife under the microscope, clean the brain tissue, being careful not 세스 damage any neural structures. Use up 세스 two zebrafish brains 세스 generate whole brain derived neurospheres.
