An improved method for the bulk isolation of spinal motoneurones.
Details are described of a procedure for the isolation of motoneurones from pig spinal cord in high yield and high purity. Mechanical dissociation, one low-speed centrifugation and two high-speed ultracentrifugations were applied to separate the motoneurons from other elements of the anterior horn. The first ultracentrifugation through a sucrose gradient resulted in a pellet consisting mainly of motoneurones, microvessels, free nuclei and red blood cells, whereas with the use of Ficoll 400 the motoneurones could be separate from the contaminating elements during a second ultracentrifugation. The high yield and purity of the motoneurones in the final fraction may permit further studies, in which the neurochemical properties of the spinal motoneurones can be elucidated.
D000110:Acetylcholinesterase / Q000032:analysis; D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002795:Choline O-Acetyltransferase / Q000032:analysis; D008855:Microscopy, Electron, Scanning; D009046:Motor Neurons / Q000032:analysis / Q000378:metabolism / Q000648:ultrastructure; D011976:Receptors, Muscarinic / Q000032:analysis; D013116:Spinal Cord / Q000166:cytology*; D013552:Swine; D014461:Ultracentrifugation
D011976:Receptors, Muscarinic; D002795:Choline O-Acetyltransferase; D000110:Acetylcholinesterase