A MICRO-SCALE PROCEDURE FOR THE PREPARATION OF SUBCELLULAR FRACTIONS FROM INDIVIDUAL AUTONOMIC GANGLIA

Abstract— A microscale modification for the preparation of subcellular fractions employing milligram and submilligram amounts of neuronal tissue (brain nuclei and autonomic ganglia) is described.
Electron microscope characterization and enzymic studies were carried out on the six subcellular fractions of sympathetic ganglia of cat thus prepared.
The synaptosomal preparations obtained from individual ganglia were poorer in their nerve ending content than those obtained from brain by previous investigators. The highest RSA for AChE was found in layer L₂ which was rich in membranes and vesicle components. ChAc activity was also highly concentrated in layers L₂ and L₃ (membranes, nerve ending-like particles, mitochondria and 'ghosts'). MAO activity was particularly high in the layers L₄ and L₅ which contained a large number of mitochondria. Layer L₁ (membrane fragments) and particularly layer L₆ which contained mainly collagen fibres, were low in activity of all three enzymes.
After preganglionic denervation, both ChAc and AChE activities were significantly reduced in the purest nerve ending fraction, L₃ while MAO activity was practically unchanged.