A simple biochemical approach to differentiate synaptosomes and non-synaptic mitochondria from rat brain.
A simple and rapid low speed density gradient centrifugation method has been described to isolate relatively pure synaptosomes and non-synaptic mitochondria from rat brain cerebral cortex. The purity of the fractions isolated were tested biochemically by considering some marker enzymes, i.e., lactate dehydrogenase, plasma membrane Na+K+ATPase and acetylcholinesterase for the synaptosomes and succinate dehydrogenase for the mitochondrial fraction. The adrenergic receptor properties in the synaptosomal membrane were evaluated by observing the effect of different beta- and alpha 2-adrenergic receptor agonists and antagonists as function of synaptic plasma membrane Na+K+ATPase activity. Isoproterenol (beta-agonist) and yohimbine (alpha 2-antagonist)-induced changes in the synaptosomal Na+K+ATPase activity were counteracted by beta-antagonist propranolol and alpha 2-agonist clonidine, respectively. In the non-synaptic mitochondria the corresponding effects were insignificant. The study illustrates an easy, rapid and low-speed preparatory method to obtain synaptosomal and non-synaptic mitochondrial fractions of high purity.
Sarkar
P K
PK
Department of Animal Physiology, Bose Institute, Calcutta, India.
Ray
A K
AK
D000818:Animals; D001921:Brain / Q000648:ultrastructure*; D002458:Cell Fractionation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D003000:Clonidine / Q000494:pharmacology; D066298:In Vitro Techniques; D007545:Isoproterenol / Q000494:pharmacology; D008928:Mitochondria / Q000201:enzymology; D011433:Propranolol / Q000494:pharmacology; D051381:Rats; D000254:Sodium-Potassium-Exchanging ATPase / Q000187:drug effects; D013574:Synaptosomes / Q000201:enzymology; D015016:Yohimbine / Q000494:pharmacology
D015016:Yohimbine; D011433:Propranolol; D000254:Sodium-Potassium-Exchanging ATPase; D007545:Isoproterenol; D003000:Clonidine