TY - JOUR
T1 - Kinetics and size of cholesterol lateral domains in synaptosomal membranes
T2 - Modification by sphingomyelinase and effects on membrane enzyme activity
AU - Muralikrishna Rao, A.
AU - Igbavboa, Urule
AU - Semotuk, Michael
AU - Schroeder, Friedhelm
AU - Wood, W. Gibson
N1 - Funding Information:
Acknowledgements--Tihnivse stigatiowna ss upporteidn part by NIAAA grant 07292 (WGW) and by the Medical ResearcShe rvicea ndt heG eriatricR, esearchE,d ucatioann d ClinicalC entero f theD epartmenotf VeteranAs ffairs.
PY - 1993/7
Y1 - 1993/7
N2 - Cholesterol domains and transport have been well-studied in non-neuronal membranes in contrast to neuronal membranes. The purpose of the experiments reported in this paper was to determine: (1) exchangeable and non-exchangeable cholesterol domains or pools were present in brain synaptosomal membranes; (2) effects of hydrolysis of sphingomyelin on cholesterol pools, that has previously been shown to alter membrane cholesterol in non-neuronal membranes and; (3) sphingomyelin hydrolysis and enzyme activity. Cholesterol pools were determined using cholesterol exchange between radiolabeled small unilamellar vesicles and mouse synaptosomes. Activity of Ca2+ + Mg2+-ATPase and Na+ + K+-ATPase were measured in synaptosomal membranes following treatment with sphingomyelinase. The size of the exchangeable pool of synaptosomal membrane cholesterol was approx 50% of total membrane cholesterol when measured at 37°C. The t 1 2 of cholesterol exchange at 37°C in synaptosomes was approx 10 h. Lowering the incubation temperature to 25°C, significantly reduced the size of the exchangeable pool and significantly increased the t 1 2 of cholesterol exchange. Sphingomyelinase treatment of synaptosomes significantly slowed cholesterol exchange but did not modify the size of the exchangeable pool of cholesterol. Ca2+ + Mg2+-ATPase activity was significantly inhibited by sphingomyelinase treatment as compared to Na+ + K+-ATPase activity. Cholesterol domains were described in neuronal tissue and the size and kinetics of those pools were altered by temperature-induced changes in fluidity and hydrolysis of sphingomyelin. Sphingomyelinase-induced changes in Ca2+ + Mg2+-ATPase activity were not affected by hydrolysis of sphingomyelin but appeared to be associated with a reduction in cytofacial phosphatidylinositol.
AB - Cholesterol domains and transport have been well-studied in non-neuronal membranes in contrast to neuronal membranes. The purpose of the experiments reported in this paper was to determine: (1) exchangeable and non-exchangeable cholesterol domains or pools were present in brain synaptosomal membranes; (2) effects of hydrolysis of sphingomyelin on cholesterol pools, that has previously been shown to alter membrane cholesterol in non-neuronal membranes and; (3) sphingomyelin hydrolysis and enzyme activity. Cholesterol pools were determined using cholesterol exchange between radiolabeled small unilamellar vesicles and mouse synaptosomes. Activity of Ca2+ + Mg2+-ATPase and Na+ + K+-ATPase were measured in synaptosomal membranes following treatment with sphingomyelinase. The size of the exchangeable pool of synaptosomal membrane cholesterol was approx 50% of total membrane cholesterol when measured at 37°C. The t 1 2 of cholesterol exchange at 37°C in synaptosomes was approx 10 h. Lowering the incubation temperature to 25°C, significantly reduced the size of the exchangeable pool and significantly increased the t 1 2 of cholesterol exchange. Sphingomyelinase treatment of synaptosomes significantly slowed cholesterol exchange but did not modify the size of the exchangeable pool of cholesterol. Ca2+ + Mg2+-ATPase activity was significantly inhibited by sphingomyelinase treatment as compared to Na+ + K+-ATPase activity. Cholesterol domains were described in neuronal tissue and the size and kinetics of those pools were altered by temperature-induced changes in fluidity and hydrolysis of sphingomyelin. Sphingomyelinase-induced changes in Ca2+ + Mg2+-ATPase activity were not affected by hydrolysis of sphingomyelin but appeared to be associated with a reduction in cytofacial phosphatidylinositol.
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U2 - 10.1016/0197-0186(93)90142-R
DO - 10.1016/0197-0186(93)90142-R
M3 - Article
C2 - 8396483
AN - SCOPUS:0027237240
SN - 0197-0186
VL - 23
SP - 45
EP - 52
JO - Neurochemistry International
JF - Neurochemistry International
IS - 1
ER -