Ethanol affects calmodulin and the calmodulin-binding proteins neuronal nitric oxide synthase and alphaII-spectrin (alpha-fodrin) in the nucleus of growing and differentiated rat astrocytes in primary culture.
The distribution of calmodulin (CaM) and the CaM-binding proteins neuronal nitric oxide synthase (nNOS) and alphaII-spectrin (alpha-fodrin) in the nucleus of growing and differentiated astrocytes was analysed using immunogold electronmicroscopy. We also analysed the effect of moderate ethanol exposure on these proteins. For this, female Wistar rat were fed with an alcoholic liquid diet and exposed to males after several weeks. Pregnant rats were fed with this diet and, after birth, the foetuses brains were used to establish primary cultures of astrocytes. Astrocytes from control and ethanol-exposed rats foetuses were cultured in the absence or presence of ethanol (30 mM) for 7 days (growing cells) and 21 days (differentiated astrocytes). Our results indicate that all the proteins studied appeared mainly on the condensed chromatin of both control- and alcohol-exposed cells and that there are significant variations in the amount of these proteins between quiescent and dividing astrocytes. Altogether, we have not found a co-localisation between CaM and the CaM-binding proteins.
Tomás
M
M
Sección de Biología y Patología Celular, Centro de Investigación, Hospital Universitario La Fe, Avda. Campanar 21, E-46009 Valencia, Spain.
Marín
M P
MP
Portolés
M
M
Megías
L
L
Gómez-Lechón
M J
MJ
Renau-Piqueras
J
J
D000437:Alcoholism / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D002147:Calmodulin / Q000378:metabolism*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D000431:Ethanol / Q000494:pharmacology*; D005260:Female; D008297:Male; D008431:Maternal-Fetal Exchange; D008840:Microfilament Proteins / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism*; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D013049:Spectrin
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D002352:Carrier Proteins; D002492:Central Nervous System Depressants; D000431:Ethanol; D008840:Microfilament Proteins; D052248:Nitric Oxide Synthase Type I; D013049:Spectrin