By Milton, Editor; Gross
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Extra resources for Alcohol Intoxication and Withdrawal: Experimental Studies II
The homogenates were centrifuged at 200 x g for 10 min. The supernatants were recentrifuged for 30 min at 12500 x g. The microsomal fractions were obtained from the recentrifuged supernatants by centrifugation for 60 min at 107000 x g. 25 M sucrose, and the radioactivities of the total microsomal fractions measured in a Packard Tri Carb 2420 liquid scintillation spectrometer from 10 ~1 samples dissolved in 10 ml of Insta-Gel (Packard) scintillation liquid. The radioactivities were measured by an external standard method with a counting efficiency of approximately 88 %.
Since earlier studies in this laboratory have dernonstrated reduced protein synthesis by both a rnixed population of ribosornes as well as free and bound polysemes in brains of ethanoldrinking anirnals (30), the present findings suggest that this reduction could involve rnRNA by affecting the poly A synthesizing systern at the prepolysornal level. Since the presence of the poly A tail in rnRNA rnolecules is also a prerequisite for successful production of this RNA (28,39,40), the severe inhibition of poly A synthesis induced by ethanol could cause an irnbalance in the brain cell's protein synthetic capacity.
To assess the effects of ethanol on this homopolynucleotide, in vitro experiments were conducted. As Tables 3 and 4 indicate, long-term ethanol administration markedly reduced poly A synthesis of brain nuclei. That this system, with its own endogenaus poly A, was partially dependent on exogenaus poly A is shown by the reduced incorporation of (8- 3H)ATP into cold TCA insoluble residue in the absence of primer poly A. It is also observed that this system could not be activated either by the addition of DNA (Table 4) or poly U (Table 5) in both the control or ethanol group.
Alcohol Intoxication and Withdrawal: Experimental Studies II by Milton, Editor; Gross