[3H]-myo-inositol uptake in rat cortical slices. Identification of Na+-dependent and Na+-independent systems
1Department of Pharmacology and Toxicology, School of Pharmacy, University of Kansas, Lawrence 66045.
Abstract:
[3H]-myo-Inositol (MI) uptake was measured in vitro using chopped rat cerebral cortical tissue. The uptake and accumulation of MI were linearly proportional to the amount of protein (0.1 to 4.0 mg) in the incubation medium. The uptake was also linear vs time for the first 20 min of incubation. When the uptake was observed at various substrate concentrations, it was found to be unsaturable up to a concentration of 0.78 M. Decreasing the concentration of NaCl or increasing the concentration of KCl in the incubation medium resulted in inhibition of the uptake and accumulation of MI. Inhibition of MI uptake was also produced by veratrine, ouabain and A23187 which alter the ionic gradients across the neuronal membranes. Inhibition of oxidative metabolism with dinitrophenol did not alter MI uptake. Sodium-independent uptake appeared to be the same as that which occurred at 0 degree. Sodium-independent uptake was still present in water-lysed homogenates and was inhibited by relatively high concentrations of ethanol. Thus, it appears that approximately one-half of the [3H]inositol uptake and accumulation in chopped rat cerebral cortex occurs by a sodium-dependent mechanism that can be altered by drugs which change the sodium gradient and the remaining occurs by a sodium-independent mechanism that can be altered by ethanol which is known to change membrane fluidity of neuronal membranes.
Insights
myo-inositol uptake in rat brain cortex involves both sodium-dependent and sodium-independent mechanisms. Ethanol affects the sodium-independent pathway by altering neuronal membrane fluidity.
Area of Science:
- Neuroscience
- Biochemistry
- Pharmacology
Background:
- myo-inositol (MI) is crucial for neuronal function and signaling.
- Understanding MI transport mechanisms is vital for neurological research.
Purpose of the Study:
- To investigate the mechanisms of [3H]-myo-inositol uptake in rat cerebral cortical tissue.
- To determine the role of sodium ions and other factors in MI transport.
Main Methods:
- In vitro uptake assays using chopped rat cerebral cortical tissue.
- Investigated linearity with protein concentration and time.
- Examined substrate concentration dependence and effects of ionic gradients, veratrine, ouabain, A23187, dinitrophenol, and ethanol.
Main Results:
- MI uptake was linear with protein concentration and time up to 20 minutes.
- Uptake was unsaturable up to 0.78 M.
- Sodium reduction inhibited uptake, while veratrine, ouabain, and A23187 also inhibited MI uptake.
- Dinitrophenol did not affect MI uptake.
- A sodium-independent uptake mechanism was identified, present at 0°C and in homogenates, inhibited by ethanol.
Conclusions:
- Rat cerebral cortex exhibits both sodium-dependent and sodium-independent mechanisms for [3H]-myo-inositol uptake.
- The sodium-dependent component is influenced by ionic gradients and related drugs.
- The sodium-independent component is affected by ethanol, suggesting a role in membrane fluidity.
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