Impaired cholinergic function in cell lines derived from the cerebral cortex of normal and trisomy 16 mice.
Murine trisomy 16 is an animal model of human Down's syndrome. We have successfully established permanently growing cell lines from the cerebral cortex of normal and trisomy 16 foetal mice using an original procedure. These lines, named CNh (derived from a normal animal) and CTb (derived from a trisomic foetus), express neuronal markers. Considering that Down's syndrome exhibits cholinergic deficits, we examined cholinergic function in these lines, using incorporation of [3H]-choline and fractional release studies. After 1, 3 and 5 min of [3H]-choline incubation, CTb cell uptake was lower by approximately 50% compared to controls. Hemicholinium-3 significantly reduced the incorporation of [3H]-choline in both CNh and CTb cells at high concentration (10 microM), suggesting high-affinity choline transport. However, CTb cells exhibited greater sensitivity to the blocker. For fractional release experiments, the cells were stimulated by K+ depolarization, glutamate or nicotine. When depolarized, CTb cells showed a 68% reduction in fractional release of [3H]-acetylcholine compared to CNh cell line, and a 45% reduction when stimulated by nicotine. Interestingly, glutamate induced similar levels of release in both cell types. The results indicate the existence of cholinergic dysfunction in CTb cells when compared to CNh, similar to that reported for primary cultures of trisomy 16 brain tissue (Fiedler et al. 1994, Brain Res., 658, 27-32). Thus, the CTb cell line may serve as a model for the study of Down's syndrome pathophysiology.
Allen
D D
DD
Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University HSC, Amarillo, TX 79106, USA.
Martín
J
J
Arriagada
C
C
Cárdenas
A M
AM
Rapoport
S I
SI
Caviedes
R
R
Caviedes
P
P
D000109:Acetylcholine / Q000378:metabolism / Q000502:physiology; D000544:Alzheimer Disease / Q000503:physiopathology; D000818:Animals; D002460:Cell Line; D002540:Cerebral Cortex / Q000166:cytology / Q000503:physiopathology*; D002794:Choline / Q000493:pharmacokinetics; D002795:Choline O-Acetyltransferase / Q000032:analysis / Q000378:metabolism; D004195:Disease Models, Animal; D004314:Down Syndrome / Q000503:physiopathology; D005260:Female; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000201:enzymology; D009538:Nicotine / Q000494:pharmacology; D018722:Nicotinic Agonists / Q000494:pharmacology; D011247:Pregnancy; D011950:Receptors, Cholinergic / Q000502:physiology*; D014314:Trisomy / Q000235:genetics / Q000503:physiopathology*; D014316:Tritium
D018722:Nicotinic Agonists; D011950:Receptors, Cholinergic; D014316:Tritium; D009538:Nicotine; D002795:Choline O-Acetyltransferase; D002794:Choline; D000109:Acetylcholine