Evidence for a noncholinergic function of acetylcholinesterase during development of chicken retina as shown by fasciculin.
Fasciculin 2 (FAS), an acetylcholinesterase (AChE) peripheral site ligand that inhibits mammalian AChE in the picomolar range and chicken AChE only at micromolar concentrations, was used in chick retinal cell cultures to evaluate the influence of AChE on neuronal development. The effects of other AChE inhibitors that bind the active and/or the peripheral site of the enzyme [paraoxon, eserine, or 1,5-bis(4-allyldimethylammoniumphenyl) pentan-3-one dibromide (BW284c51)] were also studied. Morphological changes of cultured neurons were observed with the drugs used and in the different cell culture systems studied. Cell aggregates size decreased by more than 35% in diameter after 9 days of FAS treatment, mainly due to reduction in the presumptive plexiform area of the aggregates. Eserine showed no effect on the morphology of the aggregates, although it fully inhibited the activity of AChE. In dense stationary cell culture, cluster formation increased after 3 days and 6 days of FAS treatment. However, FAS, at concentrations in which changes of morphological parameters were observed, did not inhibit the AChE activity as measured histochemically. In contrast, paraoxon treatment produced a slight morphological alteration of the cultures, while a strong inhibition of enzyme activity caused by this agent was observed. BW284c51 showed a harmful, probably toxic effect, also causing a slight AChE inhibition. It is suggested that the effect of an anticholinesterase agent on the morphological modifications of cultured neurons is not necessarily associated with the intensity of the AChE inhibition, especially in the case of FAS. Moreover, most of the effects of AChE on culture morphology appear to be independent of the cholinolytic activity of the enzyme. The results obtained demonstrate that FAS is not toxic for the cells and suggest that regions of the AChE molecule related to the enzyme peripheral site are likely to be involved with the nonclassical role of AChE.
Blasina
M F
MF
División Neuroquimica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.
Faria
A C
AC
Gardino
P F
PF
Hokoc
J N
JN
Almeida
O M
OM
de Mello
F G
FG
Arruti
C
C
Dajas
F
F
D000110:Acetylcholinesterase / Q000502:physiology*; D000818:Animals; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide / Q000494:pharmacology; D001665:Binding Sites / Q000187:drug effects; D020134:Catalytic Domain / Q000187:drug effects; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured / Q000187:drug effects; D002642:Chick Embryo; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D004546:Elapid Venoms / Q000494:pharmacology*; D005136:Eye Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D009474:Neurons / Q000187:drug effects / Q000201:enzymology; D010261:Paraoxon / Q000494:pharmacology; D010830:Physostigmine / Q000494:pharmacology; D012160:Retina / Q000166:cytology / Q000196:embryology* / Q000201:enzymology
D002800:Cholinesterase Inhibitors; D004546:Elapid Venoms; D005136:Eye Proteins; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide; C044937:fasciculin; D010830:Physostigmine; D000110:Acetylcholinesterase; D010261:Paraoxon