[Acute isolation and identification of cerebral neurons from the late third instar larvae of Drosophila melanogaster].
OBJECTIVE
To develop a new method for acute isolation of the cerebral neurons from late third instar larvae of Drosophila melanogaster.


METHODS
The dissociated cells were characterized by morphological observation and whole-cell voltage-clamp recording. The brains were dissected from late third instar larvae, torn into small fragments, and then were digested in the calcium-magnesium-free PBS solution endowed with collagenase for 45-60 minutes. Single cell was obtained by micro-shaking the digested fragments for 5 to 10 s until the clumps of the tissue were not invisible. Preparation of dispersed cells was incubated in the culture media of Drosophila for thirty minutes at room temperature (20 +/- 1) degrees C.


RESULTS
All neurons studied were categorized into three types according to morphological observation: large (> 8 microm) round type I neuroblast-like cells (7%), small (2-5 microm) type II cells (77%). and intermediate-sized type II cells (16%). Neurogliocytes were not found. The electrophysiological properties of three types of neurons were investigated by whole-cell voltage-clamp recording technique. Five types of outward potassium currents were detected readily.


CONCLUSIONS
Morphological and electrophysiological investigation showed that the method for acute isolation of Drosophila neurons is simple, available and stable.
Liu
Yang
Y
Laboratory of Anesthesiology and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
Liao
Da-Qing
DQ
Luo
Nan-Fu
NF
Zhang
Zhen-Xiong
ZX
Liu
Jin
J
D000818:Animals; D001921:Brain / Q000166:cytology* / Q000502:physiology; D002469:Cell Separation / Q000191:economics / Q000379:methods*; D004331:Drosophila melanogaster / Q000166:cytology* / Q000254:growth & development*; D004553:Electric Conductivity; D007814:Larva / Q000166:cytology; D008018:Life Cycle Stages; D008564:Membrane Potentials; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism; D015203:Reproducibility of Results
D011188:Potassium