title: [Acute isolation and identification of cerebral neurons from the late third instar larvae of Drosophila melanogaster].
abstract: 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.
authors.affiliation: Laboratory of Anesthesiology and Critical Care Medicine, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Da-Qing
authors.initials: DQ
authors.lastname: Liao
authors.forename: Nan-Fu
authors.initials: NF
authors.lastname: Luo
authors.forename: Zhen-Xiong
authors.initials: ZX
authors.lastname: Zhang
authors.forename: Jin
authors.initials: J
authors.lastname: Liu
mesh_terms: 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
chemical_list: D011188:Potassium