PMID- 10892886
OWN - NLM
STAT- MEDLINE
DCOM- 20000731
LR  - 20161124
IS  - 0146-0404 (Print)
IS  - 0146-0404 (Linking)
VI  - 41
IP  - 8
DP  - 2000 Jul
TI  - Effects of brain-derived neurotrophic factor and neurotrophin-4 on isolated 
      cultured retinal ganglion cells: evaluation by flow cytometry.
PG  - 2373-7
AB  - PURPOSE: Effects of brain-derived neurotrophic factor (BDNF) and neurotrophin 
      (NT)-4 on retinal ganglion cells (RGCs) isolated and cultured in a serum-free 
      medium are evaluated objectively by using flow cytometry. METHODS: RGCs from the 
      retinas of 2-day-old rats were isolated in a two-step panning and cultured in a 
      serum-free medium. BDNF (1, 10, and 100 pg/ml or 1, 10, and 100 ng/ml), NT-4 
      (0.1, 1, 10, and 100 ng/ml) or their vehicle, phosphate-buffered saline, were 
      individually added to aliquots of the medium to be cultured for 48 hours. Then, 
      after adding 5-chloromethylfluorescein diacetate, the survival of RGCs was 
      evaluated using flow cytometry. RESULTS: The method used allowed the authors to 
      analyze 10,000 RGCs per sample in approximately 2 minutes, so that a much larger 
      number of cells was evaluated in a shorter period than with previously reported 
      methods. RGCs were classified into either large or small RGCs, and the survival 
      of each of these groups was determined objectively by the amount of fluorescent 
      emission. BDNF improved the survival rate of RGCs concentration-dependently. In 
      particular, the survival rate of small RGCs was greatly improved. BDNF at 100 
      ng/ml increased the survival rate of small RGCs by 17.4% and that of large RGCs 
      by 7.8% in comparison to the controls. NT4 did not significantly improve the 
      survival rates of either large or small RGCs. CONCLUSIONS: BDNF improved the 
      survival rate of RGCs, particularly of small RGCs, concentration-dependently, but 
      NT-4 had little influence on the survival rate. The current method was useful in 
      evaluating the effects of neuroprotective factors or neurotoxic factors on 
      cultured RGCs.
FAU - Kashiwagi, F
AU  - Kashiwagi F
AD  - Department of Ophthalmology, Yamanashi Medical University, Tamaho Yamanashi, 
      Japan.
FAU - Kashiwagi, K
AU  - Kashiwagi K
FAU - Iizuka, Y
AU  - Iizuka Y
FAU - Tsukahara, S
AU  - Tsukahara S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - 0 (Fluoresceins)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Nerve Growth Factors)
RN  - 0 (Neuroprotective Agents)
RN  - 136832-63-8 (5-chloromethylfluorescein)
RN  - P658DCA9XD (neurotrophin 4)
SB  - IM
MH  - Animals
MH  - Animals, Newborn
MH  - Brain-Derived Neurotrophic Factor/*pharmacology
MH  - Cell Survival/drug effects
MH  - Cells, Cultured
MH  - Flow Cytometry
MH  - Fluoresceins/pharmacology
MH  - Fluorescent Dyes/pharmacology
MH  - Nerve Growth Factors/*pharmacology
MH  - Neuroprotective Agents/*pharmacology
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Retinal Ganglion Cells/cytology/*drug effects
EDAT- 2000/07/13 11:00
MHDA- 2000/08/06 11:00
CRDT- 2000/07/13 11:00
PHST- 2000/07/13 11:00 [pubmed]
PHST- 2000/08/06 11:00 [medline]
PHST- 2000/07/13 11:00 [entrez]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 2000 Jul;41(8):2373-7.