PMID- 8900474
OWN - NLM
STAT- MEDLINE
DCOM- 19961223
LR  - 20031114
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 23
IP  - 4
DP  - 1996 Apr 1
TI  - A novel method for detecting apoptosis shows that hepatocytes undergo a time 
      dependent increase in DNA cleavage and chromatin condensation which is augmented 
      after TGF-beta 1 treatment.
PG  - 312-21
AB  - This study describes a new method for quantitating apoptosis in hepatocyte 
      monolayers in which nuclei were isolated from the cells and DNA strand breaks 
      detected by in situ end-labeling and flow cytometry. Most (97%) nuclei from 
      untreated hepatocytes had low end-labelling and were derived from non-apoptotic 
      cells. Approximately 2-3% of the nuclei had high end-labelling and originated 
      from apoptotic hepatocytes. The numbers of these nuclei increased linearly from 3 
      to 85% between 0 and 48 h after treatment with transforming growth factor-beta 1 
      (TGF-beta 1). However, a morphological assessment of apoptosis with Hoechst 
      H33258 showed that the proportion of apoptotic nuclei plateaued at 18-19% between 
      24 and 48 h after TGF-beta 1 treatment. Thus, the in situ end-labeling technique 
      also detected DNA cleavage in nuclei which did not have an obvious apoptotic 
      morphology. Confocal microscopy of low and high end-labelled nuclei which had 
      been separated by fluorescent cell sorting showed that nuclei with high levels of 
      end-labeling exhibited a wide diversity of morphologies. These included nuclei 
      with little or no chromatin condensation and nuclei with characteristic apoptotic 
      morphology. In addition, nuclei from untreated hepatocytes contained low levels 
      of DNA cleavage, which were localized in areas of condensed chromatin and 
      increased according to the time in culture. Thus, hepatocytes undergo a 
      progressive and cumulative process of DNA cleavage/chromatin condensation which 
      is markedly enhanced by TGF-beta 1.
FAU - Cain, K
AU  - Cain K
AD  - MRC Toxicology Unit, Leicester, United Kingdom.
FAU - Inayat-Hussain, S H
AU  - Inayat-Hussain SH
FAU - Couet, C
AU  - Couet C
FAU - Qin, H M
AU  - Qin HM
FAU - Oberhammer, F A
AU  - Oberhammer FA
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
RN  - 0 (Chromatin)
RN  - 0 (Transforming Growth Factor beta)
SB  - IM
MH  - Animals
MH  - Apoptosis/*physiology
MH  - Cell Nucleus
MH  - Cells, Cultured
MH  - Chromatin
MH  - DNA Damage/*physiology
MH  - Flow Cytometry/*methods
MH  - Liver/cytology/*physiology
MH  - Male
MH  - Microscopy, Confocal
MH  - Rats
MH  - Rats, Inbred F344
MH  - Time Factors
MH  - Transforming Growth Factor beta/pharmacology
EDAT- 1996/04/01 00:00
MHDA- 2000/06/20 09:00
CRDT- 1996/04/01 00:00
PHST- 1996/04/01 00:00 [pubmed]
PHST- 2000/06/20 09:00 [medline]
PHST- 1996/04/01 00:00 [entrez]
AID - 10.1002/(SICI)1097-0320(19960401)23:4<312::AID-CYTO7>3.0.CO;2-I [pii]
AID - 10.1002/(SICI)1097-0320(19960401)23:4<312::AID-CYTO7>3.0.CO;2-I [doi]
PST - ppublish
SO  - Cytometry. 1996 Apr 1;23(4):312-21. doi: 
      10.1002/(SICI)1097-0320(19960401)23:4<312::AID-CYTO7>3.0.CO;2-I.