PMID- 3996139
OWN - NLM
STAT- MEDLINE
DCOM- 19850725
LR  - 20131121
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 6
IP  - 3
DP  - 1985 May
TI  - Flow cytometric analysis of mouse spermatogenic function following exposure to 
      ethylnitrosourea.
PG  - 238-53
AB  - The effects of the mutagenic agent ethylnitrosourea (ENU) on spermatogenic 
      function and sperm chromatin structure were studied by flow cytometry and the 
      results compared with sperm head morphology measurements. Groups of mice received 
      daily exposures ranging from 0 to 75 mg/kg body weight X 5 days and were 
      sacrificed 28 days later. Fresh testicular cell suspensions and epididymal sperm 
      were stained with acridine orange (AO) and measured by flow cytometry. Sperm 
      nuclei were isolated, fixed, rehydrated, and then either subjected to thermal 
      stress or not prior to staining with AO. Body weights were unaffected by the 
      chemical exposure while the testicular weights were reduced by about 50%. 
      Two-parameter (DNA, RNA) flow cytometry measurements showed a dose-response 
      relationship in the loss of certain cell types, particularly the elongated 
      spermatids, from the testes of treated animals. Flow cytometric analysis of both 
      heat-stressed and non-heat-stressed nuclei showed a relationship between dosage 
      and the coefficient of variation of alpha t [red/(red + green fluorescence)] 
      measurements of AO stained nuclei, thereby demonstrating that alterations of 
      chromatin structure occurred in response to ENU. Enzymatic digestions with RNAse, 
      DNAse, and nuclease S1 suggest that the increase in red fluorescence is due to an 
      increase of single-stranded DNA induced by heat or acid treatment of chemically 
      altered chromatin structure. The lowest daily dosage used (5 mg/kg) caused no 
      significant changes in ratios of testicular cell types, a questionable increase 
      in abnormal sperm head morphology and a detectable change in chromatin structure 
      expressed as alpha t. This report shows that our technique for assaying sperm 
      nuclear chromatin structure appears to have the same level of sensitivity to ENU 
      induced nuclear alterations as the sperm head morphology test.
FAU - Evenson, D P
AU  - Evenson DP
FAU - Higgins, P J
AU  - Higgins PJ
FAU - Grueneberg, D
AU  - Grueneberg D
FAU - Ballachey, B E
AU  - Ballachey BE
LA  - eng
GR  - R01 ES03035/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
RN  - 0 (Chromatin)
RN  - 0 (Nitrosourea Compounds)
RN  - 9007-49-2 (DNA)
RN  - F30N4O6XVV (Acridine Orange)
RN  - P8M1T4190R (Ethylnitrosourea)
SB  - IM
MH  - Acridine Orange
MH  - Animals
MH  - Chromatin/ultrastructure
MH  - DNA/metabolism
MH  - Ethylnitrosourea/*pharmacology
MH  - Flow Cytometry
MH  - Hot Temperature
MH  - Male
MH  - Mice
MH  - Mice, Inbred Strains
MH  - Microscopy, Electron
MH  - Nitrosourea Compounds/*pharmacology
MH  - Nucleic Acid Denaturation
MH  - Spermatogenesis/*drug effects
MH  - Spermatozoa/drug effects/metabolism/ultrastructure
EDAT- 1985/05/01 00:00
MHDA- 1985/05/01 00:01
CRDT- 1985/05/01 00:00
PHST- 1985/05/01 00:00 [pubmed]
PHST- 1985/05/01 00:01 [medline]
PHST- 1985/05/01 00:00 [entrez]
AID - 10.1002/cyto.990060311 [doi]
PST - ppublish
SO  - Cytometry. 1985 May;6(3):238-53. doi: 10.1002/cyto.990060311.