PMID- 11320142
OWN - NLM
STAT- MEDLINE
DCOM- 20010726
LR  - 20190513
IS  - 0267-8357 (Print)
IS  - 0267-8357 (Linking)
VI  - 16
IP  - 3
DP  - 2001 May
TI  - Evaluation of inter-scorer and inter-laboratory reliability of the mouse 
      epididymal sperm aneuploidy (m-ESA) assay.
PG  - 189-95
AB  - The mouse epididymal sperm aneuploidy (mESA) assay using 3-chromosome 
      fluorescence in situ hybridization (FISH) was recently developed for assessing 
      the aneugenic potential of chemicals on male germ cells. This study was designed 
      to identify the major technical factors that affect inter-scorer and 
      inter-laboratory variability of the mESA assay. Two laboratories participated in 
      this study (GSF and Lawrence Livermore National Laboratory, LLNL). Mice 
      (102/ElxC3H/El) F(1) were exposed in one laboratory (GSF) to vinblastine (VBL; 
      single intraperitoneal injection of 0, 0.5, 1.0 or 2.0 mg/kg), one of the 10 
      priority compounds of the Commission of the European Communities (CEC) Aneuploidy 
      Program. Twenty-two days later the mESA assay was applied to analyze sperm 
      aneuploidy. In the initial evaluation, small but statistically significant 
      differences were found between the two laboratories in baseline frequencies and 
      there was also disagreement in the determination of a VBL aneuploid effect. 
      Therefore, experiments were conducted to identify the sources of the 
      inter-laboratory differences and technical factors that affected assay 
      reliability and the VBL study was repeated. A harmonization experiment was 
      conducted by bringing the microscope scorers from both laboratories to the same 
      site (LLNL) for a cross-training exercise. Following this exercise, a second 
      group of VBL-treated and control mice were evaluated, and we concluded that VBL 
      is not a sperm aneugen. Our research has identified scoring criteria as the major 
      source of inter-laboratory variation and emphasizes the importance of strict 
      technical controls for the mESA assay, including controlling slide preparations 
      for treatment-induced reductions in sperm count, coding of slides and selection 
      of statistical tests. These considerations are particularly important for the 
      interpretation of small effects (< or =2-fold) on sperm aneuploidy. Our findings 
      suggest that 2-fold differences in frequencies can result from differences among 
      scorers, samples and treatment groups, and are readily within the normal 
      variation for the mESA assay. Such small differences should be viewed with 
      caution until independently confirmed.
FAU - Schmid, T E
AU  - Schmid TE
AD  - Institut fur Saugetiergenetik, GSF-Forschungszentrum fur Umwelt und Gesundheit 
      GmbH, Neuherberg, Ingolstadter Landstrabetae 1, D-85764, Neuherberg, Germany.
FAU - Lowe, X
AU  - Lowe X
FAU - Marchetti, F
AU  - Marchetti F
FAU - Bishop, J
AU  - Bishop J
FAU - Haseman, J
AU  - Haseman J
FAU - Wyrobek, A J
AU  - Wyrobek AJ
LA  - eng
GR  - Y01-ES-10203/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Multicenter Study
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Mutagenesis
JT  - Mutagenesis
JID - 8707812
RN  - 0 (Antineoplastic Agents, Phytogenic)
RN  - 5V9KLZ54CY (Vinblastine)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - *Aneuploidy
MH  - Animals
MH  - Antineoplastic Agents, Phytogenic/toxicity
MH  - DNA/metabolism
MH  - Genotype
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Male
MH  - Mice
MH  - Mice, Inbred C3H
MH  - Mutagenicity Tests/*methods
MH  - Observer Variation
MH  - Reproducibility of Results
MH  - Spermatozoa/*abnormalities/*metabolism
MH  - Vinblastine/toxicity
EDAT- 2001/04/26 10:00
MHDA- 2001/07/28 10:01
CRDT- 2001/04/26 10:00
PHST- 2001/04/26 10:00 [pubmed]
PHST- 2001/07/28 10:01 [medline]
PHST- 2001/04/26 10:00 [entrez]
AID - 10.1093/mutage/16.3.189 [doi]
PST - ppublish
SO  - Mutagenesis. 2001 May;16(3):189-95. doi: 10.1093/mutage/16.3.189.