Synaptic process in the rat (Rattus norvegicus): Influence of methodology on results.
Synaptonemal complex (SC) analysis is a widely used method for assessing the effects of genotoxic agents in germ cells. Although the evolution of the SCs and their related annexed structures, such as nucleoli, has been well established, sometimes it is difficult to assess whether the abnormal features observed correspond to genotoxic effects or to an artefact related to the method used to obtain the SC preparations. In this article, we describe a new method of obtaining SC preparations for electron microscopy, as well as the results of a study of the first meiotic prophase in oocytes and spermatocytes of the rat (Rattus norvegicus Sprague Dawley) in which we analysed how the methodology used can influence the results. Besides important sex-specific differences, mainly during desynapsis (diplotene), a relationship between several bivalents and nucleolar structures, that in some cases could disturb the synaptic process, was observed in oocytes. At the same time, the characteristic SC fragmentation in oocytes was verified, but this fragmentation, in addition to a sex-specific component, was influenced by the method itself. By reducing to a minimum the artefacts produced by the method, it is possible to optimise the analysis of SCs as a method of testing genotoxic effects in the germ line.
Martínez-Flores
I
I
Department of Cell Biology, Universitat Autònoma de Barcelona, E-08193 Bellaterra, Spain.
Cabero
L L
LL
Egozcue
J
J
Garcia
M
M
D000818:Animals; D002466:Cell Nucleolus / Q000648:ultrastructure; D023902:Chromosome Pairing / Q000502:physiology*; D005260:Female; D008297:Male; D008540:Meiosis; D008854:Microscopy, Electron / Q000379:methods; D009865:Oocytes / Q000648:ultrastructure; D011418:Prophase; D051381:Rats; D017207:Rats, Sprague-Dawley; D013090:Spermatocytes / Q000648:ultrastructure; D013573:Synaptonemal Complex / Q000648:ultrastructure*