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Alteration of nuclear architecture in male germ cells of chromosomally derived subfertile mice

¹ Dipartimento di Biologia Animale, Laboratorio di Biologia dello Sviluppo e Centro di Eccellenza in Biologia Applicata, Universita’ di Pavia, Piazza Botta, 9 – 27100 Pavia, Italy
² Dipartimento di Medicina Sperimentale, Sezione di Istologia ed Embriologia, Universita’ di Parma, Via Volturno, 39 – 43100 Parma, Italy
³ Department of Obstetrics and Gynaecology, Mayo Clinic, 200 First Street SW, Rochester, Minnesota 55905, USA
⁴ Laboratorio de Cariobiología y Citogenética, Programa de Genética Humana, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Casilla 70061, Santiago 7, Chile
⁵ Dipartimento di Biologia Animale e dell’Uomo, Universita’ "La Sapienza" Roma, Via Alfonso Borelli, 50 – 00161 Roma, Italy

View larger version (71K): [in a new window]   Fig. 1. Chromosome painting with probes for chromosomes 5, 11, 16, X (green) and 13, 15, 17 and Y (red). The typical hybridisation patterns (see also Materials and Methods) of germ and Sertoli cell nuclei of the three karyotypes studied are shown.

View larger version (45K): [in a new window]   Fig. 2. The frequency distributions, shown as pie charts, of the ‘typical’ pattern (those shown in Fig. 1) are 15-20 times more frequent than some of the other patterns detected among those expected, on the basis of the combined possibilities of the three categories (condensation, proximity and position) in which we placed the fluorescent signals. Blue represents condensed, overlapping signals, either central or peripheral; dark blue and purple represent condensed, opposite ends, either central or peripheral, depending on the number of hybridization signals; pink, brown and light brown represent decondensed, opposite ends, both central and peripheral, depending on the number of hybridization signals. In the Rb heterozygotes, the spatial relationship between the Rb metacentric chromosome arms and their telocentric homologues differs from that of the homozygotes (yellow).

View larger version (46K): [in a new window]   Fig. 3. Kinetics of pachytene/spermatid stages of spermatogenesis and spermiogenesis in homozygotes (all-telocentrics, C3H standard karyotype shown by O; all-Rb metacentrics, CR mice shown by ) and heterozygotes (seven trivalents in diakinesis, C3H x CR shown by ). The pachytene/spermatid ratios reach the expected 1:4 value only for the homozygous mice, whereas the C3H x CR heterozygotes show a decreased value because of germ-cell losses. On the right, the inserts show seminiferous tubules of a fully fertile (tubules with normal histoarchitectural germ cell arrangement) C3H (top) and a subfertile (tubules with different degrees of cell depletion) C3H x CR (bottom) mouse from histological sections of testes. The testicular picture of the fertile Rb homozygous CR mice is not reported as it looks like that of the C3H mice. The bar represents 100 µm.