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Transient appearance of Strongylocentrotus purpuratus Otx in micromere nuclei: cytoplasmic retention of SpOtx possibly mediated through an alpha-actinin interaction.

At the 16-cell stage, the sea urchin embryo is partitioned along the animal-vegetal axis into eight mesomeres, four macromeres, and four micromeres. The micromeres, unlike the other blastomeres, are autonomously specified to produce skeletogenic mesenchymal cells and are also required to induce the vegetal-plate territory. A long-held belief is that micromeres inherit localized maternal determinants that endow them with their cell autonomous behavior and inducing capabilities. Here, we present evidence that an orthodenticle-related protein, SpOtx appears transiently in nuclei of micromeres but not in nuclei of mesomeres and macromeres. At later stages of development, SpOtx was translocated into nuclei of all cells. To address the possibility that SpOtx was retained in the cytoplasm at early developmental stages, we searched for cytoplasmic proteins that interact with SpOtx. A proline-rich region of SpOtx resembling an SH3-binding domain was used to screen an embryo cDNA expression library, and a cDNA clone was isolated and shown to be alpha-actinin. A yeast two-hybrid analysis showed a specific interaction between the proline-rich region of SpOtx and a putative SH3 domain of the sea urchin alpha-actinin. Because micromeres lack an actin-based cytoskeleton, the results suggested that, at the vegetal pole of the 16-cell stage embryo, SpOtx was translocated into micromere nuclei, whereas in other blastomeres SpOtx was actively retained in the cytoplasm by binding to alpha-actinin. The transient appearance of SpOtx in micromere nuclei may be associated with the specification of micromere cell fate.

Actinin↗

Single-strand conformation polymorphism analysis of genetic variation in Labiostrongylus longispicularis from kangaroos.

Single-strand conformation polymorphism (SSCP) analysis was employed to screen for sequence heterogeneity in the second internal transcribed spacer (ITS-2) of ribosomal (r) DNA of Labiostrongylus longispicularis, a parasitic strongylid nematode occuring in some species of kangaroo in different geographical regions of Australia. The results showed that most of the nematodes screened had different SSCP profiles, which were subsequently shown to correspond to polymorphisms and/or an indel in the ITS-2 sequence. These variable sites related mainly to unpaired regions of the predicted secondary structure of the precursor rRNA molecule. SSCP profiles could be used to distinguish L. longispicularis in Macropus robustus robustus (New South Wales) from L. longispicularis in Macropus robustus erubescens and Macropus rufus (South Australia). This difference corresponded to a transversional change in the ITS-2 sequence at alignment position 82. The study demonstrated clearly the effectiveness of SSCP analysis for future large-scale population genetic studies of L. longispicularis in order to test the hypothesis that L. longispicularis from different geographical regions represents multiple sibling species.

Animals↗

Twins raised from separated blastomeres develop into sexually mature Strongylocentrotus purpuratus.

The first two blastomeres of Strongylocentrotus purpuratus embryos were separated and the resulting twins raised in pairs through larval life and to sexual maturity. We derive two conclusions from this study: changes in the pattern of specification of embryonic cell fates following blastomere separation result in the creation of two sets of fully functional coelomic pouches and imaginal rudiments, and sex determination in sea urchins is chromosomal, since the pairs of twins were always of the same sex.

Animals↗

Chromatin diminution and a chromosomal mechanism of sexual differentiation in Strongyloides papillosus.

Eggs obtained from feces of rabbits infected with Strongyloides papillosus were squashed and the karyotypes were determined. They contained cells with either two long and two medium sized chromosomes (2L2M), or one long, three medium and one short chromosome (L3MS). Two types of parasitic female gonad could be distinguished on the basis of oocyte chromosome morphology at prometaphase of the maturation division. All the oocytes in a gonad contained either two upaired long chromosomes and two unpaired medium sized chromosomes, or two unpaired medium sized chromosomes and two unpaired chromosomes segmented into beads in one region. At the maturation division in mitotic parthenogenesis the beads appear to be lost from one of the chromosomes. This generates a medium sized and a shorter chromosome, which together with the undiminished chromosomes make up the L3MS karyotype. Animals with beaded oocyte chromosomes lay eggs that develop into males. It is suggested that males are heteromorphic for the long homologue due to chromatin diminution, that occurs in the maturation division of mitotic parthenogenesis.

Animals↗