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Heat shock activates in somatic cells a set of genes encoding heat shock proteins which function as molecular chaperones. The basic mechanism by which these genes are activated is the interaction of the specific transcription factor HSF1 with a regulatory DNA sequence called heat shock element (HSE). In higher eukaryotes HSF1 is present in unstressed cells as inactive monomers which, in response to cellular stress, aggregate into transcriptionally competent homotrimers. In the present paper we showed that the expression of a transgene encoding mutated constitutively active HSF1 placed under the control of a spermatocyte-specific promoter derived from the hst70 gene severely affects spermatogenesis. We found the testes of transgenic mice to be significantly smaller than those of wild-type males and histological analysis showed massive degeneration of the seminiferous epithelium. The lumen of tubules was devoid of spermatids and spermatozoa and using the TUNEL method we demonstrated a high rate of spermatocyte apoptosis. The molecular mechanism by which constitutively active HSF1 arrests spermatogenesis is not known so far. One can assume that HSF1 can either induce or repress so far unknown target genes involved in germ cell apoptosis.
The annual reproductive cycle in two wild populations of three-spined stickleback was studied. Sticklebacks from the Dead Vistula River (Martwa Wisła) (brackish water) and the Oliva Stream (Potok Oliwski) (freshwater) were exposed to annual environmental changes in their natural habitats. Ovaries and livers (females), and testes and kidneys (males) were collected during 1-2 years. The gonadosomatic IG, hepatosomatic IH, nephrosomatic IN indices, kidney epithelium height (KEH) and size of oocytes were calculated.The number of mature oocytes and percentage of ovulating females were determined during the spawning season. Histological changes in the ovaries and testes were described throughout a year. Annual reproductive cycles were similar in both populations of sticklebacks. This is the first histological and morphological study carried out throughout a year, simultaneously in two wild populations of three-spined sticklebacks inhabiting different environments. An improved scale of gonadal development in conjunction with the determined indices and fecundity give a comprehensive description of the reproductive cycle. These new observations, in combination with previously reported features, provide a universal scale that can be successfully used to distinguish all phases of gametogenesis in sticklebacks in different habitats.
The Hsp70.2/Hst70 gene, which belongs to the 70 kDa heat-shock protein (HSP) family, is expressed specifically in primary spermatocytes and spermatids. The regulatory elements required for a high level of testis-specific expression of the gene are placed between the two major transcription start sites T1 and T2 (approximately 350 and 115 bp upstream of the starting ATG codon). Here we have shown that sequences proximal to the exon1/intron splicing site in the 5' untranslated region of the Hsp70.2/Hst70 gene, which include a highly conserved element called box B, are required for efficient expression of the chloramphenicol acetyltransferase reporter gene in testes of transgenic mice. However, in spite of the drastically reduced overall activity, the stage-specific expression pattern of the transgene was preserved after removal of these sequences. We have also shown that GC-box located downstream of the box B (approximately 210 bp upstream of the starting ATG codon) is indispensable for efficient expression of the Hsp70.2/Hst70 gene promoter in spermatogenic cells. The GC-box specifically binds proteins present in nuclear extracts from testes (putatively Sp1-like factors). A change in the pattern of such GC-box-interacting factors corresponds to activation of the Hsp70.2/Hst70 gene, confirming the importance of this regulatory element.
A novel approach to DNase I-sensitivity analysis was applied to examining genes of the spermatogenic pathway, reflective of the substantial morphological and genomic changes that occur during this program of differentiation. A new real-time PCR-based strategy that considers the nuances of response to nuclease treatment was used to assess the nuclease susceptibility through differentiation. Data analysis was automated with the K-Lab PCR algorithm, facilitating the rapid analysis of multiple samples while eliminating the subjectivity usually associated with Ct analyses. The utility of this assay and analytical paradigm as applied to nuclease-sensitivity mapping is presented.
Spermatogenesis in Gallegoides arfaai is similar to that described for other cestode species. Six incomplete synchronic cytokineses occur: four mitotic and two meiotic cell divisions. The primary spermatogonium divides forming two secondary spermatogonia. All further divisions occur simultaneously, resulting in a rosette of four tertiary, then eight quaternary spermatogonia and sixteen primary spermatocytes. The first meiotic division forms thirty-two secondary spermatocytes and after the second meiotic division sixty-four spermatids are formed. Spermiogenesis begins with the formation of a differentiation zone in the form of a conical projection of cytoplasm delimited by a ring of arching membranes. Within this area there are two centrioles, a centriolar adjunct and vestigial striated rootlets. During spermiogenesis, only one of the centrioles develops an axoneme that grows directly into the cytoplasmic extension. The other centriole remains oriented in a cytoplasmic bud and posteriorly aborts. The nucleus elongates and moves into the cytoplasmic extension. Granular material present in each sperm originates from electron-dense material present in the periphery of the spermatid. In the final stage of spermiogenesis two crest-like bodies appear at the Bâse of the spermatid. Finally, the ring of arching membranes constricts and the young spermatozoon detaches from the residual cytoplasm. In order to increase homogeneity in the designation of the non-typical striated rootlets previously described, in this study we propose to group them under the common designation of "vestigial striated rootlets" and its importance is discussed according to previous findings of related structures in other cyclophyllideans.
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