• Animal cells and systems
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• v.16 no.2
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• pp.104-113
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• 2012

The nucleolus is a distinct nuclear territory involved in the compartmentalization of nuclear functions. There is some evidence of a relationship between nuclear fragmentation during spermatogenesis and chromatoid body (CB) formation. The CB is a typical cytoplasmic organelle of haploid germ cells, and is involved in RNA and protein accumulation for later germ-cell differentiation. The goal of this study was to qualitatively and quantitatively describe the nucleolar cycle during the spermatogenesis of $Phrynops$ $geoffroanus$ (Reptilia Testudines), and compare this nucleolar fragmentation with CB formation in this species through the use of cytochemical and ultrastructural analysis. Qualitative analysis showed a fragmentation of the nuclear material after pachytene of the first meiotic division in the primary spermatocytes. Quantitative analysis of the nucleolar cycle revealed a significant difference in the number of nucleoli and in the size of the nucleolus between spermatogonia and early spermatids. Using ultrastructural analysis, we recorded the beginning of the CB formation process in the cytoplasm of primary spermatocytes at the same time as when nuclear fragmentation occurs. In the cytoplasm of primary spermatocytes, the CB was observed in association with mitochondrial aggregates and the Golgi complex. In the cytoplasm of early spermatids, the CB was observed in association with lipid droplets. In conclusion, our data show that the nucleolus plays a role in the CB formation process. During spermatogenesis of $P.$ $geoffroanus$, the CB is involved in some important biological processes, including acrosome formation and mitochondrial migration to the spermatozoon tail and middle piece region.

• BMB Reports
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• v.41 no.3
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• pp.248-253
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• 2008

Stresses and nutritional starvation are two main external signals for the induction of sex pheromones in the fission yeast Schizosaccharomyces pombe. In an attempt to identify the components involved in transduction of starvation signals, we screened 135 temperature-sensitive (ts) mutants and isolated 6 mutants that induced the pheromone even in the presence of a nitrogen source. These mutants exhibited two distict induction phenotypes: pheromone induction at restrictive but not at permissive temperatures; and pheromone induction at both permissive and restrictive temperatures. The times required for the maximum pheromone induction at the restrictive temperature differed slightly in each mutant. In addition to the pheromone induction phenotype, the ts243 and ts304 mutants exhibited cell-division-cycle defects. The ts304 mutant cells showed an abnormal cytoplasmic DAPI staining pattern. The nucleolus of this mutant seemed to be fragmented, a phenomenon which is typically observed in aged yeast cells. The result of our genetic analysis indicated that the pheromone induction mutants belonged to 6 separate complementation groups. We designated these mutants pws1 to pws6.

• Toxicological Research
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• v.17
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• pp.59-69
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• 2001

Arsenic exposure is associated with several human diseases, including cancers, atherosclerosis, hypertension, and cerebrovascular diseases. In cultured cells, arsenite, an inorganic arsenic com-pound, was demonstrated to interfere with many physiological functions, such as enhancement of oxidative stress, delay of cell cycle progression, and induction of structural and numerical changes of chromosomes. The objective of this study is to investigate the effects of arsenic exposure on gene expression profiles by colorimetric cDNA microarray technique. HFW (normal human diploid skin fibroblasts), CL3 (human lung adenocarcinoma cell line), and HaCaT (immortalized human keratinocyte cell line) were treated with 5 $\mu\textrm{M}$ or 10 $\mu\textrm{M}$ sodium arsenite for 6 or 16 h, respectively. By a dual-color detection system, the expression profile of arsenite-treated cultures was compared to that of control cultures. Several genes expressed differentially were identified on the microarray membranes. For example, MDM2, SWI/SNF, ubiquitin specific protease 4, MAP3K11, RecQ protein-like 5, and Ribosomal protein Ll0a were consistently induced in all three cell types by arsenite, whereas prohibitin, cyclin D1, nucleolar protein 1, PCNA, Nm23, and immediate early protein (ETR101) were apparently inhibited. The present results suggest that arsenite insults altered the expression of several genes participating in cellular responses to DNA damage, stress, transcription, and cell cycle arrest.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.5
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• pp.2291-2295
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• 2014

Although nucleolar protein nucleostemin (NS) is essential for cell proliferation and early embryogenesis and expression has been observed in some types of human cancer and stem cells, the molecular mechanisms involved in mediation of cell proliferation and cell cycling remains largely elusive. The aim of the present study was to evaluate NS as a potential target for gene therapy of human breast carcinoma by investigating NS gene expression and its effects on SKBR-3 cell proliferation and apoptosis. NS mRNA and protein were both found to be highly expressed in all detected cancer cell lines. The apoptotic rate of the pcDNA3.1-NS-Silencer group ($12.1-15.4{\pm}3.8%$) was significantly higher than those of pcDNA3.1-NS ($7.2-12.0{\pm}1.7%$) and non-transfection groups ($4.1-6.5{\pm}1.8%$, P<0.01). MTT assays showed the knockdown of NS expression reduced the proliferation rate of SKBR-3 cells significantly. Matrigel invasion and wound healing assays indicated that the number of invading cells was significantly decreased in the pcDNA3.1-NS-siRNA group (P<0.01), but there were no significant difference between non-transfected and over-expression groups (P>0.05). Moreover, RNAi-mediated NS down-regulation induced SKBR-3 cell G1 phase arrest, inhibited cell proliferation, and promoted p53 pathway-mediated cell apoptosis in SKBR-3 cells. NS might thus be an important regulator in the G2/M check point of cell cycle, blocking SKBR-3 cell progression through the G1/S phase. On the whole, these results suggest NS might be a tumor suppressor and important therapeutic target in human cancers.

• The Korean Journal of Malacology
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• v.1 no.1
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• pp.24-50
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• 1985

Five different populations of Parafossarulus manchouricus (Chongpyung, Chinju and Kunsan, Korea; and Japan and Taiwan), a population of Bitbynia (Gabbia) misella (Gongju, Korea) and two different populations of Bithynta tentaculata (Michigan, U.S.A. and Bodensee, Germany) were compared in regard to eff-laying characteristics, morphology, chromosome cytology, natural infections of parasites and ecology of habitats. A satisfactory culture method was devised for laboratory rearing of the snails. Tropical fish food (Terra SML) and powdered green leaves (Ceralife) were used as the main food sources for the snails. Benthic diatoms such as Navicula and Gomphonema from the periphyton were also essential for satisfactory growth, especially for the baby snails. The aquaria were stabilized with small stones from a local stream. Young P. manchouricus snails grew to adult size in about 54 days after hatching. They laid eggs 150-156 days after hatching. The whole cycle (birth to egg-laying) took approximately 5 months. The three species of bithyniid snails are iteroparous and lay eggs once a year. There were no major morphological differences in the shells of genera or subgenera studied here. They did exhibit the following rather minor differences. The shell of Parafossarulus has spirally raised ridges, and its apex is usually eroded; the other two genera lack these characteristics. The shell of B. (Gabbia) misella is small, nor exceeding 7.5 mm in length, while the shells of the other two species are larger, being more than 10 mm in length. Scanning electron microscopy (SEM) of the protoconch of P. manchouricus reveals nearly smooth sculpture with small, low, spiral wrinkles. This sculpture is quite different from that of the Hydrobiidae, a family to which the bithyniids are frequently assigned. Scanning electron microscopy of the radulae of the three bithyniid species showed that their radular morphologies are very similar, but there are some small differences, which may be species-specific. There were some statistical differences in shell heights between the Korean and the other populations of P. manchouricus, and between this species and the other two bithyniids as well. The shell differences between the several populations of Korean P. manchouricus may be related to environment. Edtails of the chromosome cycle of these bithyniid snails are similar to those reported for other snails. No specific differences were observed in the chromosome cycle between the various species and populations of snails employed in this study. Reporred for the first time in molluscs are two darkly stained "nucleolar organizers" during pachyterne stages of meiosis. Two different chromosome numbers were observed in the three bithyniid species: n=17 in B. tentaculata and P. manchouricus, and n=18 in B. (G.) misella. no sex chromosomes or supernumerary chromosomes were seen. There were no morphological differences in karyotypes of three Korean strains of P. manchouricus. The infection rates of cercariae of Clonorchis sinensis in Chinju and Kunsan strains of P. manchouricus were 0.14% and 1.25%, respectively. However, Clonorchis cercariae were found in Chongpyung strain of P. manchouriceu and Gongju strain of B. (G.) misella. The habitats of P. manchouricus around Jinyang Lake were relatively clean without any heavy pollution of aquatic microorganisms and organic materials during the period of this study. The levels of dissolved oxygen (D.O.) and biochemical oxygen demand (B.O.D.) of the water specimens sampled from the study areas ranged from 6.0 to 9.6 ppm and from 0.4 to 1.6 ppm, respectively. Eight metalic constituents from the water samples were also assayed, and all metalic ions detercted were remarkably low below the legal criteria. However, calcium ion in the water samples from the habitats of P. manchouricus was considerably higher than others.