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101.
Abstract: The alcohol-sensitive (ANT) rat line, developed for high behavioral sensitivity to ethanol, also exhibits enhanced sensitivity to benzodiazepines, such as diazepam. The rat line carries a point mutation in the cerebellum-specific γ-aminobutyric acid type A (GABAA) receptor subunit α6, making their diazepam-insensitive (DIS) receptors sensitive to diazepam. We now report that phenotypes of individual ANT and alcohol-insensitive rats, classified on diazepam sensitivity of cerebellar [3H]Ro 15-4513 binding, correlated well with homozygous wild-type, homozygous mutant, and heterozygous genotypes, although some heterozygotes were biased toward the parental phenotypes. GABA down-modulated DIS [3H]Ro 15-4513 binding in mutant homozygotes but tended to up-modulate it in heterozygotes and wild-type homozygotes. Slopes for GABA inhibition of cerebellar t-butylbicyclophosphoro[35S]thionate binding were larger in mutant than in wild-type homozygotes, with heterozygotes being intermediate. Diazepam displacement of [3H]Ro 15-4513 binding in heterozygotes revealed three components, with their affinities indistinguishable from those in combined wild-type and mutant homozygotes. This lack of interaction in DIS binding between wild-type and mutant α6 subunits was substantiated by experiments on recombinant receptors. The data suggest that the α6 subunit-containing GABAA receptors in the heterozygotes are formed from individual mutant and wild-type subunits with their relative expression differing from animal to animal. 相似文献
102.
Andres Mäe Riikka Heikinheimo E. Tapio Palva 《Molecular genetics and genomics : MGG》1995,247(1):17-26
The ceIV1 gene encoding a secreted cellulase (CelV1) of Erwinia carotovora subsp. carotovora SCC3193 was cloned and its nucleotide sequence determined. The gene contains an open reading frame of 1511 by and codes for an exported protein of 504 amino acids. The predicted amino acid sequence of Ce1V1 was highly similar to that of CeIV of another E. c. subsp. carotovora strain SCRI193 but completely different from the previously characterized cellulase, CelS, of the strain SCC3193. Gene fusions to the lacZ reporter were employed to characterize the regulation of celV1 and celS. Both genes are coordinately induced in a growth phase-dependent manner and are catabolite repressed. Expression of celV1 but not celS was stimulated by plant extracts. The celS gene was expressed at a much lower level than celV1 under all conditions tested. Inactivation of the celV1 gene in E. c. subsp. carotovora strain SCC3193 by marker exchange showed that celV1 encodes the major cellulase of strain SCC3193, as the resulting mutant strain SCC6001 was devoid of cellulase activity. Ce1Vl mutants exhibited reduced virulence suggesting that CelV1, although not absolutely required for pathogenicity, enhances the ability of strain SCC3193 to macerate plant tissue. Inactivation of the celS gene in the celV1 mutant did not lead to any further decrease in virulence. 相似文献
103.
Mitochondrial DNA mutations in iPS cells: mtDNA integrity as standard iPSC selection criteria? 下载免费PDF全文
Riikka H Hämäläinen 《The EMBO journal》2016,35(18):1960-1962
Somatic cells harbor random heteroplasmic mitochondrial DNA mutations, which are considered to contribute to aging. In this issue of The EMBO Journal, Perales‐Clemente et al ( 2016 ) show that mtDNA mutations, present at low levels in the starting fibroblasts, become enriched in iPS cells and lead to functional defects in iPS‐derived cells. In another recent study, Kang et al ( 2016 ) demonstrated that accumulation of mtDNA mutations of somatic origin in iPSCs is age related. 相似文献
104.
Kari T. Elo Johanna Vilkki Dirk-Jan de Koning Riikka J. Velmala Asko V. Mäki-Tanila 《Mammalian genome》1999,10(8):831-835
A multiple-marker mapping approach was used to search for quantitative trait loci (QTLs) affecting production, health, and
fertility traits in Finnish Ayrshire dairy cattle. As part of a whole-genome scan, altogether 469 bulls were genotyped for
six microsatellite loci in 12 families on Chromosome (Chr) 23. Both multiple-marker interval mapping with regression and maximum-likelihood
methods were applied with a granddaughter design. Eighteen traits, belonging to 11 trait groups, were included in the analysis.
One QTL exceeded experiment level and one QTL genome level significance thresholds. Across-families analysis provided strong
evidence (Pexperiment= 0.0314) for a QTL affecting live weight. The QTL for live weight maps between markers BM1258 and BoLA DRBP1. A QTL significant at genome level (Pgenome= 0.0087) was mapped for veterinary treatment, and the putative QTL probably affects susceptibility to milk fever or ketosis.
In addition, three traits exceeded the chromosome 5% significance threshold: protein percentage of milk, calf mortality (sire),
and milking speed. In within-family analyses, protein percentage was associated with markers in one family (LOD score = 4.5).
Received: 14 December 1998 / Accepted: 28 March 1998 相似文献
105.
Kari Törrönen Kaisa Nikunen Riikka Kärnä Markku Tammi Raija Tammi Kirsi Rilla 《Histochemistry and cell biology》2014,141(1):17-31
Hyaluronan synthases (HAS) are unique plasma membrane glycosyltransferases secreting this glycosaminoglycan directly to the extracellular space. The three HAS isoenzymes (HAS1, HAS2, and HAS3) expressed in mammalian cells differ in their enzymatic properties and regulation by external stimuli, but clearly distinct functions have not been established. To overview the expression of different HAS isoenzymes during embryonic development and their subcellular localization, we immunostained mouse embryonic samples and cultured cells with HAS antibodies, correlating their distribution to hyaluronan staining. Their subcellular localization was further studied by GFP–HAS fusion proteins. Intense hyaluronan staining was observed throughout the development in the tissues of mesodermal origin, like heart and cartilages, but also for example during the maturation of kidneys and stratified epithelia. In general, staining for one or several HASs correlated with hyaluronan staining. The staining of HAS2 was most widespread, both spatially and temporally, correlating with hyaluronan staining especially in early mesenchymal tissues and heart. While epithelial cells were mostly negative for HASs, stratified epithelia became HAS positive during differentiation. All HAS isoenzymes showed cytoplasmic immunoreactivity, both in tissue sections and cultured cells, while plasma membrane staining was also detected, often in cellular extensions. HAS1 had brightest signal in Golgi, HAS3 in Golgi and microvillous protrusions, whereas most of the endogenous HAS2 immunoreactivity was localized in the ER. This differential pattern was also observed with transfected GFP–HASs. The large proportion of intracellular HASs suggests that HAS forms a reserve that is transported to the plasma membrane for rapid activation of hyaluronan synthesis. 相似文献
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Rilla K Pasonen-Seppänen S Kärnä R Karjalainen HM Törrönen K Koistinen V Tammi MI Tammi RH Teräväinen T Manninen A 《Histochemistry and cell biology》2012,137(2):153-164
The amount of hyaluronan (HA) is low in simple epithelia under normal conditions, but during tumorigenesis, trauma or inflammation
HA is increased on the epithelial cells and surrounding stroma. Excessive HA in epithelia is suggested to interfere with cell–cell
adhesions, resulting in disruption of the epithelial barrier function. In addition, stimulated HA synthesis has been correlated
with epithelial-to-mesenchymal transition and invasion of cancer cells. However, the effects of HA overload on normal epithelial
morphogenesis have not been characterized in detail. Madin-Darby canine kidney (MDCK) cells form polarized epithelial cysts,
when grown in a 3-dimensional (3D) matrix. These cells were used to investigate whether stimulated HA synthesis, induced by
stable overexpression of GFP-HAS3, influences cell polarization and epithelial morphogenesis. GFP-HAS3 expression in polarized
MDCK cells resulted in active HA secretion at apical and basolateral membrane domains. HA-deposits interfered with the formation
of cell–cell junctions, resulting in impaired barrier function. In 3D cyst cultures, HA accumulated into apical lumina and
was also secreted from the basal side. The HAS3-expressing cysts failed to form a single lumen and instead displayed multiple
small lumina. This phenotype was correlated with aberrant mitotic spindle orientation in dividing cells. The results of this
study indicate that excess pericellular HA disturbs the normal cell–cell and cell–ECM interactions in simple epithelia, leading
to aberrant epithelial morphogenesis. The morphological abnormalities observed in 3D epithelial cultures upon stimulated HAS3
expression may be related to premalignant changes, including intraluminal invasion and deregulated epithelialization, probably
mediated by the mitotic spindle orientation defects. 相似文献
108.
The prostate has the highest level of polyamines among all tissues, and it is the only tissue in which polyamines are purposely synthesized for export. It has been suggested that the high local polyamine concentrations suppress cell growth of primary prostatic carcinomas and that this growth control is lost when cancer cells metastasize. It has also been shown that the sensitivity to polyamine-induced growth arrest correlates with antizyme induction in prostate carcinoma cell lines. In this study, we evaluated the sensitivity of poorly metastatic (LNCaP) and highly metastatic (DU145) prostate cancer cell lines to conditional antizyme 1 over-expression. Antizyme 1 induction resulted in a marked loss of ODC activity and polyamine uptake in both cell lines. However, the proliferation of LNCaP cells was repressed by antizyme 1 induction, whereas the proliferation of DU 145 cells was not affected. Neither cell line showed any reduction in polyamine pools after manipulation nor did polyamine addition into the medium save the LNCaP cells from the growth retardation. The growth inhibition of LNCaP cells was accompanied by accumulation of cells in the G1 phase and depletion of cyclin E1 protein. These results confirm that different prostate cancer cell lines show diverse sensitivities to antizyme 1 which may not be directly polyamine related. The high gene transfer capacity of the used lentiviral vector makes the present approach a useful tool to study the therapeutic potential of antizyme 1 both in cell cultures and experimental animals. 相似文献
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Virtanen S Salonen L Laukkanen-Ninios R Fredriksson-Ahomaa M Korkeala H 《Applied and environmental microbiology》2012,78(8):3000-3003
To study the origin and spread of Yersinia enterocolitica among pigs, fecal and blood samples were repeatedly taken on a fattening farm. A few piglets were found to be already infected on breeding farms. After the piglets were mixed, the infection spread through the whole unit. Eventually, all the pigs excreted the pathogen. 相似文献