• Title/Summary/Keyword: osmotic regulation

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Isolation and Characterization of Aspergillus nidulans Mutants Which Undergo Sexual Development in Light Exposure (빛의 존재하에서도 유성분화를 하는 Aspergillus nidulans의 돌연변이체 분리 및 분석)

  • Min, Jung-Youl;Kim, Hye-Ryun;Han, Kap-Hoon;Han, Dong-Min
    • Korean Journal of Microbiology
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    • v.43 no.2
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    • pp.77-82
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    • 2007
  • In a homothallic ascomycete Aspergillus nidulans, sexual development is inhibited by various environmental stresses such as acetate medium, visible light and high osmolarity conditions. In order to study the genes involved in this stress-related regulatory network, we first attempted to isolate mutants that could develop cleistothecia even in the presence of any of those stresses including intensive visible light. More than 10,000 mutants were screened and 167 mutants were analyzed. Among them, 152 mutants underwent sexual development under the single stress condition of either high osmotic, high acetate or light condition but no sexual development in more than two stresses. Six mutants can produce cleistothecia under light or acetate stress but not in salt stress. Moreover, 6 mutants showed the ability to develop cleistothecia under the light but not under the acetate or osmo-stress. The mutants were revealed to have independent single gene mutation and grouped into different complementation groups (silA-F). The mutant alleles were all recessive to that of wild type. The light responsiveness of development implies the existence of delicate regulation process including reception and translocation of light signaling and determination of development.

Suppression of Experimental Liver Tumors by Estradiol-3-Benzoate Treatment or Castration in Male Rats

  • Byeongwoo Ahn;Jin Seok Kang;Jeong-Hwan Che;Kookkyung Lee;Ki Taek Nam;Mina Choi;Seyl Kim;Na Jin Jung;Beom Seok Han
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2002.11a
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    • pp.149-149
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    • 2002
  • Epidemiologically the incidence of liver cancer is markedly sex-differentiated, with a much higher frequency in men than in women. In experimental animals, it is also higher in male than in female irrespective of carcinogen-induced or spontaneous tumors. Therefore, we tried to investigate the modulating effects of sex hormones in experimental hepatocarcinogenesis. For induction of liver tumors, mini-osmotic pump containing diethylnitrosamine at a dose level of 47.5mg was implanted into the peritoneal cavity of the rat at 6 weeks old. To remove the effects of male sex hormones, the animals of group 2 were castrated one week prior to DEN treatment. To see the effects of estrogen, pellet containing 1g or 10g of estradiol-3-benzoate was infused subcutaneously to the animals of group 3 and 4 one week prior to DEN treatment. The pellets were exchanged every 4 weeks until sacrifice. All animals were sacrificed at 26 weeks after DEN treatment. The tumor incidences in group 1 (DEN alone), group 2 (DEN +castration), group 3(DEN +EB 1g) and group 4 (DEN +EB 10g) were 100% (15/15), 93.3% (14/15), 85.7% (12/14) and 66.7% (10/15), respectively, showing that the value of group 4 is significantly different from that of group 1. Tumor multiplicity data of group 1, 2, 3 and 4 were 5.470.73, 2.800.51, 2.070.41 and 1.670.46, respectively, showing castration or EB treatment reduced number of liver tumors significantly (P<0.001). With immunohistochemistry and Western blotting of ER the expressions were detected in normal adjacent liver cells but decreased or lost in tumor cells. From these results we conclude that female sex hormone, especially estrogen, may act as a liver tumor suppressor, and it seemed that the down regulation of ER may be associated with liver tumor development.

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Detection of Carbonic Anhydrase in the Gills of Rainbow Trout (Oncorhynchus mykiss) (무지개 송어 rainbow trout, Oncorhynchus mykiss의 아가미에서의 carbonic anhydrase의 존재)

  • Kim, Soo Cheol;Choi, Kap Seong;Kim, Jung Woo;Choi, Myeong Rak;Han, Kyeong Ho;Lee, Won Kyo;Kho, Kang Hee
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1557-1561
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    • 2013
  • Carbonic anhydrase isozymes are a widespread, zinc-containing metalloenzyme family. The enzyme catalyzes the reversible inter-conversion of $CO_2$ and $HCO_3$. This reaction is the main role played by CA enzymes in physiological conditions. This enzyme has been found in virtually all organisms, and at least 16 isozymes have been isolated in mammals. Unlike mammals, there is little information available regarding CA isozymes in the tissues of non-mammalian groups, such as fish. Carbonic anhydrase is very important in the osmotic and acid-base regulation in fish. It is well-known that the gills of fish play the most important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO_3^-$, for the maintenance of systemic pH. Rainbow trout, Oncorhynchus mykiss, is the most important freshwater fish species in the aquaculture industry of Korea, with annual production increasing each year. In addition, environmental toxicology research has shown that rainbow trout is known to be the species that is most susceptible to environmental toxins. Consequently, carbonic anhydrase was detected in rainbow trout, Oncorhynchus mykiss. The isolated protein showed the specific band with a molecular weight of 30 kDa and pI of 7.0, and it was identified as being carbonic anhydrase. The immunohistochemical result demonstrated that the carbonic anhydrase was located in the epithelial cells of the gills.

Properties of Single $K^{+}$ Channels of Skeletal Muscle Incorporated into Planar Lipid Bilayer

  • Park, Jin-Bong;Kim, Hee-Jeong;Cho, Myung-Haing;Lee, Hang;Park, Hong-Ki;Lee, Mun-Han;Ryu, Pan-Dong
    • The Korean Journal of Physiology
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    • v.29 no.1
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    • pp.13-27
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    • 1995
  • single $K^{+}$ channels of skeletal muscle from the rat and frog were into planar lipid bilayers and their properties were studied. Fusion was induced by an osmotic gradient. Of the four types of $K^{+}$ channels recorded, the two most frequently observed were a voltage and $Ca^{2+}-activated$ $K^{+}$ channel and a $K^{+}$ channel with a prominent conductance substate. The first $K^{+}$ channel was identified as the large $Ca^{2+}-activated$ $K^{+}$ (BK) channel because the open-state probability was increased with depolarization (e-fold change per $10.6{\pm}3.5$ mV, n=8) and internal $Ca^{2+}$ (half-activation at $16.7{\pm}3.8$ mV, n=8, pCa 4) and its conductance was large ($247{\pm}4.9$ pS, n=24 in 0.1 M KCI). Lifetime distributions of open- and closed-states could be fitted with single exponentials of several milliseconds. The mean open- and closed-lifetimes were linearly dependent on the intracellular $[Ca^{2+}]$ and $1/[Ca^{2+}]$, respectively. The second $K^{+}$ channel showed a conductance substate at $30{\sim}60%$ of the open state. Its current-voltage relation was linear in the range of $-80\;{\sim}\;+80\;mV$. The slope conductance of the substate and open-state were 40 and 144 pS in 0.2 M KCl, respectively. The channel was highly selective for $K^{+}$ over Cl. The open-state probability was weakly voltage-dependent (e-fold change per 35 mV. The lifetime distributions of open- and closed-states were fitted with two exponentials and the major gating occurred slowly at several hundred milliseconds. Based on the above results, we think the second type of $K^{+}$ channel is the sarcoplasmic reticulum $K^{+}$ (SRK) channel. In addition, both types of channel were also incorporated into the lipids extracted from the skeletal muscle. The channel properties recorded in the bilayers termed from synthetic and extracted lipids were qualitatively similar. Our data indicate that BK and SRK channels are rich in the skeletal muscle and their properties and regulation could be effectively studied in planar lipid bilayer.

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Modulation in NMDA and $GABA_A$ Receptor Expression after Cerebroventricular Infusion of Ginsenosides

  • Oh Seikwan;Kim Hack-Seang
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.96-112
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    • 2002
  • In the present study, we have investigated the effects of centrally administered ginsenoside Rc or Rgl on the modulation of NMDA receptor and $GABA_A$ receptor binding in rat brain. The NMDA receptor binding was analyzed by quantitative autoradiography using $[^3H]MK-801$ binding, and $GABA_A$ receptor bindings were analyzed by using $[^3H]muscimol\;and\;[^3H]flunitrazepam$ in rat brain slices. Rats were infused with ginsenoside Rc or Rg1 ($10\;{\mu}g/10{\mu}l/hr$, i.c.v.) for 7 days, through pre-implanted cannula by osmotic minipumps (Alzet, model 2ML), The levels of $[^3H]MK-801$ binding were highly decreased in part of cortex and cingulated by ginsenoside Rc and Rgl. The levels of $[^3H]muscimol$ binding were strongly elevated in almost all regions of frontal cortex by the treatment of ginseoside Rc but decreased by ginsenoside Rg 1. However, the $[^3H]flunitrazepam$ binding was not modulated by ginsenoside Rc or ginsenoside Rgl infusion. These results suggest that prolonged infusion of ginsenoside could differentially modulate $[^3H]MK-801\;and\;[^3H]muscimol$ binding in a region-specific manner. Also, we investigated the influence of centrally administered ginsenoside on the regulation of mRNA levels of the family of NMDA receptor subtypes (NR1, NR2A, NR2B, NR2C) by in situ hybridization histochemistry in the rat brain. The level of NR1 mRNA is significantly increased in temporal cortex, caudate putamen, hippocampus, and granule layer of cerebellum in Rgl-infused rats as compared to control group. The level of NR2A mRNA is elevated in the frontal cortex. In contrast, it was decreased in CAI area of hippocampus in Rgl-infused rats. However, there was no significant change of NR1 and NR2A mRNA levels in Rc-infused rats. The level of NR2B mRNA is elevated in cortex, caudate putamen, and thalamus in both Rc- and Rg-infused rats. In contrast, NR2B level is decreased in CA3 in Rgl-infused rats. The level of NR2C mRNA is increased in the granule layer of cerebellum in only Rg1 but not Rc infused rats. These results show that structure difference of ginsenoside may diversely affect the modulation of expression of NMDA receptor subunit mRNA after infusion into cerebroventricle in rats.

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Characterization of the Stretch-Activated Channel in the Hamster Oocyte (햄스터난자에서 신전에 의해 활성화되는 통로의 성상)

  • Kim, Y.-M.;Hong, S.-G.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.89-99
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    • 2004
  • Stretch-activated channels (SACs) responds to membrane stress with changes in open probability (Po). They play essential roles in regulation of cell volume and differentiation, vascular tone, and in hormonal secretion. SACs highly present in Xenopus oocytes and Ascidian oocytes are suggested to be involved in the regulation of pH and fluid transport to balance the osmotic pressure, but remain unclear in mammanlian oocytes. This study was investigated to find the presence of SACs in hamster oocytes and to examine their electrophysiological properties. To infer a role of SAC in relation to the development of early stage, we followed up to the stage of two-cell zygote with patch clamp techniques. Single channels were elicited by negative pressure (lower than ­15 cm$H_2O$). Interestingly, SACs were dependent on permeable cations such as $Na^+$ or $K^+$. As permeable cation removed from both sides across the membrane, SAC activity completely disappeared. When permeable cations present only in intracellular compartment, outward currents appeared at positive potentials. In contrast to this, inward currents occurred only at the negative voltage when permeable cation absent in cell interior. These result suggests that SAC carry cations through the nonselective cation channel (NSC channel). Taken together, we found that stretch activated channels present in hamster oocyte and the channel may carry cations through NSC channels. This stretch activated-NSC channels may play physiological role(s) in oocyte growth, maturation, fertilization and embryogenesis in fertilized oocytes to two-cell zygotes of hamster.

Identification of Novel Salt Stress-responsive Genes Using the Activation Tagging System in Arabidopsis (애기장대에서 activation tagging system을 이용한 새로운 고염 스트레스 반응 유전자의 동정)

  • Seok, Hye-Yeon;Nguyen, Linh Vu;Bae, Hyoungjoon;Ha, Jimin;Kim, Ha Yeon;Lee, Sun-Young;Moon, Yong-Hwan
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1030-1041
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    • 2018
  • Abiotic stresses limit the growth and productivity of plants. Cellular adaptation to abiotic stresses requires coordinated regulation in gene expression directed by complex mechanisms. This study used the activation tagging system to identify novel salt stress-responsive genes. The study selected 9 activation tagging lines that showed salt stress-tolerant phenotypes during their germination stages. Thermal asymmetric interlaced-PCR (TAIL-PCR) was used to identify the T-DNA tagging sites on the Arabidopsis genome in selected activation tagging lines, including AT7508, AT7512, AT7527, AT7544, AT7548, and AT7556. RT-PCR analysis showed that ClpC2/HSP93-III (At3g48870), plant thionin family (At2g20605), anti-muellerian hormone type-2 receptor (At3g50685), vacuolar iron transporter family protein (At4g27870), and microtubule-associated protein (At5g16730) were activated in AT7508, AT7512, AT7527, AT7544, and AT7556, respectively. Interestingly, in AT7548, both the genes adjacent to the T-DNA insertion site were activated: Arabinogalactan protein 13 (AGP13) (At4g26320) and F-box/RNI-like/FBD-like domains-containing protein (At4g26340). All of the seven genes were newly identified as salt stress-responsive genes from this study. Among them, the expression of ClpC2/HSP93-III, AGP13, F-box/RNI-like/FBD-like domains-containing protein gene, and microtubule-associated protein gene were increased under salt-stress condition. In addition, AT7508, AT7527, and AT7544 were more tolerant to salt stress than wild type at seedling development stage, functionally validating the screening results of the activation tagging lines. Taken together, our results demonstrate that the activation tagging system is useful for identifying novel stress-responsive genes.

Genotypical Variation in Nitrate Accumulation of Lettuce and Spinach (상추와 시금치의 품종별 질산태 질소 축적 차이)

  • Chung, Jong-Bae;Lee, Yong-Woo;Choi, Hee-Youl;Park, Yong;Cho, Moon-Soo
    • Korean Journal of Soil Science and Fertilizer
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    • v.38 no.1
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    • pp.38-44
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    • 2005
  • In addition to the variation in nitrate accumulation of vegetables due to environmental conditions, there is also a distinct genetic variation. The variation of nitrate accumulation in some cultivars of lettuce and spinach commonly cultivated in Korea was investigated. Ten cultivars for both lettuce and spinach were grown in plastic containers filled with a 1:1 mixture of perlite and vermiculite with application of Hoagland No. 2 nutrient solution of high nitrate content (17.3 mM N) in a greenhouse condition. Plants were harvested four weeks after transplanting four-leaf stage seedlings. Plant growth was measured by fresh and dry matter of shoot, and contents of nitrate and other inorganic ions and organic solutes including sugar, amino acids and organic acids were measured. Large and significant genotypical variations in the nitrate content of the plants were found for both lettuce and spinach, and high negative correlations between nitrate content and fresh or dry weight were found in lettuce and spinach. Variation in nitrate accumulation of lettuce and spinach cultivars was not directly related to the differences in contents of organic and inorganic solutes, and this result indicates that photosynthesis and osmotic regulation are not directly related with the nitrate accumulation. Considering the correlations between nitrate content and plant growth of this study, it can be simply suggested that different cultivars of lettuce and spinach have their own inherited growth and physiological characteristics and also optimum nitrogen level required for the growth. Therefore when available nitrogen in root media is higher than the optimum level required for the inherited growth potential, some of the excess nitrate supplied can be accumulated in plants.