The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

This study was carried out to evaluate the effect of culture methods on development of embryos with each developmental stage after heat shock in bovine oocytes. The results obtained were as fellows. 1. The culture method after heat shock on development of embryos was better drop-culture than co-culture. 2. The medium without amino acids were not effect of heat sock on development of embryos but it was in need of amino acid during formation of blastocyst.

엽록제 small HSP의 기능을 조사하기 위하여 항상적으로 발현하는 형질전환 식물체를 작성하였다. 고온 스트레스 하에서의 형질전환 식물체의 고온내성을 chlorophyll 형광으로 측정하였다. Leaf disc를 고온조건에서 5분간 처리한 후, 광화학계 II의 불활성화를 나타내는 Fo 값의 증가 또는 Fv 값의 감소치를 조사하였다. 형질전환 식물체는 고온 스트레스 하에서의 이들 값의 증감율이 현격히 감소하였다. 또한 유식물체를 에서 45분간 처리한 후, 에서 계속적으로 배양하였을 때, 비형질전환 식물체는 전부 고사하였으나, 형질전환 식물체의 약 80%는 생존하였다. 이러한 결과는 엽록체 small HSP가 고온 스트레스 하에서 광합성기구를 보호하는데 있어서 중요한 기능을 담당하고 있음을 나타낸다.

고등식물에 있어서 엽록체에 존재하는 저 분자량 heat shock protein (smHSP)은 식물의 내열성 획득에 있어서 필수유전자임이 mutant를 이용한 유전학적인 연구에 의해 보고된 바 있다. 고온내성이 강한 작물인 벼로부터 엽록체 smHSP cDNA를 분리하고자 벼의 잎에서 분리한 mRNA로 작성한 cDNA library로부터 screening하였다. 선발된 smHSP cDNA는 1,026 bp의 염기로 구성되어 있었으며, 239개의 아미노산

목초의 내하고성을 증진시킬 목적으로 이용하고자 하는 내열성 유전자 (BcHSP17.6)의 발현양상을 배추에서 조사하였다. BcHSP17.6 단백질을 항원으로 한 항체를 생산하여 항원-항체 반응으로 확인한 결과, 생산된 항체는 항원과 결합하므로서 항체가 정상적으로 생산되었음을 확인하였다. 이 항체를 이용하여 다양한 heat shock (HS) 조건에서 15-~18-kD low molecular weight heat shock proteins (LMW HSPs)가 축적되는지를 조사하였다. 이들 LMW HSPs는 $35^{\circ}C$ 수식 이미지 처리에서 나타나기 시작하였으며, $40^{\circ}C$ 수식 이미지에서 4시간 처리하였을 때 total protein 100 mg당 $1.56{\mu}g$ 수식 이미지이 축적되어 최대치를 나타내었다. 또한 합성된 이들 LMW HSPs는 HS 처리후 24시간이 경과되어도 거의 변함이 없었으며, $40^{\circ}C$ 수식 이미지 보다 높은 온도 조건에서는 축적량이 감소하였다. 이와 같이 일단 합성된 LMW HSPs가 장시간 동안 감소하지 않는다는 사실로부터, 이들 HSPs들이 식물체가 내열성을 가지도록 하는데 관여하며, 더 나아가서 배추가 정상 생육온도보다 더 높은 치사온도에서도 살아남을 수 있도록 하는데 중요한 역할을 하는 것으로 사료된다.

본 연구는 食品衛生과 環境衛生상 주요 危害因子중의 하나인 카드뮴에 대하여 식품으로서 섭취기회가 많은 마늘성분과 비타민 A를 이용한 危害 경감 혹은 방어효과를 평가하고, 카드뮴暴露에 대한 細胞水準의 反應으로서 HSP(Heat Shock Protein, 熱衝擊蛋白質 혹은 스트레스蛋白質)의 發顯時期와 發顯程度를 관찰하고자 실시하였다. 실험동물로는 Wistar계 SPF 수컷 랫드 483마리를 사용하였으며, 對照群, 카드뮴投與群(Cd, CdCl₂20 ㎎/㎏), 카드뮴과 마늘유 (diallyl disulfide) 投與群(Cd+Dds, diallyl disulfide 50 ㎎/㎏), 카드뮴과 비타민 A(retinol acetate) 投與群(Cd+Ra, retinol acetate 50,000 I.U./㎏)으로 구분하였다. 시험물질은 시간 간격에 따라 1, 2, 4, 8, 16, 24시간, 2, 4. 7일, 2, 4, 8, 16주에 投與하였으며, 관찰한 결과는 다음과 같다. 1. 카드뮴投與 後 4시간에 血中 카드뮴含量이 0.972∼l.256 ㎍/g으로 對照群의 0.004㎍/g에 비하여 높아졌으며, 投與 2주까지는 肝臟의 카드뮴含量이 23.76∼24.84 ㎍/g으로 腎臟의 20.53∼22.03 ㎍/g보다 높았으나, 8주 후부터는 肝臟의 카드뮴含量(82.48∼86.37 ㎍/g)보다 腎臟의 카드뮴含量(98.0∼109.8 ㎍/g)이 더 높았다. 따라서, 카드뮴은 投與 後 4시간에 혈액 중에 높게 나타나고, 점차 肝臟으로 이동하며 다시 肝臟에서 腎臟으로 이동함으로써, 8주 이후에는 肝臟보다 腎臟의 카드뮴 蓄積量이 많아지는 것으로 나타났다. 2. 카드뮴投與에 의한 血淸중의 酸素 變化에서 ALT(Alanine Aminotransferase), AST(Aspartate Aminotransferase) 活性度는 投與 後 1주일부터 각자 60.3∼73.0 U/ℓ, 135.5∼149.8 U/ℓ로 증가했지만 肝臟 毒性을 일으키는 정도는 아니었으며, glucose는 投與 8주에 對照群(113.8 ㎎/㎗)에 비하여 Cd+Ra群을 제외한 모든 群에서 72.8∼77.5 ㎎/㎗의 낮은 결과를 보여 대조군과 투여군간의 통계학적 有意性(p<0.05)을 보였다. BUN(blood urea nitrogen)은 投與 4주에 對照群의 19.3 ㎎/㎗에 비하여 Cd, Cd+Dds群에서 24.8∼25.8 ㎎/㎗ 有意한 증가를 보였으나, Cd+Ra群에서는 投與 16주에, Cd群의 33.2 ㎎/㎗에 비하여 27.3 ㎎/㎗으로 有意하게(p<0.05) 낮았다. Creatinine은 모든 試驗群에서 投與 8주에 對照群의 0.77 ㎎/㎗에 비하여 0.98∼1.38 ㎎/㎗로 높은 증가를 보였으며(p<0.05), 16주에는 Cd群의 1.20 ㎎/㎗에 비하여 Cd+Ra群은 1.02 ㎎/㎗로 有意하게 낮은 결과를 보였다(p<0.05). 3. 病理組織學적 檢査 결과 Cd群과 Cd+Ra群에서는 投與 8주부터 腎臟細尿管의 內腔에 好酸性顆粒이나 尿圓柱(urinary cast)가 관찰되었으며, Cd+Dds群은 腎臟 近位細尿管 上皮細胞에 顯著한 變性壤死가 나타났다. ??丸은 카드뮴投與 8주부터 精細管(seminiferous tubule) 사이의 間質組織이 好酸性 液體의 貯留에 의해 확장되어 주위의 精細管이 심하게 萎縮되는 등의 소견을 보였다. Cd+Dds群과 Cd+Ra群에서는 Cd群보다 病變이 微弱하였다. 카드뮴을 2주동안 投與한 랫드에서 腎臟의 電子顯微鏡 所見은 近位曲細尿管 細胞의 細胞質 腫脹, 미토콘드리아의 變性, 蛋白質 小球의 증가, 毛細血管 內皮細胞의 細胞質 腫脹과 空砲形成등이 관찰되기 시작하였으며, 投與 8주에 나타난 腎臟細尿管 內腔의 無形質 小球는 탈락된 變性 上皮細胞인 것으로 확인되었다. 4. HSP_(70)은 카드뮴投與 後 2시간부터 發顯이 증가되어, 48시간까지 持續되며, 이후에 原狀態로 恢復되는 것으로 나타났다. 위와같은 HSP_(70)의 發顯은 diallyl disulfide와 retinol acetate 投與에 의해 영향을 받지 않았다. 腎臟에서의 HSP_(70) 發顯은 絲球體와 細尿管上皮細胞에서 주로 發顯되는 것으로 나타났다. 이상의 결과로부터, 랫드에 카드뮴을 投與하였을 때 HSP_(70) 發顯은 投與 後 2∼4시간 내에 나타나는 迅速한 反應임을 알 수 있었고, 마늘유(diallyl disulfide) 投與는 카드뮴에 의한 腎臟損傷을 促進시키나, 비타민 A(retinol acetate) 投與는 카드뮴에 의한 腎臟損傷을 抑制시키는 효과를 보였다. 카드뮴에 의한 ??丸손상은 마늘유(diallyl disulfide) 흑은 비타민 A(retinol acetate)를 投與함으로써 損傷 防禦效果를 보였다.

The quality and shelf life of sliced root of Platycodon grandiflorum (Doraji) treated by ozon-microbubble-heat shock (OMH) were investigated by combining modified-atmosphere packaging [MAP, (50% O2+15% CO2+35% N2)]. The study was based on microbiological (total viable bacteria, fungi, Enterobacteriaceae and coliforms numbers), physicochemical and sensory changes. OMH treatment was effective in reducing microbial populations of the sliced Doraji, especially Enterobacteriaceae and coliforms reduced by 2 log CFU/g. However OMH-MAP treatment remained the aerobe and fungi numbers. Regarding the color, OMH-MAP delayed the change of Hunter b* and the sliced Doraji by OMH-MAP treatment exhibited lower decrease of flavor and overall acceptability compared to those by polypropylene packaging after tap water treatment (Control). The OMH and 50%O2+15%CO2 MAP treatment gave better sensory quality and extended shelf-life for sliced Doraji (~3 days longer shelf-life than Control). Flavor was significantly related to overall acceptability at both Control and OMH-MAP, whereas total coliforms prevalence was associated with overall acceptability at only OMH-MAP. Therefore microbubble-heat shock treatment may improve microbial safety and sliced Doraji by OMH treatment can stored under 50%O2+15%CO2 treatment for up to 7 days. Thus, OMH and MAP treatment may be used in maintaining the storage quality and marketability of sliced Doraji.

Heat shock protein (HSP) 70, the highly conserved stress protein families, plays important roles in protecting cells against heat and other stresses in most animal species. In the present study, we identified and characterized four Hsp70 (RuHSP4, RuHSC70, RuHSP12A, RuGRP78) family proteins based on the expressed sequence tag (EST) analysis of the Korean rose bitterling R. uyekii cDNA library. The deduced RuHSP70 family has high amino acid identities of 72-99% with those of other species. Phylogenetic analysis revealed that RuHsp70 family clustered with fish groups (HSP4, HSC70, HSP12A, GRP78) proteins. Quantitative RT-PCR analysis showed the specific expression patterns of RuHsp70 family members in the early developmental stages and several tissues in Korean rose bitterling. The expression of 4 groups of Hsp70 family was detected in all tested tissue. Particularly, Hsp70 family of Korean rose bitterling is highly expressed in hepatopancreas and sexual gonad (testis and ovary). The expression of Hsp70 family was differentially regulated in accordance with early development stage of Rhodeus uyekii.

Heat shock transcription factors(HSFs) are the major heat shock factors regulating the heat stress response. They participate in regulating the expression of heat shock proteins (HSPs), which are critical in the protection against stress damage and many other important biological processes. In this study, a genome-wide analysis was carried out to identify all HSFs soybean genes. Twenty six nonredundant HSF genes(GmHsf) were identified in the latest soybean genome sequence. Chromosomal location, protein domain and motif organization of GmHsfs were analyzed in soybean genome. The phylogenetic relationships, gene duplications and expression profiles of GmHsf genes were also presented in this study. According to their structural features, the predicted members were divided into the previously defined classes A–C, as described in Arabidopsis. Using RT-PCR, the expression patterns of 26 GmHsf genes were investigated under heat stress. The data revealed that these genes presented different expression levels in response to heat stress conditions. Real-time (q)RT-PCR was performed to investigate transcript levels of five GmHsfs in response to multiple abiotic stresses. Differential expression of five GmHsfs implies their role during abiotic stresses. Subcellular localization using GFP-fusion protein demonstrated that GmHsf12 and GmHsf34 were restricted to the nucleus and GmHsf28 was localized in the nucleus and cytoplasm in plant. The results provide a fundamental clue for understanding of the complexity of the soybean HSF gene family and cloning specific function genes in further studies and applications.

Heat shock pretreatment, dark culture period and washing medium could have marked effects on microspore embryogenesis. A heat shock pretreatment of microspores at 32.5°C for 48 hours gave high production rate of microspore-derived embryo (MDE) when compare to shorter and longer period. The yield of MDE increased significantly when microspore cultured for 15 days at 25℃ in dark condition followed by heat shock pretreatment. MDE were browned and lost vitality when dark treatment period extended longer than 15 days. This is caused by an insufficient oxygen and light for growing embryo which already formed during dark treatment period. The vitality of a microspore isolated from flower bud stored at 4℃ become decreased at the very first day and the vitality of microspore stored at 4℃ in the form of flower bud itself become decreased from the 5th day after storage. This shows the possibility of getting a certain period of storage for a suitable flower bud in MDE formation. The yield of MDE was most effective when isolated microspore was had with MS medium compared to B-5 and NLN medium and also showed most effective result with sucrose 130 g􀂂L-1 in additional sucrose concentration. The above result is thought to be very useful for the development of a new cultivar of radish and other many crops including Brassica using haploid breeding technology.

We used a microarray dataset that is deposited in the public database to evaluate plant responses to heat stress and selected two genes, OsSHSP1 (Os03g16030) and OsSHSP2 (Os01g04380), that are highly expressed under heat stress in rice. OsSHSP1 and OsSHSP2 gene transcripts were highly induced in response to salt and drought. In addition, OsSHSP1 and OsSHSP2 gene transcripts were induced under ABA and SA. Subcellular localization of proteins of 35S::OsSHSP1 were associated with the cytosol, whereas those of and 35S::OsSHSP2 were associated with the cytosol and nucleus. Heterogeneous overexpression of both genes exhibited higher germination rates than those of wild-type plants under the salt treatment, but not under heat or drought stress. The network of both genes harboring 9 sHSPs as well as at least 13 other chaperone genes might support the idea of a role for sHSPs in the chaperone network. Our findings might provide clues to shed light on the molecular functions of OsSHSP1 and OsSHSP2 in response to abiotic stresses, especially heat stress.

Heat shock transcription factors (HSFs) are the major heat shock factors regulating the heat stress response. They participate in regulating the expression of heat shock proteins (HSPs), which are critical in the protection against stress damage and many other important biological processes. In this study, a genome-wide analysis was carried out to identify all HSFs soybean genes. Twenty six nonredundant HSF genes (GmHsf) were identified in the latest soybean genome sequence. Chromosomal location, protein domain and motif organization of GmHsfs were analyzed in soybean genome. The phylogenetic relationships, gene duplications and expression profiles of GmHsf genes were also presented in this study. According to their structural features, the predicted members were divided into the previously defined classes A–C, as described in Arabidopsis. Using RT-PCR, the expression patterns of 26 GmHsf genes were investigated under heat stress. The data revealed that these genes presented different expression levels in response to heat stress conditions. Real-time (q)RT-PCR was performed to investigate transcript levels of five GmHsfs in response to multiple abiotic stresses. Differential expression of five GmHsfs implies their role during abiotic stresses. Subcellular localization using GFP-fusion protein demonstrated that GmHsf12 and GmHsf34 were restricted to the nucleus and GmHsf28 was localized in the nucleus and cytoplasm in plant. The results provide a fundamental clue for understanding of the complexity of the soybean HSF gene family and cloning specific function genes in further studies and applications.

Heat shock transcription factors (HSFs) are the major heat shock factors regulating the heat stress response. They participate in regulating the expression of heat shock proteins (HSPs), which are critical in the protection against stress damage and many other important biological processes. In this study, a genome-wide analysis was carried out to identify all HSFs soybean genes. Twenty six nonredundant HSF genes (GmHsf) were identified in the latest soybean genome sequence. Chromosomal location, protein domain and motif organization of GmHsfs were analyzed in soybean genome. The phylogenetic relationships, gene duplications and expression profiles of GmHsf genes were also presented in this study. According to their structural features, the predicted members were divided into the previously defined classes A–C, as described in Arabidopsis. Using RT-PCR, the expression patterns of 26 GmHsf genes were investigated under heat stress. The data revealed that these genes presented different expression levels in response to heat stress conditions. Real-time (q)RT-PCR was performed to investigate transcript levels of five GmHsfs in response to multiple abiotic stresses. Differential expression of five GmHsfs implies their role during abiotic stresses. Subcellular localization using GFP-fusion protein demonstrated that GmHsf12 and GmHsf34 were restricted to the nucleus and GmHsf28 was localized in the nucleus and cytoplasm in plant. The results provide a fundamental clue for understanding of the complexity of the soybean HSF gene family and cloning specific function genes in further studies and applications.

The molecular responses to various abiotic stresses were investigated by the approaches with transcriptomic analysis based on an ACP system. Here we identified differentially expressed genes under abiotic stresses in alfalfa seedlings and they were mostly unknown genes and a few common stress-related genes. Among them, mitochondrial small HSP23 was responded by the diverse stress treatment such as heat, salt, As stresses and thus it could be a strong candidate that may confer the abiotic stress tolerance to plants. When expressed in bacteria, recombinant MsHSP23 conferred tolerance to salinity and arsenic stress. Furthermore, MsHSP23 was cloned in a plant expressing vector and transformed into tobacco, a eukaryotic model organism. The transgenic plants exhibited enhanced tolerance to salinity and arsenic stress under ex vitro conditions. In comparison to wild type plants, the transgenic plants exhibited significantly lower electrolyte leakage. Moreover, the transgenic plants had superior germination rates when placed on medium containing arsenic. Taken together, these overexpression results imply that MsHSP23 plays an important role in salinity and arsenic stress tolerance in transgenic tobacco. The results of the present study show that overexpression of alfalfa mitochondrial MsHSP23 in both eukaryotic and prokaryotic model systems confers enhanced tolerance to salt and arsenic stress. This indicates that MsHSP23 could be used potentially for the development of stress tolerant transgenic crops, such as forages.

HSP70 has widely been induced in in vivo hyperthermia conditions in various organisms to study gene regulation and recently neuroprotectve roles of the induced gene expression under varying conditions. We investigated different responses among various tissues in zebrafish under heat shock to evaluate whether spatial and temporal expression pattern of zebrafish (z) hsp70 in transcriptional and translational level under heat shock stress in different brain regions. Heat shock groups were given for 1 h at after recovery by transferring the treated animals back to for 1, 2 and 24 h for recovery, respectively. Control (CTRL) group was kept at . At the end of treatments, five animals were collected and used for isolation of total RNAs and peptides from the corresponding tissues. Expression of zhsp70 mRNA showed different patterns in recovery periods in the tissues including the brain, eye, intestines, muscles, heart and testis by RT-PCR. Unlike the RT-PCR analysis, Northern blot analysis demonstrated nearly 30-fold increase in zhsp70 at 1 h heat shock, suggesting that RT-PCR may not be appropriate in unmasking regulation of the time-dependent zhsp70 expression. In the experiment involving different brain regions, the cerebellum showed gradual activation at 1 h to R1h and decreases in R2h and R24h, while the medulla oblongata and optic tectum showed gradual increase at R1h and decrease at R24h, indicating that different brain tissues respond specifically to heat shock in inducing zhsp70 and recovering from the heat shock status. Western blot analysis also demonstrated that the intracellular levels of zHSP70 in three different brain regions including the cerebellum, medulla oblongata and optic tectum are differently induced and recovered to normal state. These results clearly demonstrate that different regions of the body and the brain tissues are responding differently to heat shock in the aspects of its level of expression and speed of recovery.