Heat shock proteins (HSPs) are highly conserved cellular proteins that contribute to adaptive responses of organisms to a variety of stressors. In response to stressors, cellular levels of HSPs are increased and play critical roles in protein stability, folding and molecular trafficking. The mRNA expression pattern of two well-known heat shock protein transcripts, HSP70 and HSP90 were studied in two tissues of nerve ganglia, cerebral ganglion and pleuropedal ganglion of Pacific abalone (Haliotis discus hannai). It was observed that both HSP70 and HSP90 transcripts were upregulated under heat stress in both ganglion tissues. Expression level of HSP70 was found higher than HSP90 in both ganglia whereas cerebral ganglion showed higher expression than pleuropedal ganglion. The HSP70 and HSP90 showed higher expression at Day-1 after exposed to heat stress, later decreased at Day-3 and Day-7 onwards. The present result suggested that HSP70 and HSP90 synthesize in nerve ganglion tissues and may provide efficient protection from stress.

수온의 변화는 어류의 거의 모든 생리학적 부분에 영향을 미친다. 기후 변화로 인한 수온의 상 승은 어류에게 물리적 피해를 줄 수 있다. 이 연구는 최적의 수온(15°C)보다 높은 수온(20°C)에 서의 대서양 연어의 건강상태를 평가하기 위해 수행하였다. 간 조직은 열 적응에 중요한 대사 기능을 발휘하기에 본 연구에 간 조직을 사용하였다. 생체지표유전자의 개발을 위한 분석 방법 으로는 NGS RNAseq 방법을 사용하였고, 생체지표유전자의 발현 양상을 관찰하기 위한 분석 방 법으로는 RT-qPCR을 사용하였다. NGS RNAseq 분석을 통해 1,366개의 차별적 발현 유전자를 확인하였으며, 그 중에서 880개의 증가하는 유전자와 486개의 감소하는 유전자를 확인하였다. 생체지표유전자로는 heat shock protein 90 alpha (Hsp90α), heat shock protein 90 beta (Hsp90β) 및 cytochrome P450 1A (CYP1A)을 선정하였는데 이들 유전자는 NGS RNAseq 분석에서 수온의 변화에 민감하게 반응하는 유전자들이었다. 이들 유전자의 RT-qPCR을 통한 발현 양상은 NGS RNAseq 분석과 유사하게 나타났다. 이 연구의 결과는 다른 어종에도 적용할 수 있으며, 산업 적으로도 유용하다고 생각된다.

The heat shock proteins (Hsps), one of the most highly conserved groups of proteins, play crucial roles in protecting cells against environmental stressors, such as temperature, salinity, heavy metals and pathogenic bacteria. The glutathione S-transferases (GST) have important role in detoxification of oxidative damage, environmental chemicals and environmental stress. The purpose of this study is to investigate the gene expression of Hsp70 and GST on change of temperature and salinity in Mytilus coruscus. The M. coruscus was cultured in incubator of separate temperature and salinity (8, 20, 30℃×20‰, 25‰, 30‰) for 28 days. Ten individuals in each group were selected after each 14 and 28 days exposure. Results that the expression of Hsp70 mRNA was no significant changed in M. coruscus exposed to temperature (8℃, 20℃, 30℃) and salinity (20‰, 25‰, 30‰) for 14 days. Whereas the expression of Hsp70 mRNA was increased in exposure to temperature 30℃ and salinity (20‰, 25‰, 30‰) for 28 days. The expression of GST mRNA was increased in exposure to temperature 30℃, salinity (25‰, 30‰) for 14 days and temperature (8℃, 20℃, 30℃), salinity (20‰, 25‰, 30‰) for 28 days. These results suggest that Hsp70 and GST were played roles in biomarker gene on the thermal and salinity stress.

The Apostichopus japonicus is an important species in some Asia countries including Korea, China and Japan. The purpose of the present study was to investigate the differential gene expression of heat shock protein90 (Hsp90) and ferritin as a biomarker for the thermal stress during water temperature rising in the sea cucumber, A. japonicus. The A. japonicus (1.4 g) was cultured in incubator of separate temperature (15°C, 20°C, 25°C and 30°C) for each 0, 3, 6, 12, 24, 48 hours. The mRNA expression levels of Hsp90 and ferritin were examined using RT-PCR assay. Results showed that, the expression of Hsp90 mRNA was not significantly changed at 15°C. The expression of Hsp90 mRNA was significantly increased at high temperature such as 20°C and 25°C. Furthermore, Hsp90 mRNA was early increased at 25°C than 20°C. The ferritin mRNA was similar expression pattern with Hsp90. But, Hsp90 mRNA was more sensitive than ferritin mRNA at high thermal stress. These results indicate that Hsp90 and ferritin mRNAs were involved in the temperature changes response and may be play an important role in mediating the thermal stress in A. japonicas.

Five heat shock protein 70 (hsp70) isoforms (hsc70-3, hsc70-4, hsc70-5, hsc70cb, hsp70Ab) of Apis mellifera were identified from Honeybee genome database. Specific primers for each isoform were designed for the quantitative realtime RT-PCR analysis, then analyzed transcript levels of the abdomen of adult workers (3-4 weeks old) in respond to heat shock and imidacloprid ingestion. Heat shock at 45°C for 1 h induced all 5 genes but highest in hsc70cb, hsp70Ab. Ingestion of imidacloprid pesticide did not change any hsp70 isoforms at 33°C but those bees also highly responsive to heat shock at 45°C. In addition, expression level of each hsp70 isoform was various and heat shock response of each isoform was tissue-specific. For example, hsc70-3 was highest in midgut, hsc70-4 was in hypopharyngeal gland, but hsp70Ab was in fat bodies. However, heat shock response of all 5 isoforms was the highest in the fat body than brain, hypopharyngeal gland, midgut, flight muscle and integument. Our results provide information for the understanding of complicated protective mechanism of honey bee against thermal stress.

The present study was performed to identify the relationship between plasminogen activator (PA) and Heat Shock Protein-90 (HSP-90) in porcine uterus tissues during the estrous cycle. Porcine uterus tissues were obtained from preovulatory (Pre-Ov), post-ovulatory (Post-Ov) and early to mid-luteal (Early-mid L) stages. The protein was extracted from uterus tissue by using M-PER Mammalian Protein Extraction Reagent. Proteins were refined by RIPA Buffer and quantified by BCA methods. As results, t-PA expression was significantly (p<0.05) higher from pre-ovulatory(Epithelium tissue: 29,067 μg/μl, Myometrium tissue: 30,797 μg/μl) compared to the post-ovulatory stage(Epithelium tissue: 54,357 μg/μl, Myometrium tissue: 53,270 μg/μl) and early to mid-luteal stage(Epithelium tissue: 42,380 μg/μl, Myometrium tissue: 43,139 μg/μl). On the other hand, the uPA expression indicated higher from early to mid-luteal stage (Epithelium tissue: 0.02198 μg/μl, Myometrium tissue: 0.02412 μg/μl) than pre-ovulatory stage (Epithelium tissue: 0.01577 μg/μl, Myometrium tissue: 0.01531 μg/μl) and post-ovulatory stage(Epithelium tissue: 0.01414 μg/μl, Myometrium tissue: 0.01429 μg/μl). However, expression of u-PA did not differ from each estrous cycle in the epithelium tissue and myometrium tissue(p<0.05). Expression of HSP-90 was differ t-PA and u-PA from pre-ovulatory in Epithelium tissue(25,423 μg/μl) and early to mid-luteal stage in epithelium tissue(177,922 μg/μl) and myometrium tissue(26,664 μg/μl). These results suggest that HSP-90 and u-PA were related with change of uterus cycle according to the reformation of the tissues in porcine uterus.

The objective of this study was to investigate the proteome composition in pretermand term‐derived human umbilical cord. Umbilical cord samples were collected from 6 preterm infants with gestational age less than 36 weeks or 4 full terms together with medical information during prenatal period. Several biomarkers are routinely used clinically for predicting preterm labor; however, these factors are either nonspecific or detected too late. Protein profiles were performed on samples from both preterm‐ and termderived human umbilical cord by using Two‐dimensional gel electrophoresis (2‐DE). Approximately 200 different proteins were identified between preterm‐ and term‐delivered umbilical cords. Among them, differentially expressed 34 proteins were identified in 48 protein spots. In the preterm‐derived human umbilical cords, 15 proteins were present at higher levels (2.0‐ to 9.28‐fold increases) and 19 were present at lower levels (2.0‐ to 11.8‐ fold decreases or not detectable) compared to the control term‐derived umbilical cords. Proteomics approaches such as 2‐DE could greatly facilitate the discovery of new and better biomarkers. The high sensitivity and specificity achieved by the combined use of the selected biomarkers show great potential for the early detection of Adverse pregnancy outcomes such as pre‐eclampsia, small for gestational age infants, preterm delivery and placental abruption are associated with higher mortality. Increased amount of HIF‐1 α, GAPDH and HSP27 were observed in preterm‐derived umbilical cords was due to hypoxia‐ dependant and oxidative stress‐independent manner. Moreover, we isolated HUVEC (Human umbilical vascular endothelial cells) from preterm‐ and term‐derived umbilical cords and examined LDH activity. The results of the current study may provide important insights into the molecular mechanisms underlying umbilical cord development and also these data will contribute to a better understanding of the composition of preterm‐ and term ‐ derived human umbilical cord and aid the discovery of novel biomarkers for the prenatal diagnosis of fetal abnormalities

This research was attempted to seek for a positive approach within the framework of physical therapy instead of the drug treatment in the past, with regard to the ischemic brain injury in the early stage. Accordingly, the aim of this research is to observe the change of HSP27 and HSP70, the genes that are expressed in the early stage of brain injury and to investigate the effects of needle electrode electrical stimulation(NEES), upon applying NEES after ischemia. The experimental method is to give rise to global ischemia and apply NEES to 27 SD-Pat rats with the particulars of being eight-week-old, male, around 300g, and adapted to laboratory environment for more than a week, and divide them into three groups, that is, GV20 NEES group(n=9), L14 NEES group(n=9), no applied NEES global ischemia(GI) group(n=9), and then observe their changes of HSP27 and HSP70 at the time lapse of 6, 9 hr and 12 hr after ischemia, using immunohistochemistry methods. Upon observing through the immunohistochemistry method, it was noticed that there was a significant difference between the GV20 NEES group and the L14 NEES group as for HSP27 and there were significant differences among all groups as for HSP70(p<.05). Accordingly, it is supposed that the application of NEES after the outbreak of cerebral ischemia delay the apoptosis in the early ischemic part of forebrain or protect neurons against apoptosis.

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of several cells. In our previous study, inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the pig embryonic and primary cells was reported. However, its role during early bovine embryonic development is not sufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on early bovine embryonic development. We also investigated several indicators of developmental potential, including structural integrity, gene expression (apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Bovine embryos were cultured in the CR1-aa medium with or without 17-AAG for 7 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG (33.1±9.6 vs 21.7± 8.3%). The structural integrity of the blastocysts was examined by differential staining. Blastocysts from the dbcAMP- treated group had higher numbers of ICM, TE, and total cells than those from the untreated group. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (11.2 vs 3.9, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation bovine blastocysts. The m-RNA expression of the pro-apoptotic gene (Bax) increased in 17-AAG treated group, whereas expression of the antiapoptotic gene (Bcl-XL) decreased. In conclusion, Hsp90 also appears to play a direct role in bovine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with apoptosis-related genes expression in developing bovine embryos.

프로테오믹스 기법을 이용하여 벼 고온 스트레스 관련 단백질을 분리 동정하기 위하여 에서 고온처리한 벼의 줄기로부터 단백질을 분리하였다. 분리한 단백질로부터 Rubisco 단백질을 제거하기 위해 15% PEG fractionation을 실시한 후 상등액 분획의 단백질을 이차원전기 영동한 후, CBB 염색을 통해 차별적 발현을 보이는 단백질을 분석하였다. 총 46개의 단백질 spot이 발현양에 변화를 보였으며, 그 중 24개의 단백질이 고온 스트레스에 의해

Paratlanticus ussuriensis enter prolonged diapause at an egg stage. Environmental conditions, such as temperature, can modify the diapause duration at initial diapause. Eggs enter initial diapause at 20℃, but continued early embryonic development at 30℃. Final diapause at a fully developed embryonic stage is obligatory regardless of temperature conditions. To determine temperature effects on diapause mechanism of P. ussuriensis eggs, we compared weights, DNA and RNA amounts of eggs incubated at either 20℃ or 30℃ for 50 days after oviposition. Both egg weight and total amount of DNA were constant at 20℃ but gradually increased at 30℃. However, total RNA level was rapidly increased at 15 days-old eggs at 30℃ and maintained high levels during further period whereas its level was constant at 20℃. In addition, we identified three heat shock protein 70 (hsp70a, hap70b, hsp70c) genes of P. ussuriensis and determined those expression levels at different temperature conditions. The levels of hsp70a and hsp70b was not detectable until 20 days after oviposition at both temperature conditions, but highly increased at 50 and 60 days when incubated at 30℃. In contrast, hsp70c level was rapidly peaked at 20 days after oviposition, which is the time of initial diapause entrance. Our results suggest that high temperature breakdown initial diapause and a certain hsp gene, such as hsp70c, may involve in the initial diapause mechanism of P. ussuriensis eggs.

Heat shock protein 90 (Hsp90) is ATPase-directed molecular chaperon and affects survival of cancer cell. Inhibitory effect of Hsp90 by inducing cell cycle arrest and apoptosis in the cancer cell was reported. However, its role during oocyte maturation and early embryo development is very insufficient. In this study, we traced the effects of Hsp90 inhibitor, 17-allylamino-17-demethoxygeldanamycin (17-AAG), on meiotic maturation and early embryonic development in pigs. We also investigated several indicators of developmental potential, including structural integrity, gene expression (Hsp90-, cell cycle-, and apoptosis-related genes), and apoptosis, which are affected by 17-AAG. Then, we examined the roles of Hsp90 inhibitor on viability of primary cells in pigs. Porcine oocytes were cultured in the NCSU-23 medium with or without 17-AAG for 44 h. The proportion of GV arrested oocytes was significantly different between the 17-AAG treated and untreated group (78.2 vs 34.8%, p<0.05). After completion of meiotic maturation, the proportion of MII oocytes was lower in the 17-AAG treated group than in the control group (27.9 vs 71.0%, p<0.05). After IVF, the percentage of penetrated oocytes was significantly lower in the 17-AAG treated group (25.2%), resulting in lower normal pronucleus formation (2PN of 14.6%). Therefore, the inhibition of meiotic progression by Hsp90 inhibitor played a critical role in fertilization status. Porcine embryo were cultured in the PZM-3 medium with or without 17-AAG for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without 17-AAG. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 17-AAG treated group compared with control (7.5 vs 4.4, respectively). Blastocysts that developed in the 17-AAG treated group had low structural integrity and high apoptotic nuclei than those of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. The mRNA expressions of cell cycle-related genes were down-regulated in the 17-AAG treated group compared with control. Also, the expression of the pro-apoptotic gene Bax increased in 17-AAG treated group, whereas expression of the anti-apoptotic gene Bcl-XL decreased. However, the expression of ER stress-related genes did not changed by 17-AAG. Cultured pESF cells were treated with or without 17-AAG and used for MTT assay. The results showed that viability of pESF cells were decreased by treatment of 17-AAG (2 μM) for 24 hr. These results indicated that 17-AAG decreased cell proliferation and increased cell death. Expression patterns Hsp90 complex genes (Hsp70 and p23), cell cycle-related genes (cdc2 and cdc25c) and apoptosis-related genes (Bax and Bcl-XL) were significantly changed by using RT-PCR analysis. The spliced form of pXbp-1 product (pXbp-1s) was detected in the tunicamycin (TM) treated cells, but it is not detected in 17-AAG treated cells. In conclusion, Hsp90 appears to play a direct role in porcine early embryo developmental competence including structural integrity of blastocysts. Also, these results indicate that Hsp90 is closely associated with cell cycle- and apoptosis-related genes expression in developing porcine embryos.

A glucose-regulated protein 78 (grp78) gene, which is belongs to a heat shock cognate 70 (hsc70) subfamily, was cloned from Indian meal moth, Plodia interpunctella. Its full length cDNA was 2679 bp and contains a 1980 bp open reading frame. The translated amino acid sequence consists of 660 residues with a calculated molecular mass of 72,975 Da and an isoelectronic point (pI) of 5.27. It contains several highly conserved functional motifs of the Hsp70 family and, particularly, C-terminal motif of KDEL that is characteristic for endoplasmic reticulum (ER) hsc70. Its deduced amino acid sequence shows a high identity (83-94%) with Hsc70s of other insects and grouped with Hsc70-3 among 5 Hsc70 members of Drosophila melanogaster. During development the grp78 transcript level was high in egg, feeding larval and adult stages but low in molting and wandering larval and pupal stages. Particularly its level was higher in the gut than integument and fat body of fifth instars. Furthermore its level was greatly decreased when fifth instar larvae were starved for 48 hrs but recovered at 3-6 hrs after re-fed diet. Our data suggests that grp78 is a member of hsc70 gene that belongs to ER and may have a role for energy metabolism at cellular level.

We examined the effects of cadmium exposure and various temperature stress on the expression of Pardosa astrigera heat shock protein 70 (HSP70). To do this, P. astrigera HSP70 gene was cloned and its sequence determined. Female spiders collected from non-contaminated region were exposed to 40mM CdCl2 for 2, 4 and 6 weeks by dietary uptake. At the end of every 2, 4 and 6 weeks of exposure, a batch of 5 spiders was collected and total RNA was extracted from each batch of whole bodies. Female spiders were also exposed to different temperatures (20, 25, 30 and 35℃) for 3h and RNA extracted likewise. Transcription profiles of HSP70 in response to cadmium and temperature were determined by quantitative real-time PCR using 18S rRNA as reference gene for data normalization. HSP70 transcription gradually increased during 2,4 and 6 weeks of exposure to cadmium. In particular, the expression level at 6-week exposure was 3.4-fold higher than that of untreated control. In the temperature response, an increased expression of HSP70 was also observed as temperature increased up to 30℃ and then slightly decreased at 35℃. The expression level at 30℃ was 2.3-fold higher than that of 25℃. Taken together, HSP70 gene appears to be up-regulated by general stress factors including cadmium exposure and temperature increase.

Heat shock p1'otein (Hsp) 40 acts as a co-cha perone of Hsp70 by fac ilitating the ATPase activity of Hsp70 as well as promoting the protein fo lding and rena turation by Hsp70. In the present study. Hsp40 gene was fu sed with a gene f’ragment encoding the HJV-l TAT protein transduction domain(YGRKKRRQRRR) in a bacterial exp1' ession vector pTAT-HA to produce the TAT-fused Hsp40(TAT- Hsp40). Purified TAT-Hsp40 was effectively t 1'ansduced into the HEK 293 cells in a time- and dose-dependent manne1' To examine the effec t of TAT- Hsp40 upon oxidalive stress, HEK293 cells were exposed to H2Û2. Oxidative stress induced the ra pid increase of proteasome activity foll owed by celJ death in HEK 293 cells However ‘ HEK 293 cells t 1'ansduced by TAT- Hsp40 showed r esistance against oxidative st 1'ess induced cytot oxicity, TAT- Hsp40 transduced cell s showed dec1'eased p1'oteasome activity and inhibi ted Hs p70 degradation. These results suggest that Hs p40 rnight protect cell death f rom oxida tive st1'ess by p1'ese1'ving the cellula1' level of Hs p70.