• Animal Bioscience
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• 제36권5호
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• pp.704-709
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• 2023

Objective: In tropical, subtropical and arid zones, heat stress is the main cause of productivity reduction in cattle. When climate stressors occur, animals become thermal adapted through differential expression of some genes, including heat shock proteins (HSP) family. The aim of this study was to determine levels of expression of HSP60, HSP70, and HSP90 genes in Simmental cattle raised in tropical environments of Mexico. Methods: In this study, expression of HSP60, HSP70, and HSP90 genes was analyzed in 116 Simmental cattle from three farms with tropical climate located in western Mexico. Animals were sampled twice a day, in the morning and noon. Gene expression was evaluated by quantitative polymerase chain reaction using probes marked with fluorescence. The MIXED procedure of SAS with repeated measures was used for all statistical analysis. Results: HSP60 gene expression differences were found for sex (p = 0.0349). HSP70 gene differences were detected for sampling hour (p = 0.0042), farm (p<0.0001), sex (p = 0.0476), and the interaction sampling hour×farm (p = 0.0002). Gene expression differences for HSP90 were observed for farm (p<0.0001) and year (p = 0.0521). HSP70 gene showed to be a better marker of heat stress than HSP60 and HSP90 genes. Conclusion: Expression of HSP70 gene in Simmental herds of the tropical region of western México was different during early morning and noon, but the expression of the HSP60 and HSP90 genes was similar. Identification of resilient animals to heat stress will be useful in the genetic improvement of the Simmental breed.

• Journal of Genetic Medicine
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• 제1권1호
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• pp.51-56
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• 1997

Spermatogenesis is known to be regulated by a number of genes and several factors such as hormones, growth factors, cytokines and others. This study was done to evaluate the relationship between HSPs and DAZ genes in human spermatogenesis; we observed the expression pattern of HSP gene in azoospermia men with DAZ gene that regulated the gene expression related with human spermatogenesis. RT-PCR method was used to detect DAZ, HSP70A, and HSP70B transcripts in all RNA samples. Total RNA was extracted from 21 testis tissues using TRIZOL reagent. cDNAs were synthesized with reverse transcriptase, AMV. All PCR reaction were performed on a PCR themocycler with DAZ, HSP70A, and HSP70B-specific primers. Semen analysis, karyotyping and testis histology were performed. DAZ gene, known as a candidate gene of azoospermia factor(AZF), was deleted in 2 of 21 patients. To evaluate the only effects of HSPs in this patients, 2 DAZ deleted cases were removed. We observed the mRNA of HSP70B in 5 whereas none could be seen with regard to HSP70A. Furthermore, the sperm of these 5 men were discovered to be immature. In conclusion, HSP70B as well ad DAZ gene seem to be involved causing spermatogenic failure. We suggest that HSP70B plays an important role in spermatogenesis and it is one of factors induced sperm maturation in human.

• BMB Reports
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• 제39권6호
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• pp.749-758
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• 2006

The cytosolic members of the HSP70 family of proteins play key roles in the molecular chaperone machinery of the cell. In the study we cloned and sequenced the full-length cDNA of Delia antiqua HSP70 gene, which is 2461 bp long and encodes 643 a.a. with a calculated molecular mass of 70,787 Da. We investigated gene copies of cytosolic HSP70 members of 4 insect species with complete genome available, and found that they are quite variable with species. In order to characterize this protein we carried out an alignment and a phylogenetic analysis with 41 complete protein sequences from insects. The analysis divided the cytosolic members of the family into two classes, HSP70 and HSC70, distinguishable on the basis of 15 residues. HSP70 class members were slightly shorter in length and smaller in molecular mass relative to the HSC70 class members, and the conservative and functional regions in these sequences were documented. Mainly, we investigated the expression of Delia antiqua HSP70 gene, in response to diapauses and thermal stresses. Both summer and winter diapauses elevated HSP70 transcript levels. Cold-stress led to increased HSP70 expression levels in summer- and winter-diapausing pupae, but heat-stress elevated the levels only in the winter-diapausing pupae. In all cases, the expression levels, after being elevated, gradually decreased with time. HSP70 expression was low in non-diapausing pupae but was up-regulated following cold- and heat-stresses. Heat-stress gradually increased the mRNA level with time whereas cold-stress gradually decreased levels after an initial increase.

• 생태와환경
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• 제49권1호
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• pp.22-30
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• 2016

연안의 다양한 환경변화는 서식 생물에 영향을 미치고, 양식장의 생산량 감소와 연결되고 있는 추세이다. 본 연구에서는 가막만의 대표적인 양식종인 패류 C. gigas와 M. galloprovincialis의 서식환경에 따른 스트레스 정도를 파악하고자 하였다. 이를 위해, 각 종의 체중량, 각장과 각고, 양식장 사육기간을 조사하고, 각 종의 계통학적 HSP 70 sequence를 비교한 후, 각 종의 HSP 70 유전자 발현을 분석하였다. 그 결과, C. gigas의 체중량, 각장과 각고는 C2 양식장이 높게 나타났으나, 양식장 환경 사육기간과 HSP 70 유전자 발현은 C3 양식장이 가장 높았다. M. galloprovincialis는 M1 양식장의 체중량이 높게 나타났으며 각장과 각고, 사육기간은 M2와 유사하였으나, HSP 70 유전자 발현은 M2 양식장이 통계적으로 유의한 수준으로 높게 나타났다. 그리고 C. gigas와 M. galloprovincialis의 HSP 70 sequence 분석을 통해서 다른 해양 종들과 높은 유사성이 있음을 확인하였다. 이 결과는 서식환경에 따라 생물의 외부적 형질뿐만 아니라 내부적 스트레스를 HSP 70 유전자 발현을 통하여 파악할 수 있으며 HSP 70은 외부환경 스트레스를 평가하는 지표 유전자로서 활용할 수 있을 것이다.

• International Journal of Industrial Entomology and Biomaterials
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• 제16권1호
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• pp.21-27
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• 2008

The expression of heat shock protein genes (Hsp 70, Hsp 40, Hsp 20.8 and Hsp 20.4) against thermal stress in silkworm Bombyx mori was performed through semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Upon exposure of silkworm to two temperature regimes ($38^{\circ}C$ and $42^{\circ}C$), significant change in the expression of Hsp gene was observed as compared to the control. Hsp 70 and Hsp 40 showed increased expression than the small heat shock protein genes Hsp 20.8 and Hsp 20.4. The Hsp 70 showed increased expression during the recovery period as compared to 1 hr thermal treatments ($38^{\circ}C$/1 hr and $42^{\circ}C$/1 hr). Whereas, Hsp 40, Hsp 20.8 and Hsp 20.4 genes showed higher expression level at initial stages that later gradually decrease during recovery period. Tissue specific expression of Hsp 70 showed variation in the level of expression amongst the tissues. The mid gut and fat body tissues showed higher expression than the cuticle and silk gland tissue. The Hsp 70, Hsp 40 gene expression was analyzed in thermotolerant (Nistari) and thermo susceptible silk worm strain (NB4D2) and results showed significant variation in their expression level. The Nistari showed higher expression of Hsp 70 and Hsp 40 genes than the NB4D2. These findings provide a better understanding of cellular protection mechanisms against environmental stress such as heat shock, as these Hsps are involved in an organism thermotolerance.

• 한국발생생물학회지:발생과생식
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• 제15권4호
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• pp.365-371
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• 2011

수온은 어류를 포함한 수중동물의 다양한 생물학적 사건에 영향을 미친다. 어류의 번식 및 발생도 수온의 변화와 밀접한 관련이 있다. 어류를 높은 수온에 노출시키면 HSP70 유전자 발현을 유도한다. 따라서, HSP70 유전자 발현을 효과적으로 차단할 수 있는 저해제의 개발은 온도에 민감한 생물학적 사건과 관련된 HSP70의 역할을 연구하는데 크게 기여할 수 있다. 본 연구에서는 고온($36^{\circ}C$)에 노출된 틸라피아 자어와 치어(전장 10~13 cm)에서 천연 flavonoid의 일종인 quercetin, 3,3',4',5,7-pentahydroxyflavon의 "고온 유도 HSP70 유전자 발현" 억제효과를 조사하였다. 고온 노출 전에 50 ${\mu}M$ 이상의 quercetin 용액에 6시간 동안 사전 침지한 실험어에서는 HSP70 유전자 발현이 유의하게 감소하였다(P<0.05). 특히, 100 ${\mu}M$로 침지 처리한 개체에서는 고온 처리를 하지 않은 실험어에서 만큼 낮은 HSP70 유전자 발현을 나타내었다. 한편, 0.1 $m{\ell}$씩의 0.5 mM, 5 mM 또는 20 mM의 quercetin 용액을 치어에 주사한 결과, 고수온 유도 HSP70 유전자 발현은 0.5 mM의 quercetin을 주사한 치어의 생식소와 뇌에서 모두 유의하게 감소되었다(P<0.05). 본 연구의 결과는 quercetin이 어류에서 고온에 의해 유도된 HSP70 유전자 발현을 효과적으로 차단할 수 있으며, 틸라피아 자어 침지의 경우 유효 농도 100 ${\mu}M$ 그리고 치어 주사의 경우 유효 농도 0.5 mM로 원하는 효과를 얻을 수 있음을 제시한다.

• 한국해양생명과학회지
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• 제5권1호
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• pp.17-24
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• 2020

참다슬기 아가미 조직으로부터 heat shock protein 70 유전자를 분리·동정하였다. 참다슬기 HSP70 cDNA의 open reading frame (ORF)는 1,917 bp로 639개의 아미노산을 암호화하여 분자량은 약 70 kDa으로 예측되었다. 생물정보학 배열분석에 의해 HSP 유전자 기능과 관여되어 있는 3가지 주요 signature motifs와 보존된 도메인을 확인하였다. 계통학적 분석을 통하여 참다슬기 HSP70 유전자는 왕우렁이 Pomacea canaliculate와 같은 클러스트에 포함된다는 사실을 확인하였다. 수온 및 염분 변화에 따라, 참다슬기 HSP70 mRNA 유전자 레벨은 유의적으로 증가하였으며(p < 0.05), 이는 외부자극요인을 파악할 있는 분자생물학적 마커로서 활용될 수 있을 것으로 사료된다.

• 한국양식학회지
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• 제12권2호
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• pp.91-100
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• 1999

The complete nucleotide sequence of olive flounder (Paralichthys olivaceus) hsp70-related rDNA was determined by RT- and RACE-PCR methods. A full-length of hsp70-related cDNA has an open reading frame of 1.95 kb encoding 650 amino acids with a calculated molecular weight of 71.1 kD. A corresponding hsp70-related protein contains a number of conserved elements including an ATP-binding domain, a nuclear localization signal and the carboxyl terminal motif, EEVD, which may have a role in chaperone function. Comparison of nucleotide and predicted amino acid sequence between olive flounder hsp70-related gene and hsp/hsc70 genes of other species revealed a high similarity with the cognate form of these genes. These results indicated that we recovered likely to be a olive flounder cognate hsc70 gene.

• 한국패류학회지
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• 제32권2호
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• pp.73-81
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• 2016

Heat shock proteins (HSPs), one of the most highly conserved groups of proteins characterized to date, play crucial roles in protecting cells against environmental stresses, such as heat shock, salinity and oxidative stress. The glutathione S-transferases (GST) have important role in detoxification of oxidative stress, environmental chemicals and environmental stress. GST mRNA expression have been used as biomarkers on environmental stress. The purpose of this study was to investigate the death rate and the gene expression of Hsp70 and GST during air exposure and starvation. Results showed that, the expression of Hsp70 mRNA was significantly changed in the experiment groups, such as air exposure and starvation. GST mRNA expression was significantly increased in the experimental group of starvation. These results suggest that Hsp70 and GST were played roles in biomarker gene on the air exposure and starvation.

• 한국발생생물학회지:발생과생식
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• 제14권3호
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• pp.179-184
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• 2010

Water temperature influences on various key biological events in fish, but the internal pathway of the temperature effects are not well understood. Heat shock proteins (HSPs), known to respond in the level of cells to many environmental factors including temperature, could improve our understanding on the pathway. Some biological processes such as gonadal development and sex differentiation in the Nile tilapia Oreochromis niloticus is particularly sensitive to water temperature. In this study, we have investigated the expressions of HSP70 and HSP90 genes in young tilapia at an ordinary temperature ($28^{\circ}C$) and elevated water temperature ($36^{\circ}C$). The distribution of the expressions of HSP70 and HSP90 mRNA in this species were found to be almost ubiquitous, being detected in all tissues studied here (brain, gonad, liver and muscle), suggesting the house keeping functions of these genes. Heat shock by elevating temperature from $28^{\circ}C$ to $36^{\circ}C$ significantly increased the expression of HSP70 mRNA in the gonad, liver and muscle for several hours (P<0.05) (brain tissue was not examined for this). The increased level of HSP70 gene expression recovered to the level at control temperature ($28^{\circ}C$) when fish were kept continuously at high temperature ($36^{\circ}C$) for 24 hours. Contrary to this, expression of HSP90 mRNA did not show significant increase in the gonad and muscle by the same heat shock (P>0.05), except in the liver where the expression of HSP90 mRNA increased continuously for 24 hours at $36^{\circ}C$. The results obtained in this study suggest that response to temperature change in different tissue or organ may utilize different heat shock proteins, and that HSP70 may have some importance in temperature-sensitive gonadal event in the Nile tilapia.