Identification of the tropomyosin (HL-Tm) in Haemaphysalis longicornis

Haemaphysalis longicornis tropomyosin (HL-Tm) was amplified by RT-PCR. The cDNA contained a 825 bp open reading frame coding for 274 amino acids with a predicted theoretical isoelectric point (pI) of 4.55 and molecular weight of 31.7 kDa. Real-time RT-PCR analysis showed that the expression levels of the HL-Tm in the unfed-females were significantly higher than in other tested developmental stages (eggs, unfed-larvae and unfed-nymphs). Western blot analysis showed that rabbit anti-serum against H. longicornis unfed-adult ticks recognized the recombinant HL-Tm protein (rHL-Tm). Immunization of rabbits with the rHL-Tm resulted in a statistically significant reduction of female engorgement and oviposition. Silencing of HL-Tm by RNAi showed a decrease in tick engorgement and oviposition, which is consistent with the effect of recombinant protein vaccine on the adults. These results showed that tick HL-Tm might be involved in the regulation of ticks blood-feeding, growth and oviposition.     

Identification of heat shock protein 70 in canine reticulocytes and mature erythrocytes

In the present study, we demonstrated that heat shock protein 70 (Hsp70) was present in both canine reticulocytes and mature erythrocytes, and that the canine Hsp70 in reticulocytes was decreased along with the maturation of the cells into erythrocytes. These results suggest that the Hsp70 in canine reticulocytes might act as a chaperone to remove unnecessary proteins during reticulocyte maturation. We also demonstrated that Hsp70 was present in exosomes from reticulocytes during their maturation in in vitro culture. Furthermore, the concentration of Hsp70 in reticulocyte membranes was increased in proportion to an increase of the protein in exosomes until 48 hours after the incubation of reticulocytes in vitro. At 96 hours of the incubation, however, only a trace amount of Hsp70 was detected in the membrane, while a large amount of the protein was present in the exosomes. These results suggest that Hsp70 in canine reticulocytes might play an important role for exosome formation in reticulocytes, resulting in the maturation of the cells.     

热休克蛋白70研究新进展

1962年,Ritassa在果蝇的研究中首次发现,短暂的热休克可以诱导唾液腺染色体出现3个膨突,提示这一区带转录加强,他将这一现象称为“热休克反应“(heat shock response,HSR).1974年,Tissieres发现热休克反应中转录合成的为一组特殊蛋白,而且伴随着这类蛋白的合成,细胞的其他蛋白合成却受到抑制,由于这类蛋白的合成与热休克反应有关,故命名为热休克蛋白(heat shock protein,HSP).除了高热之外,多种应激原如重金属、饥饿、缺氧、缺血等都可以诱导HSP的表达,但人们习惯上仍称为HSP或热应激蛋白(heat stress protein,HSP),有时也称为应激蛋白(stress protein,SP).……     

Identification of genes encoding cement-like antigens expressed in the salivary glands of Haemaphysalis longicornis

A cDNA expression library from the salivary glands of hard tick, Haemaphysalis longicornis, was constructed. Immunoscreening was performed using sera of the rabbit repeatedly infested with ticks and seventeen positive clones were obtained. A BLASTP search suggested that 8 sequences matched with that of hypothetical H. longicornis sequence and one clone encoded HL35 antigen U from the same tick species. Eight of 17 gave no match to any sequence reported in the database. The proteins expected from these novel sequences possess common characteristics with cement proteins which assist ticks in their attachment to the host during blood feeding. The expression of these genes in salivary glands was confirmed by RT-PCR. Four of the 8 sequences showed to be upregulated upon blood feeding. These immunodominant antigens are of particular interest as candidates for future cement protein based-tick vaccine.     

Detection of anti-Babesia gibsoni heat shock protein 70 antibody and anti-canine heat shock protein 70 antibody in sera from Babesia gibsoni-infected dogs

Antibodies that recognized either Babesia gibsoni or canine red blood cell (RBC) 70-kilodalton (kDa) protein were detected in serum from acutely and chronically B. gibsoni-infected. In those sera, antibodies that reacted with recombinant B. gibsoni and canine heat shock protein 70 (rBgHsp70 and rcHsp70) were detected; therefore, B. gibsoni and canine RBC 70-kDa proteins seemed to be BgHsp70 and cHsp70, respectively. In infected dogs, the amounts of these antibodies increased after infection. Interestingly, polyclonal antibody raised against rBgHsp70 in two rabbits reacted not only with rBgHsp70 but also with rcHsp70 and native cHsp70 from canine RBCs. Because BgHsp70 showed high homology with cHsp70 (70.8%), anti-rBgHsp70 antibody might cross-react with cHsp70. Additionally, the localizations of both BgHsp70 and cHsp70 were observed by indirect fluorescence assay. As a result, cHsp70 was not found on the membrane surface of erythrocytes, suggesting that erythrocytes would not be targets of anti-cHsp70 antibody. Meanwhile, only exoerythrocytic parasites were stained by anti-rBgHsp70 antibody. This result showed that BgHsp70 would be expressed on the surface of parasites during the exoerythrocytic stage. These results indicated that BgHsp70 was a highly immunogenic protein in canine B. gibsoni infection, and that exoerythrocytic parasites might be targets of anti-BgHsp70 antibody.     

Hepatic concentration of heat shock protein 70 kD (Hsp70) in broilers subjected to different thermal treatments.

1. The relationship between repeated thermal treatments and hepatic synthesis of Hsp 70 was studied in broiler chickens. 2. Sixty broilers were submitted to 5 different treatments (12 birds each) from day 1 to day 42. Four groups were kept in a thermoneutral environment and subjected to 0, 1, 2 and 3 heat stress episodes at 35 degrees C for 4 h per week (TN-0, TN-1, TN-2 and TN-3, respectively). The last group (HT-35) was reared at a room temperature of 35 degrees C. 3. From 39 to 42 old, the birds experienced acute heat stress at 41 degrees C. Resistance to heat stress was evaluated by the time taken for rectal temperature to increase by 3 degrees C above the pre-treatment value. Livers were collected (before and after heat stress) and Hsp70 was determined using Western Blot analysis with monoclonal anti-Hsp70 antibody. 4. Resistance to heat stress and concentration of Hsp70 were higher in those birds subjected to more heat stress episodes during the experimental period (TN-3) and HT-35. A positive correlation was observed between Hsp70 concentration and the time taken for a 3 degrees C increase in rectal temperature (r = 0.42; P<0.01). 5. Exposing birds to episodes of heat stress (35 degrees C) during rearing may improve their resistance to acute heat stress, but the previous thermal history did not seem to influence the hepatocyte Hsp70 content after exposure to more severe heat stress (41 degrees C).     

热应激蛋白70(HSP_(70))研究进展

热应激蛋白普遍存在于从原核细胞到高等真核细胞的整个生物界。本文通过HSP70 的基因特点与转录调控、HSP70 的mRNA与翻译、HSP70 的作用以及HSP70 在体内的降解等方面 ,对HSP70 进行了较为详细的介绍     

Effect of acute heat stress on heat shock protein 70 messenger RNA and on heat shock protein expression in the liver of broilers

1. The synthesis of heat shock protein 70 (Hsp70) mRNA and the expression of Hsp70 in the liver of broiler chickens submitted to acute heat stress (35°C for 5 h) was investigated.

2. Hsp70 expression was detected by SDS‐PAGE and Western blot analysis using a polyclonal antiserum against Hsp70 of Blastocladiella emersonii. The specific signal of Hsp70 mRNA was analysed by Northern blot using as probe a Hsp70 cDNA of B. emersonii.

3. An increase in the amount of Hsp70 was detected from the first up to the fifth hour of acute heat exposure. This increase in the amount of Hsp70 was accompanied by an increase in Hsp70 mRNA which peaked at 3 h.

4. This study shows that the heat induced increase in Hsp70 mRNA and protein in broiler liver, in vivo, are time dependent, similar to that in mammals.     

Effect of long-term heat exposure on peripheral concentrations of heat shock protein 70 (Hsp70) and hormones in laying hens with different genotypes

1. The effect of long-term, moderate heat stress (30 to 32 degrees C) on heat shock protein 70 (Hsp70) concentration in mononuclear blood cells and plasma concentrations of 3,5,3'-triiodothyronine (T3) and corticosterone in laying hens was investigated. 2. Three groups of 48 hens each (Ethopian line [Angete Melata, Na], New Hampshire [NH], F1 cross [Na x NH]) were divided into an experimental group (24 each) and a control group (24 each, ambient temperature 18 to 20 degrees C), respectively. All hens were kept in individual cages up to an age of 68 weeks and performance data were recorded. 3. Blood samples were taken from the wing vein of 12 hens from each group at weeks 22, 38, 51 and 65 (12 hens x 3 lines x 2 treatments). Mononuclear blood cells were isolated and Hsp70 concentrations were determined by Western Blot analysis with a monoclonal anti-Hsp70 antibody. T3 and corticosterone were measured with commercially available ELISA and RIA kits, respectively. 4. The moderate heat stress caused significantly increased Hsp70 levels compared with the control groups in weeks 51 and 65. However, the responses of the lines were not uniform at different ages. 5. In contrast, T3 levels were significantly decreased in stressed birds regardless of line and age. There was no effect of treatment and line on corticosterone levels during the experimental period. 6. Our results indicate that Hsp70 and T3 levels are affected by mild heat stress applied over a long period but are both involved in independent mechanisms of acquisition of thermotolerance. Further investigations are necessary to clarify whether the observed differences in Hsp70 response between the genotypes are indicators for differences in thermotolerance.     

双芽巴贝斯虫在长角血蜱肠内发育的形态学研究

以耳袋法将长角血蜱(Haemaphysalis longicornis)幼虫饲于实验感染双芽巴贝斯虫(Babesia bigemina)牛,幼虫饱血后24h内,其肠管内容物中红细胞内、外见有单梨子型(3.5～4.5μm×1.2～2.6μm)和双梨子型(4.1～4.8μm×1.8～3.0μm)两种裂殖体.饱血后24～48h,随着裂殖体细胞膜及核变性而发生形态变化.48～72h,绝大多数裂殖体出现溶解.72h后,这些裂殖体从肠道消失.其后,在蜱肠上皮及血淋巴中也未能找到双芽巴贝斯虫体.本实验从形态学上证明双芽巴贝斯虫在长角血蜱若虫肠道内不能发育.     

热休克蛋白70诱导剂及其应用

热休克蛋白70(HSP70)是生物体在应激原刺激下合成的一族进化上高度保守的蛋白质。HSP70家族的功能表现在多方面:作为分子伴侣,HSP70在应激状态下帮助需要折叠的蛋白质正确折叠,加快正常蛋白质合成的恢复;在细胞保护方面,HSP70表达的增加可以增强机体对应激的抵抗力,缓解细胞所受伤害。本文主要综述HSP70的生物学特性及其细胞保护作用,并讨论了HSP70诱导剂在畜牧业中潜在的应用意义。     

热休克蛋白70生物学功能及在家禽组织中的表达

热休克蛋白(Heat Shock Protein,HSP)是生物细胞在受到热应激等应激源刺激后所产生一类多肽类蛋白质,其中HSP70是HSP家族中最重要、最保守且表达最广泛的蛋白之一,其普遍存在于各种生物体之中。现对HSP70调节细胞凋亡、增强细胞耐热、细胞抗氧化、机体免疫和分子伴侣等生物学功能进行综述,并对其分子调控机制及在家禽组织中的表达水平作了阐述,为进一步探讨HSP70调控家禽的热应激反应及分子机制奠定理论基础。     

热休克蛋白70的生物学功能及应用的研究进展

热休克蛋白70(HSP 70)是热休克家族中最重要的一员,从细菌到哺乳动物中广泛存在。在正常生理状态下HSP 70表达水平很低,而理化刺激、病理刺激和生理刺激等均能显著诱导其表达。HSP 70具有较强分子伴侣、抗细胞凋亡、抗氧化、免疫调节等功能。本文就HSP 70的结构和分类、主要生物学功能及其在某些疾病治疗方面的应用作一综述。     

热应激对小鼠附植前早期胚胎HSP70的诱导表达

选择超数排卵后怀孕昆明小鼠75只,以注射人绒毛膜促性腺激素(hCG)后的胚胎发育阶段为标准,分别在合子、2细胞、4细胞、8~16细胞和囊胚期对孕鼠进行了37℃、39℃、41℃和43℃热应激4 h。热应激后取胚,用免疫荧光细胞化学法检验胚胎热休克蛋白70(HSP70)的诱导表达。结果表明,HSP70在4细胞以上胚胎内呈阳性表达,相对高温时其表达量增加,囊胚时最显著。可见,附植前早期胚胎HSP70的热诱导表达与细胞期和温度密切相关。     

长角血蜱肠道细菌的分离与鉴定

为深入研究肠道细菌在蜱的生理学和传播病原上的作用,本研究进行了长角血蜱肠道细菌的分离鉴定。按照细菌传统分离方法并结合16S rDNA测序鉴定,获得7个分离株,这些菌株分别属于显核菌属(Caryophanon sp.)、考克氏菌属(Kocuria sp.)、葡萄球菌属(Staphylococcus sp.)、微杆菌属(Microbacterium sp.)和短杆菌属(Brevibacterium sp.)5个菌属。