RESUMO
Tibial dyschondroplasia (TD) is a type of bone deformity found in fast-growing chickens, which induce inflammatory responses. Prostaglandins (PGs) implicate in bone formation and bone resorption, associated with inflammation in an autocrine/paracrine manner. This study used qRT-PCR and immunohistochemistry analysis to identify the expression patterns of PG-related genes in the erythrocytes of broiler chickens and explore the effects of thiram-induced TD and the recombinant glutathione-S-transferase A3 (rGSTA3) protein on the expression of PG-related genes: GSTA3, cyclooxygenase 2 (COX-2), prostaglandin D2 synthase (PTGDS), prostaglandin E synthase (PTGES), prostaglandin E2 receptor (PTGER) 3, PTGER4 and prostaglandin reductase 1 (PTGR1). Interestingly, the results showed that these seven PG-related genes expression was identified in the erythrocytes of broiler chicken, and thiram-induced TD suppressed the expression of these PG-related genes in the initial stage of TD and promoted their expression in TD recovery. These findings demonstrated that the immunoregulatory function of erythrocytes can be inhibited in the early stage of TD and promoted in the recovery stage by modulating the expression of PG-related genes. Further, the rGSTA3 protein can modulate the expression of PG-related genes in erythrocytes and participate in the recovery of TD.
Assuntos
Galinhas , Glutationa Transferase/farmacologia , Osteocondrodisplasias/veterinária , Doenças das Aves Domésticas/genética , Prostaglandinas/genética , Tíbia/patologia , Animais , Proteínas Aviárias/farmacologia , Eritrócitos/metabolismo , Mutagênicos/farmacologia , Osteocondrodisplasias/induzido quimicamente , Osteocondrodisplasias/genética , Doenças das Aves Domésticas/induzido quimicamente , Prostaglandinas/metabolismo , Proteínas Recombinantes/farmacologia , Tiram/farmacologiaRESUMO
Thiram, a carbamate pesticide, is known to induce tibial dyschondroplasia (TD) in broiler chickens. This study used a thiram-induced TD model to explore whether apoptosis-related genes were expressed in erythrocytes of broiler chickens and the impacts of thiram-induced TD and the recombinant GSTA3 protein in regulating these genes expression. In this study, mRNA and protein expression of six types of apoptosis-related genes (Bcl-2, Bax, Murine double minute MDM2, Bcl-2-associated athanogene BAG-1, BAG-3, STAT3) were identified in erythrocytes of broiler chickens by real-time PCR and immunohistochemistry, and we also found that thiram-induced TD induced the decreased expression of these antiapoptotic genes in the initial stage of TD and promoted their expression in TD recovery, which suggested that the expression of these apoptosis-related genes in erythrocytes is highly related to the development of TD. Further, the recombinant GSTA3 protein promoted the expression of all apoptosis-related genes in the initial stage of TD and recovered the normal expression of these genes in the recovery stage of TD, which indicated that the recombinant GSTA3 protein may participate in the recovery of TD. Further studies are needed to elucidate the mechanism of the response of erythrocytes to thiram-induced TD and the recombinant protein GSTA3 in broiler chickens.
Assuntos
Apoptose/fisiologia , Glutationa Transferase/farmacologia , Osteocondrodisplasias/veterinária , Doenças das Aves Domésticas/genética , Tiram/toxicidade , Animais , Apoptose/genética , Galinhas/genética , Eritrócitos/metabolismo , Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica , Predisposição Genética para Doença , Glutationa/metabolismo , Osteocondrodisplasias/genética , Doenças das Aves Domésticas/metabolismo , RNA Mensageiro/genética , Proteínas Recombinantes/farmacologia , TransferasesRESUMO
Based on the prediction of the hydrophilicity, epitopes, secondary structure and flexibility of the CS3 subunit, a novel vector pCSX72 which permits the insertion of foreign epitopes into CS3 at the position of 72nd aa was constructed. Two epitopes, the VP1 of FMDV and a ten-peptides epitope of C-myc, were displayed with it respectively. Compared with the two previously-constructed vectors, the vector pCSX72 expressed the hybrid fimbriae in higher level. Mice produced dual immune response against the CS3 and the inserted epitopes when they were immunized by injecting the live recombinant bacteria intraperitoneally.
Assuntos
Proteínas de Bactérias/genética , Proteínas de Escherichia coli , Escherichia coli/genética , Proteínas de Fímbrias , Fímbrias Bacterianas/genética , Vetores Genéticos , Animais , Proteínas de Bactérias/imunologia , Epitopos , Feminino , Humanos , Imunização , Camundongos , Camundongos Endogâmicos BALB CRESUMO
To confirm the possibility of pCSB136 and pCSX72 as vectors for displaying heterologous epitopes, the gene fragments coding for C3 epitope of poliovirus and a ten-peptides epitope of C-myc were synthesized and inserted into pCSB136 and pCSX72 respectively. The recombinants were screened by whole-strain PCR. The expression of recombinant proteins were detected by whole-cell ELISA and electronic microscopy. The results indicated the recombinant proteins were expressed as hybrid fimbriae, and the antigenicity of both CS3 and inserted epitopes kept. All results above showed vectors pCSB136 and pCSX72 could be used to display the foreign epitopes.