RESUMEN
Mastitis is the most important disease in the global dairy industry, and causes large economic losses. Staphylococcus aureus is one of most common pathogens that cause bovine mastitis. CXCR1 has been implicated as a prospective genetic marker for mastitis resistance in dairy cows; CXCR1 expression significantly increases when cows have mastitis. To investigate the mechanisms involved in its increased expression, bisulfite sequencing polymerase chain reaction (PCR) was used to detect the methylation status of CXCR1 CpG island, and quantitative fluorescence PCR was used to detect CXCR1 expression in bovine mammary tissue induced with S. aureus in three Chinese Holstein cows. No CpG island was found for bovine CXCR1 in the upstream 2-kb region, whereas one CpG island that contained 13 CpG sites was found in exon 1 of CXCR1. All of the CpG sites were under hypermethylation from 90 to 100% in the mammary tissues. When the mammary gland mRNA expression of CXCR1 was 12.10-fold higher in infected cow quarters than in uninfected quarters, the methylation levels of the CpG site at position 519 were significantly lower in the infected quarters than in the uninfected quarters. Pearson correlation analysis showed that the methylation level at position 519 was significantly negatively correlated with the CXCR1 mRNA expression level (P < 0.05). These results indicate that the methylation of the CpG site at position 519 may regulate CXCR1 expression in cows with mastitis induced by S. aureus, but further studies are needed to elucidate the mechanisms involved.
Asunto(s)
Metilación de ADN , Glándulas Mamarias Animales/metabolismo , Mastitis Bovina/genética , Receptores de Interleucina-8A/genética , Infecciones Estafilocócicas/veterinaria , Staphylococcus aureus/aislamiento & purificación , Animales , Bovinos , Islas de CpG , Femenino , Mastitis Bovina/metabolismo , Mastitis Bovina/microbiología , Reacción en Cadena de la Polimerasa , Estudios Prospectivos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptores de Interleucina-8A/metabolismo , Infecciones Estafilocócicas/metabolismoRESUMEN
MicroRNAs (miRNAs) may be important mediators of the profound molecular and cellular changes that occur after traumatic brain injury (TBI). However, the changes and possible roles of miRNAs induced by voluntary exercise prior to TBI are still not known. In this report, the microarray method was used to demonstrate alterations in miRNA expression levels in the cerebral cortex of TBI mice that were pretrained on a running wheel (RW). Voluntary RW exercise prior to TBI: i) significantly decreased the mortality rate and improved the recovery of the righting reflex in TBI mice, and ii) differentially changed the levels of several miRNAs, upregulating some and downregulating others. Furthermore, we revealed global upregulation of miR-21, miR-92a, and miR-874 and downregulation of miR-138, let-7c, and miR-124 expression among the sham-non-runner, TBI-non-runner, and TBI-runner groups. Quantitative reverse transcription polymerase chain reaction data (RT-qPCR) indicated good consistency with the microarray results. Our microarray-based analysis of miRNA expression in mice cerebral cortex after TBI revealed that some miRNAs such as miR-21, miR-92a, miR-874, miR-138, let-7c, and miR-124 could be involved in the prevention and protection afforded by voluntary exercise in a TBI model.
Asunto(s)
Humanos , Antiinfecciosos/uso terapéutico , Revisión de la Utilización de Medicamentos , Antiinfecciosos/economía , Costos de los Medicamentos , Revisión de la Utilización de Medicamentos/organización & administraciónRESUMEN
MicroRNAs (miRNAs) may be important mediators of the profound molecular and cellular changes that occur after traumatic brain injury (TBI). However, the changes and possible roles of miRNAs induced by voluntary exercise prior to TBI are still not known. In this report, the microarray method was used to demonstrate alterations in miRNA expression levels in the cerebral cortex of TBI mice that were pretrained on a running wheel (RW). Voluntary RW exercise prior to TBI: i) significantly decreased the mortality rate and improved the recovery of the righting reflex in TBI mice, and ii) differentially changed the levels of several miRNAs, upregulating some and downregulating others. Furthermore, we revealed global upregulation of miR-21, miR-92a, and miR-874 and downregulation of miR-138, let-7c, and miR-124 expression among the sham-non-runner, TBI-non-runner, and TBI-runner groups. Quantitative reverse transcription polymerase chain reaction data (RT-qPCR) indicated good consistency with the microarray results. Our microarray-based analysis of miRNA expression in mice cerebral cortex after TBI revealed that some miRNAs such as miR-21, miR-92a, miR-874, miR-138, let-7c, and miR-124 could be involved in the prevention and protection afforded by voluntary exercise in a TBI model.
Asunto(s)
Lesiones Encefálicas/terapia , Corteza Cerebral/lesiones , Terapia por Ejercicio , MicroARNs/metabolismo , Actividad Motora/fisiología , Animales , Lesiones Encefálicas/metabolismo , Lesiones Encefálicas/mortalidad , Regulación hacia Abajo , Masculino , Ratones Endogámicos C57BL , Análisis por Micromatrices , Reflejo de Enderezamiento/fisiología , Tasa de Supervivencia , Transcripción Genética/fisiología , Regulación hacia ArribaRESUMEN
Complete coding DNA sequences of a closely related chlorogenic acid synthetase gene (LjCCoAOMT1) were isolated from Lonicera japonica Thunb. by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). LjCCoAOMT1 was subsequently overexpressed in Escherichia coli and a 25-kD protein was detected by electrophoresis and western blot analysis. High-performance liquid chromatography (HPLC) analysis showed that recombinant LjCCoAOMT1 methylates the caffeic acid substrate to generate ferulic acid. Further analysis showed that the chlorogenic acid content was significantly correlated with the expression level of LjCCoAOMT1 in various tissues of L. japonica Thunb. at different developmental stages. A plant expression vector containing LjCCoAOMT1 was constructed and Agrobacterium-mediated transgenic rice was successfully obtained. Light treatment analysis showed that LjCCoAOMT1 transgenic rice was more sensitive than wild-type rice in responding to the changes in lighting conditions. Although gibberellic acid (GA3) could promote the growth of both wild-type and LjCCoAOMT1 transgenic rice, LjCCoAOMT1 transgenic rice appeared to be more sensitive to GA3. Furthermore, high concentrations of GA3 significantly facilitated the growth of LjCCoAOMT1 transgenic rice.
Asunto(s)
Aciltransferasas/genética , Clonación Molecular , Expresión Génica , Genes de Plantas , Lonicera/genética , Oryza/genética , Aciltransferasas/metabolismo , ADN Complementario/genética , Flores/genética , Flores/metabolismo , Giberelinas/metabolismo , Luz , Oryza/metabolismo , Oryza/efectos de la radiación , Plantas Modificadas GenéticamenteRESUMEN
Exogenous gibberellins (GAs) are widely applied to increase crop yields, with knowledge about the physiological functioning and biochemistry mechanisms of these phytohormones improving; however, information remains limited about the effect of GAs on seed filling. In this study, the siliques (containing the seeds) of oilseed rape (Brassica napus L.) were treated with GA3 at 3 stages of seed filling. We confirmed that GA3 regulates the deposition of storage reserves in developing seeds. The percentage of crude fat in the seeds increased during the early stage, but remained stable during the middle and late stages. In comparison, the percentage of total protein decreased during the early and middle stages, but significantly increased during the late stage. In addition, Q-PCR was employed to analyze the expression level of related genes in response to GA3. It was found that the expression of WRI and ABI3 transcription factors corresponded to crude fat content and total protein content, respectively. The expression of storage reserve related genes DGAT, MCAT, SUC2, and GPT was consistent with crude fat content, whereas the expression of Napin corresponded to total protein content. The results of this study indicate that exogenous GA3 has a different effect on storage reserve deposition in seed during different stages of seed filling, and the effect might be achieved via changing the expression of related genes.