RESUMO
Ovarian-specific promoter 1 (OSP-1) is a retrovirus-like element isolated from the complementary DNA library of rat that has been thought to be specifically expressed in ovary. To exploit this promoter in dairy goat ovary granulosa cells (GCs), OSP-1 from rat was used to construct the reporter vector pOSP-1-EGFP, in which egfp coding for enhanced green fluorescent protein (EGFP) was used as a reporter to examine the activity of OSP-1 in GCs. EGFP was successfully expressed in dairy goat GCs transfected with pOSP-1-EGFP. Reverse transcriptase-polymerase chain reaction analysis confirmed the tissue-specific transcription of EGFP messenger RNA in dairy goat GCs transfected with pOSP-1-EGFP. We concluded that OSP-1 promoter from rat can specifically drive foreign gene expression in dairy goat GCs. Thus, we obtained a tissue-specific regulation element and provided a potential tool for the research of regulation and development of the ovary in dairy goats.
Assuntos
Claudinas/genética , Cabras/genética , Células da Granulosa/fisiologia , Ovário/metabolismo , Animais , Células Cultivadas , DNA Complementar/genética , Feminino , Expressão Gênica/genética , Genes Reporter/genética , Vetores Genéticos/genética , Cabras/metabolismo , Células da Granulosa/metabolismo , Proteínas de Fluorescência Verde/genética , Regiões Promotoras Genéticas , RNA Mensageiro/genética , Ratos , Retroelementos , Transcrição Gênica , Transfecção/métodosRESUMO
In this study, suppressive subtractive hybridization (SSH) was performed to screen differentially expressed genes in ovarian tissues between polytocous and monotocous goats. From the SSH cDNA library, we obtained 29 differentially expressed sequence tags (ESTs) that have high similarity with known genes in the public database, which were involved in signal transduction, transcriptional regulation, cellular molecular dynamics, cytoskeleton, metabolism and oxidation-reduction. In addition, one novel EST that has no similarity with known genes in the public database was obtained. Eight ESTs, TIMP1, NUCB1, OAZ1, CXXC5, CAPNS2, ATP5A1, Z and RPL5, were further examined for the reproducibility of the SSH data by the real-time quantitative PCR. The results confirmed an increased expression of respective mRNA in ovarian tissues of polytocous goats compared with those of monotocous goats. The study has identified several genes (known or unknown) that may have effect on follicular development, ovulation and egg activation in goats.
Assuntos
Perfilação da Expressão Gênica/veterinária , Regulação da Expressão Gênica/fisiologia , Cabras/metabolismo , Animais , Indústria de Laticínios , Etiquetas de Sequências Expressas , Feminino , Cabras/genética , Ovário/metabolismo , Gravidez , Gravidez Múltipla , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
The use of epidural ropivacaine may result in significant haemodynamic fluctuations during combined epidural and general anaesthesia. We designed this study to investigate whether epidural anaesthesia with a goal-directed approach, when combined with general anaesthesia, improved haemodynamic stability in elderly patients undergoing major abdominal surgery. Seventy-five elderly patients undergoing major abdominal surgery were randomly and evenly assigned to one of three groups receiving intraoperative epidural anaesthesia with either ropivacaine 0.1% (Group 1), ropivacaine 0.375% (Group 2) or ropivacaine 0.375% for abdominal wall pain and ropivacaine 0.1% for visceral pain (Group 3). General anaesthesia was induced using a target-controlled infusion of combined propofol and remifentanil. The remifentanil target concentration was adjusted according to the mean arterial pressure and heart rate, and vasoactive agents were administered to maintain stable haemodynamics. The need for vasoactive drug administrations was 1.4 (standard deviation 0.9) in Group 3 (n=24), representing a significantly lower frequency of administration compared with Groups 1 (n=24) and 2 (n=24) (P <0.05 versus Group 1; P <0.01 versus Group 2). The total intraoperative dose of remifentanil was significantly greater in Group 1 (P <0.01 versus Group 2; P <0.05 versus Group 3) but did not differ significantly between Groups 2 and 3. Goal-directed epidural anaesthesia with different ropivacaine concentrations can improve haemodynamic stability when combined with general anaesthesia for elderly patients undergoing major abdominal surgery.