RESUMO
In this study, chicken adipocytes were cultured to evaluate RNA interference by the leptin receptor gene. A small interfering RNA of the leptin receptor gene was synthesized, with a suppression rate of 60% being generated (P < 0.01). After the knockdown of the leptin receptor, the expression levels of certain genes decreased significantly; specifically, peroxisome proliferator-activated receptor γ, fatty acid synthase, adipose triglyceride lipase, and lipoprotein lipase. In addition, a significant increase in the expression of the adiponectin gene was documented. These results demonstrate that the leptin receptor gene might contribute to lipid metabolism by influencing the expressions of the peroxisome proliferator-activated receptor γ, fatty acid synthase, adipose triglyceride lipase, lipoprotein lipase, and adiponectin genes.
Assuntos
Adipócitos/metabolismo , Interferência de RNA , Receptores para Leptina/genética , Animais , Galinhas , Regulação da Expressão Gênica , Metabolismo dos Lipídeos/genética , RNA Interferente Pequeno/síntese química , RNA Interferente Pequeno/genética , TransfecçãoRESUMO
Estrogen regulates reproductive behavior and drives the proliferation and differentiation of several cell types. These physiological functions of estrogen are mediated by estrogen receptors (ERs), and each ER isoform plays a distinct role. To clarify the molecular mechanism of estrogen action and to evaluate the effect of ERs on the secretion of ovalbumin (OVA) in pigeon oviduct epithelial cells (POECs), we determined the complete coding sequences encoding ER alpha (ERα) and ER beta (ERß) in pigeons. The abundance of pigeon ERα and ERß mRNA was detected using quantitative polymerase chain reaction. These results revealed that pigeon ERα is highly expressed in the oviduct, while pigeon ERb is highly expressed in the ovary and kidney. We hypothesize that ERα mRNA predominates over that of ERß in the oviduct. The expression of ERα can be down-regulated by 17ß-estradiol, and the knockdown of ERα promoted OVA mRNA expression in cultured POECs, indicating that ERα may play an important role in OVA secretion.
Assuntos
Células Epiteliais/metabolismo , Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/genética , Animais , Clonagem Molecular , Columbidae/genética , Estradiol/metabolismo , Receptor alfa de Estrogênio/biossíntese , Receptor beta de Estrogênio/biossíntese , Feminino , Expressão Gênica , Oviductos/metabolismo , RNA Mensageiro/biossínteseRESUMO
The full-length pigeon ovalbumin (OVA) gene cDNA was cloned and sequenced by reverse transcription-polymerase chain reaction (RT-PCR) and rapid-amplification of cDNA ends. A 386-amino acid protein was predicted for the obtained sequence, which had 67% identity with the chicken protein. Similar to chicken OVA, the pigeon OVA gene is a non-inhibitory serine protease inhibitor. Quantitative PCR analysis revealed that pigeon OVA mRNA was highly expressed in the oviduct, and trace amounts were detected in other tissues. During the reproductive cycle, pigeon oviduct OVA mRNA expression reached its peak during the egg-laying stage, decreased with brooding, and then increased again during the squab-feeding period. Moreover, the relative OVA expression level in pigeon oviduct epithelial cells could be upregulated by a constant concentration of steroid hormones.
Assuntos
Columbidae/genética , Células Epiteliais/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Ovalbumina/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Columbidae/crescimento & desenvolvimento , Columbidae/fisiologia , Feminino , Hormônios Esteroides Gonadais/metabolismo , Dados de Sequência Molecular , Especificidade de Órgãos , Ovalbumina/metabolismo , Oviductos/citologia , Oviductos/metabolismo , Oviductos/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
Several studies have documented the process of early embryonic development in poultry; however, the molecular mechanisms underlying its developmental regulation are poorly understood, particularly in ducks. In this study, we analyzed differential gene expression of embryos 6 and 25 h following oviposition to determine which genes regulate the early developmental stage in ducks. Among 216 randomly selected clones, 39 protein-encoding cDNAs that function in metabolism, transcription, transportation, proliferation/apoptosis, cell cycle, cell adhesion, and methylation were identified. Additionally, the full-length cDNA of the Nanog gene, encoding a 302-amino acid protein, was obtained. Quantitative real-time polymerase chain reaction analyses were performed to detect expression levels of the selected genes during early and late embryonic stages, which revealed that these genes are expressed in a particular spatial and temporal pattern. These results indicate that these genes may play pivotal roles in the process of area pellucida formation through a complex and precise regulatory network during development in duck embryos.