Assuntos
Arthrodermataceae/isolamento & purificação , Dermatomicoses/microbiologia , Doenças dos Genitais Masculinos/microbiologia , Escroto/microbiologia , Adolescente , Adulto , Arthrodermataceae/patogenicidade , Arthrodermataceae/ultraestrutura , Infecções Assintomáticas , Dermatomicoses/diagnóstico , Dermatomicoses/patologia , Doenças dos Genitais Masculinos/diagnóstico , Doenças dos Genitais Masculinos/patologia , Humanos , Masculino , Microscopia Eletrônica de Varredura , Escroto/patologia , Adulto JovemRESUMO
It has been reported that calcium hydroxide can induce proliferation, migration, and mineralization in dental pulp stem cells (DPSCs), but the underlying molecular mechanisms are still unclear. In this study, we sought to explore the role of calcium hydroxide in the cell proliferation and directional differentiation of DPSCs and to study the regulatory effect of NF-κB, p38MAPK, and Wnt signaling on differentiation of DPSCs. CCK8 cell assay, Wound Healing Assay, and Alkaline Phosphatase Staining Assay were respectively used to determine the proliferation rate, migration and ALP expression of DPSCs. Alizarin Red Staining Assay was used to observe the mineralization of DPSCs. RT-PCR analysis and Western Blot Analysis displayed the expression of related fators at mRNA and protein level, respectively. In the present study, we found that NF-κB, p38MAPK, and Wnt signaling could abolish calcium hydroxide-induced proliferation of DPSCs. The inhibition of NF-κB, p38MAPK, and Wnt signaling suppressed the migration, ALP expression, and mineralization of DPSCs. NF-κB, p38MAPK, and Wnt signaling involved in directional differentiation of DPSCs. Moverover, calcium hydroxide could activate NF-κB, p38MAPK, and Wnt pathway by regulating TNF-α. Our study showed that NF-κB, p38MAPK, and Wnt signaling pathway were involved in calcium hydroxide-induced proliferation, migration, mineralization, and osteogenic differentiation in DPSCs. Calcium hydroxide affected NF-κB, p38MAPK, and Wnt pathway by regulating TNF-α.
Assuntos
Hidróxido de Cálcio/farmacologia , Polpa Dentária/efeitos dos fármacos , Células-Tronco/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismo , Adolescente , Diferenciação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Polpa Dentária/citologia , Humanos , NF-kappa B/metabolismo , Osteogênese/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células-Tronco/citologia , Via de Sinalização Wnt/efeitos dos fármacos , Adulto Jovem , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismoRESUMO
Cyclin B is a regulatory subunit of maturation-promoting factor (MPF), which has a key role in the induction of meiotic maturation of oocytes. MPF has been studied in a wide variety of animal species; however, its expression in crustaceans is poorly characterized. In this study, the complete cDNA sequence of Cyclin B was cloned from the red claw crayfish, Cherax quadricarinatus, and its spatiotemporal expression profiles were analyzed. Cyclin B cDNA (1779 bp) encoded a 401 amino acid protein with a calculated molecular weight of 45.1 kDa. Quantitative real-time PCR demonstrated that Cyclin B mRNA was expressed mainly in the ovarian tissue and that the expression decreased as the ovaries developed. Immunofluorescence analysis revealed that the Cyclin B protein relocated from the cytoplasm to the nucleus during oogenesis. These findings suggest that Cyclin B plays an important role in gametogenesis and gonad development in C. quadricarinatus.
Assuntos
Astacoidea/genética , Ciclina B/genética , Regulação da Expressão Gênica no Desenvolvimento , Fator Promotor de Maturação/genética , Oócitos/metabolismo , Oogênese/genética , Sequência de Aminoácidos , Animais , Astacoidea/citologia , Astacoidea/crescimento & desenvolvimento , Sequência de Bases , Núcleo Celular/metabolismo , Clonagem Molecular , Ciclina B/metabolismo , Citoplasma/metabolismo , DNA Complementar/genética , DNA Complementar/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Feminino , Fator Promotor de Maturação/metabolismo , Meiose , Dados de Sequência Molecular , Peso Molecular , Oócitos/citologia , Oócitos/crescimento & desenvolvimento , Fases de Leitura Aberta , Ovário/citologia , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Transporte Proteico , Reação em Cadeia da Polimerase em Tempo Real , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de SequênciaRESUMO
To better understand the reproductive transformation mechanism of Daphnia carinata, a Doublesex (Dsx) gene was cloned based on rapid amplification of cDNA ends (RACE), and was designated DapcaDsx2. Next, we compared similarities and assumed homology based on deduced amino acid sequences. It showed 97.52, 87.94, and 85.11% identity to orthologous genes in D. magna, D. pulex, and D. galeata respectively. Phylogenetic analysis revealed that DapcaDsx2 clustered in the same class, and was evolutionarily more distant to sequences from other species. qRT-PCR showed that DapcaDsx2 was most abundantly expressed during sexual reproduction (P < 0.05). Using digoxigenin-labeled RNA probes, we studied DapcaDsx2 expression in parthenogenetic and sexual females by whole-mount in situ hybridization. The results revealed that DapcaDsx2 was mainly expressed in the second antennae and several sites of the ventral carapace, whereas higher expression levels were found in sexual than in parthenogenetic females. This suggests that the DapcaDsx2 gene is involved in switching modes of reproduction and in sexual differentiation.
