RESUMEN
KEY MESSAGE: The rust resistance genes Lr53 and Yr35 were introgressed into bread wheat from Aegilops longissima or Aegilops sharonensis or their S-genome containing species and mapped to the telomeric region of chromosome arm 6BS. Wheat leaf and stripe rusts are damaging fungal diseases of wheat worldwide. Breeding for resistance is a sustainable approach to control these two foliar diseases. In this study, we used SNP analysis, sequence comparisons, and cytogenetic assays to determine that the chromosomal segment carrying Lr53 and Yr35 was originated from Ae.longissima or Ae. sharonensis or their derived species. In seedling tests, Lr53 conferred strong resistance against all five Chinese Pt races tested, and Yr35 showed effectiveness against Pst race CYR34 but susceptibility to race CYR32. Using a large population (3892 recombinant gametes) derived from plants homozygous for the ph1b mutation obtained from the cross 98M71 × CSph1b, both Lr53 and Yr35 were successfully mapped to a 6.03-Mb telomeric region of chromosome arm 6BS in the Chinese Spring reference genome v1.1. Co-segregation between Lr53 and Yr35 was observed within this large mapping population. Within the candidate region, several nucleotide-binding leucine-rich repeat genes and protein kinases were identified as candidate genes. Marker pku6B3127 was completely linked to both genes and accurately predicted the absence or presence of alien segment harboring Lr53 and Yr35 in 87 tetraploid and 149 hexaploid wheat genotypes tested. We developed a line with a smaller alien segment (< 6.03 Mb) to reduce any potential linkage drag and demonstrated that it conferred resistance levels similar to those of the original donor parent 98M71. The newly developed introgression line and closely linked PCR markers will accelerate the deployment of Lr53 and Yr35 in wheat breeding programs.
Asunto(s)
Aegilops , Mapeo Cromosómico , Resistencia a la Enfermedad , Genes de Plantas , Puccinia , Aegilops/genética , Aegilops/microbiología , Cromosomas de las Plantas/genética , Resistencia a la Enfermedad/genética , Introgresión Genética , Ligamiento Genético , Marcadores Genéticos , Fitomejoramiento , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Polimorfismo de Nucleótido Simple , Puccinia/fisiología , Triticum/genética , Triticum/microbiologíaRESUMEN
The embryonic tectum displays an anteroposterior gradient in development and produces the superior colliculus and inferior colliculus. Studies suggest that partition of the tectum is controlled by different strengths and durations of FGF signals originated from the so-called isthmic organizer at the mid/hindbrain junction; however, the underlying mechanism is unclear. We show that deleting Ptpn11, which links FGF with the ERK pathway, prevents inferior colliculus formation by depleting a previously uncharacterized stem cell zone. The stem-zone loss is attributed to shortening of S phase and acceleration of cell cycle exit and neurogenesis. Expression of a constitutively active Mek1 (Mek1DD), the known ERK activator, restores the tectal stem zone and the inferior colliculus without Ptpn11. By contrast, Mek1DD expression fails to rescue the tectal stem zone and the inferior colliculus in the absence of Fgf8 and the isthmic organizer, indicating that FGF and Mek1DD initiate qualitatively and/or quantitatively distinctive signaling. Together, our data show that the formation of the inferior colliculus relies on the provision of new cells from the tectal stem zone. Furthermore, distinctive ERK signaling mediates Fgf8 in the control of cell survival, tissue polarity and cytogenetic gradient during the development of the tectum.
Asunto(s)
Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Factores de Crecimiento de Fibroblastos/metabolismo , Colículos Inferiores/citología , Colículos Inferiores/metabolismo , Células Madre/citología , Células Madre/metabolismo , Animales , Apoptosis/genética , Apoptosis/fisiología , Axones/metabolismo , Tipificación del Cuerpo/genética , Tipificación del Cuerpo/fisiología , Ciclo Celular/genética , Ciclo Celular/fisiología , Muerte Celular/genética , Muerte Celular/fisiología , Quinasas MAP Reguladas por Señal Extracelular/genética , Factor 8 de Crecimiento de Fibroblastos/genética , Factor 8 de Crecimiento de Fibroblastos/metabolismo , Factores de Crecimiento de Fibroblastos/genética , Regulación del Desarrollo de la Expresión Génica/genética , Regulación del Desarrollo de la Expresión Génica/fisiología , MAP Quinasa Quinasa 1/genética , MAP Quinasa Quinasa 1/metabolismo , Mesencéfalo/citología , Mesencéfalo/metabolismo , Ratones Noqueados , Neurogénesis/genética , Neurogénesis/fisiología , Proteína Tirosina Fosfatasa no Receptora Tipo 11/genética , Proteína Tirosina Fosfatasa no Receptora Tipo 11/metabolismoRESUMEN
Neurofibromatosis type 1 (NF1) is caused by pathogenic variants or mutations in the NF1 gene that encodes neurofibromin. We describe here a new approach to determining the functional consequences of NF1 genetic variants. We established a heterologous cell culture expression system using a full-length mouse Nf1 cDNA (mNf1) and human cell lines. We demonstrate that the full-length murine cDNA produces a > 250 kDa neurofibromin protein that is capable of modulating Ras signaling. We created mutant cDNAs representing NF1 patient variants with different clinically relevant phenotypes, and assessed their ability to produce mature neurofibromin and restore Nf1 activity in NF1-/- cells. These cDNAs represent variants in multiple protein domains and various types of clinically relevant predicted variants. This approach will help advance research on neurofibromin structure and function, determine pathogenicity for missense variants, and allow for the development of activity assays and variant-directed therapeutics.
Asunto(s)
Variación Genética/genética , Neurofibromatosis 1/genética , Neurofibromina 1/genética , Relación Estructura-Actividad , Animales , Línea Celular , ADN Complementario/genética , Regulación de la Expresión Génica/genética , Técnicas de Silenciamiento del Gen , Pruebas Genéticas , Humanos , Ratones , Mutación/genética , Neurofibromatosis 1/diagnóstico , Neurofibromatosis 1/patología , Transducción de Señal/genéticaRESUMEN
Neurofibromatosis type 1 (NF1) is a common neurogenetic condition characterized by significant clinical heterogeneity. A major barrier to developing precision medicine approaches for NF1 is an incomplete understanding of the factors that underlie its inherent variability. To determine the impact of the germline NF1 gene mutation on the optic gliomas frequently encountered in children with NF1, we developed genetically engineered mice harboring two representative NF1-patient-derived Nf1 gene mutations (c.2542G>C;p.G848R and c.2041C>T;p.R681X). We found that each germline Nf1 gene mutation resulted in different levels of neurofibromin expression. Importantly, only R681X(CKO) but not G848R(CKO), mice develop optic gliomas with increased optic nerve volumes, glial fibrillary acid protein immunoreactivity, proliferation and retinal ganglion cell death, similar to Nf1 conditional knockout mice harboring a neomycin insertion (neo) as the germline Nf1 gene mutation. These differences in optic glioma phenotypes reflect both cell-autonomous and stromal effects of the germline Nf1 gene mutation. In this regard, primary astrocytes harboring the R681X germline Nf1 gene mutation exhibit increased basal astrocyte proliferation (BrdU incorporation) indistinguishable from neo(CKO) astrocytes, whereas astrocytes with the G848R mutation have lower levels of proliferation. Evidence for paracrine effects from the tumor microenvironment were revealed when R681X(CKO) mice were compared with conventional neo(CKO) mice. Relative to neo(CKO) mice, the optic gliomas from R681X(CKO) mice had more microglia infiltration and JNK(Thr183/Tyr185) activation, microglia-produced Ccl5, and glial AKT(Thr308) activation. Collectively, these studies establish that the germline Nf1 gene mutation is a major determinant of optic glioma development and growth through by both tumor cell-intrinsic and stromal effects.
Asunto(s)
Astrocitos/patología , Mutación de Línea Germinal/genética , Neurofibromatosis 1/complicaciones , Neurofibromina 1/genética , Glioma del Nervio Óptico/patología , Nervio Óptico/patología , Animales , Astrocitos/metabolismo , Células Cultivadas , Modelos Animales de Enfermedad , Femenino , Proteína Ácida Fibrilar de la Glía/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microglía/metabolismo , Microglía/patología , Nervio Óptico/metabolismo , Glioma del Nervio Óptico/etiologíaRESUMEN
Folding of the cortex and the persistence of radial glia (RG)-like cells called Bergmann glia (BG) are hallmarks of the mammalian cerebellum. Similar to basal RG in the embryonic neocortex, BG maintain only basal processes and continuously express neural stem cell markers. Past studies had focused on the function of BG in granule cell migration and how granule cell progenitors (GCP) regulate cerebellar foliation. The molecular control of BG generation and its role in cerebellar foliation are less understood. Here, we have analyzed the function of the protein tyrosine phosphatase Shp2 in mice by deleting its gene Ptpn11 in the entire cerebellum or selectively in the GCP lineage. Deleting Ptpn11 in the entire cerebellum by En1-cre blocks transformation of RG into BG but preserves other major cerebellar cell types. In the absence of BG, inward invagination of GCP persists but is uncoupled from the folding of the Purkinje cell layer and the basement membrane, leading to disorganized lamination and an absence of cerebellar folia. In contrast, removing Ptpn11 in the GCP lineage by Atoh1-cre has no effect on cerebellar development, indicating that Shp2 is not cell autonomously required in GCP. Furthermore, we demonstrate that Ptpn11 interacts with Fgf8 and is essential for ERK activation in RG and nascent BG. Finally, expressing constitutively active MEK1 rescues BG formation and cerebellar foliation in Shp2-deficient cerebella. Our results demonstrate an essential role of Shp2 in BG specification via fibroblast growth factor/extracellular signal-regulated protein kinase signaling, and reveal a crucial function of BG in organizing cerebellar foliation.
Asunto(s)
Cerebelo/citología , Cerebelo/fisiología , Sistema de Señalización de MAP Quinasas/fisiología , Neuroglía/fisiología , Proteína Tirosina Fosfatasa no Receptora Tipo 11/fisiología , Animales , Células Ependimogliales/fisiología , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Proteína Tirosina Fosfatasa no Receptora Tipo 11/deficienciaRESUMEN
Combinatorial expression of transcription factors forms transcriptional codes to confer neuronal identities and connectivity. However, how these intrinsic factors orchestrate the spatiotemporal expression of guidance molecules to dictate the responsiveness of axons to guidance cues is less understood. Thalamocortical axons (TCAs) represent the major input to the neocortex and modulate cognitive functions, consciousness and alertness. TCAs travel a long distance and make multiple target choices en route to the cortex. The homeodomain transcription factor Gbx2 is essential for TCA development, as loss of Gbx2 abolishes TCAs in mice. Using a novel TCA-specific reporter, we have discovered that thalamic axons are mostly misrouted to the ventral midbrain and dorsal midline of the diencephalon in Gbx2-deficient mice. Furthermore, conditionally deleting Gbx2 at different embryonic stages has revealed a sustained role of Gbx2 in regulating TCA navigation and targeting. Using explant culture and mosaic analyses, we demonstrate that Gbx2 controls the intrinsic responsiveness of TCAs to guidance cues. The guidance defects of Gbx2-deficient TCAs are associated with abnormal expression of guidance receptors Robo1 and Robo2. Finally, we demonstrate that Gbx2 controls Robo expression by regulating LIM-domain transcription factors through three different mechanisms: Gbx2 and Lhx2 compete for binding to the Lmo3 promoter and exert opposing effects on its transcription; repressing Lmo3 by Gbx2 is essential for Lhx2 activity to induce Robo2; and Gbx2 represses Lhx9 transcription, which in turn induces Robo1. Our findings illustrate the transcriptional control of differential expression of Robo1 and Robo2, which may play an important role in establishing the topography of TCAs.
Asunto(s)
Axones/fisiología , Proteínas de Homeodominio/fisiología , Proteínas con Homeodominio LIM/genética , Proteínas del Tejido Nervioso/genética , Tálamo/embriología , Animales , Axones/metabolismo , Células Cultivadas , Corteza Cerebral/embriología , Corteza Cerebral/metabolismo , Corteza Cerebral/fisiología , Embrión de Mamíferos , Femenino , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Proteínas con Homeodominio LIM/metabolismo , Ratones , Ratones Transgénicos , Proteínas del Tejido Nervioso/metabolismo , Neurogénesis/genética , Neurogénesis/fisiología , Embarazo , Receptores Inmunológicos/genética , Receptores Inmunológicos/metabolismo , Tálamo/metabolismo , Tálamo/fisiología , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas RoundaboutRESUMEN
In vertebrates, the common expression border of two homeobox genes, Otx2 and Gbx2, demarcates the prospective midbrain-hindbrain border (MHB) in the neural plate at the end of gastrulation. The presence of a compartment boundary at the MHB has been demonstrated, but the mechanism and timing of its formation remain unclear. We show by genetic inducible fate mapping using a Gbx2(CreER) knock-in mouse line that descendants of Gbx2(+) cells as early as embryonic day (E) 7.5 do not cross the MHB. Without Gbx2, hindbrain-born cells abnormally populate the entire midbrain, demonstrating that Gbx2 is essential for specifying hindbrain fate. Gbx2(+) and Otx2(+) cells segregate from each other, suggesting that mutually exclusive expression of Otx2 and Gbx2 in midbrain and hindbrain progenitors is responsible for cell sorting in establishing the MHB. The MHB organizer gene Fgf8, which is expressed as a sharp transverse band immediately posterior to the lineage boundary at the MHB, is crucial in maintaining the lineage-restricted boundary after E7.5. Partial deletion of Fgf8 disrupts MHB lineage separation. Activation of FGF pathways has a cell-autonomous effect on cell sorting in midbrain progenitors. Therefore, Fgf8 from the MHB may signal the nearby mesencephalic cells to impart distinct cell surface characteristics or induce local cell-cell signaling, which consequently prevents cell movements across the MHB. Our findings reveal the distinct function of Gbx2 and Fgf8 in a stepwise process in the development of the compartment boundary at the MHB and that Fgf8, in addition to its organizer function, plays a crucial role in maintaining the lineage boundary at the MHB by restricting cell movement.
Asunto(s)
Factor 8 de Crecimiento de Fibroblastos/metabolismo , Proteínas de Homeodominio/metabolismo , Mesencéfalo/embriología , Mesencéfalo/metabolismo , Rombencéfalo/embriología , Rombencéfalo/metabolismo , Animales , Linaje de la Célula , Factor 8 de Crecimiento de Fibroblastos/genética , Eliminación de Gen , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/genética , Mesencéfalo/citología , Ratones , Ratones Transgénicos , Rombencéfalo/citología , Transducción de SeñalRESUMEN
This study evaluated the effect of glutamate supplementation on the physiological parameters of heat-stressed Hu sheep. Forty-eight male Hu sheep with an average initial body weight of 17.74 ± 0.17 kg were randomly divided into two groups: The control group (CON) was fed a basal diet and a treatment group (GLU) was fed a basal diet + 3 g/head/day of L-glutamate. There were six replications in each group with four sheep in each replication for a 90 days feeding test. Growth performance, serum biochemistry, and serum hormones were measured during phase 1 (1-30 days), phase 2 (31-60 days), and phase 3 (61-90 days) of the experiment; rumen fermentation characteristics, nutrient digestibility, and slaughter performance were measured at the end of the experimental periods. There were no differences in growth performance, serum biochemical indices, and immune indices between CON and GLU during phases 1 and 2. However, a higher average daily gain (ADG), a lower average daily feed intake (ADFI), and a lower F:G ratio (ADFI/ADG) were observed in GLU during phase 3 (p < 0.05). Serum levels of glutamate, globulin, immunoglobulin A, immunoglobulin G, immunoglobulin M, and growth hormone in GLU were higher than those in CON only on day 90 (p < 0.05). Serum levels of heat shock protein 70, adrenocorticotrophic hormone, corticosterone, triiodothyronine, and tetraiodothyronine in GLU were lower than those in CON on day 90 (p < 0.05). At the end of the experiment, ruminal pH, microbial crude protein, ammonia nitrogen, and isovalerate concentrations in GLU were higher than those in CON (p < 0.05). The apparent digestibility of dry matter, organic matter, and crude protein in GLU was higher than those in CON (p < 0.05). There were no differences in carcass traits and organ indices but spleen weight and spleen index tended to be higher in GLU. In conclusion, dietary glutamate supplementation improved rumen fermentation, increased nutrition digestibility and metabolism, enhanced immunity, and promoted growth performance of heat-stressed Hu sheep. This suggests that a longer period of glutamate supplementation (not less than 60 days) at a level of 3 g/head/day is beneficial to Hu sheep under heat stress.
RESUMEN
The single Fgf8 gene in mice produces eight protein isoforms (Fgf8a-h) with different N-termini by alternative splicing. Gain-of-function studies have demonstrated that Fgf8a and Fgf8b have distinct activities in the developing midbrain and hindbrain (MHB) due to their different binding affinities with FGF receptors. Here we have performed loss-of-function analyses to determine the in vivo requirement for these two Fgf8 spliceforms during MHB development. We showed that deletion of Fgf8b-containing spliceforms (b, d, f and h) leads to loss of multiple key regulatory genes, including Fgf8 itself, in the MHB region. Therefore, specific inactivation of Fgf8b-containing spliceforms, similar to the loss of Fgf8, in MHB progenitors results in deletion of the midbrain, isthmus, and cerebellum. We also created a splice-site mutation abolishing Fgf8a-containing spliceforms (a, c, e, and g). Mice lacking Fgf8a-containing spliceforms exhibit growth retardation and postnatal lethality, and the phenotype is variable in different genetic backgrounds, suggesting that the Fgf8a-containing spliceforms may play a role in modulating the activity of Fgf8. Surprisingly, no discernable defect was detected in the midbrain and cerebellum of Fgf8a-deficient mice. To determine if Fgf17, which is expressed in the MHB region and possesses similar activities to Fgf8a based on gain-of-function studies, may compensate for the loss of Fgf8a, we generated Fgf17 and Fgf8a double mutant mice. Mice lacking both Fgf8a-containing spliceforms and Fgf17 display the same defect in the posterior midbrain and anterior cerebellum as Fgf17 mutant mice. Therefore, Fgf8b-containing spliceforms, but not Fgf8a, are essential for the function of Fgf8 during the development of the midbrain and cerebellum.
Asunto(s)
Cerebelo/crecimiento & desarrollo , Factor 8 de Crecimiento de Fibroblastos/fisiología , Mesencéfalo/crecimiento & desarrollo , Empalme Alternativo/fisiología , Animales , Factor 8 de Crecimiento de Fibroblastos/genética , Factores de Crecimiento de Fibroblastos , Ratones , Ratones Noqueados , MutaciónRESUMEN
Soluble growth factors play an important role in the coordination and integration of cell proliferation, differentiation, fate determination, and morphogenesis during development of multicellular organisms. Fibroblast growth factors (FGFs) are a large family of polypeptide growth factors that are present in organisms ranging from nematodes to humans. RNA alternative splicing of FGFs and their receptors further enhances the complexity of this ligand-receptor system. The mouse Fgf8 gene produces eight splice variants, which encode isoform proteins with different N-termini and distinct receptor-binding affinity and biological activity. In this article, we review the roles of Fgf8 in vertebrate development and summarize the recent findings on the in vivo function of different Fgf8 splice variants. We propose that multiple Fgf8 isoform proteins act in concert to regulate the overall function of Fgf8 and account for the diverse and essential role of Fgf8 during vertebrate development.
Asunto(s)
Encéfalo/metabolismo , Factor 8 de Crecimiento de Fibroblastos/metabolismo , Transducción de Señal , Animales , Encéfalo/embriología , Encéfalo/crecimiento & desarrollo , Diferenciación Celular , Proliferación Celular , Factor 8 de Crecimiento de Fibroblastos/genética , Factores de Crecimiento de Fibroblastos/metabolismo , Humanos , Ratones , Morfogénesis , Unión Proteica , Isoformas de Proteínas , Receptores de Factores de Crecimiento de Fibroblastos/metabolismo , Proteínas de Pez Cebra/metabolismoRESUMEN
Being supportive cells for neurons in the central nervous system, astrocytes have recently found to be associated with neurogenesis. Ventral mesencephalon (VM) astrocytes were also detected being instructive for VM dopaminergic (DA) neurogenesis, but the underling mechanisms are still unclear. This research is to figure out whether VM astrocytes are more efficient than those from other brain regions in inducing VM DA neurons from their precursors and whether transforming growth factor-betas (TGF-betas) are the underlying molecules. We found that, compared with astrocytes preparations from striatum and hippocampus, VM astrocytes preparations displayed markedly higher efficacy in inducing DA neurogenesis. Besides, they also expressed higher level of TGF-beta3 than those of two other regions. When TGF-beta3 gene expression in astrocytes preparations was inhibited by its antisense oligonucleotide, the induction of DA neurons decreased to a similar level among these three astrocytes preparations. Thus, our experiment indicates that VM astrocytes preparations which contained highly purified astrocytes are more efficient in inducing DA neurogenesis than those from other regions. Furthermore, it also suggests that the regional differences are regulated by different expression levels of TGF-beta3 in those astrocytes preparations from different derivations.
Asunto(s)
Astrocitos/fisiología , Dopamina/metabolismo , Mesencéfalo/citología , Factor de Crecimiento Transformador beta3/metabolismo , Animales , Astrocitos/citología , Células Cultivadas , Técnicas de Cocultivo , Femenino , Mesencéfalo/fisiología , Neurogénesis/fisiología , Embarazo , Ratas , Ratas Sprague-Dawley , Factor de Crecimiento Transformador beta2/genética , Factor de Crecimiento Transformador beta2/metabolismo , Factor de Crecimiento Transformador beta3/genéticaRESUMEN
Silkworms are economically important insects because of the value of their silk. After finishing silk spinning, silkworms begin another important physiological process, vitellogenesis. In this study, we explored the relationship between silk spinning and vitellogenin (BmVg) expression in silkworms. In silkworms with the silk fibroin heavy chain gene knocked-out, the concentration of amino acids in the hemolymph was found to be significantly higher than that in the wild type, and the expression of BmVg was advanced at day 7 of the fifth instar stage and 0â¯h after spinning. Furthermore, through culturing fat body in vitro with different substances treatment including glucose, trehalose, amino acids, 20-hydroxyecdysone, and insulin, we found that only amino acids could induce BmVg expression. RNA interference of BmTOR1 in female silkworms could down-regulate BmVg transcription, resulting in shortened egg ducts and smaller eggs relative to the control. Therefore, these results showed that amino acids could induce BmVg expression through the TOR signaling pathway. Fat body cultured with amino acids in vitro and experiments involving amino acids injected into the silkworm showed that the majority of main amino acids of silk protein could induce BmVg expression. These results suggested that BmVg expression is related to silk spinning and this study would lay a foundation for elucidating the stage specificity expression of BmVg.
Asunto(s)
Aminoácidos/metabolismo , Bombyx/metabolismo , Vitelogeninas/metabolismo , Aminoácidos/farmacología , Animales , Bombyx/genética , Bombyx/crecimiento & desarrollo , Cuerpo Adiposo/efectos de los fármacos , Cuerpo Adiposo/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Genitales Femeninos/metabolismo , Hemolinfa/química , Proteínas de Insectos/metabolismo , Estadios del Ciclo de Vida , Interferencia de ARN , Seda/química , Técnicas de Cultivo de Tejidos , Vitelogeninas/genéticaRESUMEN
GATA transcription factors (GATAs) are widely expressed among various organisms and belong to the zinc finger protein family. GATA transcription factors play important roles in the proliferation, differentiation, and development of eukaryotes. Previous studies have shown that GATA participates in oogenesis by selective splicing in silkworms. In this study, we investigated the function of GATAs during vitellogenesis using female silkworms (Bombyx mori). Six types of GATA transcription factors were successfully cloned in the fat body of silkworms during the wandering stage and only BmGATAß4 induced the activity of the Bombyx mori vitellogenin (BmVg) promoter. Furthermore, BmVg and BmGATAß4 exhibited similar expression patterns in the fat body of female silkworms during the wandering stage. Electrophoretic mobility shift assays, cell transfection assays, and chromatin immunoprecipitation showed that BmGATAß4 was involved in regulating the transcription of BmVg by directly binding to the GATA cis-response element 1 (CRE1) and GATA cis-response element 2 (CRE2) in the promoter of the BmVg gene. RNA interference of BmGATAß4 in female silkworms downregulated BmVg transcription, resulting in a decrease in egg size and shortening of the length of egg tubes relative to the control. In summary, our results indicated that BmGATAß4 bound to the GATA CRE1 and CRE2 motifs in the BmVg promoter to upregulate BmVg expression in the fat body of female silkworms.
Asunto(s)
Bombyx/genética , Factores de Transcripción GATA/genética , Proteínas de Insectos/genética , Transcripción Genética , Vitelogeninas/metabolismo , Secuencia de Aminoácidos , Animales , Bombyx/crecimiento & desarrollo , Bombyx/metabolismo , Factores de Transcripción GATA/química , Factores de Transcripción GATA/metabolismo , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Óvulo/crecimiento & desarrollo , Filogenia , Alineación de SecuenciaRESUMEN
Microglia plays an important role in mediating neuroinflammation in Alzheimer's disease (AD). Intervention in microglia activation may exert a neuroprotective effect. In the present study, we reported that oligomeric Abeta1-42 dramatically increased the level of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta compared to monomeric and fibrillar Abeta1-42 in rat microglial cultures. Pretreatment of the cultures with triptolide, an anti-inflammatory reagent, alleviated the elevation of TNF-alpha and IL-1beta level induced by oligomeric Abeta1-42. Our results showed that oligomeric Abeta played an important role in mediating neuroinflammation and triptolide was able to suppress the production of pro-inflammatory cytokines from microglia.
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Péptidos beta-Amiloides/farmacología , Diterpenos/farmacología , Inmunosupresores/farmacología , Interleucina-1beta/metabolismo , Microglía/efectos de los fármacos , Fragmentos de Péptidos/farmacología , Fenantrenos/farmacología , Factor de Necrosis Tumoral alfa/metabolismo , Análisis de Varianza , Animales , Animales Recién Nacidos , Encéfalo/citología , Células Cultivadas , Relación Dosis-Respuesta a Droga , Ensayo de Inmunoadsorción Enzimática/métodos , Compuestos Epoxi/farmacología , Ratas , Ratas Sprague-Dawley , Factores de TiempoRESUMEN
OBJECTIVE: To investigate the possible influencing factors of pulmonary dysfunction in coal worker's pneumoconiosis (CWP). METHODS: A total of 141 patients with CWP and 200 control miners with similar exposure histories but without apparent pulmonary disease or inflammation were interviewed with the detailed questionnaires (including histories of coal dust-exposure, smoking habits, alcohol consumption, protective mask uses, et al). Lung function examinations were performed at the same time. Predicted formula of lung function index were established by the local healthy residents characters and the pulmonary dysfunction was classified by the ratios between tested and predicted values. RESULTS: All parameters of lung function were significantly lower in CWP cases when compared with that of control miners and the healthy controls (P < 0.05). The main types of pulmonary dysfunction were restrictive and mixed ventilation disorders in CWP patients. The factors such as the category of CWP, the mask worn, the smoking quantity and exposure to coal mine dust were included in the unconditional logistic regression model. CONCLUSIONS: The category of CWP, the usage of mask, the smoking and long duration exposure to coal mine dust may be the main possible influencing factors of pulmonary dysfunction of CWP. Influencing factor analyses were given to inform choice of pertinence preventive measures.
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Antracosis/fisiopatología , Anciano , Estudios de Casos y Controles , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Ventilación Pulmonar , Factores de RiesgoRESUMEN
Current gene therapy models for Parkinson's disease (PD) have adapted two treatment strategies. One is to restore dopamine (DA) production by delivering the genes of DA-synthesizing enzymes such as tyrosine hydroxylase (TH) to the striatum to relieve motor symptoms of PD. Another is to block or slow down progressive degenerative changes by delivering neurotrophic factors such as glial cell line-derived neurotrophic factor (GDNF) to protect the remained neurons. To test the assumption that the combination of the two strategies may have a compound or synergistic effect, we had constructed tetracycline-inducible (tet-off) AAV vector carrying GDNF and TH. After co-transfection of PC12 cells with this vector and the inducer plasmid, the expression of GDNF and TH protected these cells from 1-methyl-4-phenyl-pyridinium-induced injury, and significantly increased the content of dopamine in GDNF/TH-expressing cells compared with the control. Furthermore, mRNA expression of GDNF and TH could be effectively and reversibly regulated by doxycycline (Dox) and the function of GDNF and TH could be repressed by Dox. These results suggest that the tet-off AAV vector carrying GDNF and TH may be a useful tool for gene therapy in the treatment of PD.
Asunto(s)
Doxiciclina/farmacología , Regulación Viral de la Expresión Génica/efectos de los fármacos , Vectores Genéticos/efectos de los fármacos , Factor Neurotrófico Derivado de la Línea Celular Glial/genética , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Tirosina 3-Monooxigenasa/genética , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina/farmacología , Animales , Antibacterianos/farmacología , Dopamina/metabolismo , Regulación Viral de la Expresión Génica/genética , Terapia Genética/métodos , Vectores Genéticos/fisiología , Factor Neurotrófico Derivado de la Línea Celular Glial/metabolismo , Degeneración Nerviosa/genética , Degeneración Nerviosa/metabolismo , Degeneración Nerviosa/terapia , Células PC12 , Enfermedad de Parkinson/genética , Enfermedad de Parkinson/metabolismo , Enfermedad de Parkinson/terapia , Ratas , Transfección/métodos , Tirosina 3-Monooxigenasa/metabolismoRESUMEN
Neurofibromatosis type 1 (NF1) is a common genetic disorder characterized by the occurrence of nerve sheath tumors and considerable clinical heterogeneity. Some translational studies have been limited by the lack of animal models available for assessing patient-specific mutations. In order to test therapeutic approaches that might restore function to the mutated gene or gene product, we developed mice harboring NF1 patient-specific mutations including a nonsense mutation (c.2041C>T; p.Arg681*) and a missense mutation (c.2542G>C; p.Gly848Arg). The latter is associated with the development of multiple plexiform neurofibromas along spinal nerve roots. We demonstrate that the human nonsense NF1(Arg681*) and missense NF1(Gly848Arg) mutations have different effects on neurofibromin expression in the mouse and each recapitulates unique aspects of the NF1 phenotype, depending upon the genetic context when assessed in the homozygous state or when paired with a conditional knockout allele. Whereas the missense Nf1(Gly848Arg) mutation fails to produce an overt phenotype in the mouse, animals homozygous for the nonsense Nf1(Arg681*) mutation are not viable. Mice with one Nf1(Arg681*) allele in combination with a conditional floxed Nf1 allele and the DhhCre transgene (Nf1(4F/Arg681*); DhhCre) display disorganized nonmyelinating axons and neurofibromas along the spinal column, which leads to compression of the spinal cord and paralysis. This model will be valuable for preclinical testing of novel nonsense suppression therapies using drugs to target in-frame point mutations that create premature termination codons in individuals with NF1.
Asunto(s)
Codón sin Sentido/genética , Mutación Missense/genética , Neurofibromatosis 1/genética , Neurofibromina 1/genética , Animales , Modelos Animales de Enfermedad , Embrión de Mamíferos/patología , Humanos , Integrasas/metabolismo , Ratones , Neurofibroma/patología , Fenotipo , Nervio Ciático/patología , Nervio Ciático/ultraestructura , Médula Espinal/patología , Médula Espinal/ultraestructuraRESUMEN
Unlike the laminar arrangement of neurons in the neocortex, thalamic neurons aggregate to form about dozens of nuclei, many of which make topographic connections with specific areas in the neocortex. The molecular mechanisms underlying the formation of thalamic nuclei remain largely unknown. Homeodomain transcription factor Gbx2 is specifically expressed in the developing thalamus. Deleting Gbx2 leads to severe disruption of the histogenesis of the thalamus in mice, demonstrating an essential role of Gbx2 in this brain structure. Using inducible genetic fate mapping, we have previously shown that the neuronal precursors for different sets of thalamic nuclei have distinctive onset and duration of Gbx2 expression, suggesting that the dynamic expression of Gbx2 plays an important role in the specification and differentiation of thalamic nuclei. Here, we showed that the Gbx2 lineage exclusively gives rise to neurons but not glia in the thalamus. We performed conditional deletion to examine the temporal requirements of Gbx2 in the developing thalamus in mice. Corresponding to the dynamic and differential expression of Gbx2 in various thalamic nucleus groups, deleting Gbx2 at different embryonic stages disrupts formation of distinct sets of thalamic nuclei. Interestingly, different thalamic nuclei have remarkably different requirements of Gbx2 for the survival of thalamic neurons. Furthermore, although Gbx2 expression persists in many thalamic nuclei until adulthood, only the initial expression of Gbx2 following neurogenesis is crucial for the differentiation of thalamic nuclei. Our results indicate that the dynamic expression of Gbx2 may act as an important determinant in coupling with other developmental programs to generate distinct thalamic nuclei.
Asunto(s)
Proteínas de Homeodominio/metabolismo , Neuronas/citología , Neuronas/metabolismo , Núcleos Talámicos/citología , Núcleos Talámicos/metabolismo , Animales , Supervivencia Celular/genética , Supervivencia Celular/fisiología , Corteza Cerebral/citología , Corteza Cerebral/metabolismo , Femenino , Técnica del Anticuerpo Fluorescente , Regulación del Desarrollo de la Expresión Génica/genética , Regulación del Desarrollo de la Expresión Génica/fisiología , Proteínas de Homeodominio/genética , Hibridación in Situ , Masculino , RatonesRESUMEN
Triptolide (T10), an extract from the traditional Chinese herb, Tripterygium wilfordii Hook F (TWHF), has been shown to attenuate the rotational behavior induced by D: -amphetamine and prevent the loss of dopaminergic neurons in the substantia nigra in rat models of Parkinson's disease. To examine if the neuroprotective effect is mediated by its stimulation of production of neurotrophic factors from astrocytes, we investigated the effect of T10 on synthesis and release of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) in rat astrocyte cultures. T10 did not affect the synthesis and release of either BDNF or GDNF. However, it significantly increased NGF mRNA expression. It also increased both intracellular NGF and NGF level in culture medium. These results indicate that the neuroprotective effect of T10 might be mediated, at least in part, via a stimulation of the production and release of NGF in astrocytes.