RESUMO
A rat biological model of septoplasty was used to study the effect of surgery on passive avoidance conditioning (PAC). Septoplasty was shown to increase anxiety and to reduce exploratory activity in rodents during PAC. A neurochemical analysis of the hypothalamus was carried out immediately after the end of the experiment and showed an increase in norepinephrine (NE) metabolism after septoplasty. The finding was tentatively associated with activation of the hypothalamic-pituitary-adrenal axis.
Assuntos
Aprendizagem da Esquiva , Hipotálamo , Animais , Ratos , Aprendizagem da Esquiva/fisiologia , Hipotálamo/metabolismo , Masculino , Norepinefrina/metabolismo , Ratos Wistar , Cavidade Nasal/metabolismo , Cavidade Nasal/cirurgia , Sistema Hipotálamo-Hipofisário/metabolismo , Septo Nasal/cirurgia , Septo Nasal/metabolismoRESUMO
Genetic and environmental factors may lead to abnormal growth of the orofacial skeleton, affecting the overall structure of the face. In this study, we investigated the craniofacial abnormalities in a mouse model for Keutel syndrome, a rare genetic disease caused by loss-of-function mutations in the matrix Gla protein (MGP) gene. Keutel syndrome patients show diffuse ectopic calcification of cartilaginous tissues and impaired midface development. Our comparative cephalometric analyses of micro-computed tomography images revealed a severe midface hypoplasia in Mgp-/- mice. In vivo reporter studies demonstrated that the Mgp promoter is highly active at the cranial sutures, cranial base synchondroses, and nasal septum. Interestingly, the cranial sutures of the mutant mice showed normal anatomical features. Although we observed a mild increase in mineralization of the spheno-occipital synchondrosis, it did not reduce the relative length of the cranial base in comparison with total skull length. Contrary to this, we found the nasal septum to be abnormally mineralized and shortened in Mgp-/- mice. Transgenic restoration of Mgp expression in chondrocytes fully corrected the craniofacial anomalies caused by MGP deficiency, suggesting a local role for MGP in the developing nasal septum. Although there was no up-regulation of markers for hypertrophic chondrocytes, a TUNEL assay showed a marked increase in apoptotic chondrocytes in the calcified nasal septum. Transmission electron microscopy confirmed unusual mineral deposits in the septal extracellular matrix of the mutant mice. Of note, the systemic reduction of the inorganic phosphate level was sufficient to prevent abnormal mineralization of the nasal septum in Mgp-/-;Hyp compound mutants. Our work provides evidence that modulation of local and systemic factors regulating extracellular matrix mineralization can be possible therapeutic strategies to prevent ectopic cartilage calcification and some forms of congenital craniofacial anomalies in humans.
Assuntos
Calcinose , Proteínas de Ligação ao Cálcio/deficiência , Condrócitos , Anormalidades Craniofaciais , Proteínas da Matriz Extracelular/deficiência , Septo Nasal , Animais , Calcinose/embriologia , Calcinose/genética , Calcinose/metabolismo , Calcinose/patologia , Condrócitos/metabolismo , Condrócitos/patologia , Anormalidades Craniofaciais/embriologia , Anormalidades Craniofaciais/genética , Anormalidades Craniofaciais/metabolismo , Anormalidades Craniofaciais/patologia , Humanos , Camundongos , Camundongos Knockout , Septo Nasal/embriologia , Septo Nasal/metabolismo , Septo Nasal/patologia , Proteína de Matriz GlaRESUMO
Streptococcus pneumoniae is an opportunistic pathogen that colonizes the nasopharynx. Herein we show that carbon availability is distinct between the nasopharynx and bloodstream of adult humans: glucose is absent from the nasopharynx, whereas galactose is abundant. We demonstrate that pneumococcal neuraminidase A (NanA), which cleaves terminal sialic acid residues from host glycoproteins, exposed galactose on the surface of septal epithelial cells, thereby increasing its availability during colonization. We observed that S. pneumoniae mutants deficient in NanA and ß-galactosidase A (BgaA) failed to form biofilms in vivo despite normal biofilm-forming abilities in vitro Subsequently, we observed that glucose, sucrose, and fructose were inhibitory for biofilm formation, whereas galactose, lactose, and low concentrations of sialic acid were permissive. Together these findings suggested that the genes involved in biofilm formation were under some form of carbon catabolite repression (CCR), a regulatory network in which genes involved in the uptake and metabolism of less-preferred sugars are silenced during growth with preferred sugars. Supporting this notion, we observed that a mutant deficient in pyruvate oxidase, which converts pyruvate to acetyl-phosphate under non-CCR-inducing growth conditions, was unable to form biofilms. Subsequent comparative transcriptome sequencing (RNA-seq) analyses of planktonic and biofilm-grown pneumococci showed that metabolic pathways involving the conversion of pyruvate to acetyl-phosphate and subsequently leading to fatty acid biosynthesis were consistently upregulated during diverse biofilm growth conditions. We conclude that carbon availability in the nasopharynx impacts pneumococcal biofilm formation in vivo Additionally, biofilm formation involves metabolic pathways not previously appreciated to play an important role.
Assuntos
Biofilmes/crescimento & desenvolvimento , Metabolismo dos Carboidratos/fisiologia , Carboidratos/farmacologia , Galactose/farmacocinética , Neuraminidase/fisiologia , Infecções Pneumocócicas/microbiologia , Streptococcus pneumoniae/fisiologia , Análise de Variância , Animais , Biofilmes/efeitos dos fármacos , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Feminino , Galactose/metabolismo , Galactose/farmacologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Ácido N-Acetilneuramínico/metabolismo , Líquido da Lavagem Nasal/química , Septo Nasal/metabolismo , Septo Nasal/microbiologia , Nasofaringe/metabolismo , Nasofaringe/microbiologia , Neuraminidase/metabolismo , Infecções Pneumocócicas/metabolismo , Streptococcus pneumoniae/efeitos dos fármacos , beta-Galactosidase/deficiência , beta-Galactosidase/metabolismoRESUMO
Tissue Engineering is an important method for generating cartilage tissue with isolated autologous cells and the support of biomaterials. In contrast to various gel-like biomaterials, human demineralized bone matrix (DBM) guarantees some biomechanical stability for an application in biomechanically loaded regions. The present study combined for the first time the method of seeding chondrocyte-macroaggregates in DBM for the purpose of cartilage tissue engineering. After isolating human nasal chondrocytes and creating a three-dimensional macroaggregate arrangement, the DBM was cultivated in vitro with the macroaggregates. The interaction of the cells within the DBM was analyzed with respect to cell differentiation and the inhibitory effects of chondrocyte proliferation. In contrast to chondrocyte-macroaggregates in the cell-DBM constructs, morphologically modified cells expressing type I collagen dominated. The redifferentiation of chondrocytes, characterized by the expression of type II collagen, was only found in low amounts in the cell-DBM constructs. Furthermore, caspase 3, a marker for apoptosis, was detected in the chondrocyte-DBM constructs. In another experimental setting, the vitality of chondrocytes as related to culture time and the amount of DBM was analyzed with the BrdU assay. Higher amounts of DBM tended to result in significantly higher proliferation rates of the cells within the first 48 h. After 96 h, the vitality decreased in a dose-dependent fashion. In conclusion, this study provides the proof of concept of chondrocyte-macroaggregates with DBM as an interesting method for the tissue engineering of cartilage. The as-yet insufficient redifferentiation of the chondrocytes and the sporadic initiation of apoptosis will require further investigations.
Assuntos
Técnica de Desmineralização Óssea , Matriz Óssea/citologia , Cartilagem/citologia , Condrócitos/citologia , Septo Nasal/citologia , Engenharia Tecidual/métodos , Apoptose/fisiologia , Materiais Biocompatíveis , Matriz Óssea/metabolismo , Cartilagem/metabolismo , Caspase 3/metabolismo , Agregação Celular/fisiologia , Diferenciação Celular/fisiologia , Proliferação de Células , Células Cultivadas , Condrócitos/metabolismo , Colágeno Tipo II/metabolismo , Humanos , Septo Nasal/metabolismo , Fatores de TempoRESUMO
Marsupials are born very immature and crawl on their mother's belly to attach to teats. Sensory information is required to guide the newborn and to induce attachment to the teat. Olfaction has been classically proposed to influence neonatal behaviors, but recent studies suggest that the central olfactory structures are too immature to account for them. In the newborn opossum, we previously described a fascicle of nerve fibers expressing neurofilament-200 (NF200, a marker of fiber maturity) from the olfactory bulbs to the rostral telencephalon. The course of these fibers is compatible with that of the terminal nerve that, during development, is characterized by the presence of neurons synthetizing gonadotropin hormones (GnRH). To evaluate if these fibers are related to the terminal nerve and if they play a role in precocious behaviors in opossums, we used immunohistochemistry against NF200 and GnRH. The results show that NF200-labeled fibers are present between P0 and P11, but do not reach much further caudally than the septal region. Only a few NF200-labeled fibers were found near the olfactory and vomeronasal epitheliums and they did not penetrate the olfactory bulbs. NF200-labeled fibers follow the same path as fibers labeled for GnRH. In contrast to the latter, NF200-labeled fibers are no longer visible at P15. These results suggest that these fibers are neither from the olfactory nor from the vomeronasal nerves but may be part of the terminal nerve. Their limited caudal extension does not support a role in the sensorimotor behaviors of the newborn opossum.
Assuntos
Monodelphis , Animais , Telencéfalo/metabolismo , Bulbo Olfatório , Septo Nasal/metabolismo , Hormônio Liberador de Gonadotropina/metabolismoRESUMO
Mucus on the nasal mucosa is translocated to the pharynx by ciliary beating, which is an important nonspecific defense mechanism called mucociliary clearance (MC). MC is one of the important factors determining the rate and extent of drug absorption after nasal application. The purpose of this study is to evaluate MC using rat nasal septum under physiological condition in an in vitro system. The nasal septum was excised from rats anesthetized with urethane and the movement of fluorescent microspheres (FMS) applied on the nasal septum was observed with a fluorescence microscope. FMS were transported at a constant velocity in the same direction for a few minutes, but addition of 4% mucin solution on the nasal septum maintained MC for at least 90 min after excision. With our evaluation system established by modifying the method of Saldiva, MC was determined to be around 1 mm/min. Furthermore, the ciliostatic effect of benzalkonium chloride was observed, and it was confirmed that ß-adrenergic antagonists and a cholinergic antagonist decreased MC, and that ß-adrenergic agonists and a cholinergic agonist tended to increase MC, indicating that our system is valid and useful for evaluating MC function and the effect of drugs and pharmaceutical additives for nasal application on MC.
Assuntos
Depuração Mucociliar , Mucosa Nasal/metabolismo , Septo Nasal/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Antagonistas Adrenérgicos beta/farmacologia , Anestésicos/farmacologia , Animais , Anti-Infecciosos Locais/farmacologia , Compostos de Benzalcônio/farmacologia , Agonistas Colinérgicos/farmacologia , Antagonistas Colinérgicos/farmacologia , Técnicas In Vitro , Masculino , Microesferas , Mucinas/farmacologia , Depuração Mucociliar/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
BACKGROUND: When assessing the efficacy of gene transfer agents (GTAs) for cystic fibrosis (CF) gene therapy, we routinely evaluate gene transfer in the mouse nose and measure transfection efficiency by assessing transgene-specific mRNA using the real-time (TaqMan) quantitative reverse transcriptase-polymerase chain reaction. TaqMan is traditionally used to quantify expression in whole tissue homogenates, which in the nose would contain many cells types, including respiratory and olfactory epithelium. Only the respiratory epithelium is a satisfactory model for human airway epithelium and therefore CFTR gene transfer should be specifically assessed in respiratory epithelial cells (RECs). METHODS: We have compared laser microdissection, pronase digestion and nasal brushing for: (i) the ability to enrich RECs from the wild-type mouse nose and (ii) the length of time to perform the procedure. Using TaqMan, we subsequently assessed gene transfer in enriched RECs after nasal perfusion of GL67A/pCF1-CFTR complexes in a CF mouse model. RESULTS: Laser microdissection successfully isolated RECs; however, time-consuming sample preparation made this technique unsuitable for high-throughput studies. Pronase digestion was sufficiently rapid but only yielded 19% (range = 13%) RECs (n = 6). The nasal brushing method was superior, yielding 92% (range = 15%) RECs (n = 8) and was equally effective in CF knockout mice (91%, range = 14%, n = 10). Importantly, gene transfer was detectable in brushed RECs from 70% of perfused mice and the number of vector-specific transcripts was comparable to 3.5% of endogenous wild-type Cftr levels. CONCLUSIONS: Isolation of RECs by brushing allows accurate assessment of GTA transfection efficiency in an experimental system that is relevant for CF gene therapy.
Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/genética , Regulação da Expressão Gênica , Cavidade Nasal/patologia , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologia , Transgenes/genética , Animais , Separação Celular , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Proteínas de Ligação a Ácido Graxo/metabolismo , Técnicas de Transferência de Genes , Vetores Genéticos/genética , Humanos , Lasers , Camundongos , Camundongos Endogâmicos C57BL , Microdissecção , Cavidade Nasal/metabolismo , Septo Nasal/metabolismo , Septo Nasal/patologia , Pronase/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismoRESUMO
An eosinophilic substance (ES) is usually observed in the mouse nasal septum. In contrast to textbooks and one report describing ES as amyloid, a previous study by the authors revealed that ES is not amyloid but consists of collagen and an amorphous material. Furthermore, it was suggested that the amorphous material was produced by clear HE-stained nasal gland epithelial cells present at the dorsal portion directly above the vomeronasal organ. In this histological examination, ES deposition showed sex difference (more intense in males than in females). ES increased with age but not in seniles, suggesting that the increase has a limit. In the detailed examination using subserial HE-stained nasal sections, it was revealed that the clear HE-stained nasal glands continued to the vomeronasal glands, which communicated with the lumen of the vomeronasal organ, and the vomeronasal gland epithelial cells contained strongly periodic acid-Schiff (PAS) positive granules, similar to the clear HE-stained nasal gland epithelial cells. ES also deposited in the interstitium of the vomeronasal glands. The results suggested a possibility that ES deposition may be related to vomeronasal organ.
Assuntos
Eosinófilos/metabolismo , Septo Nasal/citologia , Órgão Vomeronasal/citologia , Animais , Feminino , Masculino , Camundongos , Cavidade Nasal/citologia , Cavidade Nasal/metabolismo , Septo Nasal/metabolismo , Distribuição por Sexo , Órgão Vomeronasal/metabolismoRESUMO
The airway epithelium is exposed to an acidic environment in certain conditions. The acid-sensing ion channel 2 (ASIC2) belongs to the epithelial amiloride-sensitive sodium channel and degenerin (ENaC/DEG) family and is expressed on cilia of the respiratory epithelium. The aim of this study was to detect the expression of ASIC2 in the nasal septum in the embryonic stage of the rat. ASIC2 expression was not observed in the primary cilium but was found in some cilia on embryonic day 17 (E17). After E18, all cilia showed ASIC2 immunoreactivity. RT-PCR analysis revealed that ASIC2b, a subtype of ASIC2, was expressed in the nasal septum while ASIC2a was not. Quantitative Real-time RT-PCR studies indicated that the expression level of ASIC2 mRNA was highest on E21, just before birth. These results imply that ASIC2 plays little part in the development of the nasal septum epithelium. On the other hand, ASIC2, especially ASIC2b, may function for the survival and retention of ciliated cells of the nasal septum against dynamic changes in the pH environment at birth.
Assuntos
Cílios/metabolismo , Septo Nasal/citologia , Septo Nasal/embriologia , Proteínas do Tecido Nervoso/metabolismo , Canais de Sódio/metabolismo , Canais Iônicos Sensíveis a Ácido , Animais , Regulação da Expressão Gênica no Desenvolvimento , Imuno-Histoquímica , Mucosa Nasal/citologia , Mucosa Nasal/metabolismo , Septo Nasal/metabolismo , Septo Nasal/ultraestrutura , Proteínas do Tecido Nervoso/genética , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Canais de Sódio/genéticaRESUMO
BACKGROUND: Articular cartilage injury has a poor repair ability and limited regeneration capacity with therapy based on articular chondrocytes (ACs) implantation. Here, we validated the hypothesis that human nasal septum-derived chondrocytes (hNCs) are potent therapeutic agents for clinical use in cartilage tissue engineering using an injectable hydrogel, type I collagen (COL1). METHODS: We manufactured hNCs incorporated in clinical-grade soluble COL1 and investigated their clinical potential as agents in an articular defect model. RESULTS: The hNCs encapsulated in COL1 (hNC-collagen) were uniformly distributed throughout the collagen and showed much greater growth rate than hACs encapsulated in collagen for the 14 days of culture. Fluorescent staining of hNC-collagen showed high expression levels of chondrocyte-specific proteins under clinical conditions. Moreover, a negative mycoplasma screening result were obtained in culture of hNC-collagen. Notably, implantation of hNC-collagen increased the repair of osteochondral defects in rats compared with implantation of collagen only. Many human cells were detected within the cartilage defects. CONCLUSION: These results provide reliable evidences supporting for clinical applications of hNC-collagen in regenerative medicine for cartilage repair.
Assuntos
Doenças das Cartilagens/terapia , Condrócitos/metabolismo , Colágeno/metabolismo , Septo Nasal/metabolismo , Engenharia Tecidual/métodos , Animais , Doenças das Cartilagens/metabolismo , Cartilagem Articular/lesões , Proliferação de Células , Sobrevivência Celular , Colágeno Tipo I , Humanos , Hidrogéis , Masculino , Mycoplasma , Ratos , Ratos Sprague-Dawley , Alicerces TeciduaisRESUMO
An eosinophilic substance is usually observed in the mouse nasal septum, and its volume increases with age. In contrast to descriptions in textbooks defining the eosinophilic substance as amyloid, our previous report revealed that the observed eosinophilic substance is not amyloid, but consisted of collagen and an amorphous material. Furthermore, it was suggested that the amorphous material was produced by the clear hematoxylin and eosin (HE)-stained nasal gland epithelial cells. In this study, we investigated the deposition process of the amorphous material produced by nasal gland epithelial cells in the interstitium morphologically. In most cases, the amorphous materials in the clear HE-stained nasal gland epithelial cells accumulated at the basal portion. Collagen fibers surrounding the nasal glands partially disappeared, whereas the amorphous material in contact with the rough endoplasmic reticulum of the nasal gland epithelial cells continued to the amorphous material in the interstitium. These findings suggested that the amorphous material produced by the clear HE-stained nasal gland epithelial cells migrated to the interstitium through the partial opening of the basement membrane.
Assuntos
Eosinófilos , Septo Nasal/citologia , Septo Nasal/metabolismo , Envelhecimento , Animais , Feminino , Masculino , CamundongosRESUMO
Nasoseptal cartilage has been assumed to be isotropic, unlike the well-defined zonal organization of articular cartilage attributed to postnatal biomechanical loading. We know from clinical experience that malrotation of surgical nasoseptal cartilage grafts can lead to increased graft absorption. Other studies have also suggested directionally dependent compressive stiffness suggesting anisotropy, but morphological investigations are lacking. This study characterizes immature and mature native bovine nasoseptal cartilage using a combination of immunohistochemistry, biomechanical testing and structural imaging. Our findings indicate that there is extensive postnatal synthesis and reorganization of the extracellular matrix in bovine nasoseptal cartilage, independent of joint loading forces responsible for articular cartilage anisotropy. Immature nasoseptal cartilage is more cellular and homogenous compared to the zonal organization of cells and extracellular matrix of mature cartilage. Mature samples also exhibited greater glycosaminoglycan content and type II collagen fibre alignment compared to immature cartilage and this correlates with greater compressive stiffness. Engineered neocartilage often consists of immature, isotropic, homogenous tissue that is unable to meet the functional and mechanical demands when implanted into the native environment. This study demonstrates the importance of anisotropy on biomechanical tissue strength to guide future cartilage tissue engineering strategies for surgical reconstruction.
Assuntos
Cartilagem/citologia , Cartilagem/metabolismo , Colágeno Tipo II/metabolismo , Força Compressiva , Septo Nasal/citologia , Septo Nasal/metabolismo , Animais , Anisotropia , Cartilagem/cirurgia , Bovinos , Septo Nasal/cirurgiaRESUMO
Intranasal administration of nano-drug-delivery systems emerged as an appealing strategy to surpass the blood-brain barrier and thus increase drug bioavailability in the central nervous system. However, a systematic study of the effect of the structural properties of the nanoparticles on the nose-to-brain transport is missing. In this work, we synthesized and characterized mixed amphiphilic polymeric nanoparticles combining two mucoadhesive graft copolymers, namely, chitosan- g-poly(methyl methacrylate) and poly(vinyl alcohol)- g-poly(methyl methacrylate), for the first time. Chitosan enables the physical stabilization of the nanoparticles by ionotropic cross-linking with tripolyphosphate and confers mucoadhesiveness, while poly(vinyl alcohol) is also mucoadhesive and, owing to its nonionic nature, it improves nanoparticle compatibility in nasal epithelial cells by reducing the surface charge of the nanoparticles. After a thorough characterization of the mixed nanoparticles by dynamic light scattering and nanoparticle tracking analysis, we investigated the cell uptake by fluorescence light and confocal microscopy and imaging flow cytometry. Mixed nanoparticles were readily internalized at 37 °C, while the uptake was inhibited almost completely at 4 °C, indicating the involvement of energy-dependent mechanisms. Finally, we assessed the nanoparticle permeability across liquid-liquid and air-liquid monolayers of a nasal septum epithelial cell line and studied the effect of nanoparticle concentration and temperature on the apparent permeability. Overall, our findings demonstrate that these novel amphiphilic nanoparticles cross this in vitro model of intranasal epithelium mainly by a passive (paracellular) pathway involving the opening of epithelial tight junctions.
Assuntos
Nanopartículas/química , Mucosa Nasal/metabolismo , Septo Nasal/metabolismo , Polímeros/química , Astrócitos/metabolismo , Transporte Biológico , Barreira Hematoencefálica/metabolismo , Linhagem Celular , Quitosana/química , Sistemas de Liberação de Medicamentos/métodos , Humanos , Espectroscopia de Ressonância MagnéticaRESUMO
Functional reconstruction of large cartilage defects in subcutaneous sites remains clinically challenging because of limited donor cartilage. Tissue engineering is a promising and widely accepted strategy for cartilage regeneration. To date, however, this strategy has not achieved a significant breakthrough in clinical translation owing to a lack of detailed preclinical data on cell yield and functionality of clinically applicable chondrocytes. To address this issue, the current study investigated the initial cell yield, proliferative potential, chondrogenic capacity, and regenerated cartilage type of human chondrocytes derived from auricular, nasoseptal, and costal cartilage using a scaffold-free cartilage regeneration model (cartilage sheet). Chondrocytes from all sources exhibited high sensitivity to basic fibroblast growth factor within 8 passages. Nasoseptal chondrocytes presented the strongest proliferation rate, whereas auricular chondrocytes obtained the highest total cell amount using comparable cartilage sample weights. Importantly, all chondrocytes at fifth passage showed strong chondrogenic capacity both in vitro and in the subcutaneous environment of nude mice. Although some significant differences in histological structure, cartilage matrix content and cartilage type specific proteins were observed between the in vitro engineered cartilage and original tissue; the in vivo regenerated cartilage showed mature cartilage features with high similarity to their original native tissue, except for minor matrix changes influenced by the in vivo environment. The current study provides detailed preclinical data for choice of chondrocyte source and thus promotes the clinical translation of cartilage regeneration approach.
Assuntos
Separação Celular , Condrócitos , Condrogênese , Cartilagem Costal/citologia , Cartilagem da Orelha/citologia , Septo Nasal/citologia , Animais , Condrócitos/citologia , Condrócitos/metabolismo , Condrócitos/transplante , Cartilagem Costal/metabolismo , Cartilagem da Orelha/metabolismo , Humanos , Camundongos Nus , Septo Nasal/metabolismoRESUMO
Exposure to lipopolysaccharides (LPS) causes extensive neutrophilic inflammation in the airways followed by mucous cell hyperplasia (MCH) that is sustained by the anti-apoptotic protein, Bcl-2. To identify inflammatory factor(s) that are responsible for Bcl-2 expression, we established an organ culture system consisting of airway epithelial tissue from the rat nasal midseptum. The highest Muc5AC and Bcl-2 expression was observed when organ cultures were treated with brochoalveolar lavage (BAL) fluid harvested from rats 10 h post LPS instillation. Further, because BAL harvested from rats depleted of polymorphonuclear cells compared to controls showed increased Bcl-2 expression, analyses of cytokine levels in lavages identified IL-13 as an inducer of Bcl-2 expression. Ectopic IL-13 treatment of differentiated airway epithelial cells increased Bcl-2 and MUC5AC expression in the basal and apical regions of the cells, respectively. When Bcl-2 was blocked using shRNA or a small molecule inhibitor, ABT-263, mucous cell numbers were reduced due to increased apoptosis that disrupted the interaction of Bcl-2 with the pro-apoptotic protein, Bik. Furthermore, intranasal instillation of ABT-263 reduced the LPS-induced MCH in bik +/+ but not bik -/- mice, suggesting that Bik mediated apoptosis in hyperplastic mucous cells. Therefore, blocking Bcl-2 function could be useful in reducing IL-13 induced mucous hypersecretion.
Assuntos
Inflamação/metabolismo , Interleucina-13/metabolismo , Lipopolissacarídeos/efeitos adversos , Septo Nasal/citologia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Animais , Apoptose , Líquido da Lavagem Broncoalveolar/imunologia , Hiperplasia , Masculino , Mucina-5AC/metabolismo , Septo Nasal/metabolismo , Septo Nasal/patologia , Técnicas de Cultura de Órgãos , RatosRESUMO
Previously, we identified PHR1 as an abundantly expressed gene in photoreceptors and showed that it encodes four isoforms, each with N-terminal pleckstrin homology (PH) and C-terminal transmembrane domains. To better understand PHR1 function and expression, we made a Phr1 null mouse by inserting a beta-galactosidase/neor cassette into exon 3. In addition to photoreceptors, we found abundant expression of specific Phr1 splice forms in olfactory receptor neurons and vestibular and cochlear hair cells. We also found Phr1 expression in cells with a possible sensory function, including peripheral retinal ganglion cells, cochlear interdental cells, and neurons of the circumventricular organ. Despite this discrete expression in known and putative sensory neurons, mice lacking PHR1 do not have overt sensory deficits.
Assuntos
Proteínas de Membrana/metabolismo , Neurônios Aferentes/fisiologia , Isoformas de Proteínas/metabolismo , Sequência de Aminoácidos , Animais , Encéfalo/citologia , Encéfalo/metabolismo , Orelha Interna/citologia , Orelha Interna/metabolismo , Eletrofisiologia , Feminino , Masculino , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Dados de Sequência Molecular , Septo Nasal/citologia , Septo Nasal/metabolismo , Neurônios Aferentes/citologia , Fenótipo , Isoformas de Proteínas/genética , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Retina/citologia , Retina/metabolismo , Sensação/fisiologia , Alinhamento de SequênciaRESUMO
INTRODUCTION: Although the role of immunoglobulin E-mediated hypersensitivity reactions in allergic rhinitis is well known, the relative contribution of sensory nerves to the symptoms of rhinitis is uncertain. This study looked at the level of specific neuronal markers including the nerve marker protein gene product 9.5 (PGP 9.5), sensory and autonomic neuropeptides, the capsaicin/heat receptor TRPV1, and nerve growth factor (NGF) in patients with allergic rhinitis and controls and their correlation with nasal sensitivity. MATERIALS AND METHODS: Forty patients (23 controls, 17 rhinitis) having nasal surgery were recruited. Nasal sensitivity was tested using graded monofilaments. Inferior turbinate biopsies were collected and studied using immunohistology, with measurement of nerve fibers by direct observation or computerized image analysis. RESULTS: Nerve fibers (PGP 9.5) in the epithelium, subepithelium, and glandular/vascular regions were significantly increased in allergic rhinitis (P=.037, <.01, and .04, respectively), as were subepithelial and glandular/vascular fibers immunoreactive for neuropeptide substance P (P=.04 subepithelium; .02 glandular/vascular) and neuropeptide tyrosine (P<.01 glandular/vascular), markers for sensory and sympathetic nerves, respectively. TRPV1 epithelial fiber counts were higher in rhinitis, but this was not statistically significant. Epithelial NGF immunoreactivity (% area) was significantly increased in rhinitis (P=.027). Nasal sensitivity was correlated significantly with PGP 9.5 subepithelial innervation (control touch P=.023, irritation P=.046; rhinitis touch P=.042, irritation P=.043). A correlation was also observed between epithelial NGF and subepithelial PGP 9.5 innervation, which included all subjects (P=.044). CONCLUSION: The increased number and specific phenotypical changes of sensory nerves may play a role in nasal hypersensitivity and provide new targets for the treatment of rhinitis.
Assuntos
Mucosa Nasal/inervação , Fatores de Crescimento Neural/genética , Fatores de Crescimento Neural/metabolismo , Neurônios Aferentes/metabolismo , Rinite Alérgica Perene/genética , Rinite Alérgica Perene/metabolismo , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismo , Adolescente , Adulto , Idoso , Biópsia , Feminino , Humanos , Imunoglobulina E/genética , Imunoglobulina E/metabolismo , Masculino , Pessoa de Meia-Idade , Mucosa Nasal/metabolismo , Mucosa Nasal/patologia , Septo Nasal/metabolismo , Septo Nasal/patologia , Septo Nasal/cirurgia , Fibras Nervosas/metabolismo , Cuidados Pré-Operatórios , Rinite Alérgica Perene/patologia , Rinoplastia , Substância P/metabolismoRESUMO
The olfactory system in rodents and many other mammals is classically divided into two anatomically separate, and morphologically distinct, sensory systems: the main olfactory system and the accessory olfactory system. We have now identified a novel third population of olfactory marker protein-expressing sensory neurons that is located in a discrete pocket of the rostral nasal septum, which we refer to as the septal organ of Grüneberg (SOG). Neurons in this region of the septum are located in the submucosa, in small grape-like clusters, rather than in a pseudostratified neuroepithelium, as seen in both the olfactory and vomeronasal neuroepithelia. Despite their unusual location, axons projecting from the SOG neurons fasciculate into several discrete bundles and terminate in a subset of main olfactory bulb glomeruli. These glomeruli most likely represent a subset of atypical glomeruli that are spatially restricted to the caudal main olfactory bulb. The unique rostral position of the SOG suggests that the SOG may be functionally specialized for the early detection of biologically relevant odorants.
Assuntos
Gânglios Sensitivos/citologia , Neurônios Aferentes/citologia , Bulbo Olfatório/citologia , Proteína de Marcador Olfatório/metabolismo , Condutos Olfatórios/citologia , Animais , Feminino , Gânglios Sensitivos/embriologia , Gânglios Sensitivos/metabolismo , Masculino , Camundongos , Camundongos Transgênicos , Septo Nasal/citologia , Septo Nasal/embriologia , Septo Nasal/metabolismo , Vias Neurais/citologia , Vias Neurais/embriologia , Vias Neurais/metabolismo , Neurônios Aferentes/metabolismo , Bulbo Olfatório/embriologia , Bulbo Olfatório/metabolismo , Proteína de Marcador Olfatório/genética , Mucosa Olfatória/citologia , Mucosa Olfatória/embriologia , Mucosa Olfatória/metabolismo , Condutos Olfatórios/embriologia , Condutos Olfatórios/metabolismo , Ratos , Ratos Sprague-Dawley , Septo do Cérebro/citologia , Septo do Cérebro/embriologia , Septo do Cérebro/metabolismoRESUMO
To investigate the effects of welding fumes on the histological structure and properties of mucins of the nasal respiratory mucosa, Sprague-Dawley rats were exposed to manual metal arc-stainless steel (MMA-SS) welding fumes at a concentration of 56-76 mg/m3 total suspended particulates for 2 h per day in an inhalation chamber for 90 days. Experimental animals were sacrificed at 2 h, 15, 30, 60 and 90 days after exposure. Loss of cilia, desquamation of epithelial cells, mucigenous epithelial cells and destruction of nasal septal glands were observed frequently in the welding fumes-exposed groups. These changes became more severe as the exposure continued. The amount of neutral mucins in goblet cells in the welding fumes-exposed group had a tendency to increase, the amount of sulfomucins decreased, while the sialomucins increased as the exposure continued. Mucinogenic epithelial cells, not visible in the control group, contained minimal to small amounts of neutral mucins. In the dorsal septal glands, neutral mucins not visible in the control group appeared, and neutral mucins in the ventral septal glands increased slightly. These results indicate that the observed changes in the properties of mucins due to inhalation of welding fumes may play roles in protection against toxicants.
Assuntos
Gases/toxicidade , Exposição por Inalação/efeitos adversos , Mucinas/biossíntese , Mucosa Nasal/metabolismo , Soldagem , Animais , Gases/análise , Concentração de Íons de Hidrogênio , Imuno-Histoquímica , Masculino , Mucinas/química , Mucosa Nasal/efeitos dos fármacos , Mucosa Nasal/patologia , Septo Nasal/metabolismo , Septo Nasal/patologia , Ratos , Ratos Sprague-DawleyRESUMO
OBJECTIVE: To investigate the difference in mucociliary clearance and surface mucosal structure of the nasal septum and lateral nasal wall in patients with and without septal deviation. METHOD: The saccharine-dye test was used to measure the mucociliary clearance time in both nasal cavities of 20 patients with nasal septal deviation (study group) and was compared with that of 30 patients without septal deviation (control group). Bilateral septal and lateral nasal wall mucosal biopsies were taken from the study group during septoplasty, and unilateral biopsies were taken from 10 of the control group. These biopsies were studied under the scanning electron microscope. RESULTS: In the study group, mucociliary clearance on the side opposite the septal deviation was significantly slower than on the other side. Mucociliary clearance on both sides of the deviated septum of the study group was significantly slower than clearance in the control group. There was no statistically significant difference in the distribution of mucosal cilia of the cavities on either side of the deviated septum in the study group, nor between the distribution in the study group and controls. CONCLUSION: Patients with septal deviation display no change in mucosal surface anatomy but have decreased mucociliary activity on both sides of the deviation, the least activity being on the side opposite the deviation.