RESUMO
Arabidopsis thaliana CEL1 protein was detected in young expanding tissues. Immunostaining revealed that CEL1 accumulated mostly in xylem cells. The primary, as well as the secondary xylem showed considerable CEL1 staining. CEL1 was also observed in young epidermal cells, in which the thicker lateral and tangential walls stained more intensely than the inner walls. In newly formed cell walls, the lateral tangential walls were labeled more intensively than the inner walls. Cellulase activity was found to be significantly higher in growing tissue compared to mature parts of the plant. Cel1 expression concurrently with cellulase activity could be restored in detached matured leaves by sucrose treatment after 48 h in the culture medium.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/anatomia & histologia , Arabidopsis/enzimologia , Parede Celular/metabolismo , Celulase/metabolismo , Celulose 1,4-beta-Celobiosidase/metabolismo , Anticorpos/imunologia , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Western Blotting , Glucuronidase/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/enzimologia , Raízes de Plantas/citologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/enzimologia , Caules de Planta/citologia , Caules de Planta/efeitos dos fármacos , Caules de Planta/enzimologia , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas/genética , Transporte Proteico/efeitos dos fármacos , Sacarose/farmacologiaRESUMO
Development of effective therapies for multiple sclerosis (MS) is dependent on the advancement of improved tools for evaluation of progression of this disease in animal models. We present a novel technique utilizing scanning electron microscopy (SEM) for imaging wet biological specimens thus enabling rapid and high-resolution imaging of myelin in mouse spinal cord (SC). We demonstrate the advantages of using the wet SEM technique to image myelin in a murine model of MS, experimental autoimmune encephalomyelitis (EAE) induced in the Biozzi (antibody-high) mouse, by sensitization with spinal cord homogenate (SCH) in adjuvant. Our studies show that the methodology allows easy identification of normal and pathological components with great clarity, which is then correlated with light microscopy (LM) and validated thereby. Furthermore, we demonstrate gold immunolabeling of specific epitopes. We conclude that the new technique provides a quick, accurate, and detailed structural evaluation of the SC that can be applied to advance the research of MS.