In the peripheral nervous system (PNS), Schwann cells ensheathe and myelinate axons to promote saltatory conduction of nerve impulses. Close interactions between Schwann cells and axons, and Schwann cells and the basal lamina are essential for the regulation of Schwann cell development and function. Myelinating Schwann cells are highly polarized radially and longitudinally for specifying distinct domains in the axon, which is required for fast action potential propagation. In addition, the Schwann cell cytoplasm is organized into discrete compartments, called Cajal bands, which contain different dystrophin-glycoprotein complexes that are believed to segregate the Schwann cell plasma membrane into appositions between the outer surface of the myelin sheath and the cytoplasmic face of the Schwann cell plasma membrane. Periaxin is expressed in myelinating Schwann cells, and homodimerizes at its PDZ domain to form a transmembrane complex with dystrophin-related protein 2 (DRP2) and dystroglycan. This PDG complex is concentrated at the appositions, and is essential for myelin sheath maintenance and stability in the mature PNS. In mice lacking Periaxin, an intact myelin sheath is formed but subsequently becomes unstable. Periaxin-null Schwann cells are also shorter, which has been proposed to result in a reduction in nerve conduction velocity. This thesis is a study of how Periaxin PDZ domain dimerization contributes to the regulation of PDG complex stability, apposition maintenance, Schwann cell internodal distance and myelin stability. I have studied the function of Periaxin by generating a conditional mutant mouse that lacks the PDZ domain, which is predicted to abrogate dimerization. In these mutants, DRP2 is severely depleted and appositions containing DRP2 fail to form. Mutant Schwann cells also have disrupted Cajal bands and shorter internodal lengths. In the mature peripheral nerves, mutant mice display a peripheral neuropathy characterized by hypermyelination with focally folded myelin. Nerve conduction velocity, motor coordination and sensory function were also studied in these mutant mice. Taken together, these data suggest that dimerization of the Periaxin PDZ domain is required for the stabilization of the PDG and appositions, and regulation of Schwann cell elongation and myelin maintenance. By analyzing a tamoxifen-inducible conditional mouse lacking Periaxin’s PDZ domain in mature myelinating Schwann cells, this work also shows that Periaxin dimerization is essential for maintaining Schwann cell compartmentalization and myelin stability in adult nerves. Finally, studies of single amino acid mutations of the Periaxin PDZ reveal that subtle changes in the structure of the PDZ domain can abrogate dimerization,and a possible mechanism for PDZ-PDZ homodimerization of Periaxin is proposed.

Submitted by Rafaela Prosdocini Parmeggiani (rafaela.prosdocini@gmail.com) on 2016-02-16T16:01:51Z No. of bitstreams: 1 Atividade cambial em ramos de Kielmeyera grandiflora (Wawra) Saddi (Calophyllaceae) ocorrentes no cerrado paulista.pdf: 2086514 bytes, checksum: d4efa2e5aa9b442218c364e59bc53a9a (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-02-17T15:53:14Z (GMT) No. of bitstreams: 1 parmeggiani_rp_me_bot.pdf: 2086514 bytes, checksum: d4efa2e5aa9b442218c364e59bc53a9a (MD5) / Made available in DSpace on 2016-02-17T15:53:14Z (GMT). No. of bitstreams: 1 parmeggiani_rp_me_bot.pdf: 2086514 bytes, checksum: d4efa2e5aa9b442218c364e59bc53a9a (MD5) Previous issue date: 2015-12-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / No cerrado, as estações são bem demarcadas pela sazonalidade pluvial, portanto, espera-se que haja uma sazonalidade na atividade do câmbio vascular. Kielmeyera grandiflora é considerada uma espécie modelo para estudos de atividade cambial em ramos, pois possui módulos de crescimento bem demarcados, formados por cicatrizes da gema terminal. Nosso objetivo foi de entender qual a relação entre a atividade cambial de Kielmeyera grandiflora (Wawra) Saddi (Calophyllaceae), os fatores ambientais e a fenologia e responder se a atividade cambial varia entre três módulos de crescimentos consecutivos. A área de estudo é um fragmento de cerrado em regeneração no município de Botucatu, estado de São Paulo, Brasil (S 22º53’11.0’’, W 48º29’17.3’’). Foram amostrados mensalmente os três últimos módulos de crescimento dos ramos de três indivíduos durante o período de fevereiro de 2012 a fevereiro de 2013. No mesmo período, a fenologia (brotação, folhas novas, folhas adultas, folhas senescentes e queda foliar) foram avaliadas. O material foi fixado em CRAF III para conservação do conteúdo celular. As amostras foram desidratadas em série alcoólica e incluídas em resina metacrilato para posterior corte em micrótomo rotativo. As análises foram feitas em fotomicroscópio de luz. Utilizamos Modelo Generalizado Linear Misto (GLMM) com erro Binomial para a variável resposta presença/ausência da atividade cambial e erro de Poisson para a variável resposta de contagem de paredes recém formadas. O início da atividade cambial, a brotação e o desenvolvimento de folhas novas ocorreram durante o mês de agosto no período seco. O período de atividade cambial sobrepôs-se à estação chuvosa, temperaturas elevadas e comprimento do dia entre 12 e 13,6 h. O câmbio vascular entrou em dormência em maio, mês que ainda chovia, contudo a temperatura e o comprimento do dia passavam por quedas acentuadas. A variação da atividade cambial entre os módulos de crescimento foi representativa somente no período de reativação cambial, mostrando que o módulo 1, mais próximo às fontes de auxina, apesar de ter iniciado atividade junto ao módulo 3, produziu em todo o período analisado, maior quantidade de células novas, possivelmente devido à ação de hormônios produzidos nos tecidos jovens e em desenvolvimento. / In the cerrado, the seasons are well marked by rain seasonality; therefore, there is a seasonality in the vascular cambium activity. Kielmeyera grandiflora is considered a model for branch cambial activity studies because it has well marked growth modules formed by terminal bud scars. Our goal was to understand what the relationship between cambial activity of Kielmeyera grandiflora (Wawra) Saddi (Calophyllaceae), environmental factors and the phenology and answear wheter cambial activity varies between three consecutive sections of internodal growth. The study area is a cerrado fragment in regeneration in the city of Botucatu, São Paulo State, Brazil (22° 53 ' S 11.0 '', W 48° 29 ' 17.3 ''). Were sampled on a monthly basis the last three modules of growth of branches of three individuals during the period between February 2012 to February 2013. The material was fixed in CRAF III for preservation of cell content. The samples were dehydrated in alcohol series and included in historesin for further cut in rotary microtome. Analyses were made on light microscope. We used Generalized Linear Mixed Model (GLMM) with Binomial error for the variable reply presence/absence of cambial activity and Poisson error for the variable count response of newly formed walls. The beginning of cambial activity, the sprouting and the development of new leaves occurred during the month of August in the dry period. The period of cambial activity overlapped the rainy season, high temperatures and day length between 12 and 13, 6 h. The vascular cambium dormancy was observed in May, a raining month; however, temperature and day length was decreasing. The variation of cambial activity between the internodal growth sections was representative just in reactivation period, showing that the younger internodal section, closest to the sources of auxin, has produced in the whole study period a larger number of new cells that the older, possibly due to the action of hormones produced in young and development tissues.

Potato is one of the main targets for genetic improvement by gene transfer. The aim of this study was to compare the efficacy of genetic transformation of potato, cultivar Bintje, using two methods: Agrobacterium tumefaciens mediated transformation and microprojectile bombardment. The same plasmid p35SGUSint, which cosists of 35S CaMV promoter, gus and nptII genes, was used for both transformations of internodal potato explants. Kamamycin selection, transient and stable expressions of {$\beta$}-glucuronidase and PCR amplification of gus and nptII transgenes were used for transgenic plant selection, identification and analysis.