Transient transgenesis of the tapeworm XXXX .

@entity1 @entity6583 @entity3993 @entity19 @entity6475 @entity8785 @entity2834 @entity21150
@entity1 and porcine @entity3993 is caused by the larval stage of the flatworm @entity8785 (Cestoda). Infestation of the @entity1 brain, also known as @entity6583 , is the most common parasite disease of the central nervous system worldwide. Significant advances in the understanding of the disease have been achieved using the @entity21150 @entity19 model. We describe here a successful transfection protocol of @entity21150 cysticerci as the first step to approach a number of currently inaccessible biological questions on @entity3993 . @entity21150 cysticerci ( @entity6475 ) were microinjected with the plasmid pcDNA3.1/NT-GFP-TOPO, encoding the green fluorescent protein (GFP) driven by a cytomegalovirus promoter ( @entity2834 ). Twelve hours after the microinjection, GFP fluorescence gradually developed in patches associated to bud structures in the bladder wall of cysts. Fluorescence reached a peak at 24-48  h and lasted up to 72  h after the microinjection. Immunohistochemical studies on tissue sections of transfected cysts using an anti-GFP antibody, demonstrated co-localization of the antibody and the GFP fluorescence in the tegumentary cytoplasm and subtegumentary cytons. To validate at the mRNA level the expression of GFP, we carried out RT-PCR using two pairs of nested primers. Results showed expression of GFP-mRNA at 24  h post-transfection. Moreover, western blot assays of crude extracts of transfected cysts, carried out using the anti-GFP specific antibody, showed the expected protein band of 27  kDa, demonstrating that the GFP expression started at 24 after plasmid microinjection and was maintained up to 72  h. These findings will facilitate the development of functional genomics approaches applied to this model of @entity3993 .
