No abstract

After the spinal cord injury, inflammation and cytotoxicity cause further damage to neural cells. The progression of this secondary injury might be reduced by the administration of anti-inflammatory drugs. To allow the local delivery of such drugs while minimizing dural opening, we have created a polypyrrole (PPy)-coated microneedle array using a microscale three-dimensional (3D) printing technology that facilitates electronically controlled encapsulation and the transdural release of drugs. PPy microneedles demonstrated an electronically controlled release of steroid dexamethasone (Dexa) in a novel in vitro transdural model and in vivo. The biological activity of the device was then tested by the electronic release of Dexa into an in vitro model of neuroinflammation, using activated microglia. Following electrically activated Dexa release, inflammation was reduced, as demonstrated by a decrease in nitric oxide and proinflammatory cytokines Il-6 and MCP-1. These results demonstrate the feasibility of PPy-coated microneedles for the transdural delivery of anti-inflammatory drugs to the central nervous system.

Background The cadherin‐associated protein p120 catenin regulates convergent extension through interactions with cadherin proteins, Cdc42, and Rac1, as we previously showed in zebrafish (Danio rerio). Phosphorylation of p120 catenin changes the nature of its activity in vitro but is virtually unexplored in embryos. We used our previously developed antisense RNA splice‐site morpholino targeted to endogenous p120 catenin‐δ1 to cause defects in axis elongation probing the functions of three p120 catenin tyrosine‐phosphorylation sites in gastrulating zebrafish embryos. Results The morpholino‐induced defects were rescued by co‐injections with mouse p120 catenin‐δ1‐3A mRNAs mutated at residues Y228 and Y217 to a non‐phosphorylatable phenylalanine (F) or mutated at residue Y335 to a phosphomimetic glutamic acid (E). Co‐injection of the complementary mutations Y228E, Y217E, or Y335F mRNAs partially rescued embryos whereas dual mutation to Y228E‐Y217E blocked rescue. Immunopurification showed Y228F mutant proteins preferentially interacted with Rac1, potentially promoting cell migration. In contrast, the phosphomimetic Y228E preferentially interacted with E‐cadherin increasing adhesion. Both Y228F and Y335F strongly bind VAV2. Conclusions p120 catenin serves dual roles during gastrulation of zebrafish. Phosphorylation and dephosphorylation of tyrosine residues Y217, Y228, and Y335 precisely balance cell adhesion and cell migration to facilitate somite compaction and axis elongation.

Background: Cadherin-associated protein p120 catenin regulates cell adhesion and migration in cell cultures and is required for axial elongation in embryos. Its roles in adhesion and cell migration are regulated by phosphorylation. We determined the effects of phosphorylation of six serine and three threonine residues in p120 catenin during zebrafish (Danio rerio) embryogenesis. Results:We knocked down endogenous p120 catenin-δ1 with an antisense RNA-splice-site morpholino (Sp-MO) causing defects in axis elongation. These defects were rescued by co-injections of mRNAs for wildtype mouse p120 catenin-δ1-3A or various mutated forms. Several mRNAs containing serine or threonine codons singly or doubly mutated to phosphomimetic glutamic acid rescued, and some nonphosphorylatable mutants did not. Conclusions: We discovered that phosphorylation of serine residue S252 or S879 is required for convergent extension of zebrafish embryos, since rescue Ariana Kupai, Hiroko Nakahara, and Kathleen M. Voss contributed equally to this study.