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Duchenne muscular dystrophy (DMD) is an intractable genetic muscular disorder characterized by the loss of DYSTROPHIN. The restoration of DYSTROPHIN is expected to be a curative therapy for DMD. Because muscle stem cells (MuSCs) can regenerate damaged myofibers with full-length DYSTROPHIN in vivo, their transplantation is being explored as such a therapy. As for the transplanted cells, primary satellite cells have been considered, but donor shortage limits their clinical application. We previously developed a protocol that differentiates induced pluripotent stem cells (iPSCs) to MuSCs (iMuSCs). To ameliorate the respiratory function of DMD patients, cell transplantation to the diaphragm is necessary but difficult, because the diaphragm is thin and rapidly moves. In the present study, we explored the transplantation of iMuSCs into the diaphragm. First, we show direct cell injection into the diaphragm of mouse was feasible. Then, to enhance the engraftment of the transplanted cells in a rapidly moving diaphragm, we mixed polymer solutions of hyaluronic acid, alginate and gelatin to the cell suspension, finding a solution of 20% dissolved hyaluronic acid and 80% dissolved gelatin improved the engraftment. Thus, we established a method for cell transplantation into mouse diaphragm and show that an injectable hyaluronic acid-gelatin solution enables the engraftment of iMuSCs in the diaphragm.

[Resumen]: En el presente trabajo se ha estudiado la localización y distribución de la proteína HuC/D en el diencéfalo del pez cebra (Danio rerio). Esta proteína de unión al ARN ha permitido la detección de somas neuronales en el encéfalo de este teleósteo gracias al empleo de técnicas inmunohistoquímicas indirectas e inmunofluorescencia, revelando una abundante expresión positiva a la HuC/D en las diferentes regiones y principales núcleos que conforman el diencéfalo., [Resumo]: No presente traballo estudouse a localización e distribución da proteína HuC/D no diencéfalo do peixe cebra (Danio rerio). Esta proteína de unión ao ARN permitiu a detección de somas neuronais no encéfalo deste teleósteo grazas ao empleo de técnicas inmunohistoquímicas indirectas e inmunofluorescencia, revelando unha abundante expresión positiva á HuC/D nas diferentes rexións e principais núcleos que conforman o diencéfalo., [Abstract]: In the present work, we have studied the location and distribution of the HuC/D protein in the zebrafish diencephalon (Danio rerio). This RNA binding protein has allowed the detection of neuronal somas in the brain of this teleost, thanks to the use of indirect immunohistochemical techniques and immunofluorescence, revealing an abundant positive expression of HuC/D in different regions and main nuclei that form the diencephalon.

The M22.8 monoclonal antibody (mAb) developed against an antigen expressed at the mussel larval and postlarval stages of Mytilus galloprovincialis was studied on adult samples. Antigenic characterization by Western blot showed that the antigen MSP22.8 has a restricted distribution that includes mantle edge tissue, extrapallial fluid, extrapallial fluid hemocytes, and the shell organic matrix of adult samples. Other tissues such as central mantle, gonadal tissue, digestive gland, labial palps, foot, and byssal retractor muscle did not express the antigen. Immunohistochemistry assays identified MSP22.8 in cells located in the outer fold epithelium of the mantle edge up to the pallial line. Flow cytometry analysis showed that hemocytes from the extrapallial fluid also contain the antigen intracellularly. Furthermore, hemocytes from hemolymph have the ability to internalize the antigen when exposed to a cell-free extrapallial fluid solution. Our findings indicate that hemocytes could play an important role in the biomineralization process and, as a consequence, they have been included in a model of shell formation. This is the first report concerning a protein secreted by the mantle edge into the extrapallial space and how it becomes part of the shell matrix framework in M. galloprovincialis mussels