Spatial and temporal expression profile of pinin during mouse development.

SR and SR-related proteins are splicing regulators involved in embryo development in higher eukaryotes. Pinin (pnn) is a SR-related protein localized both within nucleus (nuclear pnn, N-pnn) and at desmosome of cell-cell adhesion (desmosomal pnn, D-pnn). To investigate the role of N-pnn during mouse embryo development, we examined its expression using Northern blot, real-time RT-PCR, immunostaining, and mRNA in situ hybridization (ISH). On Northern analysis, we found that pnn transcripts display two isoforms due to differential utilization of a polyadenylation site and exhibit tissue variable expression with thymus expressing the highest level of transcript. Analysis of pnn expression in mouse embryos revealed N-pnn expression starts from the two-cell fertilized egg stage and is ubiquitous at all stages of mouse embryo development. ISH and immunofluorescent staining of embryo cryosections showed that during mouse organogenesis N-pnn is highly expressed in the central nervous system. In addition, N-pnn was found to be highly expressed in the cortex region of thymus of E16.5 mouse fetus, while in the hepatic primordium the strongest signals were noted at E13.5 to E14.5 rather than at later developmental stages. Finally, we also determined the subcellular location of N-pnn in photoreceptors of developing retinas by nuclear fractionation and Western blot, because N-pnn displayed a staining pattern reminiscent of cytoplasmic proteins at the microscopic level in developing mouse photoreceptors. Altogether these data provide us with a better understanding of the tissue distribution pattern of N-pnn during mouse development.

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