The T11 Ab recognizes an epitope that lies 0.05 µm laterally to the A–I junction along the thin filaments. In longitudinal sections, this Ab showed a pattern of transverse fluorescent elements, which, at higher magnification, were composed by doublets lying astride unstained Selleckchem Ku 0059436 bands. The latter represent the I bands, while intervals in between doublets are occupied by the A bands, as shown by comparative evaluation of IF and corresponding phase contrast images in isolated myofibrils [39]. When sections

incubated with both anti-ZNF9 and anti-T11 Abs were examined by confocal microscopy, the merged image for the two fluorochromes showed a complete separation of the two fluorescent patterns, with that of ZNF9 occupying the internal space of the T11 doublets, that is the I bands (Figure 2B). When similar experiments of double IF were conducted using anti-K20

and anti-T12, the merged images revealed a co-localization of ZNF9 and T12, which showed a less restricted localization within the I bands as compared with T11 (not shown). The immunogold staining of ultrathin longitudinal Z-VAD-FMK datasheet muscle sections showed a clear association of ZNF9 with thin filaments in the I bands while the A bands were not immunodecorated (Figure 2C). No immunolocalization was observed in mitochondria or in other intracellular organelles. In DM2 patients’ muscles, localization of ZNF9 was comparable with that of normal muscles (Figure 2D). In intramuscular nerve twigs, as in neuromuscular junctions, nerve axons and terminals were intensely marked by anti-ZNF9 Ab, the immune reaction being more intense than in myofibres. On the other hand, myelin sheaths and Schwann cell bodies were not immunoreactive (Figure 2E). In coronal sections of rat brain we observed a marked staining for ZNF9 in the white matter, corresponding to axonal localization, and in neurites and cytoplasm of pyramidal neurones in the telencephalic cortex

(Figure 2F). Other neuronal populations, such as small cortical neurones and striatal neurones, were not immunostained. Zinc finger motifs are present in numerous proteins that bind DNA or RNA [40]. The function of most Ureohydrolase zinc finger proteins is still unknown, although some of them may act as transcription factors or activators. In particular, several zinc finger proteins act as regulators of muscle development and muscle-specific gene expression [41]. The extraordinary sequence conservation of ZNF9, reaching 99% in the coding region of chicken, mouse, rat and human cDNAs, suggests an important physiological role for this protein [30]. Tissue-specific expression of ZNF9 in chicken shows a ubiquitous pattern, further indicating that this protein may play a role in basic cellular processes [28]. Subcellular fractionation studies of adult mouse liver have shown that ZNF9 is present in the cytoplasmic and endoplasmic reticulum fractions, but not in the nuclear fractions [29].