Ryazantsev et al. in addition to conventional enzyme replacement therapy to preserve the CNS in patients with MPS II. [17]. Autophagy is considered to be highly involved in the neuropathy of LSD. In 1997, Elleder et al. recognized that immunostaining of subunit c of mitochondrial ATP synthetase (SCMAS) was positive in Mouse monoclonal to CD32.4AI3 reacts with an low affinity receptor for aggregated IgG (FcgRII), 40 kD. CD32 molecule is expressed on B cells, monocytes, granulocytes and platelets. This clone also cross-reacts with monocytes, granulocytes and subset of peripheral blood lymphocytes of non-human primates.The reactivity on leukocyte populations is similar to that Obs the neurons of patients with neuronal ceroid lipofuscinosis (NCL), which is an LSD. In addition, they showed the accumulation of SCMAS in the brain neurons of patients with MPS I, II, and IIIA; NiemannCPick disease A and C; and GM1 and GM2 gangliosidoses, and they suggested that the accumulation of SCMAS may be a common mechanism of neuropathy in LSDs [18]. Another marker of autophagy, p62, localizes to protein aggregates and abnormal mitochondria via a ubiquitin signal and is degraded by autophagy along with these structures [19]. To investigate the role of autophagy in the mechanisms of brain damage, we performed SCMAS and p62 antibody staining in iduronic acid-2-sulfatase (IDS) knockout (KO) mice, a model of MPS II. As in humans, the IDS gene in mice is located on the X chromosome, and X-linked recessive traits are seen in IDS-KO mice; 50% of males that are homozygous for the KO gene develop the disease. At three to four months after birth, bone deformity, joint disorders, and hepatomegaly are observed; neurologic signs appear; and movement becomes sluggish; the mice die 12 to 14 months after birth. We also examined the possible therapeutic effects of chloroquine as a treatment for neurodegeneration in NFAT Inhibitor our mouse model. We showed enhanced levels of p62 and SCMAS, suggestive of autophagy, as a result of enhanced initiation of autophagy, and/or reduced elimination of autophagic material in the neurons of the MPS II model mouse brain, and that administration of chloroquine was effective for treating neuronal degeneration. 2. Results 2.1. Vacuoles in Neurons, Microglia, and Pericytes in the CNS of IDS-KO Mice The appearance of IDS-KO mice was not markedly different from that of the wild-type mice (Figure 1A), and there was no difference in weight between the two genotypes. Hematoxylin and eosin (HE) and toluidine blue (TB) staining of the cerebral cortex of IDS-KO mice revealed neurons and glial cells with small vacuoles in the cytoplasm (Figure 1B). Electron microscopy revealed a layered structure, with spiral inclusion bodies resembling zebra bodies [20] and large vacuoles in the cytoplasm of neurons of the cerebral cortex. Similar vacuoles were found in the microglia and pericytes of the blood vessel walls (Figure 1C eCh). However, these abnormalities were not observed in the cerebral cortex of wild-type mice (Figure 1C aCd). Open in a separate window NFAT Inhibitor Figure 1 The appearance of wild-type and iduronic acid-2-sulfatase (IDS)-knockout (KO) mice. (A) The appearance of IDS-KO mice was not markedly changed. (B) Hematoxylin and eosin (HE) and toluidine blue (TB) staining of cerebral cortex from six-month-old mice. The neurons of the IDS-KO mice contained numerous vacuoles (arrows). Bar = 100 m. (C) Electron micrographs of the cerebral cortex of wild-type (aCd) and IDS-KO (eCh) mice: (a,b,e,f) show neurons; (c,g) show microglia; (d,h) show pericytes. Panel b shows a higher magnification of the boxed area in (a). Numerous spiral structures and layered structures (e) and autophagy-like vacuoles (f) were found in the cytoplasm of the neurons in IDS-KO mice, and numerous vacuoles were observed in the cytoplasm of the microglia NFAT Inhibitor (g) and pericytes (h). Arrows indicate vacuoles. Bar = 1 m. 2.2. Autophagy-Like Changes in the Cerebral Cortex of IDS-KO Mice Anti-p62 and anti-SCMAS antibodies were used to examine.