Pathogenic LRRK2 compromises the subcellular distribution of lysosomes in a Rab12-RILPL1-dependent manner.

Mutations in leucine-rich repeat kinase 2 (LRRK2) cause familial Parkinson's disease (PD). Recent studies have shown that LRRK2 physiologically phosphorylates several Rab family proteins including Rab12 and that this phosphorylation is accelerated by the pathogenic mutations in LRRK2, although the significance in the PD pathogenesis remains unknown. Here we examined the effect of the overexpression of LRRK2 on the distribution of organelles in cultured cells and found that lysosomes become clustered in a perinuclear region upon the overexpression of pathogenic mutant LRRK2 in a manner dependent on its kinase activity. The perinuclear clustering of lysosomes was abolished by knocking out RAB12 as well as its effector protein RILPL1. Re-expression of Rab12 in RAB12 knockout cells suggested that the phosphorylation at Ser106 of Rab12 is required for the perinuclear clustering of lysosomes. Moreover, phosphorylated Rab12 was also accumulated on the clustered lysosomes, and the phosphorylation of Rab12 increased its interaction with RILPL1, leading us to conclude that the increase in the phosphorylation of Rab12 by pathogenic LRRK2 compromised intracellular lysosomal transport via the enhanced interaction of Rab12 with RILPL1. These data suggest the involvement of abnormal regulation of lysosomal transport in the LRRK2-mediated pathogenesis of PD.

BORCS6 is involved in the enlargement of lung lamellar bodies in Lrrk2 knockout mice.

Leucine-rich repeat kinase 2 (LRRK2) has been implicated in the pathogenesis of Parkinson disease. It has been shown that Lrrk2 knockout (KO) rodents have enlarged lamellar bodies (LBs) in their alveolar epithelial type II cells, although the underlying mechanisms remain unclear. Here we performed proteomic analyses on LBs isolated from Lrrk2 KO mice and found that the LB proteome is substantially different in Lrrk2 KO mice compared with wild-type mice. In Lrrk2 KO LBs, several Rab proteins were increased, and subunit proteins of BLOC-1-related complex (BORC) were decreased. The amount of surfactant protein C was significantly decreased in the bronchoalveolar lavage fluid obtained from Lrrk2 KO mice, suggesting that LB exocytosis is impaired in Lrrk2 KO mice. We also found that the enlargement of LBs is recapitulated in A549 cells upon KO of LRRK2 or by treating cells with LRRK2 inhibitors. Using this model, we show that KO of BORCS6, a BORC subunit gene, but not other BORC genes, causes LB enlargement. Our findings implicate the LRRK2-BORCS6 pathway in the maintenance of LB morphology.

Complete Genome Sequence of a Genotype 3 Atypical Porcine Pestivirus Strain (OKN/2021) from Okinawa Prefecture, Japan.

We report the complete genome sequence of atypical porcine pestivirus (APPV) OKN/2021, which was sampled in the Okinawa Prefecture, Japan. The sequence bears the closest resemblance to another previously detected Japanese genotype 3 APPV sequence. This genome sequencing will help researchers in Japan learn more about the virus epidemiology and evolutionary characteristics.

Isolation of Culicoides- and Mosquito-Borne Orbiviruses in the Southwestern Islands of Japan Between 2014 and 2019.

The circulation of arboviruses in livestock ruminants has often gone unrecognized owing to the fact that a significant percentage of arboviruses probably induce subclinical infections and/or negligible symptoms in infected animals. To determine the current situation of arbovirus circulation in the Yaeyama Islands, attempts to isolate viruses from bovine blood samples collected between 2014 and 2019 have been made. In total, 308 blood samples were collected during the study period, and 43 of them induced cytopathic effects (CPEs) in cell cultures. The identification of the CPE agents was performed by reported RT-PCR assays and a high-throughput analysis with a next-generation sequencing platform. The obtained viruses consisted of an orthobunyavirus (Peaton virus), Culicoides-borne orbiviruses (bluetongue virus serotypes 12 and 16, epizootic hemorrhagic disease virus [EHDV] serotypes 5, 6, and 7, D'Aguilar virus, and Bunyip Creek virus), and potential mosquito-borne orbiviruses (Yunnan orbivirus, Guangxi orbivirus, and Yonaguni orbivirus). Most of the orbiviruses were recovered from washed blood cells with mosquito cell cultures, suggesting that this combination was more efficient than other combinations such as plasma/blood cells and hamster cell lines. This marked the first time that the isolation of EHDV serotypes 5 and 6 and three potential mosquito-borne orbiviruses was recorded in Japan, showing a greater variety of orbiviruses on the islands than previously known. Genetic analysis of the isolated orbiviruses suggested that the Yaeyama Islands and its neighboring regions were epidemiologically related. Some of the viruses, especially the potential mosquito-borne orbiviruses, were isolated during several consecutive years, indicating their establishment on the islands.

Prevalence of granulomatous pleuropneumonia associated with Actinobacillus pleuropneumoniae serotype 2 in slaughter pigs.

A total of 14,818 slaughtered pigs were examined macroscopically. Of these, 25 pigs with porcine pleuropneumonia were collected and the relations among Actinobacillus spp. and granulomatous lesions in organs (lungs and tonsils) were evaluated. In the lungs, only Actinobacillus pleuropneumoniae serotype 2 was isolated from 20 of the pigs. Histologically, granulomatous pneumonia with A. pleuropneumoniae antigen was detected in 8 of the pigs. The antigen was visible in the centers of the lesions along with asteroid bodies, epithelioid cells and multinucleated giant cells. In the tonsils, granulomatous lesions were not detected, although A. pleuropneumoniae serotype 2 (5 pigs), serotype 7 (1 pig), Actinobacillus porcitonsillarum (1 pig) and Actinobacillus minor (1 pig) were isolated. The present survey suggests that multifocal granulomatous pneumonia in slaughter pigs could be highly associated with A. pleuropneumoniae serotype 2 infection.

Physiological and pathological functions of LRRK2: implications from substrate proteins.

Leucine-rich repeat kinase 2 (LRRK2) encodes a 2527-amino acid (aa) protein composed of multiple functional domains, including a Ras of complex proteins (ROC)-type GTP-binding domain, a carboxyl terminal of ROC (COR) domain, a serine/threonine protein kinase domain, and several repeat domains. LRRK2 is genetically involved in the pathogenesis of both sporadic and familial Parkinson's disease (FPD). Parkinson's disease (PD) is the second most common neurodegenerative disorder, manifesting progressive motor dysfunction. PD is pathologically characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta, and the presence of intracellular inclusion bodies called Lewy bodies (LB) in the remaining neurons. As the most frequent PD-causing mutation in LRRK2, G2019S, increases the kinase activity of LRRK2, an abnormal increase in LRRK2 kinase activity is believed to contribute to PD pathology; however, the precise biological functions of LRRK2 involved in PD pathogenesis remain unknown. Although biochemical studies have discovered several substrate proteins of LRRK2 including Rab GTPases and tau, little is known about whether excess phosphorylation of these substrates is the cause of the neurodegeneration in PD. In this review, we summarize latest findings regarding the physiological and pathological functions of LRRK2, and discuss the possible molecular mechanisms of neurodegeneration caused by LRRK2 and its substrates.

Occurrence of cerebrocortical necrosis in a goat in Okinawa prefecture, Japan.

A 6-month-old male Nubian goat suddenly showed dullness, tachypnea, recumbency and opisthotonus in August 2011 in Okinawa, Japan. The goat was consequently necropsied: gross lesions in the brain included slightly swollen foci of yellowish discoloration on the cerebral hemisphere. Histopathologically, necrosis of the cortex with ischemic changes of neuronal cells was present, and swelling of the vascular endothelium, thickening of the basement membrane and diffuse infiltration of macrophages were observed in the cerebral lesion. Autofluorescence of the cerebral cortex was confirmed by an ultraviolet light test. The thiamine levels of the blood serum and tissue samples (brain, liver and heart) of the goat were low compared with goats from the same herd. The goat was diagnosed with cerebrocortical necrosis (CCN). This is the first case report of caprine CCN in Japan.

Chronic hepatitis C associated with monoclonal gammopathy of undetermined significance.

BACKGROUND: Two patients were admitted to Mie Prefectural General Medical Center and diagnosed as chronic hepatitis C complicated with monoclonal gammopathy of undetermined significance (MGUS). METHODS: The MGUS class were immunoglobulin (Ig)G. The hepatitis C virus (HCV) RNA genotype was Ib. Based on these findings, they were diagnosed as chronic hepatitis C complicated with MGUS. RESULTS: Histological studies showed chronic hepatitis in the liver and a mild rise in plasma cells without dysplasia and abnormalities in the bone marrow. Serum examination for cryoglobulin was negative. CONCLUSION: Chronic HCV infection might play a pathogenic role in the multistage process leading to lymphoproliferative disorders.