Inducible nitric oxide synthase expression in cerebellum of dogs naturally infected with canine distemper virus

Abstract

The aim of this study is to investigate of inducible nitric oxide synthase (iNOS) and nitrotyrosine (NT) expression in the cerebellum of dogs naturally infected with canine distemper virus (CDV), a disease characterized by demyelination in the white matter of the cerebellum. Cerebellum samples collected from naturally infected twenty-three which were 14 acute and 9 chronically infected. with CDV and six non-infected dogs. The presence of CDV infection was confirmed using histopathology and immunohistochemical localization of CDV antigen in glial cells of the white matter of the cerebellum. The CDV infected cerebella were also categorized Luxol Fast Blue staining according to the severity of demyelination as mild (n=7), moderate (n=9), and severe (n=7). The streptavidin-biotin-peroxidase technique was used to detect immunolocalisation of the CDV antigen, iNOS, NT and glial fibrillary acidic protein (GFAP). CDV antigens, immunopositive astrocytes, oligodendrocytes and granular neurons were seen in the white matter of the infected dogs. In the control dogs, iNOS and NT immunopositivity were not seen in the white matter of the cerebellum. In CDV infected dogs, iNOS immunopositivity was observed especially in astrocytes in white matter of cerebellum. The number of iNOS immunopositive astrocytes were significantly higher in chronic cases compared to acute cases of CDV infected dogs (p < 0.05). iNOS immunopositive gemistocytic astrocytes were also immunopositive for NT. iNOS and NT immunopositivity was not detected outside the region of the demyelinated areas. The iNOS immunopositive total cell number was found to be significantly higher in severely affected demyelination areas (p < 0.05). These data indicated that there was a relationship between degrees of the CDV associated with demyelination and the level of iNOS immunopositivity in the glial cells

Keywords

• Canine Distemper Virus
• Cerebellum
• Demyelination
• iNOS

Köpek gençlik hastalığı virusu ile doğal olarak enfekte köpeklerin serebellumunda indüklenebilir nitrik oksit sentaz ekspresyonu

Abstract

Bu çalışmanın amacı, köpek gençlik hastalığı virusu (KGHV) ile doğal olarak enfekte köpek serebellumlarının beyaz maddesinde demyelinasyonla karakterize alanlarda indüklenebilir nitrik oksit sentaz (iNOS) ve nitrotirozin (NT) ekspresyonunun immunohistokimyasal olarak incelenmesidir. Çalışmada, KGHV ile 14 adedi akut ve 9 adedi kronik olmak üzere toplam 23 adet doğal enfekte ve 6 adet enfekte olmayan kontrol köpek serebellumu kullanıldı. KGHV enfeksiyonu histopatolojik ve immunohistokimyasal olarak serebellumun beyaz maddesindeki glial hücrelerde viral antijen varlığına dayanılarak doğrulandı. KGHV ile enfekte serebellumlar Luxol Fast Blue boyama ile demyelinasyonun şiddetine göre hafif (n=7), orta (n=9) ve şiddetli (n=7) olarak sınıflandırıldı. KGHV, iNOS, NT ve glial fibriler asidik proteinin (GFAP) immunpozitifliği ve immunolokalizasyonunu belirlemek için streptavidin-biotin-peroksidaz tekniği kullanıldı. KGHV immunopozitifliği enfekte köpek serebellumlarının beyaz maddesindeki astrositlerde, oligodendrositlerde ve granüler nöronlarda görüldü. Kontrol serebellumlarda iNOS ve NT immun pozitifliğinin bulunmadığı, enfekte köpek serebellumlarında ise glial hücrelerde özellikle astrositlerde immunopozitiflik olduğu belirlendi. iNOS immunopozitif astrosit sayısının kronik olgularda akut olgulara göre önemli derecede yüksek olduğu saptandı (p<0.05). Hem iNOS ve hem de NT immunopozitifliği gemistositik astrositlerde görüldü. Demyelinasyon alanlarının dışındaki bölgelerde iNOS ve NT immunopozitifliğine rastlanılmadı. Demyelinasyon alanlarında iNOS immunopozitif hücre sayısı önemli derecede yüksekti (p<0.05). Bu bulgular, KGH’daki demyelinasyonun derecesi ile iNOS immunopozitifliğinin düzeyi arasında bir ilişkinin olduğunu göstermektedir

Keywords

• Demyelinasyon
• iNOS
• köpek gençlik hastalığı virusu
• serebellum

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