Albendazole Sensitive vs. Resistant Nematodes – The Mitochondrial Ultra-Structural Changes

Öz

Studies on microtubule inhibitors have shown that the loss of equilibrium between tubulin and microtubules can generate a multitude of histochemical changes in mitochondria. This disruption of balance is also considered the basis of benzimidazole anthelmintic (BZ) activity. Studies have shown that BZ does not bond to the tubulin of the BZ-resistant Haemonchus contortus, as opposed to sensitive ones. This affinity alteration can be easily recognised by changes in the optical density and can help in the classification of H. contortus mitochondria, into sensitive (dark) and resistant (clear, unmodified zones). In order to confirm this hypothesis, we started our study from albendazole (ABZ) resistant and sensible H. contortus individuals, collected from the intestinal tract of sheep, aiming towards the identification of mitochondrial features, using the Electron Microscopy Transmission (EMT) technique. The EMT has confirmed that the structure of sensitive trichostrongilian populations was affected rapidly, only four hours after ABZ treatments. The main changes that appeared in the intestinal mitochondria of sensitive helminths were: cristae thickening and decreasing in number and cellular membrane thickening. Twelve hours after anthelmintic administration, a total blocking of metabolic functionality was observed, and finally, these changes completely altered the optical density of the mitochondria. In ABZ resistant populations, the optical density has remained normal; and the cristae number, size or functionality of resistant nematode mitochondria has remained unchanged.

Anahtar Kelimeler

• Albendazole, mitochondria, ultra-structural, examination, H. contortus

Albendazol Duyarlıya Karşı Albendazol Dirençli Nematodlar - Mitokondrial Ultra-Strüktürel Değişiklikler

Öz

Mikrotübül inhibitörleri üzerine yapılan çalışmalar, tübülin ve mikrotübüller arasındaki denge kaybının mitokondriyada birçok histokimyasal değişikliğe neden olabileceğini göstermiştir. Bu denge bozulmasının benzimidazol anthelmentik (BZ) aktivitesinin de temeli olduğu düşünülmektedir. Çalışmalarda BZ’nin duyarlı olanların aksine BZ-dirençli Haemonchus contortus tübülinine bağlanmadığı gösterilmiştir. Bu eğilim değişikliği optik dansitedeki değişmeler ile kolaylıkla belirlenebilir ve H.contortus mitokondriyasının duyarlı (koyu) ve dirençli (berrak, değişmemiş) zonlar şeklinde sınıflandırılmasını kolaylaştırabilir. Bu hipotezi teyit edebilmek amacıyla, biz çalışmamıza koyun intestinal sisteminden toplanan albendazol (ABZ) dirençli ve duyarlı H.contortus bireyleri ile başladık ve Elektron Mikroskopisi Transmisyonu (EMT) tekniği kullanarak mitokondriyal özellikleri tanımlamayı hedefledik. EMT, ABZ uygulamalarından sadece dört saat gibi bir süre sonra duyarlı trikostrongilianların yapısının hızla etkilenmiş olduğunu doğruladı. Duyarlı helmintlerin intestinal mitokondriyalarında görülen başlıca değişiklikler; “krista kalınlaşması ve sayıda ve hücresel membran kalınlaşmasında azalma” olarak belirlendi. Antihelmentik uygulamadan on iki saat sonra metabolik işlevsellikte tam bir blokaj gözlemlendi ve son olarak da, bu değişiklikler mitokondriyanın optik dansitesini tamamen değiştirmiştir. ABZ dirençli popülasyonlarda, optik dansite normal kalmış; krista sayısı, dirençli nematod mitokondriya işlevselliği ya da boyutu değişmemiştir.

Anahtar Kelimeler

• Albendazol, mitokondria, ultra-strüktürel inceleme, H. contortus

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