Argyrophilic nucleolar organizer regions staining for cytology smears in dogs and cats

The argyrophilic nucleolar organizer regions (AgNORs) are cellular proliferation markers, crucial for predicting the clinical course and aggressiveness of tumors. The purpose of this study was to establish an easy and practical AgNOR staining method in the cytology of dogs and cats. Air-dried cytological slides were prepared from dogs (n=14) and cats (n=12). Acetone, formalin, ethanol and methanol were tested as fixatives for AgNOR staining. Subsequently, various methods of Romanowsky-based counterstains were tested before and after AgNOR staining. Clear and strong AgNOR spots were observed with all fixatives, and post-May–Grünwald staining was the best counterstaining method. The established method showed clear AgNOR spots even in the long-term storage samples and Romanowsky-stained ones.

[1]  M. Kiupel,et al.  Diagnosis and Prognosis of Canine Cutaneous Mast Cell Tumors. , 2019, The Veterinary clinics of North America. Small animal practice.

[2]  A. Srivastava,et al.  AgNOR Pleomorphic Count as a Tumor Marker in Cervical Carcinogenesis and Feasibility of Its Introduction in Cervical Cancer Screening Programs to Discriminate High-Risk Cases of Squamous Intraepithelial Lesions of the Cervix , 2019, Acta Cytologica.

[3]  N. Miyoshi,et al.  Rapid multiple immunofluorescent staining for the simultaneous detection of cytokeratin and vimentin in the cytology of canine tumors. , 2018, Veterinary clinical pathology.

[4]  A. M. Grégio,et al.  Identification of AgNORs and cytopathological changes in oral lichen planus lesions. , 2017, Acta histochemica.

[5]  R. Annigeri,et al.  Evaluation of diagnostic value of AgNOR and PAP in early detection of dysplastic changes in leukoplakia and lichen planus – a preliminary case–control study , 2017, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[6]  N. Shunyu,et al.  Brush Cytology and AgNOR in the Diagnosis of Oral Squamous Cell Carcinoma , 2016, Acta Cytologica.

[7]  O. Yamato,et al.  Development and application of multiple immunofluorescence staining for diagnostic cytology of canine and feline lymphoma. , 2015, Veterinary clinical pathology.

[8]  P. Caldeira,et al.  Oral leukoplakias with different degrees of dysplasia: comparative study of hMLH1, p53, and AgNOR. , 2011, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[9]  N. Malathi,et al.  Argyrophilic nucleolar organizer regions in inflammatory, premalignant, and malignant oral lesions: a quantitative and qualitative assessment. , 2008, Indian journal of dental research : official publication of Indian Society for Dental Research.

[10]  A. Moritz,et al.  Argyrophilic nucleolar organizing regions and Ki67 equally reflect proliferation in fine needle aspirates of normal, hyperplastic, inflamed, and neoplastic canine lymph nodes (n = 101). , 2007, Journal of veterinary internal medicine.

[11]  P. Jeannesson,et al.  Improvement in the staining and in the visualization of the argyrophilic proteins of the nucleolar organizer region at the optical level , 2005, The Histochemical Journal.

[12]  P. Vajdovich,et al.  Use of the Argyrophilic Nucleolar Region Method for Cytologic and Histologic Examination of the Lymph Nodes in Dogs , 2004, Veterinary pathology.

[13]  G. Ogilvie,et al.  Application of rapid CD3 immunophenotype analysis and argyrophilic nucleolar organizer region (AgNOR) frequency to fine needle aspirate specimens from dogs with lymphoma. , 1997, Veterinary clinical pathology.

[14]  G. Ogilvie,et al.  Frequency of argyrophilic nucleolar organizer regions in fine-needle aspirates and biopsy specimens from mast cell tumors in dogs. , 1996, Journal of the American Veterinary Medical Association.

[15]  K. Plate,et al.  Nucleolar organizer regions (NORs). Basic concepts and practical application in tumor pathology. , 1989, Pathology, research and practice.

[16]  J. Crocker,et al.  Nucleolar organizer regions in lymphomas , 1987, The Journal of pathology.