Karaca ve Yörük YYU Vet Fak Derg. 2004, 15 (1-2):115-121 A Morphological and Histometrical Study on Distribution and Heterogeneity of Mast Cells of Chicken’s and Quail’s Digestive Tract * Turan KARACA Mecit YÖRÜK Department of Histology-Embryology, Faculty of Veterinary Medicine, University of Yüzüncü Yıl, Van-Turkey SUMMARY This study was carried out to determine distribution and heterogeneity of mast cells in the digestive tract of the both chicken and quail. Ten Leghorn chickens and ten Japan quails were use during the investigation. Having been sacrified by decapitation, the chickens and quails’ tongue, crop, esophagus, proventriculus, gizzard, duodenum, jejunum, ileum, caecum and colon tissue samples were taken. Those samples were processed and blocked following the fixation with the BLA (basic lead acetate - Mota), Carnoy’s and IFAA (isotonic formaldehyde acetic acid) solutions. The histologic sections were stained with toluidine blue and alcian blue – safranine in order to determine the mast cells distribution and heterogenity in the digestive tract. Both Mast cells distribution and heterogeneity were found to be different between the digestive tract organs. In both animal species, the mast cells were found to be at very high number in the tongue, crop, esophagus and proventriculus whereas they were very few in the gizzard.In additition, it was found that BLA and Carnoy’s solutions were better relative to IFAA to determine the distribution of mast cells. Positive reactions were observed against alcian blue in the staining of all mucosal mast cells in both animal species with alcian blue - safranine mix. However, safranine was reactive only to esophagus, proventriculus, gizzard, colon, and tongue mast cells. In addition, alcian blue (+) granules beside the safranine (+) granules could be observed rarely, in those mentioned organs. Key Words: Quail, chicken, light microscope, mast cells, digestive tract Tavuk ve Bıldırcın Sindirim Kanalında, Mast Hücrelerinin Da ğılım ve Heterojenitesi Üzerine Morfolojik ve Histometrik Bir Çalı şma ÖZET Bu çalı şma, tavuk ve bıldırcın sindirim kanalındaki mast hücrelerinin da ğılım ve heterojenitesini belirlemek amacıyla yapıldı. Çalı şmada, 10’ar adet Leghorn ırkı tavuk ve Japon ırkı bıldırcın kullanıldı. Tavuk ve bıldırcınlar dekapite edilerek dil, kursak, özofagus, ön mide, mide, duodenum, jejunum, ileum, sekum ve kolondan doku örnekleri alındı. Alınan örnekler BLA (basic lead asetate – Mota), Carnoy ve IFAA (izotonic formaldehyde asetic acid) tespit solüsyonlarında tespit edilerek takip ve blokajı yapıldı. Hazırlanan histolojik kesitlere mast hücrelerinin da ğılım ve heterojenitesini belirlemek amacıyla toluidine blue ve alcian blue - safranin boyamaları uygulandı. Mast hücrelerinin da ğılımı ve heterojenitesinin sindirim kanalı organlarında farklılıklar gösterdi ği ve bu organlar içerisinde her iki hayvan türünde de dil, kursak, özofagus ve ön midenin sayısal olarak en fazla, müsküler midenin ise en az olan organ oldu ğu belirlendi. Kullanılan tespit sıvılarından BLA ve Carnoy’un IFAA’ya göre mast hücrelerinin metakromatik özelliklerinin belirlenmesi ve sayısal da ğılımın ortaya konmasında daha iyi sonuç verdi ği kanısına varıldı. Alcian blue-safranin boyamasında, her iki hayvan türünde de, mukozal mast hücreleri alcian blue (+) reaksiyon verirken, safranin ile özofagus, ön mide, mide, kolon ve dilde pozitif reaksiyonlar gözlendi. Ayrıca bu organlarda seyrek olarak hem safranin (+) hem de alcian blue (+) hücreler de saptandı. Anahtar Sözcükler: Bıldırcın, ı şık mikroskobu, mast hücresi, sindirim kanalı, tavuk INTRODUCTION the form of oval (13, 16), thin, long or spindle shaped (7, 26). The oval ones are generally observed in loose tissues, Since the digestive tract is in direct relationship whereas those in the form of spindle shaped are found in with outer environment, it encounters with many foreign compact tissues where collagen fibers are intensive (26). material throughout the life. Many of those foreign Mast cells can be divided into two groups on the materials have antigen properties and they are eliminated basis of their origins, locations, responses to applied in tunica mucosa by special and non-special defense fixative solutions, types of gylcoaminoglycans they have, mechanisms. Mast cells together with humoral and type of intragranuler serine proteinases (1), histochemical cellular defense cells have important role within that differences, functional criteria and morphological shapes. system. Those groups are Connective Tissue Mast Cell (CTMC) They are found as small groups within connective and Mucosal Mast Cell (MMC) (12). Nomenclature of tissues especially adjacent to the blood vessels (7,16). Enerabck (8) in rat intestines is used commonly for Mast cells having different locations have different description of heterogeneity between mast cells histochemical, cytochemical, ultra structural and populations. Not only staining properties and type of functional properties (18). Light microscopical fixations but also their locations and functional appearances of those cells may change with respect to differences within various parts of the same organ have their types and locations. They are generally observed in important role in heterogeneity of mast cells (10,14). Staining properties of mast cells change according to type * This study is the summary of a Ph.D thesis with same of fixative solutions (3,6,9), pH value and concentration title 115 YYU Vet Fak Derg. 2004, 15 (1-2):115-121 Karaca ve Yörük of dye and duration of staining are effective on staining area covered by 100 square ocular micrometer was properties (9). determined. The area of 100 square ocular micrometer By this study, it was aimed to make a was calculated by means of micrometrical lam by 40 morphological and histometrical research on distribution objective enlargement. Then the mast cell density in each of mast cells in chicken and quail’s digestive tract organs site was found and recorded as mast cell numbers/mm2 and in various parts of those organs on the basis of (17). Variance analyses of mast cell numbers for both different fixation solutions and different staining methods. types were conducted by using SAS v.12.0 package Findings obtained from tissue preparations of chicken for programs. Inter-groups and intra-group differences were each digestive track organ were compared. The same determined by Duncan test (21). procedure was applied to quail as well. RESULTS MATERIALS and METHODS Mast cells in cross sections of 6 µm thickness In the study, 10 adult white Leghorn chicken and taken from tissue samples belonging to various regions 10 Japanese quails (Coturnix coturnix Japonica) obtained and fixed with three different fixation solutions were from commercial farms were used. After they were distinguished easily by metachromatic staining method. It decapitated, their abdominal region was opened and their was determined for both animals that, mast cells were digestive tracts were examined. Tissue samples from dense especially near blood vessels in lamina propria tongue, crop, esophagus, proventriculus, gizzard, (Figure 1), submukosa and tunica serosa. They were duodenum, jejunum, ileum, caecum and colon were taken located around glands and both inside and around nerve in required size. Those samples were examined according fibers in lamina propria and submucosa. They were also to histological methods mentioned below: observed around blood vessels between muscle groups in tunica muscularis. For the Examination of Light Microscopy Mast cell in sections stained with toluidine blue Three different fixative solutions, IFAA (isotonic had various size and appearance. They were flat, oval or formaldehyde), Carnoy and BLA (basic lead acetate), in the form of spindle shaped (Figure 2). Cytoplasms of were used in the present study. Some tissue samples were mast cells taken from almost all organs and stained with fixed in IFAA fixative solution (formaldehyde 1.5 ml, metachromatic dye were homogenous. Hence, their distilled water 98 ml, glacial acetic acid 0.5 ml, pH 2.9) granular structure was hardly observed by light for 24 hours (22), some of others were fixed in Carnoy microscope. Heterochromatic appearance in nucleus of solution (absolute ethanol 60 ml, chloroform 30 ml, those cells was obvious (Figure 2). It was found that mast glacial acetic acid 10 ml) for 12 hours. Fixed samples 0 cells were intensive near blood vessels and between were kept in alcohol solution of 70 for 12 hours. The glands in microscopic papilla and were intensive in remaining tissue samples were fixed in BLA (Mota) compact tissues under upper surface epithelium of tongue. (basic lead acetate 1g, ethanol 50 ml, distilled water 50 Mast cells in esophagus were observed around gland ml, glacial acetic acid 0.5 ml) for 24 hours. Then, samples esophagus in a close relationship with nerve fibers and fixed in three different solutions were blocked in paraplast generally in the position of subepithelial. Mast cells found by means of routine histological techniques (8). For mast between muscle groups in tunica muscularis layers of cells count and idendification; serial cross sections taken digestive tract organs were generally in spindle shaped. µ from prepared blocks and having thickness of 6 m were Large cells showing good metachromasia were observed stained with 0.5 % toluidine blue solution, which was around blood vessels in tunica seroza layer of the tract prepared in Mac Ilvaine’s citric acid disodiumphosphate wall and its subserosa