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http:// www.jstage.jst.go.jp / browse / jpsa doi:10.2141/ jpsa.011038 Copyright Ⓒ 2012, Japan Poultry Science Association. Morphological and Histological Studies on the Adrenal Gland of the Chicken (Gallus domesticus) Kober A.K.M. Humayun1, 2, Masato Aoyama1 and Shoei Sugita1 1 Department of Animal Science, Faculty of Agriculture, Utsunomiya University, 350 Minemachi, Utsunomiya, Tochigi 321-8505, Japan 2 United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, 3-5-8 Saiwaichou, Fuchu 183-8509, Japan Morphological and histological studies of the adrenal gland of the adult White Leghorn chicken (Gallus domesticus) were performed. Macro- and microscopic observations of azan- or formal-dichromate stained (to observe the chromaffin reaction of adrenal medulla) sections were conducted. The right and left adrenal glands differed in shape, position, weight, length, width and thickness. The adrenal glands were encapsulated with a thin connective tissue containing blood vessels. The adrenal parenchyma was composed of two main tissue types, the cortex and medulla; these were mixed throughout the organ. The adrenal gland could be divided into a subcapsular layer, peripheral zone, and central zone. The whole subcapsular layer was occupied by medulla, and the proportion of medulla in the central zone (49.7%) was larger than that in the peripheral zone (24.8%). Approximately 60% of the adrenal gland was cortex, 39% was medulla, and the remaining 1% was sinusoids. The adrenal cortico-medullary ratio in the adult chicken was approximately 1.6:1. Cortical cells were arranged in columns with a small, round to slightly oval, eccentric nucleus approximately 4 μm in diameter. Medullary cells were polygonal in shape with a large, spherical, centrally placed nucleus approximately 5 μm in diameter. Key words: adrenal cortex, adrenal gland, adrenal medulla, chicken, cortico-medullary ratio J. Poult. Sci., 49: 39-45, 2012 Introduction Adrenal glands are important organs in all animals. They help to maintain homeostasis as well as play important roles in all types of stress response (Freeman, 1985; Randall et al., 2002). They function by producing a variety of hormones that are needed in many facets of everyday life. Hormone production within the avian adrenal gland occurs in a divided cortex that produces glucocorticoids (corticosterone) and mineralocorticoids (aldosterone) and an adrenal medulla that produces catecholamines (norepinephrine and epinephrine). Experimental removal of the adrenal gland in birds leads to eventual death (Peng et al., 2005). In birds, the right and left adrenal glands are small and yellowish (Carsia and Harvey, 2000). Müller (1929) reported that the right adrenal gland in adult chicken is roughly pyramidal in shape with a caudally pointing apex, while the left adrenal gland is less constant in shape. The weight, length, width, and thickness of adrenal Received: April 30, 2011, Accepted: August 29, 2011 Released Online Advance Publication: September 25, 2011 Correspondence: S. Sugita, Department of Animal Science, Faculty of Agriculture, Utsunomiya University, 350 Mine-machi, Utsunomiya, Tochigi 321-8505, Japan. (E-mail: [email protected]) glands vary depending upon a number of factors, such as species, breed, age, health, and environment (Wells and Wight, 1971). The avian adrenal gland is unique in that it consists of a complete intermingling of cortical (interregnal) and medullary (chromaffin) tissues with no definite cortex and medulla (Ghosh et al., 200l). The adrenal gland in the chicken consists of three regions: subcapsular layer, peripheral zone, and central zone (Hodges, 1974). Several studies on the avian adrenal gland have provided basic description of this gland, but none have described them in detail (e. g., chicken: Hodges, 1974; duck: Holmes and Cronshaw, 1984; pigeon: Bhattacharyya, 1975; quail: Basha et al., 2004; and African ostrich: Tang et al., 2009). Little is currently known about the “normal” structure of the chicken adrenal gland. Prompted by lack of information on the size differences between the right and left adrenal glands, the cortico-medullary ratio in the different adrenal regions as well as cortico-medullary ratio in the whole adrenal gland and variation in the diameters of cell nuclei of cortical and medullary tissues among the different regions and this will require further study. The cortico-medullary ratio will aid in understanding the functional significance of the chicken adrenal gland. Therefore, the present study was undertaken Journal of Poultry Science, 49 (1) 40 to demonstrate in detail morphological and histological studies on the adrenal gland of the chicken. Materials and Methods Collection of Chickens Twelve healthy adult White Leghorn chickens (6 males, 6 females) weighing 1.8 to 2.3 kg were used to conduct the present stud. The chickens were obtained from the Tochigi Prefectural Livestock Experiment Station in Tochigi, Japan. All chickens were cared for according to guidelines suggested in the care and use of laboratory animals at Utsunomiya University. All procedures involving animals were carried out in accordance with the Animal Protection Regulations of Japan. Tissue Processing and Morphometric Measurements of the Adrenal Gland The chickens were sacrifice by an overdose of pentobarbital sodium (30 mg/kg body weight). The chickens were perfused transcardially with a Ringer’s solution (0.9% NaCl, 0. 042% KC1, and 0. 025% CaCl2 in distilled water) and Zamboni’s solution (0.1M phosphate buffer, pH 7.4 contain ing 2. 0% paraformaldehyde and 0. 2% picric acid). After sacrifice, the adrenal glands were carefully removed, weighed on a balance, and the length, width, and thickness of each gland was measured using calipers. The length of each adrenal gland was measured along the longitudinal axis; the width was measured at the thickest portion of the gland, and the maximum thickness was measured dorsoventrally. Each adrenal gland was then immersed in 10% formalin in 0.1 M phosphate buffer (pH 7.4) for 1 week. Tissue blocks were subsequently prepared and processed conventionally for paraffin wax embedment. Serial sections were cut at thickness of 5 μm in the coronal plane using a microtome and mounted on glass slides. Randomly selected sections were stained with azan. Light Microscopy with Formal-dichromate Staining and Calculation of Adrenal Cortico-medullary Ratio Six adult chickens (3 males, 3 females) were used to assess the proportions between cortical and medullary tissue and to examine patterns of medullary tissue in the adrenal gland. The adrenal medulla demonstrates the chromaffin reaction (Hartman et al., 1947) and it has also been suggested (Hartman et al., 1947) that an identical histological picture can be produced by fixing the adrenal medulla in formal-dichromate. After collection, the adrenal glands were incubated with potassium dichromate for 48 hours. Thereafter, the glands were incubated for 24 hours in 10% formalin and for 24 hours in 30% sucrose. Finally, coronal serial sections were cut at 30 μm thickness with a freezing microtome and mounted on glass slides. The relative proportions of the different tissue components (cortical, medullary, and sinusoids) were measured using ImageJ software (National Institutes of Health, Bethesda, MD) in each selected section that included the whole adrenal gland. The values for the proportions of each area (cortical, medullary, and sinusoids) were averaged from two sections for each chicken and expressed as the mean±standard error (SE). The proportions between cortical and medullary tissue in the peripheral and central zones of adrenal gland were obtained by using six chickens (3 males, 3 females). Adrenal cortico-medullary ratio in the peripheral and central zones data was obtained from ten photomicrographs taken randomly from the peripheral and central zone of ten cross-sections of each bird using a photomicroscope. Cortex, medulla, and sinusoids were measured using ImageJ software and averaged. Values were expressed as the mean±SE. Measurement of the Diameter of the Cell Nuclei in Cortical and Medullary Tissues Six adult chickens (3 males, 3 females) were used to measure the diameters of cell nuclei of cortical and medullary tissues of the adrenal gland. The diameters of cell nuclei of cortical and medullary tissues were quantified using 100× photographs (Olympus BX51) and ImageJ software. The diameters were calculated from 50 randomly selected nuclei of cortical and medullary cells from three regions (subcapsular layer, peripheral zone, and central zone) of each chicken and subsequently averaged. Values were expressed as the mean±SE. Data Analysis Two-way repeated measures analysis of variance and the paired t-test were used to analyze the differences in weight, length, width, and thickness between right and left adrenals. Differences were considered significant if P<0.05. Results Topographic Location and Morphology of the Adrenal Gland The two adrenal glands of the chicken were small and brownish-yellow to creamy-yellow in color. The glands were situated on each side of the median line just anterior to the bifurcation of the caudal vena cava, close to the gonads (Fig. 1a). The right adrenal gland differed from the left one in the shape and the location. The right adrenal gland was often roughly pyramidal in shape with a caudally pointing apex, partly adherent to the anterior cranial region of the right kidney, and located close to the anterior aspect of the right testis. The left adrenal gland appeared elongated in outline and was situated on the medial border of the left kidney, close to the left testis (Fig. 1a, b). Morphometry of the Adrenal Gland Two-way repeated measures analysis of variance shows that there were no significant differences between males and females adrenal gland morphometric data (P>0.05), while significant variation was observed between sides (P<0.05). Hence, data were pooled together and averaged. In addition, data was also categorized by sexes in Table 1 for better understanding. The average body weight was 2071±35.2 g (n=12) in both sexes. Morphometric data on each adrenal gland are summarized in Table 1. The mean weights of the right and left adrenal glands were 97.2±1.2 mg and 104.1± 1.4 mg, respectively, indicating that the right adrenal gland was significantly (P<0. 01) lighter than the left adrenal gland. The left adrenal gland was significantly longer than the right (0. 9±0. 0 cm vs 0. 8±0. 0 cm, respectively; P< Humayun et al.: Chicken Adrenal Gland Anatomical position and general morphology of the adrenal gland of the normal chicken (non-perfused). (a) Glands are shown in situ. The two adrenal glands of the chicken are situated on each side of the median line just anterior to the bifurcation of the caudal vena cava, close to the gonads. Arrowhead and arrow indicate right and left adrenal gland, respectively; black-and-white and black star indicate right and left testis, respectively; black arrow indicates vena cava. Scale bar indicates 1 cm. (b) Shape of the adrenal gland. The right adrenal gland (R) was often roughly pyramidal in shape with a caudally pointing apex, and the left adrenal gland (L) appeared elongated. Scale bar indicates 0.5 cm. Fig. 1. 0.01). The right adrenal gland was significantly wider than the left one (0.6±0.0 cm vs 0.5±0.0 cm, respectively; P< 0.01). The right adrenal gland was significantly thicker than the left one (0.5±0.0 cm vs 0.4±0.0 cm, respectively; P< 0.01). Histology of the Adrenal Gland The adrenal gland of the chicken was encapsulated with thin connective tissue containing blood vessels (Fig. 2a). Adrenal glands were composed of two morphologically and physiologically different endocrine glands: the cortex and medulla (Fig. 2a). Cortical and medullary tissues were intimately mixed together throughout the gland (Fig. 2a). The adrenal gland was divided into a subcapsular layer (SZ), 41 peripheral zone (PZ), and central zone (CZ) (Fig. 2a). The SZ consisted of medullary cells immediately below the capsule. The PZ consisted mainly of cortical cells, but the two cell types were evenly intermingled in the CZ. The medullary tissue formed an almost complete meshwork of islets, including some larger irregular islets, some with slight anastomosis, and some elongated islets (Fig. 3a, b). Medullary cells were polygonal in shape and larger than cortical cells, with basophilic cytoplasm and spherical, centrally placed nuclei (Fig. 2b). The diameters of cell nuclei of medullary cells in the SZ, PZ, and CZ were 5.1±0.0, 5.0± 0.0, and 5.1±0.0 μm, respectively (Table 2). However, on average, the diameter of the cell nuclei of medullary cells was 5.1±0.0 μm (Table 2). Microscopic examination of the chicken adrenal gland showed that cortical tissue occupied a major part of the adrenal gland. The adrenal cortex consisted of solid, irregular, cylindrical cords of cells (Fig. 2a). In cross-section, the cords usually appeared round to oval in shape. The cortical cords in the peripheral zone were larger; these cords were not regularly arranged, but intimately intertwined with the medullary strands (Fig. 2a). The cells of the cortical tissue were arranged in columnar cells with a small, round to slightly oval and eccentrically placed nucleus (Fig. 2b). The diameters of cell nuclei of cortical tissue in the PZ and CZ were 4. 1±0. 0 and 4. 0±0. 0 μm, respectively (Table 2). However, on average, the diameter of cell nuclei of cortical tissue cells was 4.1±0.0 μm (Table 2). These cells stained eosinophilic. Capillaries as well as sinusoids were present throughout the adrenal (Fig. 2). Usually, sinusoids were more frequent inside the glandular tissue and often seemed to create larger sinus venosi in the center of the gland (Fig. 2). The Adrenal Cortico-medullary Ratio Because there was no remarkable difference in the adrenal cortico-medullary percentages between males and females, data were pooled together. In addition, data was also categorized by sexes in Table 3 and Table 4 for better understanding. In the PZ, the overall mean percentages of cortical tissue, medullary tissue, and cortico-medullary ratio were 74.0±0.1%, 24.8±0.1%, and 3:1, respectively (Table 3), and the remaining 1.1±0.0 was sinusoids. In the CZ, the overall mean percentages of cortical tissue, medullary tissue, and cortico-medullary ratio were 48.7±0.0%, 49.7±0.0%, and 1:1, respectively (Table 3), and the remaining 1.5±0.0 was sinusoids. However, in the whole adrenal gland, the overall mean percentages of cortical tissue and medullary tissue were 59.7%±0.1%, 39.1%±0.2%, respectively (Table 4), and the remaining 1.1%±0.4% was sinusoids. The cortico-medullary ratio was approximately 1.6:1 in the whole adrenal gland. Discussion In this study, we examined the general morphology and histology of the chicken (Gallus domesticus) adrenal gland. We found that the location of the adrenal gland is similar to that described by Wells and Wight (1971), who reported that the two adrenal glands of chicken were situated on each side Journal of Poultry Science, 49 (1) 42 Mean values (±SE) of weight, length, width, and thickness of the chicken adrenal gland Table 1. Sex Parameters Side Overall mean (n=12) Male (n=6) Female (n=6) Weight (mg) Right Left 97 . 2±1 . 2* 104 . 1±1 . 4 98 . 3±1 . 9* 107 . 0±1 . 4 96 . 0±1 . 4* 101 . 3±0 . 4 Length (cm) Right Left 0 . 8±0 . 0* 0 . 9±0 . 0 0 . 8±0 . 0* 0 . 9±0 . 0 0 . 8±0 . 0* 0 . 9±0 . 0 Width (cm) Right Left 0 . 6±0 . 0* 0 . 5±0 . 0 0 . 6±0 . 0* 0 . 5±0 . 0 0 . 6±0 . 0* 0 . 5±0 . 0 Thickness (cm) Right Left 0 . 5±0 . 0* 0 . 4±0 . 0 0 . 5±0 . 0* 0 . 4±0 . 0 0 . 5±0 . 0* 0 . 4±0 . 0 Asterisk (*) indicates significant difference (Paired t-test, * P<0.05) from the organ of the left side. of the median line just anterior to the bifurcation of the caudal vena cava, close to the gonads and the anterior division of the kidneys. The shape and color of the adrenal glands in our study were similar to those in a previous study of the chicken (Hodges, 1974). Hodges (1974) suggested that the shape of the right adrenal gland in the adult chicken is roughly pyramidal in shape with a caudally pointing apex and that the left adrenal gland is less constant in shape. He stated that the color of adrenal glands in chicken varies between grayish-yellow and reddish-yellow. The present study indicated significant variations in weight, length, width, and thickness of the right and left adrenal glands of the chicken. In this study, the weight of the right adrenal gland was significantly lighter than that of the left one (P<0.01). Wells and Wight (1971) reported that the weight of the adrenal gland was 100 to 200 mg in the chicken and 0.45±0.04 g in the ostrich chick (Tang et al., 2009). The weight of adrenal glands varies considerably according to species, breed, age, health, and various environmental factors (Wells and Wight, 1971). In terms of length, our present result indicated that the left adrenal gland was significantly (P<0.01) longer than the right one. In relevant studies in other species, the length was recorded as 2.0 to 3.0 cm in the ostrich (Tang et al., 2009) and 13 mm in the chicken (Hodges, 1974). Regarding weight, the right adrenal gland was significantly (P<0.01) wider than the left in our study. Tang et al. (2009) found the ostrich chick adrenal gland to be 0.6 cm in width, while Müller (1929) found an average 8 mm width in the domestic hen. Finally, the right adrenal gland was, on average, significantly thicker (P< 0.01) than the left one. Similar studies reported that the adrenal gland thickness was 4 to 5 mm in the chicken (Hodges, 1974) and 0.3 cm in the ostrich chick (Tang et al., 2009). To the best of our knowledge, no publication exists on this topic in the literature; thus, no comparisons of the right and left adrenal glands could be made with other studies. Further studies are needed to clarify the reasons for the differences in size between the right and left adrenal glands. Structurally, chicken adrenal glands appear to be encapsulated with thin connective tissue containing blood vessels; cortical and medullary tissues are completely intermingled and consist of three regions: the subcapsular layer, peripheral zone, and central zone (Hodges, 1974). These results are consistent with those of our study. In birds, the medullary tissue forms a pattern that varies from scattered islets to an interwoven network, depending upon the amount of tissue present. The pattern of the medullary tissue in our study was similar to those of Hartman et al. (1947) and Holmes and Cronshaw (1980). They stated that medullary tissue forms an attractive meshwork pattern, seeming to take the form of islets in the Florida bobwhite. Moreover, the present results correspond with those of Hodges (1974) and Ohmori (1998); similarly, they suggested that adrenal medulla cells in the chicken are arranged in a polygonal shape, with a large, spherical, centrally placed nucleus. The basophilic staining of the cytoplasm is due to its content of numerous small, basophilic granules. In this study, we measured, for the first time, the diameters of cell nuclei of medullary cells in the three different zones of the chicken adrenal gland. There were a few variations in the diameters of cell nuclei within the SZ, PZ, and CZ of the adrenal gland of the chicken. As far as we know, no data exist on this topic in the literature; no comparisons could be made with other studies. Further study is needed to clarify the reasons for the differences in the diameters of cell nuclei of medullary cells in the three different zones of the chicken adrenal gland. There is one possible explanation that the nuclei of cells in the CZ are more highly developed than those in the SZ and PZ. However, on average, the diameters of cell nuclei of medullary cells in our study were inconsistent with those of Tang et al. (2009), who found that the medullary cell nuclei in ostrich chicks were approximately 8 μm in diameter. Moreover, Hodges (1974) found the mean diameter of the nuclei in medullary cells in domestic hens to be 6.7 μm. The present results correspond with those of Aire (1980) and Hodges (1974), who suggested that adrenal cortical cells in the chicken are arranged in columns with a small, round to Humayun et al.: Chicken Adrenal Gland Histology of the adrenal gland of the normal chicken. Sections were stained with azan. (a) Prominent eosinophilic cortical (light staining) and basophilic medullary tissues (darker staining) are arranged in islets. The cortical (arrow) and medullary (arrowhead) tissues are intermingled. The adrenal gland could be divided into a peripheral zone (PZ), consisting mainly of cortical cells but with a subcapsular layer of medullary cells (SZ), and a central zone (CZ) in which the two cell types were evenly intermingled. C, adrenal capsule; VS, venous sinus. Scale bar indicates 100 μm. (b) Characteristics of two different cell types in the adrenal parenchyma. (N) Strand of cortical cells in which the cells are columnar in type with a small, round to slightly oval, eccentric nucleus. (M) Group of medullary cells are polygonal with a large, spherical, centrally placed nucleus. Scale bar indicates 10 μm. Fig. 2. 43 Fig. 3. Cross-sections of adrenal gland of the normal chicken. The adrenal gland was treated with formal-dichromate (to observe the chromaffin reaction of adrenal medulla). (a and b) Distribution patterns of medullary tissue (arrow) and cortical (arrowhead) tissue. The medullary tissue formed an almost complete meshwork of islets, including some larger irregular islets, some with slight anastomosis, and some elongated islets. Scale bars indicate 1 mm (a) and 60 μm (b). slightly oval, eccentrically placed nucleus with eosinophilic staining. In this study, we measured, for the first time, the diameters of cell nuclei of cortical cells in the different zones of the chicken adrenal gland. There were variations in the diameters of cell nuclei within the PZ and CZ. The PZ cells were characterized for their larger diameters of cell nuclei compared to that of the cells nuclei of CZ. However, there might have certain structural differences between the cortical cells in PZ and CZ of the chicken adrenal gland. To the best Journal of Poultry Science, 49 (1) 44 Comparisons of diameters of cell nuclei of cortical and medullary tissues in the three different regions of the chicken adrenal gland (n=6) Table 2. Regions Component SZ (Mean±SE) PZ (Mean±SE) CZ (Mean±SE) Overall mean±SE Medulla (μm) Cortex (μm) 5 . 1±0 . 0 ─ 5 . 0±0 . 0 4 . 1±0 . 0 5 . 1±0 . 0 4 . 0±0 . 0 5 . 1±0 . 0 4 . 1±0 . 0 (−) indicate that SZ was no cortical tissue Table 3. Mean values (±SE) of cortex, medulla, and cortico-medullary (CM) ratios in the three different regions of the chicken adrenal gland Regions Overall cortex mean (%) (n=6) Overall medulla mean (%) (n=6) SZ PZ CZ 0 . 0±0 . 0 74 . 0±0 . 1 48 . 7±0 . 0 100±0 . 0 24 . 8±0 . 1 49 . 7±0 . 0 Sex Male (n=3) Cortex Medulla (%) (%) 0 . 0±0 . 0 73 . 7±0 . 1 48 . 7±0 . 0 100±0 . 0 25 . 0±0 . 0 49 . 7±0 . 0 Female (n=3) Cortex Medulla (%) (%) 0 . 0±0 . 0 74 . 2±0 . 1 48 . 7±0 . 0 100±0 . 0 24 . 5±0 . 0 49 . 7±0 . 0 Overall CM ratio 0:1 3:1 1:1 Mean values (±SE) of cortex, medulla, and cortico-medullary ratio in the chicken whole adrenal gland Table 4. Parameters Overall mean (n=6) Cortex (%) Medulla (%) 59 . 7±0 . 2 39 . 1±0 . 2 Sex Male (n=3) Female (n=3) 59 . 9±0 . 0 38 . 5±0 . 1 59 . 5±0 . 4 39 . 7±0 . 2 of our knowledge, no data exist on this topic in the literature; no comparisons could be made with other studies. The functional significance of these few variations in the diameters of cell nuclei in chicken adrenal glands is currently unknown. It is possible that adrenal cortical tissue synthesizes and secretes at least two types of cells. However, on average, the diameters of cell nuclei of cortical cells in our study were smaller than those found in ostrich chicks (5-7 μm) (Tang et al., 2009) and domestic fowl (5 μm) (Hodges, 1974). Hodges (1974) also reported that the few large venous sinuses that were found in the middle of the chicken adrenal gland correspond with our results. Notably, there is no true central vein as in man. Ultimately, the sinuses lead to an adrenal vein that joins the caudal vena cava (Wells and Wight, 1971). In this study, we report, for the first time, the relative proportions of cortical and medullary tissues in the different zones of the chicken adrenal gland. There was a variation in the relative proportions of cortical and medullary tissues in the PZ and CZ of the adrenal gland of the chicken. The cortico-medullary ratios in the PZ and CZ were approximately 3:1 and 1:1, respectively. The highest cortico-medul- Overall CM ratio 1.6 : 1 lary was found in the PZ of the adrenal gland. The functional significance of differences in cortico-medullary ratios of PZ and CZ is not completely understood. However, one possible reason is that the higher ratio (large cortex) in PZ may be required for more production of corticosteroid. Further study is needed to clarify the reason for such a difference of cortico-medullary ratio in the PZ and CZ of the adrenal gland of the chicken. The results of the current study correspond with those of Carsia et al. (1985), who described that adrenal cortical and medullary tissues comprise 60% and 40%, respectively, of the adrenal gland in the chicken. Similarly, Aire (1980) indicated that approximately 52% of tissue is adrenal cortical in the Rhode Island Red chicken. This report indicates an adrenal cortical percentage lower than that in our study, which may be attributable to breed differences. Undoubtedly, the most striking variations in the cortex and medulla percentage are those due to age, sex, breed, health, and environment (Siller et al., 1975). The contention by Sivaram (1965) that the proportion of medullary tissue is consistently higher than that of cortical tissue in the domestic fowl is not supported by the present findings or by those of Aire (1980) and Carsia et al. (1985), who stated that cortical Humayun et al.: Chicken Adrenal Gland tissue was normally greater in proportion than medullary tissue. The functional significance of this higher proportion of cortex is not completely understood, but one possible explanation could be the increased need to regulate water and electrolyte balance. It is known that the adrenal cortex produces hormones (glucocorticoids and mineralocorticoids) that aid in water and electrolyte balance; thus, a larger cortex may be needed for increased production of these hormones in the chicken, which lives in areas in which water conservation is extremely important. In this study, the cortico-medullary ratio was approximately 1.6:1 in the whole adrenal gland, which probably relates to the physiological status of the chickens. For example, as we know, these two cell types are responsible for hormone secretion (the cortex, which produces glucocorticoids [corticosterone] and mineralocorticoids [aldosterone], and the adrenal medulla, which produces catecholamines [norepinephrine and epinephrine]). When a bird is exposed to stress, the adrenal glands release corticosterone and catecholamines into the blood, which helps to adapt the bird to its new set of environmental conditions. Consequently, a distinct difference was observed in the relative proportions of cortical and medullary tissues. Acknowledgments This research was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (No. 083281). References Aire TA. Morphometric study of the avian adrenal gland. Journal of Anatomy, 131: 19-23. 1980. Basha SH, Vijayaragavan C and Ramesh G. Light and electron microscopic studies on the interrenal tissue of the adrenal gland in Japanese quail (Coturnix coturnix japonica). Indian Journal of Animal Sciences, 74: 1021-1023. 2004. Bhattacharyya TK. Fine structure of the interrenal cell in the quail and the pigeon. Anatomy and Embryology (Berl.), 146: 301310. 1975. Carsia RV, Scanes CG and Malamed S. 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Canadian Journal of Zoology, 43: 1021-1031. 1965. Tang L, Peng KM, Wang JX, Luo HQ, Cheng JY, Zhang GY, Sun YF, Liu HZ and Song H. The morphological study on the adrenal gland of African ostrich chicks. Tissue and Cell, 41: 231-238. 2009. Wells JW and Wight PAL. The adrenal glands. In: Physiology and biochemistry of the domestic fowl (Bell DJ and Freeman GM eds.). pp. 489-520. Academic Press, London. 1971.