Okajimas Folia Anat. Jpn., 75(2-3): 141-154, August, 1998 Fine Structure of the Parathyroid and Ultimobranchial Glands of the Snake, Elaphe quadrivirgata By Kishiko YOSHIDA-TERASAWA, Daisuke HAYAKAWA, Huayue CHEN Shoichi EMURA, Akira TAMADA, Toshihiko OKUMURA and Shizuko SHOUMURA Department of Anatomy,Laboratory of Electron Microscope, Gifu University School of Medicine, 40 Tsukasa- machi, Gifu 500-8705, Japan College of Medical Sciences, Gifu University, 70-1, Kitano-machi, Gifu 500-8807, Japan -Received for Publication, June 30, 1998- Key Words: Parathyroid gland, Ultimobranchial gland, Snake, Electron microscope Summary: The ultrastructure of the parathyroid gland and ultimobranchial gland of the snake, Elaphe quadrivirgata, was studied. The parenchyma of the parathyroid gland was consisted of chief cells arranged in cords. Oxyphil cells and water- clear cells were not recognized. The chief cells contained irregular shaped nudeus, moderately-developed mitochon- dria, granular endoplasmic reticulum and Golgi complexes. A few secretory granules of 100-300 nm in diameter dis- tributed in the cytoplasm. Small aggregations of glycogen particles and lipid droplets were dispersed widely in the cytoplasm. The ultimobranchial glands consisted of follides and interfollicular aggregates of C-cells. Follicles were invaginated and composed of C-cells, goblet cells and ciliated cells. C-cells were located in the basal position of the follicle and possessed variety of secretory granules of 100-300 nm in diameter in the basal region of the cell. C-cells presented various secretory cycles. Goblet cells projected microvilli into follicular lumen. At the apical region the goblet cells had large granules of 300-1,200 nm in diameter. Ciliated cells projected cilia intermixed with micovilli from the apical sur- face of the cell into the follicular lumen. Parathyroid hormone (PTH), ultimobranchial 1976). As there have been a few detailed papers gland hormone (calcitonin) and vitamin D are on the ultrastructure of Elaphe quadrivirgata para- known as major calcium regulating hormone. PTH thyroid glands and ultimobranchial glands, we ob- elevates serum calcium level, whereas calcitonin served these two glands of Elaphe quadrivirgata lowers antagonistically. It is mentioned when the with electron microscopy. ultimobranchial body became active, the para- The snake, Elaphe quadrivirgata (class Reptilia, thyroid glands exhibited reduced activity (Srivastav order Squamata, suborder Ophidia) is the Japanese and Rani, 1992; Srivastav et aL, 1994). proper species that lives in Hokkaido, Honshu, The parathyroid glands are recognized in all Shikoku, Kyushu, Kunashiri Islands, Izu Islands vertebrates higher than fishes. In mammals C-cells and Osumi Islands (Matsui, 1987). are called parafollicular cells in the thyroid. In birds, reptiles, amphibian and fish C-cells form the independent organ, ultimobranchial gland. Materials and Methods The ultrastructure of the reptile parathyroid gland has been studied in turtles (Clark and Khai- Twenty snakes of both sexes weighing 95-440 g rallah, 1972; Chin, 1974; Grandi, 1982), iguanas were used. The snakes were collected from spring (Anderson and Capen, 1976), Japanese lizards to autumn. The caudal parathyroid glands and (Isono et al., 1979) and geckos (Isono et al., 1986). ultimobranchial glands were excised under ether The ultrastructure of the reptile ultimoranchial anesthesia. Both glands were fixed with Millonig's glands has been studied in turtles (Khairallah and fixative and dehydrated through ascending concen- Clark, 1971) and iguanas (Anderson and Capen, trations of acetone. The specimens were embedded 141 142 K. Yoshida-Terasawa et al. in Epon 812. Thin sections were cut on a PORTER- follicles and interfollicular cell aggregates (Fig. 7). BLUM MT-1 ultramicrotome, stained with uranyl Follicular lumen was filled with the moderately acetate and lead salts, and examined with a Hitachi electron dense substance (Fig. 8). There were three H-700H or Hitachi H-800 electron microscope. For types of cells in the follicle walls (Fig. 8). light microscopy 1-2 gm sections were cut and stained with 0.1% Azur II. (1) C-cells The C-cells were found not only in the follicular walls but also in interfollicular connective tissues Observations forming the cell aggregates (Figs. 8, 9, 10). In the follicle the C-cells occupied the basal position. The Parathyroid gland nucleus was irregular in shape with deep indenta- Elaphe quadrivirgata possessed two pairs of tions. Mitochondria and granular endoplasmic reti- parathyroid glands, the rostra' and caudal glands. culum distributed throughout the cytoplasm. Well- The parathyroid glands existed along the carotid developed Golgi complex was seen near the nucleus artery. The caudal parathyroid glands were situated (Fig. 11). Many secretory granules of 100-300 nm dorsal surface of the thymus (Fig. 1). The para- in diameter of various electron densities were rec- thyroid glands were fusiform measuring 0.3-0.4 mm ognized near the basal region of the cell. Number in short diameter and 0.4-1.2 mm in long diameter. and density of the secretory granules varied from Connective tissue capsule enclosed the parathyroid cell to cell. Secretory granules attached to the lat- gland and divided the gland into the cell cords (Fig. eral and basal cell membranes were often observed 1). (Fig. 10). Lysosomes 300-500 nm in diameter con- The parenchyma consisted of chief cells alone. tained heterogeneously electron dense materials Oxyphil cells and water-clear cells were absent. The (Fig. 9). chief cells were round, spindle or polygonal in shape and arranged in cell cords (Figs. 2, 3). (2) Goblet cells The nucleus was irregular in shape (Fig. 3). The apical surface of the goblet cell was covered Plasma membranes of adjacent chief cells pursued with numerous microvilli (Fig. 12). Mitochondria a straight course except for occasional interdigita- and granular endoplasmic reticulum were distrib- tions (Figs. 4, 5). Opposing plasma membranes uted throughout the cytoplasm. The goblet cell had were connected mainly by intermediate junctions a lot of large granules 300-1,200 nm in diameter at (Fig. 5). the apical region of the cell. The large granules Mitochondria and granular endoplasmic reticu- contained moderate electron dense, flocculent lum were scattered throughout the cytoplasm (Fig. materials and dense cores. Some of the large gran- 5). Golgi complex was moderatelydeveloped (Fig. ules attached to the apical surface of the follicular 6). lumen (Fig. 12). The follicular lumen was filled with The chief cells had a few secretory granules moderately dense materials (Figs. 8, 12). containing homogeneously dense materials 100-- 300 nm in diameter. Most secretory granules oc- (3) Ciliated cells cured singly (Fig. 5). They appeared near the Abundant cilia were observed at the luminar plasma membrane or in the Golgi region. Occa- surface (Figs. 8, 13). Numerous mitochondria were sional secretory granule attached to the plasma observed just beneath the basal bodies of the cilia membrane (Fig. 6). Lysosomes were round or irre- (Fig. 13). Microvilli intermixed among cilia (Fig. gular in shape measuring 250-1,000 nm. Some lyso- 13). somes contained the lipid droplets, dense granules or vacuoles (Fig. 4). Small aggregations of glycogen particles dispersed in the cytoplasm (Figs. 4, 5). Discussion Lipid droplets measuring 140-1,000 nm occured in the cytoplasm (Fig. 4). Parathyroid gland As to the parathyroid function of reptile, experi- Ultimobranchial gland mental studies have been reported mainly through The ultimobranchial gland was located near the parathyroidectomy (Sidky, 1966; Oguro, 1970; caudal parathyroid gland and was oval or spindle in Singh and Kar, 1979). Parathyroidectomized fresh- shape measuring about 0.7 mm in length and 0.4 water turtles do not show any particular changes mm in width. The connective tissue capsule envel- (Clark, 1965), whereas parathyroidectomized lizards oped the ultimobranchial gland (Fig. 7). The ulti- die of hypocalcemia and tetany (Sidky, 1966;Clark mobranchial gland was composed of invaginated et al., 1969). Though the parathyroidectomized Parathyroid and Ultimobranchial Gland of Snake 143 Elaphe quadrivirgata and another species of snakes seem to be less active than those of the gecko and show the decrease in serum calcium, only few Japanese lizard. These observations are in agree- animals showed tetanic convulsions (Oguro, 1970; ment with the report on the function of the snake Singh and Kar, 1979). parathyroid gland (Oguro, 1970). The parenchyma of the parathyroid gland of Elaphe quadrivirgata consists of only one type chief Ultimobranchial gland cell same as Japanese lizard (Isono et al. , 1979) It has been reported that snakes have one pair and gecko (Isono et aL, 1986). Oxyphil cells are of the ultimobranchial glands and that the glands observed in the mammalian parathyroid gland such are located midway between the rostral and caudal as, human, ape, horse, cow and bat (Shoumura et pairs of the parathyroid glands (Singh and Kar, aL, 1993). In reptile, Clark and Khairallah (1972) 1983; Padgaonkar et aL, 1992). In the present study has reported the existence of oxyphil cells in the Elaphe quadrivirgata ultimobranchial glands are fresh-water turtles. In the present study no oxyphil found near the caudal parathyroid glands. cells are observed in Elaphe quadrivirgata para- In amphibia the ultimobranchial gland mainly thyroid glands. Emura et aL (1990, 1992) have re- consists