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A Scanning Electron Microscope Study of Myoepithelial Cells in the Intercalated Ducts of Rat Parotid and Exorbital Lacrimal Glands

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A Scanning Electron Microscope Study of Myoepithelial Cells in the Intercalated Ducts of Rat Parotid and Exorbital Lacrimal Glands Okajimas Folia Anat. Jpn., 67(5): 309-314, October 1990 A Scanning Electron Microscope Study of Myoepithelial Cells in the Intercalated Ducts of Rat Parotid and Exorbital Lacrimal Glands By Michitaka MURAKAMI, Toshikazu NAGATO and Hiroaki TANIOKA Department of Oral and Maxillofacial Surgery, School of Medicine, Ehime University, 454 Shitsukawa, Shigenobu, Onsen-gun, Ehime 791-02, Japan -Received for Publication, June 27, 1990- Key Words: Myoepithelial cell, Exocrine gland, Intercalated duct, Scanning electron microscopy, Rat Summary: Myoepithelial cells in the intercalated ducts of rat parotid and exorbital lacrimal glands were examined by scanning electron microscopy. The basal surface of the intercalated ducts revealed myoepithelial cells running parallel with its long axis. These myoepithelial cells were linked with one another, forming a well-developed network, and numerous wrinkles running transversely were observed on the surface of the myoepithelial cells. Also, some myoepithelial cells in the terminal portion linked with those in the intercalated duct. Based on these findings, it is suggested that myoepithelial cells in the intercalated duct may function as a protective wall against constriction of the narrow lumen of the intercalated duct when it is subjected to pressure by the surrounding tissues. Since Henle (1841) first reported the presence of in order to observe the non-free surface of cells and smooth muscle-like cells around the ducts of the human tissues. As reported previously, our study clarified the mammary gland, numerous morphological studies on three-dimensional ultrastructure of myoepithelial cells myoepithelial cells have been carried out by iron- in the terminal portion of each gland and demonstrated hematoxylin staining (Zimmermann, 1898), silver that a close correlation exists between the degree of staining (Richardson, 1949) and histochemical tech- development of the myoepithelial cells and the viscosity niques employing ALPase (Silver, 1954; Leeson, 1956; of the secretory products (Nagato et al., 1980). Garrett, 1973) and ATPase (Shear, 1964; Garrett and Although the presence of myoepithelial cells in ducts Harrison, 1970). Also, attempts have been made to is well known (Richardson, 1949; Tamarin, 1966; elucidate the overall structure of myoepithelial cells in Alexander et al., 1973), studies based on scanning a number of exocrine glands, including the salivary electron-microscopic observations have not been gland. The ultrastructure of myoepithelial cells has been reported except for one investigation on the ducts of shown to resemble closely that of smooth muscle cells the rat mammary gland (Yoshida et al., 1980). in the walls of blood vessels and the digestive tract (Ellis, In the present study, the basal surface of intercalated 1965; Tamarin, 1966). It is also well known that ducts of rat parotid and exorbital lacrimal glands was myoepithelial cells contain the same contractile protein visualized under the scanning electron microscope that is present in smooth muscle, as has been employing a slightly modified extracellular connective demonstrated by investigations using an actin- and tissue component removal technique. As a result, myosin-specific fluorescent antibody method (Archer several interesting findings concerning the three- and Kao, 1968; Drenckhahn et al., 1977). In an earlier dimensional architecture of the myoepithelial cells in study, we observed certain exocrine glands, i.e., three the intercalated duct were obtained. Also, an evalua- major salivary, mammary and exorbital lacrimal glands tion of the functions of the myoepithelial cells in the in the rat under the scanning electron microscope intercalated duct was made based on these findings. utilizing a procedure which selectively removed extra- cellular connective tissue components and basal laminae Address all correspondence to: Dr. Michitaka Murakami, Department of Oral and Maxillotacial Surger^, School of Medicine. Ehime j ni versity, 454 Shitsukawa, Shigenobu, Onsen-gun, Ehime 791-02, Japan 309 310 M. Murakami et al. Materials and Methods tight network on the basal surface of the duct (Fig. 2). On the other hand, wrinkles running at right angles to Parotid and exorbital lacrimal glands from Sprague- the long axis of the duct were observed on the surface Dawley rats of both sexes, aged 3 to 6 months, were of the body and the myoepithelial cells. The area used for the study. covered by the myoepithelial cells accounted for about Glandular tissues removed from the rats under 60°7o of the total basal surface area of the intercalated general anesthesia induced by intraperitoneal ad- duct. ministration of pentobarbital sodium (Nembutal; 50 mg/kg body weight) were immediately placed in Eagle's II. Exorbital lacrimal gland minimum essential medium (MEM), and then cut into The intercalated ducts of the exorbital lacrimal small blocks with each side measuring less than 1 mm. glands were longer than those of the parotid gland, According to the method of Nagato et al. (1980), these measuring 200-300 pm on average, but sometimes unfixed fresh specimens were transferred to Eagle's markedly long ones of about 500y m in length were seen MEM containing 300 U/ml of collagenase and 10 (Fig. 3). Since the specimens tended to detach from the TRU/ml of hyaluronidase, and allowed to undergo interlobular duct, it was difficult to observe the transi- digestion at 37°C for 10-20 min. The specimens were tional area from the intercalated duct to the interlobular subsequently fixed in 3 °7o phosphate-buffered duct. The intercalated duct was 15-20 pm in diameter glutaraldehyde at room temperature for 1 hr, and then near the interlobular duct, and thereafter gradually hydrolyzed with 8 N hydrochloric acid at 60°C for 30 became smaller while branching a few times, finally min. The bottle containing the specimens was shaken reaching about 7 pm at the transitional area to the vigorously to break the tissues into small pieces. After terminal portion. a second hydrolysis at 37°C for 10-20 min, the The basal surface of the intercalated duct revealed specimens were washed thoroughly in water, and treated myoepithelial cells running parallel with its long axis. for post-fixation in 13/4 osmic acid. According to the The myoepithelial cell body near the terminal portion usual method, the specimens were dehydrated through was observed to be about 4 pm in width, and from both a graded series of ethanol followed by substitution of ends of the cell body approximately two primary pro- isoamyl acetate, critical point dried, coated with jections of about 1-2 ium in width extended parallel to platinum, and observed under a Hitachi S-500A the long axis of the duct. Moreover, secondary projec- scanning electron microscope. Also, the entire basal tions of about 0.5-1 pm in width extended from the surface area of the intercalated ducts and the area primary projections. These secondary projections covered by myoepithelial cells were measured morpho- branched a few times, and then linked with those of metrically with a Leitz texture analysis system to obtain adjacent myoepithelial cells to form a well-developed a guide which would enable us to compare the degree network on the basal surface of the duct (Fig. 4). Also, of development of the myoepithelial cells in the inter- some myoepithelial cells in the terminal portion ex- calated ducts of each gland. tended their projections to the intercalated duct and linked with projections of the myoepithelial cells in the intercalated duct (Fig. 5). Near the terminal portion, Observations the area covered by the myoepithelial cells accounted for about 55% of the total basal surface area of the I. Parotid gland intercalated duct. On the other hand, near the inter- Most of the intercalated ducts were detached from lobular duct, the projections of the myoepithelial cells the striated ducts, since the specimens tended to break were slightly wider, while the numbers of primary and easily at the boundary between the intercalated and secondary projections were somewhat smaller than striated ducts during the extracellular connective tissue those observed near the terminal portion (Fig. 6). The component removal process. The intercalated ducts area covered by the myoepithelial cells accounted for measured 6-8 p m in diameter and approximately about 60°7o of the total basal surface area, a ratio not 20-25 pm in length (Fig. 1), but some were as long as very different from that near the terminal portion. Also, about 50 pm. The basal surface of the intercalated ducts wrinkles arranged at right angles to the long axis of the revealed myoepithelial cells running along the long axis. duct were observed on the surface of the myoepithelial The bodies of the myoepithelial cells were 4-5 pm in cell body and the projections (Figs. 4 and 6). width, and from both ends of the cell body a few primary processes of about 2 pm in width extended parallel to the long axis of the duct. Additionally, a few Discussion secondary processes of about 1 pm in width extended from the primary processes. These processes were linked Since Evan et al. (1976) inttodti( i.(1 ploccduic with those of adjacent myoepithelial cells, forming a which selectively removed extraecllulai Conliecti c tissue SEM of Myoepithelial Cells in Intercalated Ducts 311 components and basal laminae in the field of scanning epithelial cell network and the viscosity of the secretory electron microscopy for the observation of non-free products. On the other hand, the present study surfaces of tissues and cells, several researchers have demonstrated that the myoepithelial cells in the inter- clarified the three-dimensional ultrastructure of calated duct had a spindle-shaped cell body which was myoepithelial cells in the terminal portion of various arranged parallel to the long axis of the duct. From both exocrine glands, including the salivary gland (Nagato, ends of the cell body, a few processes extended which 1978; Abe, 1979; Brocco and Tamarin, 1979; Nagato ran almost parallel with the long axis of the duct, and et al., 1980; Nagai and Nagai, 1985).
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