Anat Cell Biol.  2013 Jun;46(2):101-112. 10.5115/acb.2013.46.2.101.

Heterogeneity of glandular cells in the human salivary glands: an immunohistochemical study using elderly adult and fetal specimens

Affiliations
  • 1Division of Otorhinolaryngology, Sendai Municipal Hospital, Sendai, Japan.
  • 2Medical Education Center, Aichi Medical University School of Medicine, Nagakute, Japan. sho5-884@umin.ac.jp
  • 3Department of Otolaryngology-Head and Neck Surgery, Tohoku University Graduate School of Medicine, Sendai, Japan.
  • 4Department of Anatomy, Chonbuk National University College of Medicine, Jeonju, Korea.
  • 5Department of Anatomy, Tokyo Dental College, Chiba, Japan.
  • 6Division of Internal Medicine, Iwamizawa Kojin-kai Hospital, Iwamizawa, Japan.

Abstract

Using immunohistochemical staining for alpha-smooth muscle actin (alpha-SMA), glial fibrillary acidic protein (GFAP), S100 protein (S100), p63, cytokeratin 14 (CK14), and cytokeratin 19 (CK19), we studied acinar and myoepithelial cells of major and minor salivary glands obtained from 14 donated cadavers (78-92 years old) and 5 donated fetuses (aborted at 15-16 weeks of gestation). CK and p63 expression was investigated only in the adult specimens. SMA was detected in all adult glands as well as in fetal sublingual and pharyngeal glands. GFAP expression was seen in a limited number of cells in adult glands, but was highly expressed in fetal pharyngeal glands. S100-positive myoepithelial-like cells were present in adult minor glands as well as in fetal sublingual and pharyngeal glands. Expression of p63 was evident in the ducts of adult glands. CK14 immunoreactivity was observed in a limited number of glandular cells in adults, in contrast to consistent expression of CK19. In both adults and fetuses, a mosaic expression pattern was usually evident for each of the examined proteins. A difference in immunoreactivity for the nerve markers GFAP and S100 was observed between the major and minor glands. Thus, in the present histologic study, we distinguished between the specific gland types on the basis of their immunohistochemical staining. A mosaic expression pattern suggested that the immunoreactivity against nerve protein markers in myoepithelial cells could not be due to the persistence of neural crest remnants or the physiological status of the gland, such as age-related degeneration.

Keyword

Salivary glands; Myoepithelial cells; Immunohistochemistry; Adult; Fetus

MeSH Terms

Actins
Adult
Aged
Cadaver
Fetus
Glial Fibrillary Acidic Protein
Humans
Immunohistochemistry
Keratin-14
Keratin-19
Muscles
Neural Crest
Population Characteristics
Proteins
Salivary Glands
Salivary Glands, Minor
Actins
Glial Fibrillary Acidic Protein
Keratin-14
Keratin-19
Proteins

Figure

  • Fig. 1 Immunohistochemistry of the parotid gland (78-year-old woman). (A) p63, (B) alpha-smooth muscle actin (α-SMA), (C) glial fibrillary acidic protein (GFAP), (D) S100 protein (S100), (E) cytokeratin 19 (CK19), (F) cytokeratin 14 (CK14). (A) p63 is expressed in the thick ducts (arrows). (B) α-SMA is expressed in some of the myoepithelial-like cells. (C) GFAP is weakly expressed in ducts (arrows). (D) Fatty tissues as well as nerve terminals around the acinus express S100. (E) CK19 is expressed in all the ducts. (F) CK14 is expressed in cells included in thick ducts (arrows). All micrographs were taken at the same magnification. Scale bar in (A)=0.1 mm (A-F).

  • Fig. 2 Immunohistochemistry of the submandibular gland (78-year-old woman). The specimen was obtained from the same cadaver as in Fig. 1. (A) p63, (B) alpha-smooth muscle actin (α-SMA), (C) glial fibrillary acidic protein (GFAP), (D) S100 protein (S100), (E) cytokeratin 19 (CK19), (F) cytokeratin 14 (CK14). (A) p63 is positive in thick ducts (arrows). (B) SMA is strongly expressed in all or most of the myoepithelial-like cells. (C) Reactivity toward GFAP is evident in some nerve-like structures. (D) S100 expression appears to be limited to nerves. (E) CK19 is expressed in some acinar cells as well as in all the ducts. (F) CK14 expression was not observed. All micrographs were taken at the same magnification. Scale bar in (A)=0.1 mm (A-F).

  • Fig. 3 Immunohistochemistry of a small lingual gland at the dorsal aspect of the tongue (81-year-old man). (A) p63, (B) alpha-smooth muscle actin (α-SMA), (C) glial fibrillary acidic protein (GFAP), (D) S100 protein (S100), (E) cytokeratin 19 (CK19), (F) cytokeratin 14 (CK14). (A) p63 is very weakly expressed in the thick ducts (arrows). (B) SMA is expressed in some myoepithelial-like cells (arrows). (C) Reactivity toward GFAP is evident in a limited number of myoepithelial-like cells (arrows). (D) All or most of the myoepithelial cells express S100 (arrows). (E) CK19 is expressed in some acinar cells as well as in all ducts. (F) CK14 expression was not observed. All micrographs were taken at the same magnification. Scale bar in (A)=0.1 mm (A-F).

  • Fig. 4 Immunohistochemistry of a small pharyngeal gland near the tonsillar fossa (85-year-old man). (A) p63, (B) alpha-smooth muscle actin (α-SMA), (C) glial fibrillary acidic protein (GFAP), (D) S100 protein (S100), (E) cytokeratin 19 (CK19), (F) cytokeratin 14 (CK14). (A) p63 is weakly expressed in some thick ducts (arrows). (B) SMA is strongly expressed in all myoepithelial-like cells. (C) GFAP is negative. (D) S100 expression is seen in some acinus-like and myoepithelial-like cells (arrows) as well as in nerves. (E) CK19 is expressed in acinar cells as well as in all ducts. (F) CK14-positive cells are included in some thick ducts (arrows). All micrographs were taken at the same magnification. Scale bar in (A)=0.1 mm (A-F).

  • Fig. 5 Immunohistochemistry of a small palatal gland at the soft palate (88-year-old man). (A) p63, (B) alpha-smooth muscle actin (α-SMA), (C) glial fibrillary acidic protein (GFAP), (D) S100 protein (S100), (E) cytokeratin 19 (CK19), (F) cytokeratin 14 (CK14). (A) p63 is weakly expressed in some thick ducts (arrows). (B) SMA is expressed in a limited number of myoepithelial-like cells (arrows). (C) GFAP is weakly expressed in some ducts (arrows). (D) S100 expression is seen in myoepithelial-like cells (arrows) as well as in nerves. (E) CK19 is expressed in all the ducts. (F) CK14 reactivity is seen in the debris in the thick ducts (stars) as well as in some myoepithelial-like cells (arrows). All micrographs were taken at the same magnification. Scale bar in (A)=0.1 mm (A-F).

  • Fig. 6 Immunohistochemistry of a small gland in the soft palate (78-year-old woman). The specimen was obtained from the same cadaver as that shown in Figs. 1 and 2. (A) Alpha-smooth muscle actin (α-SMA), (B) glial fibrillary acidic protein (GFAP), (C) S100 protein (S100). (A) α-SMA is expressed in most of the myoepithelial-like cells. In contrast to Fig. 5, GFAP is expressed in some myoepithelial-like cells (arrows, B). (C) S100 is expressed in some acinus-like cells and most of the myoepithelial-like cells (arrows). All micrographs were taken at the same magnification. Scale bar in (A)=0.1 mm (A-C).

  • Fig. 7 Immunohistochemistry of the fetal parotid and submandibular glands (15 weeks). (A-C) Parotid gland, (D, E, G) submandibular gland. (F) Glial fibrillary acidic protein (GFAP) immunostaining of the brain (same specimen) as a positive control for GFAP. (A, D) Alpha-smooth muscle actin (α-SMA), (B, E, F) GFAP, (C, G) S100 protein (S100). α-SMA reactivity is restricted to vessels. GFAP is expressed in the brain (F) but not in these glands. (G) S100 is expressed in developing nerves, which encircle most of the acinus of the submandibular gland (arrows). All micrographs were taken at the same magnification Scale bar in (A)=0.1 mm (A-G).

  • Fig. 8 Immunohistochemistry of fetal sublingual and pharyngeal glands (15 weeks). A specimen obtained from the same fetus as that shown in Fig. 7. Upper panels (A-C) displays the sublingual gland and the lower panels (D, E, G) shows the pharyngeal gland. The lower panels at a magnification higher than that of the upper panels (scale bar in A and D). Panel (F) shows glial fibrillary acidic protein (GFAP) immunostaining of the brain (same specimen) as a positive control for GFAP. (A, D) Alpha-smooth muscle actin (α-SMA), (B, E, F) GFAP, (C, G) S100 protein (S100). α-SMA reactivity is seen not only in vessels, but also in some myoepithelial-like cells (arrowheads in A and D). (E) GFAP is expressed in some myoepithelial-like cells of the pharyngeal gland (arrowheads) and in the brain (F). (C, G) S100-positive nerves encircle most of the acinus of the submandibular and pharyngeal glands (arrows). In addition, some acinus-like cells in both glands also express S100 (arrowheads, C and G). Panels (A-C) (panels D-G) were prepared at the same magnification. Scale bar in (A)=0.1 mm (A-C), in (D)=(D-G).


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