Changes in the number and distribution of myoepithelial cells in the rat parotid gland during postnatal development

The mature rat parotid gland shows hardly any cell bodies of myoepithelial cells around the acini, only a few cell processes being visible. However, in the early postnatal period, the rat parotid gland shows many myoepithelial cell bodies around the acini, including the intercalated ducts. In order to clarify the reason for the disappearance of myoepithelial cells from the area around the acinus during postnatal development, changes in the number and distribution of myoepithelial cells in the rat parotid gland were examined histochemically and chronologically, with particular reference to cell proliferation and cell death. From day 7 to day 14, many myoepithelial cells showing a positive reaction with anti-actin antiserum were found around the acini and intercalated ducts, but thereafter the number of such cells decreased gradually, particularly around the acini, and had almost disappeared after day 35. BrdU/PCNA-positive myoepithelial cells surrounding the acini were easily detected on day 14, but disappeared by day 21, whereas BrdU/PCNA-positive acinar cells remained numerous even after day 21. TUNEL/ISEL staining showed no positive myoepithelial cells throughout the observation period. Transmission electron microscopy also demonstrated no myoepithelial cells with chromatin condensation characteristic of apoptosis through the observation period. These findings suggest that the main reason for the disappearance of myoepithelial cells from the area around the acinus during postnatal development is the large difference between the number of myoepithelial cells and that of acinar cells, because the acinar cells retain their proliferative activity even after myoepithelial cells have become quiescent.     

Origin of acinar cell regeneration after atrophy of the rat parotid induced by duct obstruction

Acinar cell regeneration in the rat parotid gland after atrophy induced by a one week period of duct obstruction was examined using histology, immunohistochemistry and transmission electron microscopy (TEM). For immunohistochemistry, antibodies to 5-bromo-2'-deoxyuridine (BrdU), injected one hour before tissue collection, and cytokeratin were employed. When clips were removed from the duct, only ductal epithelial cells remained; all acinar cells had been deleted. Some duct cells were BrdU positive. After three days, newly-formed acini comprising immature acinar cells had appeared; many of the cells were BrdU positive and mitotic figures were readily identified. Thereafter progressive acinar cell maturation and proliferation occurred, parotid gland weight returning to control levels by 7 days. Peak BrdU labelling indices for duct and acinar cells were on days 0 and 4, respectively. By TEM, cytoplasmic organelles in epithelial cells of transitional duct-acinar structures seen at 2 days were poorly developed. Immature acinar cells seen on day 3 contained zymogen granules and had increased endoplasmic reticulum and mitochondria. By day 5, maturing acinar cells had abundant endoplasmic reticulum and zymogen granules, resembling acinar cells in control glands. These observations indicated origin of acinar cell precursors from duct cells during regeneration of the acinar cell-free atrophic gland. Subsequent expansion of the acinar cell population was dependent on maturation and proliferation of these newly-formed cells.     

Regeneration of myoepithelial cells in rat submandibular glands after yttrium aluminium garnett laser irradiation

The regeneration of myoepithelial cells in rat submandibular salivary gland after partial irradiation with yttrium aluminium garnett (YAG) laser was investigated. The irradiated glands were examined immunohistochemically for actin, histochemically for alkaline phosphatase (ALP), and by transmission electron microscopy (TEM). In control glands, myoepithelial cells were positive for actin and ALP. Electron microscopically, the positive reaction for actin was associated with the myofilaments of myoepithelial cells, and the plasma membrane of myoepithelial cells was positive for ALP. One day after YAG laser irradiation, the irradiated region was necrotic. By 5 days, duct-like structures and epithelial clusters were observed at the interface between the necrotic zone and the remaining undamaged glands; immature acini appeared after 7 days. No reaction in duct-like structures or epithelial clusters to actin or ALP was recognizable by 5 days. However, at 7 days, actin and ALP-positive spindle cells appeared at the periphery of the duct-like structures and immature acini. After 10 days, both actin-positive and ALP-positive cells increased in number. These observations indicate that during regeneration, actin-positive and ALP-positive cells regenerate myoepithelial cells, and it is suggested that this differentiation to myoepithelial cells is closely related to that of luminal to acinar cells. In addition, TEM observations indicate that regenerated myoepithelial cells originated from the basal cells of duct-like structures.     

Regeneration of rat submandibular gland following partial extirpation. A light and electron microscopic study

A wedge of parenchymal tissue was excised from the left submandibular gland of six week old male Sprague-Dawley rats. The animals were randomly grouped by body weight and killed at intervals of one day to five weeks following the operation. Tissue from and adjacent to the site of injury was removed and prepared for routine light and electron microscopy. Light microscopic findings consisted of degeneration and necrosis of the parenchymal tissue during the first 24 hours, followed by hyperemia and endothelial as well as epithelial proliferation from one to three days. Extensive epithelial proliferation occurred during the next two weeks, followed by regeneration of new lobules, beginning at the periphery of the injured lobes. Ultrastructurally, the new parenchymal tissue appeared to have regenerated from residual duct cells. Dedifferentiated epithelial cells gave rise to two different cell lines: one line which transformed into terminal tubules and acinar cells, and another which became striated ducts. These differentiating cells were organizing into lobules as early as three weeks after the operation. Because of their proximity to cells of regenerating striated ducts as well as intercalated ducts and acini, the myoepithelial cells appeared to be of epithelial origin.     

Immunohistochemical and Immunocytochemical Localization of Amylase in Rat Parotid Glands and von Ebner's Glands by Ion Etching-Immunoscanning Electron Microscopy

The distribution of amylase in rat parotid glands and von Ebner's glands was examined using ion etching-immunoscanning electron microscopy, which enables both light and electron microscopic observations of identical semi-thin resin sections immunolabeled with anti-α-amylase and immunogold in association with silver enhancement. At the light microscopic level, most acinar secretory granules (SG) and striated duct secretions of parotid glands were strongly stained dark brown. In von Ebner's glands, acinar SG and duct secretions were weakly to strongly stained light to dark brown. At the electron microscopic level, labeling was observed as bright gold-silver particles. The labeling intensity of acinar SG of parotid glands was higher than that of von Ebner's glands. In parotid glands, weak labeling of SG in transitional cells between acini and intercalated ducts, very weak labeling of SG in intercalated ducts, and strong labeling of striated duct secretions were observed. In von Ebner's glands, the secretions and some SG of interlobular ducts were strongly labeled compared to those of intralobular ducts and SG of acini. Less amylase was synthesized in von Ebner's acini compared to parotid acini, whereas von Ebner's ducts may secrete significantly more amylase to modify saliva than parotid ducts.     

Ultrastructural aspects of calf submandibular glands

Submandibular gland biopsies from four calves were examined by electron microscopy. Most of the parenchyma consists of mucous acini capped by seromucous demilunes. Secretory product of the demilunes reaches the acinar lumen via intercellular canaliculi located between adjacent demilunar cells or by narrow apical extensions of demilunar cells bordering the lumen in common with acinar cells. Intercellular canaliculi are absent between mucous acinar cells, but intercellular space is present at junctions of demilunar cells, acinar cells, and intercalated duct cells. Intercalated ducts are short and connect mucous acini with striated ducts. Striated ducts show more basal infoldings and mitochondria than those of bovine parotid glands. Nuclear bodies are present in most epithelial cell types of the gland but are larger and more easily recognized in nuclei of striated duct cells. Attempts are made to correlate the structure of bovine submandibular glands with its secretion of small amounts of hypotonic saliva relative to the larger volume of isotonic saliva secreted by parotid glands of the same animal.     

On the ultrastructure of the parotid and submandibular gland of the mongoose Herpestes edwardsi (Viverridae) (author's transl)

The parotid and submandibular glands of the mongoose are described. Essential differences between the 2 glands were recognized in the acini; however, the intra- and interlobular ducts are built up similarly. The parotid gland is acinar. Its secretory cells are filled with distinct types of granula, which show a considerable variation of size and structure of their secretory material. Organelles are found sparsely. The submandibular gland, however, is tubuloacinar. Its tubuli are capped with cells which belong to the demilunes of v. EBNER, but because of their pale granules they occupy an exceptional position. As the acinar cells of the parotid gland, they form intercellular canaliculi by their plasmalemmata. In the secretory cells of the tubules an intimate contact between the rER and the granules is observed. The intralobular duct surface is built up by an onelayered epithelial cell formation. The cytoplasm of the intercalated duct cells is rich in bundles of filaments, and these cells contain mitochondria with a particular dense matrix. Some microvilli cover the apical surface. In the cells of the striated ducts several populations of granules differing in size and electron density are found. The material of the dense granules shows a marginal plate-like condensation, sometimes it cristallizes. It is supposed that they were released by an apocrine extrusion mechanism. Terminal axons innervate the acini, the duct cells, and also the myoepithelial cells. The findings are compared with the well-known morphology of the salivary glands of the cat.     

Telocytes in Parotid Glands

The parotid histological structure includes acinar, ductal, and myoepithelial cells, surrounded by a connective stromal component. The parotid stroma is mostly regarded as an inert shell, consisting of septa, which divide the parenchyma. Telocytes were recently identified as a new stromal cell type in various organs, including exocrine pancreas. We aimed to evaluate telocytes presence in parotid stroma and whether their topographical features might support an involvement in parotid function modulation. Serial ultrathin sections of human and rat parotid glands were studied and compared by transmission electron microscopy. Two‐dimensional concatenation of sequenced micrographs allowed the ultrastructural identification of parotid telocytes, with their specific long, thin, and moniliform prolongations (telopodes). Telocyte location appeared frequently as a strategic one, in close contact or vicinity of both secretory (acini and ducts) and regulatory (nerves and blood vessels) apparatuses. They were also found in the interacinar and the subductal stroma. Two previously reported telocyte markers (c‐kit/CD117 and vimentin) were assayed by immunohistochemistry. Actin expression was also evaluated. Telocytes are making a network, especially by branching of their long telopodes. Elements of this telocyte network are interacting with each other (homocellular connections) as well as with other cell types (heterocellular connections). These interactions are achieved either by direct contact (stromal synapse), or mediated via shed microvesicles/exosomes. Since telocyte connections include both neurovascular and exocrine elements (e.g., acini and ducts), it is attractive to think that telocytes might mediate and integrate neural and/or vascular input with parotid function. Anat Rec, 2012. © 2011 Wiley Periodicals, Inc.     

腮腺主导管结扎诱导萎缩后的组织转归

背景：人体大唾液腺常因受到头颈部肿瘤放射治疗、舍格伦综合征及涎腺炎等因素的影响发生腺体萎缩,目前对长期萎缩性腮腺内组织形态变化的观察仍较少。 目的：观察腮腺主导管结扎诱导腮腺萎缩后的组织转归。 方法：通过结扎SD大鼠右侧腮腺主导管诱导腺体萎缩,采用苏木精-伊红染色观察正常腮腺及导管结扎后0(对照),1,3,7,14,30,60 d萎缩性腮腺组织内腺泡、导管细胞的面积;免疫组织化学染色定量分析肌上皮细胞在腮腺萎缩不同时间点的数量分布变化。 结果与结论：结扎腮腺主导管后腺泡细胞出现快速凋亡,至14 d时已基本消失。随着腺体萎缩,间质逐渐纤维化并伴随炎性细胞浸润,组织内形成大量导管样结构,导管面积逐渐增加,到14 d时达到顶峰,随后逐渐减少,导管样结构呈典型的双套层结构,结扎各时间点腺泡、导管面积与对照组比较差异均有显著性意义(P < 0.05)。结扎后7 d内肌上皮细胞数量快速增加,随后肌上皮细胞数量增长缓慢,维持在一定的范围。表明腮腺主导管结扎诱导腺体萎缩早期腺泡细胞快速消失,出现大量导管样结构,肌上皮反应性增殖,随着腺体的萎缩由导管样结构及肌上皮细胞组成“双套层”结构可能抑制腺体的进一步萎缩。     

Differential distribution of sialic acid in exocrine pancreas and parotid gland

The sialic acid-specific lectin limulin (LPA, from Limulus polyphemus hemolymph) was used for the investigation of the distribution of accessible sialoglycoconjugates on the surface of cells from rat and rabbit parotid gland and exocrine pancreas. Fluorescence microscopy with rhodamine-conjugated LPA on fresh-frozen and fixed-frozen sections of the parotid gland revealed lectin binding sites on acinar and ductular epithelial cells. The staining was localized only at the periphery of the acini and ducts and was absent from the apical and lateral surface of epithelial cells. This staining pattern contrasted with that found in epithelial cells of acini and ducts in exocrine pancreas, where the luminal surface was intensely labeled by the fluorescent lectin. The luminal content of ductular tract and acini in parotid gland and pancreas was devoid of lectin-reactive sialoglycoconjugates. Connective tissue surrounding ducts and blood vessels bound the lectin heavily in both glands. These results outline that cells with similar structure and function, but constituents of different exocrine glands, exhibit differential distribution of sialoglycoconjugates on their corresponding plasmalemmal domains.     

Secretory protein expression patterns during rat parotid gland development

The rat parotid gland produces a number of well-characterized secretory proteins. Relatively little is known, however, about the onset of their synthesis and cellular localization during gland development. Secretory protein expression was studied in parotid glands of fetal and postnatal rats using light and electron microscopic immunocytochemistry and Northern blotting. Amylase, parotid secretory protein (PSP), common salivary protein-1 (CSP-1), and SMGB were first detected by immunofluorescence in parotid glands of 18 day fetuses. By 5 days after birth, light and electron microscopic immunolabeling localized all of these proteins to the secretory granules of developing acinar cells. Labeling of acinar cells for DNAse I, however, was not observed until 18 days after birth. Between 9 and 25 days, CSP-1 and SMGB reactivity of acinar cells declined, but increased in intercalated duct cells. After 25 days, CSP-1 and SMGB were found only in intercalated ducts, and amylase, PSP, and DNAse I were restricted to acinar cells. Levels of CSP-1 and SMGB mRNA were relatively constant through 21 postnatal days, but declined significantly after that. Amylase and PSP mRNA increased rapidly and continuously from five days after birth to the adult stage. In contrast, DNAse I mRNA was not detectable until 18 days after birth. The immunocytochemical and molecular analyses define three basic patterns of protein expression in the rat parotid gland: proteins whose synthesis is initiated early in development and is maintained in the acinar cells, such as amylase and PSP; proteins that are initially synthesized by immature acinar cells but are restricted to intercalated ducts in the adult gland, such as CSP-1 and SMGB; and proteins that are synthesized only by mature acinar cells and first appear during the third postnatal week, such as DNAse I. The parotid gland exhibits four distinct developmental stages: prenatal, from initiation of the gland rudiment until birth; neonatal, from 1 day up to about 9 days postnatal; transitional, from 9 days to 25 days of age; and adult, from 25 days on. Although differences exist in timing and in the specific proteins expressed, these developmental stages are similar to those seen in the rat submandibular gland. Additionally, the results support the suggestion that intercalated ducts may differentiate from the neonatal acini. Anat. Rec. 252:485–497, 1998. © 1998 Wiley-Liss, Inc.     

Regeneration in chronic sialadenitis: an analysis of proliferation and apoptosis based on double immunohistochemical labelling

The understanding of regeneration in salivary glands as a finely tuned balance of cellular proliferation, differentiation and apoptosis has been limited by the difficulty of identifying proliferating cells. This has been overcome in the present investigation by double immunohistochemical labelling for the proliferation-associated antigen Ki67 and for different cell-type-specific antigens applied to 8 specimens of normal parotids and 16 specimens of chronic parotid sialadenitis with particular reference to acini and intercalated ducts. In comparison with low baseline rates of proliferation in normal parotids, proliferative indices were significantly increased in chronic sialadenitis in mature acinar cells, intercalated ductal cells and myoepithelial cells without evidence of proliferation by an additional population of cells. In accordance with findings in glands of experimental animals, the present data do not support the previously postulated concept of regeneration of acini and intercalated ducts by a hypothetical population of uncommitted ductal stem cells. The demonstration of a profound capacity for intrinsic glandular regeneration from differentiated cells represents a biological basis for the good results obtained from conservative therapy of chronic sialadenitis and offers hope for novel therapies designed to reconstitute impaired salivary flow.     

A scanning electron microscope study of myoepithelial cells in the intercalated ducts of rat parotid and exorbital lacrimal glands

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.     

Hamartoma of the parotid gland: A case report with immunohistochemical and electron microscopic study

Summary A case of a solid parotid tumour in a 16-year-old boy is presented. Histologically, the tumour demonstrated some peculiar findings. An acinar pattern was predominant although every component seen in the normal salivary gland was present, namely, serous and mucous gland acini, ducts, myoepithelial cells, adipose and lymphoid tissue. Large eosinophilic granules were abundant in the large acinar cell cytoplasm. Immunohistochemically, the tumour demonstrated the proteins which are present in the normal parotid gland, for example, amylase, lactoferrin and lysozyme. Electron microscopic features were quite similar to those of normal parotid tissue except for accumulation of a large number of cytoplasmic granules in the acinar cells. There has been no previous report of a tumour with the same features as seen in this case. Our pathological diagnosis is hamartoma, although the possibility of hyperplasia or neoplasia can not be excluded.     

Ultrastructure of the anterior medial glands of the rat nasal septum

The anterior medial glands lying in the submucosa of the rat nasal septum were studied by light and electron microscopy. The glands consist of a single long duct, which is studded with numerous solitary acinar formations connected perpendicularly to the main duct by short intercalated ducts. Proximal acini (those furthest from the stoma of the main duct) consist of typical serous cells with many dense secretory granules and an extensive rough endoplasmic reticulum. The most distal acini consist of cells whose major feature is the enwrapment of each mitochondrion by a cisternal profile of rough endoplasmic reticulum. Myoepithelial cells are absent from proximal acini, but are abundant on distal acini. Intracellular nerve terminals are extremely common, particularly in distal acini. The main ducts resemble, to a degree, the striated ducts of salivary glands.     

Further evidence for AQP8 expression in the myoepithelium of rat submandibular and parotid glands

Previously (Wellner et al., Pflugers Arch 441:49–56, 2000) we suggested that the localization of the aquaporins (AQPs) AQP5 and AQP8 in the apical and basolateral membranes of rat submandibular gland (SMG) acinar cells, respectively, provides for transcellular water flow during saliva formation. While the localization of AQP5 in this gland has been verified in several laboratories, there have been differing reports regarding AQP8 localization. Other investigators subsequently reported that AQP8 is not expressed in the acinar or ductal cells of the major salivary glands of the rat, but in the myoepithelium of each gland. Thus, we have carried out additional studies: (1) to reassess the localization of AQP8 in the rat SMG and (2) to assess the localization of AQP8 in the rat parotid gland (PG). Initially, we compared the localizations of AQP8 with recognized basolateral markers in acinar cells [the Na⁺,K⁺-ATPase and the Na⁺–K⁺–2Cl^– cotransporter (NKCC1)]. Our results indicated that Na⁺,K⁺-ATPase localized in both the basal and lateral membranes of rat SMG acinar cells, whereas AQP8 was detected only in the basal regions of the acini. In the rat PG, AQP8 was invested near intercalated ducts and adjacent acini, whereas NKCC1 localized in the basolateral membranes of acinar cells. As these results were suggestive of myoepithelial localization in both glands, we compared AQP8 localization with the localization of smooth muscle actin, a myoepithelial marker. We found that AQP8 and smooth muscle actin colocalized in both the rat SMG and PG, providing additional strong support for a myoepithelial localization of AQP8. Thus, in agreement with an earlier report by other investigators (Elkjaer et al., Am J Physiol Renal Physiol 281:F1047–F1057, 2001), we report that AQP8 is expressed in the myoepithelial cells, but not in the acinar cells, of both the rat SMG and PG.     

An electron microscope study of the development of the exocrine and endocrine pancreas with special reference to intercellular junctions

Intercellular junctions in pancreatic acinar, duct and endocrine cells were studied by thin section and freeze-fracture methods in developing rats and mice. Undifferentiated cells were joined by the zonula occludens and isolated fragments of tight junctional strands. Small gap junctions were either occasionally associated with tight junctional strands or appeared independent of them. During the morphological differentiation of acinar cells, strands of the zonula occludens developed to form a complicated meshwork while gap junctions rapidly increased in size. Duct cells were joined by the less-developed zonula occludens but gap junctions were rarely seen. In the neonate, intercellular junctions were similar to those in adult acini and intercalated ducts. Endocrine cells were joined by maculae occludentes and small gap junctions. During late prenatal days, the macula occludens increased in size and gap junctions in number. Sometimes tight junctional strands disappeared to leave membrane elevations, some of which were associated with small gap junctions. Maculae occludentes on endocrine cells were gradually fragmentized and diminished during postnatal development. They were completely lost in the rat. These results suggest that intercellular junctions play important roles in pancreatic development. In particular, the transient development of maculae occludentes is associated with endocrine cell development, and intercellular communication mediated by gap junctions may be important for the differentiation of acinar and endocrine cells.     

Ultrastructure of baboon parotid gland

The parotid gland of the olive baboon, Papio anubis, was examined by electron microscopy. The acini are all serous in nature, and consist of pyramidal cells with abundant secretory granules of varying size. These granules consist of a dense matrix in which a denser spherule or lenticular body is present. Granules linked by a short isthmus are observed in the apical cytoplasm, and granules in the process of discharging their contents to the acinar lumen may be connected to the luminal plasma membrane by a neck-like protrusion. Intercalated duct cells contain granules reminiscent of those found in the rat submandibular acinar cells. The striated ducts consist of tall cells interloked in a complex fashion near their bases, with numerous vertically-oriented mitochondria lodged in their basal crenulations. Small vesicles whose contents vary in density are present in the apical cytoplasm as are large deposits of lipofuscin. The striated duct cells display a proclivity for ballooning into the duct lumen. Excretory ducts consist of simple to pseudostratified columnar epithelium, and lack basal striations or apical blebs.     

Intercalated duct cells in the rat parotid gland may behave as tissue stem cells

This study was conducted to determine whether intercalated duct cells in the rat parotid gland have the properties of tissue stem cells. After induction of cellular proliferation by repeated administration of isoproterenol (IPR), a β-adrenergic agonist, proliferation activity in acinar, intralobular, and intercalated ductal cells was quantified using Ki-67 immunohistochemistry. The total number of each type of component cell in a gland was also estimated in the course of IPR treatment. IPR was found to induce proliferation of acinar and intercalated duct cells, but not intralobular duct cells. The total number of acinar cells in a gland on day 5 of IPR treatment was 1.6 times of that at day 0 (baseline). In contrast, the total numbers of intercalated and intralobular duct cells did not change from baseline, indicating a high possibility that the proliferated intercalated duct cells differentiated into acinar cells. On days 2 to 3 of IPR treatment, intercalated duct cells with amylase-positive secretory granules were recognized in a region very close to the acini, and were suspected of being transitional cells from intercalated duct to acinar cells. This quantitative study indicates that intercalated duct cells may have the properties of tissue stem cells upon IPR stimulation.     

Contributions of intercalated duct cells to the normal parenchyma of submandibular glands of adult rats

The parenchyma of the submandibular gland in the adult male rat is self-renewing, with most newly formed acinar and granular duct cells believed to differentiate from the rapidly proliferating intercalated duct (ID) compartment. Since the ID cells are phenotypically diverse, based on their different expression of perinatal secretory proteins, we systemically injected tritiated thymidine for 24 hours, and followed the pattern of thymidine distribution in cells by autoradiography and immunocytochemistry of defined cellular phenotypes over a 1-month chase period. Proliferating cells were found within all parenchymal cell compartments; they were most numerous in ID, and primarily in those cells lacking immunoreactivity for the perinatal proteins SMG-B1, -C, and -D. The labeling index (LI) of the ID cells reached a peak at 7 days postinjection, and then decreased over the next 3 weeks. Concurrently, the LI increased significantly in those cells at the junctions of ID with both acini and granular ducts, and also within these larger parenchymal elements. We conclude that the ID cells not reactive for perinatal proteins proliferate to expand the ID compartment, and that ID cells at the ends of the ducts differentiate into both acinar and granular duct cells. Our data provide no evidence for the differentiation of ID cells into cells of striated ducts (SD); however, the small number of excretory duct (ED) profiles seen in our preparations showed extremely high LI (>25%), suggesting that more extensive data might reveal a precursor role for the ED in replacement of SD cells. In addition to the stepwise passage of cells from ID to other parenchymal elements at their junctions, the reported occurrence of occasional clusters of B1-positive acini (BAC) among the typical B1-negative acini had suggested an alternate pathway, in which entire segments of newly expanded ID might develop directly into a recapitulated perinatal stage of B1-reactive cell, pursuant to becoming mature acinar cells. Consistent with this suggestion, the BAC had a fourfold greater LI than typical adult acini; moreover, when analyzed by electron microscopic immunocytochemistry, they appeared similar to the novel perinatal Type III cells both ultrastructurally and in their pattern of B1-immunogold labeling. In contrast, the less common acini showing a sublingual gland phenotype had no significant difference in LI from typical acinar cells. Overall, our results emphasize the importance of the nonimmunoreactive ID cells in normal cellular replacement, and the possibility that ID can undergo en bloc differentiation into replacement acini as well as incremental addition of single cells at the boundaries of ID with acini and with granular ducts.