The stomachs of adult CD1 mice were investigated by anatomical examination and light microscopy. Serial sections were prepared of entire stomachs; the various types of gastric glands were characterized; and, using every 30th or 60th serial section, maps of gland distribution were obtained through point-plotting serial reconstruction. Gross examination shows that the cephalic third of the stomach consists of a thin-walled, domelike structure, the forestomach. The rest of the organ, or stomach proper, is subdivided into two parts, the thick-walled corpus, which approximately occupies the middle third, and the less vascular pyloric antrum which forms the remaining caudal third of the organ. Histologically, glands are absent from the forestomach mucosa but are numerous throughout the stomach proper. They are of two main types, namely, zymogenic glands, which contain, among others, zymogenic cells, and mucous glands, which lack these cells but contain mucous cells. Both gland types show a few enteroendocrine cells. Moreover, some of the mucous glands include parietal cells (mucoparietal glands), while others do not (pure mucous glands). Mucosal maps reveal that the glands of each type are located in distinct areas of the mucosa. Thus a compact zymogenic region may be defined, occupying 56% of the glandular mucosa and containing only zymogenic glands. The mucous region, on the other hand, composed only of mucous glands, is extensive and divided into a narrow cephalic band (5.2% of the glandular mucosa) and a large caudal part (38.8%). Along the lesser curvature these parts are continuous, and together they encapsulate the zymogenic region. In proximity to the border of this region, and only there, do the mucous glands include parietal cells. A comparison of mucosal maps and gross features indicates that the corpus includes both the zymogenic region and the cephalic band of mucous glands, whereas the antrum is composed entirely of the mucous glands of the caudal part.

The isthmus of typical mucous units of the pyloric antrum was investigated in 3- to 4-month-old CD1 mice using light and electron microscopy as well as 3H-thymidine radioautography. On the average, the isthmus measured 25 microns in length and was composed of 36 isthmal cells and two enteroendocrine cells. Isthmal cells generally displayed features found in embryonic cells, such as many free ribosomes, scant organelles, and a large reticulated nucleolus, and were, therefore, at an immature stage of development. Isthmal cells could be devoid of secretory granules ("granule-free cells," 2%) or contain a few small, spherical, PA-Schiff-positive, mucous granules in their apex. The granules in some of the cells had a variegated appearance and a diameter averaging 235 nm ("mottled granule cells," 39%); in other cells, the granules had a large diameter, 278 nm, with a pale background and a dense core ("core granule cells," 28%); while in still others they were homogeneously dark and measured 264 nm ("dense granule cells," 12%). Finally, some cells included a mixture of core and dense granules ("mixed granule cells," 14%). One hour after a single injection of 3H-thymidine, 37% of the isthmal cells were labeled. Each of the five isthmal cell types could acquire label and, therefore, divide. After one or more days of continuous 3H-thymidine infusion, all isthmal cells were labeled. Their turnover time was estimated to be 16.1 hr (t1/2 = 11.2 hr). The isthmus is thus composed of several cell types which are turning over rapidly. While all are relatively immature, the various types are thought to represent different developmental stages in the life history of an isthmal cell. A model devised on this basis proposes that the granule-free cells are stem cells, from which mottled granule cells are derived. These in turn evolve into either the dense granule cells of the upper isthmus or the core granule cells of the lower isthmus, or into the mixed granule cells (which are believed to develop eventually into dense granule cells or core granule cells). Maintenance of a steady state requires that the rapid production of isthmal cells be associated with rapid emigration; the dense granule cells presumably going to the pit and the core granule cells to the gland. The turnover of isthmal cells is accordingly described as following a "bidirectional pattern" of renewal.

The renewal of gland cells was investigated by three-dimensional reconstruction of typical mucous units of the pyloric antrum using electron microscopy and 3H-thymidine radioautography in 3 to 4 month-old CD1 mice. Based on analysis of 42 units, the average gland measured 31 micron in length and was composed of 37 (mucous) gland cells with eight enteroendocrine cells scattered among them. The gland neck cells located close to the isthmus showed the cytoplasmic and nuclear features characteristic of differentiating cells. The mid-gland cells occupying the central portion of the gland appeared to be at a more advanced stage of development and completing differentiation. The gland base cells comprising the blunt end of the gland were fully mature. To quantify the renewal process, the percent of gland cell nuclei carrying label was determined at several times following 3H-thymidine administration. The rate of proliferation was found to be greatest in the gland neck, lower in the mid-gland, and even lower within the gland base. Furthermore, the isthmus contributed to gland-cell renewal by providing an estimated 12.4 cells per day. Labeled cells migrated toward the blunt end of the gland. The migration rate became progressively slower with their descent, and many cells were lost along the migration pathway, mainly in the gland neck. The loss took place without being preceded by gradual cell degeneration, but occurred as a result of rapid extrusion to the lumen or, less frequently by pyknosis, which could be followed by phagocytosis. It is concluded that the rapid rate of mitosis within the isthmus and gland neck generates a pressure causing downward migration of the cells toward the blunt end of the gland. The rate of migration, however, gradually diminishes as cells descend into the gland, presumably owing both to decreasing proliferation rate and to cell loss. Thus, while cells migrate down toward the gland base, many are lost before reaching it. This sequence is described as "the cascade pattern" of renewal.

The pit (foveola) of typical mucous units of the pyloric antrum was investigated in 3- to 4-month-old CD1 mice, using light and electron microscopy, sometimes combined with 3H-thymidine radioautography. Reconstruction of units from serial sections revealed that, on the average, the pit measured 151 microns in length and was lined by 184 mucus-containing pit cells. Of these, 164 were located along the wall of the pit, whereas 20 surrounded its opening on the free surface. For ultrastructural examination the pit was divided into equal thirds. The proximal third, located next to the isthmus and referred to as pit base, was composed of cells showing electron-dense mucous granules greater in number but similar in density and diameter to those of isthmal dense granule cells. Nucleoli were rather large, irregular, and reticulated; these and other features were indicative of partial differentiation. The appearance of the cells gradually changed with the distance from the isthmus. In the middle third or mid pit, cells had small, fairly rounded nucleoli, while mucous granules were more numerous than in the pit base but similar in appearance and size; these cells were considered to be mature. In the distal third or pit top-surface, granules became elongated and nucleoli shrank, and lysosomes and vacuoles greatly increased in number, indicating that cells were at a terminal stage. Indeed, some of the cells were extruded into the stomach lumen while others were phagocytosed by adjacent cells. Following a single injection of 3H-thymidine, labeling was found only in a small cohort of cells in the pit base. At the end of 1 day of continuous infusion, the cohort of labeled cells had reached the mid pit; by 2 days, the pit top; and by 3 days, the free surface, where cells were eventually lost. The renewal time of pit cells was assessed at 2.98 days (t1/2 = 1.8 days), giving a turnover rate of 33.5% per day. It is estimated that the divisions of pit base cells provide two-thirds of the cells needed daily for pit-cell renewal, while the other third is supplied by an influx of dense granule cells from the isthmus. These cells enter the pit and continuously migrate toward the gastric lumen, while differentiating in the pit base, maturing in the mid pit, and reaching a terminal stage at the pit top-surface. The progressive and orderly migration of pit cells is described as a "pipeline pattern" of renewal. It is completed in about 3 days when terminal cells are lost at the pit top-surface.

RNA synthesis was examined in the epithelial cells of the mouse pyloric antrum using radioautography 20 min after injection of either 3H-uridine or 3H-orotic acid. The epithelium of the mouse antrum was known to invaginate into blind tubular units composed of mucous cells arranged from base to top into a gland, an isthmus, and a pit. These were subdivided into segments and, after radioautography, silver grains were counted over cell nuclei in each segment. Following 3H-uridine injection, silver grains were present over all nuclei but were more abundant over those of the isthmus than of the gland or the pit. When nuclei were examined in the electron microscope, nucleoplasmic as well as nucleolar silver grains were more numerous in the isthmus than in the pit or gland. Following 3H-orotic acid injection, silver grains were again present over all nuclei; but maximal incorporation appeared to be in pit cell nuclei where, by electron microscopy, it was mainly assigned to the nucleoplasm. When the incorporation was calculated per whole nucleus, however, it was less in pit cell than in isthmal cell nuclei. Even so, the proportion of label in pit cell nuclei was much greater than after 3H-uridine injection. The interpretation of these findings is based on the fact that isthmal cells are immature, whereas cells migrating from the isthmus to become gland or pit cells show increasing differentiation. The immature cells of the isthmus incorporate both uridine and orotic acid more effectively than do the differentiated cells of pit and gland. Since silver grain counts over nuclei provide an index of the rate of RNA synthesis, this synthesis proceeds more actively in the isthmus than in the pit or gland. This is true of ribosomal RNA synthesis, as shown by nucleolar grain counts, and of other RNA's synthesis, as shown by nucleoplasmic grain counts. It seems, however, that while uridine is involved in the synthesis of all types of RNA, orotic acid is mainly implicated in the synthesis of the heterogeneous RNA from which the messenger RNA arises.