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PubMed1989

Photoablation of ocular melanoma with a high-powered argon endolaser.

We studied the use of a 15-W argon blue-green laser in the treatment of choroidal melanoma in a rabbit model. Greene melanoma cells were used to produce 2- to 4-mm thick tumors posteriorly in the suprachoroidal space in pigmented rabbits. Endophotocoagulation delivered through a 600-micron fiberoptic probe was performed to ablate the tumor tissue and a surrounding margin of normal tissue. A vitreous cutter was used simultaneously to remove liberated necrotic debris. The effect of the laser on tumor and normal ocular tissue was evaluated by light microscopy and the extent of the proliferative response by tritiated thymidine radioautography. Application of 100 to 400 pulses of laser energy using treatment parameters of 12 to 14 W of power and 0.1-s pulses resulted in complete ablation of melanoma tissue, overlying retina, and choroid. There was no substantial intraoperative or postoperative hemorrhage. Material liberated during the laser treatment was found to be nonviable. The effect of the laser on tissue appeared localized to within approximately 1.25 mm of the margin of the central lesion. The high-energy argon laser seems to offer a means of effectively ablating melanoma tissue via an internal resection approach.

Animals↗
PubMed1993

Evidence of abnormal stromelysin mRNA expression in suspected carriers of otosclerosis. A possible molecular marker.

OBJECTIVE: There is strong evidence that otosclerosis is a genetic disease affecting bone remodeling. We propose that, if otosclerosis is genetic, it will manifest itself universally, particularly at the mRNA transcription level. DESIGN: Skin biopsy specimens were taken in a single blind from human subjects who had been clinically and surgically identified as having otosclerosis. SETTING: Subjects were volunteers from the community, identified through hospital records. All procedures were carried out in a clinical research facility. PATIENTS: Twenty-one volunteers underwent a biopsy, including those positively identified as having otosclerosis (n = 4), their blood relatives (n = 8), or nonrelatives with normal hearing and no known history of otosclerosis (n = 9). INTERVENTION: Three connective tissue remodeling factors, procollagenase, prostromelysin, and tissue inhibitor of metalloprotease, were analyzed. The mRNA was extracted from each biopsy specimen, hybridized against radiolabeled cDNA, and quantitatively measured by radioautography. MAIN OUTCOME MEASURE: We expected to see significant differences in the pattern of mRNA expression for one or more of the three measured bone remodeling factors in otosclerotics, compared with age- and sex-matched negative controls. RESULTS: Two of the otosclerotic subjects had abnormally low levels of mRNA for prostromelysin and two had higher than normal levels. In three (75%) of the four, variability of mRNA expression among procollagenase, prostromelysin, and metalloprotease tissue inhibitor was higher than normal. Three (38%) of the eight relatives showed a similar pattern and two (22%) of the nine control subjects also tested as abnormal. CONCLUSION: This observed variability in otosclerotic subjects might be a manifestation of a genetic control defect, and abnormal stromelysin mRNA expression could serve as a genetic marker for otosclerosis.

Adolescent↗
PubMed1996

Cementogenesis reviewed: a comparison between human premolars and rodent molars.

BACKGROUND: Cementum continues to be the least-known mineralized tissue. Although recent advances in the field of molecular biology have contributed to an understanding of the involvement of molecular factors in cementum formation during development and regeneration, cementogenesis on a cell biological basis is still poorly understood. Virtually nothing is known about cementoblast origin, differentiation, and the cell dynamics during normal development, repair, and regeneration. This review describes the recent findings of cementogenesis on roots of human premolars and opposes them to those of teeth from other mammals, particularly the rodent molar. METHODS: Using light and electron microscopy, light microscopic radioautography, and various measurements, a comprehensive insight into the development and repair of cementum during and after root formation and tooth eruption has been achieved for human premolars. RESULTS: Cementum is a highly responsive mineralized tissue. This biological activity is necessary for root integrity and for bringing and maintaining the tooth in its proper position. With regard to cementum formation and periodontal fiber attachment, considerable species-particularities exist that are mainly based on differences in growth rates and tooth sizes. Since root development and initial cementogenesis last on the average 5-7 years in human premolars, cementum formation in these teeth is characterized by along-lasting phase of prefunctional development, with occurs independent of principal periodontal fiber attachment to the root and which may take 5 years or more. The first molar of the rat, however, is in functional occlusion 3 1/2 weeks after the onset of root formation. Since initial cementum formation and periodontal fiber attachment to the root occur almost at the same time in this tooth, the distinction between cells associated with one or the other process is very difficult to achieve, and cementogenesis cannot be described independent of periodontal fiber attachment to the root. Therefore, the determination of cementoblast origin in the rodent molar may be intricate. CONCLUSIONS: Taking into account these species differences, the current description on the origin and differentiation of cementoblasts is inconsistent and the description of cementogenesis is still incomplete. This review calls into question the currently held concept of cementogenesis and offers a possible alternative.

Animals↗
PubMed1998

Changes in the rate of RNA synthesis during the cell cycle.

BACKGROUND: Although the rate of RNA synthesis is known to drop at mitosis, the recent identification of 11 stages in the cell cycle (El-Alfy et al., 1994) makes it possible to measure the rate of this synthesis at each one of the stages and thus find out how it varies throughout the cell cycle. METHODS: Mice were injected intravenously with the RNA precursor, 3H-uridine; the duodenum was fixed 5-15 minutes later for embedment in Epon, and the duodenal crypts were cut in semithin serial sections for study of the rapidly dividing crypt columnar cells. Using Feulgen-stained sections, each cell nucleus was assigned to one of the 11 stages described in the cell cycle, and the same nucleus was identified in the next serial section that had been processed for radioautography, so that the overlying silver grains were enumerated. The count was taken as an index of the rate of RNA synthesis by this nucleus. RESULTS: Starting from stage I of the cell cycle (the period defined by the presence of a minimal amount of chromatin during which the S phase begins) and up to stage IV (when the S phase ends and the G2 phase begins), all or nearly all nuclei are synthesizing RNA with the rate peaking at stage III. During stages V to VIII (the period comprising the mitotic steps), the percentage of RNA-synthesizing nuclei decreases to over half at stage V (prophase), -10% at stages VIa (prometaphase) and VIb (metaphase) and none at stages VII (anaphase) and VIII (telophase). During stages IX-XI (which correspond to the G1 phase), the percentage rises sharply at stage XI to reach up to 100% at stages X and XI. Finally, on the average, 35% of nuclear silver grains are over the nucleolus (presumably representing ribosomal RNA precursors), whereas 65% are over the nucleoplasm (presumably representing mainly heterogeneous RNA precursors). CONCLUSIONS: Cells synthesize RNA during the interphase, but at a variable rate with a peak in S. The synthesis proceeds in a majority of the cells at prophase, but only in a few of them at prometaphase and metaphase, and in none at anaphase and telophase.

Animals↗
PubMed1999

Agonist- and antagonist-induced plasticity of rat 5-HT1A receptor in hippocampal cell culture.

We examined the response and regulation of 5-HT1A receptor on hippocampal cultured fetal neurons grown in the absence of serotonin and steroids using three experimental designs: 1) functional response using an antibody against phosphorylated cyclic adenosine monophosphate response element binding protein (pCREB); 2) transcriptional regulation using in situ hybridization; and 3) translational expression using antipeptide 5-HT1A receptor antibody. Pretreatment of cultured hippocampal cells with the agonist 8-hydroxy-2-(di-N-propylamino)-tetralin (8-OH-DPAT) (10(-8) M) or ipsapirone (IPS) (10(-9) M) for 10 min blocked the forskolin-stimulated increase in pCREB immunoreactivity. In situ hybridization radioautography revealed that IPS (10(-9) M) decreased the 5-HT1A receptor mRNA expression (-33%) after a 24-h treatment. The decrease in 5-HT1A receptor mRNAwas accompanied by a change in protein immunoreactivity using a 5-HT1A receptor antipeptide antibody. Computer-assisted morphometric analyses showed a reduction in the 5-HT1A receptor immunoreactive (IR) intensity as compared to control 24 h after treatment with 8-OH-DPAT (10(-7)-10(-12) M) and IPS (10(-9) M). Thus, fetal hippocampal neurons have a functional 5-HT1A receptor that is downregulated at both the transcription and translation levels. In addition, we found increased 5-HT1A receptor-IR intensity (+17% approximately +39%) 24 h after treatment with the antagonist N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl) cyclohexane carboxamide (WAY 100635) (10(-7)-10(-12) M). Our results indicate that the 5-HT1A receptor is sensitive to both agonists (downregulation) and antagonists (upregulation) in hippocampal fetal neurons grown in the absence of serotonin and steroids.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1980

A study of drug-induced kinetic perturbations in the marrow of a patient with neuroblastoma.

The kinetic perturbations induced by vincristine, cyclophosphamide, and adriamycin were studied in the bone marrow of a patient with neuroblastoma. Serial marrow samples were studied by dual parameter (Coulter volume and DNA content), flow cytometry, and radioautography, and findings were correlated with cytomorphologic changes. Multiparameter studies were useful in sorting out the various sequences of drug-induced kinetic changes. Distinctive effects of vincristine were observed at 2 and 24 hours after drug administration. An early drug-induced proliferative response was observed between 2 and 48 hours after initiation of therapy. Cell damage was apparent on day 4, with recovery in cell proliferative rate observed on days 6 through 9. The kinetics of perturbation may be useful in identifying temporal windows for optimum scheduling of chemotherapeutic agents in man.

Adolescent↗
PubMed1980

DNA synthesis of benign human breast tumors in the untreated athymic "nude" mouse. An in vivo model to study hormonal influences on growth of human breast tissues.

DNA synthesis in 33 human benign breast tissue biopsy specimens (HBT) was determined by 3H-thymidine radioautography 0, 15, 30, and 60 days after s.c. transplantation to female nude mice. A third of the grafts was removed from each mouse at each time period. Grafts and 0-day slices were exposed to 3H-thymidine in vitro. Labeling indices (LI) [number of radiolabeled epithelial cells per unit area epithelium (0.1 mm2)] were calculated for 0-day slices and grafts from each mouse at each time period. The LI of high 0-day HBT grafts (LI greater than 15) decreased significantly between 0 and 15 and 0 and 30 days. The LI of low 0-day HBT grafts (LI less than 15) increased significantly at 15 days then decreased to 0-day levels at 30 days. No significant differences in LI of HBT grafts were observed between 30 and 60 days. Thus, a model has been proposed for studies of factors effecting the growth of human breast tissues transplanted to nude mice in which (1) each mouse is used as its own control by measuring DNA synthesis of grafts before and after treatment, and (2) the treatment is initiated 30 days or more after transplantation of breast tissues when graft DNA synthesis is stabilized. Using this model, we have observed marked increases in HBT graft DNA snythesis induced by human placental lactogen treatment of host nude mice from day 42 to day 60 after HBT transplantation.

Adenofibroma↗
PubMed1981

Regulation and localization of lymphocyte production in the bone marrow.

The homeostatic mechanisms which control B lymphocyte renewal in the bone marrow are unknown. Mouse bone marrow produces many small lymphocytes which develop surface IgM and other B lymphocyte properties. Putative precursors show cytoplasmic mu chains but earlier progenitors have been characterized. Some marrow small lymphocytes are long-lived recirculating B and T cells. [3H]Thymidine and IgM labelling in femoral marrow sections suggest that recirculating lymphocytes migrate mainly through the marrow periphery while indigenous lymphocytes may be formed peripherally and migrate centrally as they mature. Thus, the localization of lymphocytes appears to be non-random. The effects of possible regulatory factors on bone marrow small lymphocytes production have been examined by [3H]thymidine labelling and radioautography. Administration of anti-IgM antibodies in vivo eliminates all B lymphocytes but the marrow lymphocyte production rate remains unchanged. After sublethal X-irradiation the marrow shows an over-shoot B lymphocyte production, while the lymphocyte numbers in shielded marrow remain stable. In neonatally thymectomized or congenitally athymic mice marrow lymphocyte production is unaffected. Studies in germ-free and antigen-stimulated mice reveal a basal level of marrow lymphocyte production, normally stimulated non-specifically by environmental factors. Thus, marrow lymphocyte production appears to be basically independent of feedback control from the peripheral B lymphocyte pool or of specific humoral factors, but fluctuates widely after perturbation or when amplified by exogenous stimuli. These findings suggest the importance of microenvironmental factors, as yet undefined, in the regulation of bone marrow lymphocytes.

Animals↗
PubMed1975

Fetal monkey absorption of 55Fe from amniotic fluid.

A single dose of 0.5 mCi radioiron (55Fe) was injected directly into the amniotic cavity of each of four pregnant monkeys near term. At 6 or 24 hours after 55Fe adminstration, samples of fetal jejunum, ileum, blood, spleen, liver, lung and fetal membranes were taken for radioautography and scintillation counts. Counts were also made on corresponding maternal tissues. Intestinal radioiron levels and localization indicated that fetuses swallowed and absorbed 55Fe within 6 to 24 hours after injection. Silver grains over the meconium bodies of fetal ileum suggested that intestinal epithelium retained at least some of the absorbed radioiron. However, elevated 55Fe levels in fetal blood and hematopoietic organs indicated that transport of radioiron to the circulation had occurred, presumably through fetal intestine, extraembryonic membranes and/or fetal lungs. Transfer of 55Fe across the placenta from fetus to mother occurred at a slow rate, i.e., samples of maternal organs obtained at 24 hours gave low counts.

Amniotic Fluid↗
PubMed1975

Morphological effects of a large single dose of cycloheximide on the intestinal epithelium of the rat.

Adult male rats received 15 mg/kg cycloheximide and the subsequent morphological effects at three and six hours after injection were evaluated using histometry, light and electron microscopy, histological demonstration of terminal web and acid phosphatase, and radioautography with tritiated thymidine. Rapid atrophy of the villi took place, progressing from the villus tip by premature exfoliation of epithelial cells. The crypts also diminished by random exfoliation of many crypt cells and by partial or complete disintegration. Mitosis and epithelial cell migration were absent. By six hours, the area occupied by the villi and the crypts per unit length of histological section was decreased by about 70-90% in most of the small intestine but only by about 40-60% in the duodenum and the terminal ileum. In the upper half of the villi, the epithelium was strongly positive for acid phosphatase and contained large numbers of round bodies resembling primary lysosomes. In the lower half, the microvillous border and terminal web were found to be disrupted. Animals receiving only 5 mg/kg cycloheximide also showed the atrophy of villi and crypts, and the round bodies resembling lysosomes. Evidence from several sources has indicated that protein synthesis in normal villus epithelial cells subsides toward the villus tip and becomes minimal at exfoliation. At exfoliation, proteins responsible for epithelial cohesion probably fail because they are no longer replenished. Cycloheximide appears to accelerate this process.

Animals↗
PubMed1977

Morphological aspects of type II alveolar pneumocytes following treatment with puromycin in vivo.

Ultrastructural modifications of type II pneumocytes (PNM-II) in mice were analysed 125 and 155 minutes after puromycin treatment (12 mg/100 gm at 0, 30, 60 and 90 minutes). A quantitative evaluation of the cell compartments was carried out and the inhibition of protein synthesis in PNM-II was monitored by light microscopic radioautography, following 3H-leucine injection. In electron micrographs, following a 125-minute puromycin treatment, the number and size of lamellar bodies, the precursors of lung surfactant material appeared markedly reduced. The multivesicular bodies (MVB), which are normally very frequent in PNM-II, had almost completely disappeared, as had composite bodies. Golgi saccules were dilated, while the area occupied by Golgi vesicles was enlarged. Observations following the 155-minute puromycin treatment showed a strong enhancement of these modifications. Smooth and coated vesicles of the Golgi area, as well as peroxisomes, did not appear modified by puromycin. Elongated zones of autophagy were more prevalent after 125-minute treatment than after the 155-minute one. Small bodies were frequently observed in the cytoplasm, near the Golgi zone. They were bounded by a smooth membrane and contained tiny vesicles and/or electron-dense lamellae similar to those present within the lamellar bodies. Parallel membranes formed folds, some of them in continuity with lamellar bodies, thus encircling portions of cytoplasm. These structures, which were few in number in controls, were very frequently observed in treated cells, mainly after the 125-minute treatment. These extensive alterations of PNM-II morphology appeared to be related to a disturbed production of pulmonary surfactant.

Animals↗
PubMed1979

Increase in free cholesterol content of the adrenal cortex after stress: radioautographic and biochemical study.

The purpose of this investigation was to quantify free cholesterol biochemically and in radioautographs of 3H-digitonin cholesterol complex in fasciculata cells of control and stressed rat adrenal cortex. Stress was induced by ether, laparotomy, and adrenal and intestinal handling. Control rats were anesthetized with nembutal. All animals were killed ten minutes from the beginning of anesthesia. The adrenals were excised and either fixed in glutaraldehyde containing 3H-digitonin or homogenized for biochemical determination of free cholesterol. The plasma corticosterone level of each animal was measured. The fixed adrenals were processed, using different methods of dehydration and embedment, for light and electron microscopic radioautography. The mean number of silver grains (mean) per unit area of zona fasciculata was counted from light microscopic radioautographs. Crystals of cholesterol-digitonide complex were more numerous in stressed fasciculata cells, particularly over SER. Silver grains were localized over or close to the crystals. The mean for stressed rats was significantly higher than control values, indicating more free cholesterol in fasiculata cells of stressed rats. The results were not affected by either the method of dehydration or the type of embedding medium used. The morphologic results were substantiated by biochemical findings of increase in free cholesterol in adrenals of stressed rats. Plasma corticosterone was significantly high in stressed rats. The increase in free cholesterol in stimulated fasciculata cells is consistent with a previously reported increase in cholesterol esterase activity after ACTH stimulation.

Adrenal Cortex↗
PubMed1980

Specific uptake of tritiated serotonin in the adult rat pancreas: evidence for the presence of serotonergic fibers.

Fragments of adult rat pancreas were incubated in vitro with tritiated serotonin at concentrations from 10(-8) to 10(-7) M. The pancreas exhibited an uptake of serotonin which was saturable, with an uptake constant (Km) of 8.75 x 10(-7) M, and a Vmax of 873 pmoles per gram. Specificity was determined by the addition of fluoxetine or norepinephrine to the reaction mixture, both at 10(-5) M. Fluoxetine significantly reduced the 3H-5HT uptake, whereas norepinephrine was without effect. Metergoline (10(-6) M), a specific 5-HT postsynaptic receptor blocker, similarly had no effect on the serotonin uptake in the pancreas. Radioautography of the fragments following uptake of tritiated serotonin (5 x 10(-8) M) revealed silver- grain aggregates dispersed along blood vessels in the interstitial spaces of the exocrine and endocrine pancreas, areas known to be traversed by nerve fibers. There were no silver- grain aggregates over the exocrine or islet parenchymal cells. These data support the hypothesis that the pancreas is innervated by serotonergic fibers. Further evidence for this hypothesis was provided by a preliminary study demonstrating the presence of tryptophan hydroxylase in pancreatic homogenates. These serotonergic fibers may be involved in the regulation of pancreatic secretion.

Animals↗
PubMed1981

The stem-cell zone of the small intestinal epithelium. I. Evidence from Paneth cells in the adult mouse.

Stem cells in the small intestinal epithelium are known to differentiate into columnar, mucous, enteroendocrine, and Paneth cells. However, the site of initiation of stem-cell differentiation has been unknown. To approach this problem we determined the site of stem-cell differentiation along the Paneth cell line, using light microscopic morphometry and radioautography. The smallest Paneth cells containing the smallest granules were in positions 6 and 7, while the largest ones containing the largest granules were in positions 1 and 2 at the base of the crypt. Paneth cell death was less prevalent above position 3 than it was in position 1. Since cell size, granule size, and cell death are indicators of Paneth cell age, it was deduced that there is a gradient of Paneth cell age in the crypt base, with the oldest Paneth cells at the bottom, and the youngest at the top. After single injection or continuous infusion of 3H-thymidine, the first labeled Paneth cells to appear were the highest Paneth cells in their crypt column. Later, labeled Paneth cells became more prevalent in lower positions, and, eventually, appeared in position 1. The size of granules in labeled Paneth cells increased with time. It was concluded that Paneth cells originate in position 5 or above and then migrate downward. These results are consistent with a stem-cell zone hypothesis, which proposes that stem cells in positions 1-4 receive no inducement to differentiate. Only those stem cells that migrate up out of the stem-cell zone into position 5 will be induced and then begin to differentiate.

Animals↗
PubMed1981

The stem-cell zone of the small intestinal epithelium. III. Evidence from columnar, enteroendocrine, and mucous cells in the adult mouse.

In the first two articles of this series we demonstrated restriction of Paneth cell formation to positions 5 and above. Restriction was independent of the Paneth cell population-density gradient in the crypt base. We concluded that our results were consistent with the presence in the adult of a stem-cell zone in positions 1-4 in which stem cells received no inducement to differentiate. To further test the stem-cell zone hypothesis we determined the site of stem-cell differentiation along mucous, enteroendocrine, and columnar cell lines using radioautography with 3H-thymidine as a label. One hour after injection of 3H-thymidine, labeled mucous cells were not observed below position 5. Only later did they appear in lower positions and not until 4 days after injection were they observed in position 1. Labeled enteroendocrine cells first appeared above, and then were seen in the top of, and finally in the middle and bottom of the Paneth cell distribution. Thirty hours after injection there were two populations of labeled columnar cells in the crypt base, a heavily labeled population and a lightly labeled one. At this time interval the heavily labeled columnar cells were only observed in positions 5 and above, but they appeared in positions 1-4 by 66 hours after injection. The above evidence led us to conclude that all differentiated offspring of the common epithelial stem cell originate in positions 5 and above. Most columnar, mucous, and enteroendocrine cells originating in positions 5 and above migrate upward. However some of these cells migrate down. All differentiated cells found in positions 1-4 migrated down from their origin in position 5 or above. We also found that only stem cells proliferate in positions 1-4. We concluded that in the adult, there is a stem-cell zone in positions 1-4 where stem cells are not induced to differentiate and persist as stem cells throughout life.

Animals↗
PubMed1983

Morphological observations on mucus-secreting nongoblet cells in the deep crypts of the rat ascending colon.

In the ascending colon of adult male rats, large accumulations of differentiated, mucous-type cells were observed which formed the deep one-half to one-third of the crypts almost exclusively. Mitotic activity was localized to the midcrypt, to columnar-type cells. The mucous-type cells were designated as "deep crypt secretory" (DCS), and their histological study was initiated. In the light microscope, the apical cytoplasm was distended with mucous vacuoles which stained differently from goblet cells after using periodic acid-Schiff (PAS), alcian blue, or a Masson stain. In electron-micrographs, a well-developed Golgi complex and rough edoplasmic reticulum in addition to a large number of electron-opaque vacuoles indicated active production of a glycoprotein. The midcrypt mitotic activity was further demonstrated by using the colchicine technique and 3H-thymidine radioautography. A gradual increase of mucus content and a decrease of nuclear and nucleolar size in the DCS cells from midcrypt to crypt base indicated that these cells originate in midcrypt and mature as they progress toward the crypt base. Cell counts showed that the number of cells constituting the crypts was about the same all along the colon, but the percentage of DCS cells varied: 33% in ascending and 21% in transverse colon. Only 5% and 8% deep-crypt mucous-type cells were found in descending colon and cecum, respectively; but these cells differed in appearance from the mature DCS cells. In conclusion, the presence of a large number of nonmitotic end cells with intense secretory activity has been ascertained for the deep crypts of the ascending colon. The exact relation of these cells to the vacuolated or other epithelial cell types remains for further study.

Animals↗
PubMed1985

Dynamic histology of the antral epithelium in the mouse stomach: II. Ultrastructure and renewal of isthmal cells.

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.

Animals↗
PubMed1985

Dynamic histology of the antral epithelium in the mouse stomach: III. Ultrastructure and renewal of pit cells.

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.

Animals↗