Non-granulated Cells Research Articles

Granulation Rescue and Developmental Marking of Juxtaglomerular Cells Using “Piggy-BAC” Recombination of the Mouse RenLocus

Mice lacking a functional Ren-1(d) gene exhibit a complete lack of renal juxtaglomerular cell granulation and atypical macula densa morphology. Transgenic mice carrying a 145-kilobase BAC clone encompassing the Ren-1(d) and Ren-2 loci were generated, characterized, and backcrossed with Ren-1(d-/-) mice. Homozygous Ren-1(d)-null mice expressing the BAC clone exhibited complete restoration of normal renal structure. Homologous recombination in Escherichia coli was used to generate a modified version of the BAC clone, in which an IRESbeta-geo cassette was inserted specifically into the Ren-1(d) gene. When introduced into the germline, the modified clone provided a marker for juxtaglomerular cell differentiation and beta-geo was expressed appropriately in juxtaglomerular cells throughout development. Parallel backcross experiments onto the Ren-1(d)-null background demonstrated that the juxtaglomerular cells expressed the modified Ren-1(d) locus in the absence of regranulation. These data demonstrate that the nongranulated cells constitute bona fide juxtaglomerular cells despite their altered morphology, that overexpression of renin-2 cannot compensate for the loss of renin-1(d), and that primary structural differences between the two isoforms are responsible for the differences in granulation. The use of BAC modification as part of functional complementation studies illustrates the potential for in vivo molecular dissection of key physiological mechanisms.

Colloid in the pituitary pars distalis of viscacha (Lagostomus maximus maximus): ultrastructure and occurrence in relation to season, sex, and growth.

Randomly distributed extracellular colloidal accumulations were observed in the pars distalis of viscacha (Lagostomus maximus maximus). They were preferentially located in the peripheral zone of the gland and showed variability in shape and size. Two different types of colloidal accumulations were found by electron microscopy: 1) those surrounded by nongranulated follicular cells that correspond to characteristic follicles, and 2) those surrounded by granulated cells. In the follicles lined by nongranulated follicular cells, long, prominent microvilli and cytoplasmic processes protruded into the lumen. The frequency of these accumulations varies during the year in adult male animals, showing an increase in number during summer and a decrease during winter. The lowest value was registered in August (winter). The mean follicular diameter did not vary seasonally. The number of colloidal accumulations did not vary seasonally in adult female viscachas, but a significant difference in the mean follicular diameter between pregnant and non-pregnant females was observed. Pituitaries of immature animals contain fewer colloidal accumulations than those of adults. In fetuses, these accumulations were absent. The administration of melatonin provoked a decrease in the number of these structures. The numeric changes of the colloidal accumulations observed in this study are associated with: 1) the seasonal reproductive activity in adult males, and 2) the reproductive condition, body weight and sexual maturity in males and females. The fact that melatonin administration decreases the population of colloidal accumulations in males suggests participation of the pineal gland in these changes.

Immunohistochemical, morphological and ultrastructural resemblance between dendritic cells and folliculo-stellate cells in normal human and rat anterior pituitaries.

Immunolabeling of cryo-sections of human anterior pituitaries obtained at autopsy, and of cryo-sections of freshly prepared rat anterior pituitaries, with a panel of monoclonal antibodies against markers of the monocyte/dendritic cell/macrophage lineage, reveals in both species a characteristic pattern of immunopositive cells, among which many cells with dendritic phenotype are found. Cells characterized by marker expression of MHC-class II determinants and a dendritic morphology are present in both human and rat anterior pituitary. Markers characteristic of dendritic cells such as the L25 antigen and the OX62 antigen were present in anterior pituitaries from human and rat respectively. The population of MHC-class II expressing dendritic cells of the rat anterior pituitary is compared at the ultrastructural level with the folliculo-stellate cell population, which cell type has been previously characterized by its distinctive ultrastructure and immunopositivity for the S100 protein. Using immuno-electron microscopy of rat anterior pituitaries fixed with periodate-lysine-paraformaldehyde, we were able to distinguish non-granulated cells expressing MHC-class II determinants, whereas no MHC-class II expression was found in the granulated endocrine cells. Using double immunolabeling of cryo-sections of these rat AP with 25 nm and 15 nm gold labels, we demonstrated an overlap between the populations of MHC-class II-expressing and S100 protein-expressing cells. Furthermore, MHC-class II-expressing and S100-positive cells showed ultrastructural characteristics that have been previously ascribed to folliculo-stellate cells. At the light microscopical level in the rat AP, a proportion of 10 to 20% of the S100-positive cells was found immunopositive for the MHC-class II marker OX6. In the human AP, S100-positive folliculo-stellate cells and cells expressing the leukocyte common antigen CD45 were found to occupy predominantly different tissue compartments in the human anterior pituitary, namely the epithelial parenchyme cords and perivascular compartments respectively. A proportion of CD45+ cells was found in the parenchyme compartment and, vice versa, indicating an overlap of the tissue compartments in which both cell types occur. However, at the light microscopical level we could not find cells expressing both the S100 and CD45 marker. The present finding of a proportion of S100-positive pituitary cells with ultrastructural and immunohistochemical characteristics of both dendritic cells and folliculo-stellate cells, confirms the suggested heterogeneity of the latter cell group with respect to their ultrastructural phenotype and putative function. The possibility of a myeloid origin of part of the folliculo-stellate cell group in the AP, is discussed and might elucidate some of the discrepancies in the literature concerning the embryological origin of this cell group.

Non-granulated peripolar cells exist in the rat glomerulus

The peripolar cell is a unique cell type in the mammalian glomerulus. Peripolar cells are said to be identifiable during light microscopy by their cytoplasmic granules and by their position at the vascular pole; and during scanning electron microscopy by their distinctive surface morphology. We used both techniques to count peripolar cells in 6 normal rat kidneys. Scanning microscopy revealed that 55(+/- 5)% of glomeruli contained at least one peripolar cell whereas light microscopy revealed granulated peripolar cells in only 4(+/- 2)% of glomeruli. Vascular poles which contained peripolar cells previously identified by scanning were then examined by light and by transmission electron microscopy. Serial sections through these peripolar cells demonstrated the absence of cytoplasmic granules. Our observations suggest that the majority of peripolar cells in the rat contain no granules.

Expression of c-kit by mesenteric lymph node cells from Nippostrongylus brasiliensis-infected mice and by mast cell colonies developing from these cells in response to 3T3 fibroblast-conditioned medium.

Mast cell committed progenitors are nongranulated cells found in mesenteric lymph nodes of mice infected with Nippostrongylus brasiliensis (Nb-MLN) but not from normal mice. Mast cell committed progenitors can respond to either IL-3 or to a factor(s) present in 3T3 fibroblast conditioned media (F-CM) by formation of mast cell colonies. Previous studies from ours and other laboratories suggested that mast cell differentiation involved the W allele product, c-kit, as a receptor and Sl allele product, stem cell factor, as a growth factor. We report here that Nb-MLN cells, which can respond to F-CM by mast cell colony formation, also contain cells that express message for c-kit, and that c-kit message cannot be detected in naive mesenteric lymph node cells, which cannot respond to F-CM. Antisense oligonucleotides to c-kit inhibit mast cell colony formation by Nb-MLN cells in response to F-CM, but not to conditioned medium of PWM-stimulated spleen cells as a source of IL-3. The antisense oligonucleotides also inhibit the degree of granulation by mast cells derived from culture. The results suggest that c-kit and its ligand, stem cell factor, are necessary for mast cell-committed progenitors to proliferate and granulate in response to F-CM but not IL-3.

Folliculostellate cells of the pituitary.

In addition to the hormone-producing granulated cells, the so-called folliculostellate (FS) cells of the adenohypophysis represent a population of nongranulated cells extensively described in a large number of species. They show distinctive morphological features including a star shape with thin cytoplasmic projections extending between granulated cells and well-developed junctional complexes. FS cells are joined together surrounding irregular microcavities and project microvilli into the lumina. The immunocytochemical localization of S-100 protein, glial fibrillary acidic protein, and vimentin constitutes a reliable and easy method for investigating their presence and distribution in the normal pituitary gland and in pituitary adenomas. Although the expression of glial cell markers raised the hypothesis of a neuroectodermal origin of FS cells, most evidence supports that they derive from the epithelium of the Rathke's pouch, as do granulated adenohypophyseal cells. Morphological studies indicate that FS cells are involved in phagocytosis and possess sustentacular functions. Investigations using cell cultures show that FS cells play important roles in the paracrine regulation of adenohypophy-seal secretion by their ability to liberate several growth factors and regulate the ionic composition of the extracellular fluid. Further research using novel immunocytochemical markers and ceil culture techniques may clarify the origin and the role of this enigmatic cell type in the normal and pathological pituitary gland.

Ultrastructural morphometry of gastric endocrine cells before and after omeprazole

Abstract Long-term toxicological experiments with inhibitors of acid secretion were found to induce hyperplasia and eventually carcinoid tumors of the enterochromaffinlike cells of the oxyntic mucosa. To evaluate the effects of 6 months' treatment with omeprazole in humans, the oxyntic endocrine cells were morphometrically investigated at the ultrastructural level in five patients with active duodenal ulcer. No omeprazole-induced changes were found in the volume density of the total endocrine cell population and specific cell types (including the enterochromaffinlike cell) as well as in the other cytological parameters investigated (number of cell profiles per unit area, mean cross-sectional area of cell profiles, nuclear-cytoplasmic ratio, and density of cytoplasmic secretory granules). Both pretreatment and posttreatment values in our patients with duodenal ulcer significantly differed from those of a previous investigation of healthy volunteers with regard to the volume density of enterochromaffinlike cells and nongranulated cells, which increased, and of D cells, which markedly decreased. The latter result may provide a cellular basis for impairment in the paracrine release of fundic somatostatin in peptic ulcer disease. Finally, morphometric data on endocrine cell volume density provided by electron microscopy were found to correlate with those obtained in the same patients using light microscopy techniques (Grimelius silver impregnation and chromogranin A immunostaining). It is concluded that 6 months' treatment with pharmacological doses of omeprazole is devoid of appreciable trophic effect on endocrine cells of human oxyntic mucosa.

Ultrastructural morphometry of gastric endocrine cells before and after omeprazole: A study in the oxyntic mucosa of duodenal ulcer patients

Long-term toxicological experiments with inhibitors of acid secretion were found to induce hyperplasia and eventually carcinoid tumors of the enterochromaffinlike cells of the oxyntic mucosa. To evaluate the effects of 6 months' treatment with omeprazole in humans, the oxyntic endocrine cells were morphometrically investigated at the ultrastructural level in five patients with active duodenal ulcer. No omeprazole-induced changes were found in the volume density of the total endocrine cell population and specific cell types (including the enterochromaffinlike cell) as well as in the other cytological parameters investigated (number of cell profiles per unit area, mean cross-sectional area of cell profiles, nuclear-cytoplasmic ratio, and density of cytoplasmic secretory granules). Both pretreatment and posttreatment values in our patients with duodenal ulcer significantly differed from those of a previous investigation of healthy volunteers with regard to the volume density of enterochromaffinlike cells and nongranulated cells, which increased, and of D cells, which markedly decreased. The latter result may provide a cellular basis for impairment in the paracrine release of fundic somatostatin in peptic ulcer disease. Finally, morphometric data on endocrine cell volume density provided by electron microscopy were found to correlate with those obtained in the same patients using light microscopy techniques (Grimelius silver impregnation and chromogranin A immunostaining). It is concluded that 6 months' treatment with pharmacological doses of omeprazole is devoid of appreciable trophic effect on endocrine cells of human oxyntic mucosa.

Ultrastructure of the adenohypophysis in the anadromous sea lamprey, Petromyzon marinus, during metamorphosis.

The ultrastructural changes occurring in the adenohypophysis (AH) of the anadromous sea lamprey, Petromyzon marinus, during metamorphosis (stages one through seven) were examined. The rostral pars distalis initially contains one granulated (secretory) cell type A and one nongranulated type I cell. A second granulated cell (type B) appears during the later stages (stages six and seven) of metamorphosis. The most pronounced ultrastructural changes take place in the caudal pars distalis (CPD). Initially, most cells (80-90%) are nongranulated cells type II and some type I. Granulated type C and D cells form the remainder of the CPD. Almost all cells during stages three and four demonstrate a marked increase in synthetic activity evident by conspicuous Golgi regions, abundant rough endoplasmic reticulum (RER), and increased cell volume. Most cells are sparsely granulated. Secretory cell types C and D and, two new cell types, E and F, are present. Synthetic activity subsides by stage five. Most cells (80-90%) during stages five through seven are granulated. Type E are most prevalent with variable numbers of types C and D and few type F. Nongranulated cells now represent only 10-20% of the CPD. The increase in granulated cells occurs at the expense of type II cells that differentiate into granulated cell types. The fine structure of the pars intermedia throughout metamorphosis remains similar to that of the larva. Most cells are granulated, highly vesiculated type G cells. A few nongranulated type I cells are also present. The functional significance of the secretory cells in the AH is related to the requirement for an intact pituitary gland for the initiation and completion of metamorphosis.

The mast cell-committed progenitor. I. Description of a cell capable of IL-3-independent proliferation and differentiation without contact with fibroblasts.

We have identified a late, committed stage in the differentiation of the mast cell progenitor just before granulation. Mast cell committed progenitors (MCCP) are nongranulated cells with a density of 1.060 to 1.070 g/ml which can be harvested from the mesenteric lymph node of mice infected with Nippostrongylus brasiliensis. Mast cell-committed progenitors are able to proliferate and differentiate in the absence of IL-3 or IL-4 when cultured on a monolayer of embryonic skin or 3T3 fibroblasts and can form colonies in methylcellulose supplemented with fibroblast conditioned medium. Fibroblast conditioned medium appears to contain a soluble MCCP proliferation factor that maintains biologic activity when heated to 56 degrees C for 45 min but is destroyed by incubation with either trypsin or chymotrypsin. It can be selectively precipitated with 60 to 70% saturated ammonium sulfate. The factor is not absorbed by immobilized antibodies to nerve growth factor. The MCCP proliferation activity of the factor could not be mimicked by IL-1, IL-2, IL-4, granulocyte-macrophage-CSF, granulocyte-CSF, macrophage-CSF, IFN-alpha/beta, IFN-gamma, nerve growth factor, epidermal growth factor, serum fibronectin, heparin, or a number of glycosaminoglycans. At high salt concentrations, the factor passes through a 50-kDa membrane and can be concentrated above a 5-kDa membrane. MCCP acquire a connective tissue phenotype when cultured on a fibroblast monolayer and a mucosal phenotype when cloned in the presence of conditioned medium from PWM-stimulated spleen cells. When cultured in the absence of IL-3 on a monolayer of embryonic skin or 3T3 fibroblasts, mast cell-committed progenitors produce mast cells which stain with berberine sulfate suggesting a connective tissue phenotype; however, the mast cells that develop when mast cell-committed progenitors are cultured in the presence of IL-3 or conditioned media from PWM-stimulated spleen cells do not stain with berberine sulfate. MCCP intercalate into monolayers of embryonic skin or 3T3 fibroblasts, but T cells are not able to associate with the monolayer and can be completely washed away. Attempts to enrich mast cell-committed progenitors by intercalation and elution from embryonic skin monolayers proved unsuccessful, but some enrichment of mast cell-committed progenitors could be achieved by discontinuous Percoll gradients. Thus, we have identified a way to obtain late-stage, mast cell-committed progenitors in an environment that is virtually uncontaminated with other hematopoietic progenitors.

Expression of leucocyte antigens by cells from the metrial gland of the pregnant rat

The function of the metrial gland of the rat, and particularly of its characteristic population of granulated cells, remains unknown. However, several lines of evidence suggest that the granulated cells may derive from lymphocytes, and play a role in the immunology of pregnancy. In this study, antigen expression by granulated and other cell populations from the metrial glands of rats at Days 13 and 14 of pregnancy was studied by an indirect immunoperoxidase method. Acetone-fixed frozen sections, and cytocentrifuge preparations of collagenase-dispersed metrial gland tissue in which numbers of granulated cells had been increased by density-gradient centrifugation, were used. The primary antibodies used recognised, inter alia, B lymphocytes (MRC OX-3, MRC OX-6, MRC OX-12), T lymphocytes (MRC OX-8, W3/25, MRC OX-19), neutrophils (MRC OX-42) and cells of the monocyte/macrophage series (MRC OX-3, MRC OX-6, MRC OX-42, MRC OX-43). The majority of the granulated cells, including smaller, "immature" forms, were unlabelled by any of these antibodies. Some lymphocytes, and varying numbers of larger, non-granulated cells, were labelled by OX-6, OX-12, W3/25, OX-42 and OX-43. In addition to lymphocytes, labelled cells included neutrophils (OX-42), endothelial cells (OX-43), and probably some macrophages (OX-6, OX-43). OX-12, which recognises the kappa chain of rat IgG, labelled some large cells which may have been stromal cells. These findings do not support the concept that the granulated cells are derived from lymphocytes.

Changes with age in the number and size of anterior pituitary cells in female mice from suckling to adulthood.

Changes with age in the number and size of anterior pituitary cells in female mice were calculated during their postnatal development by using a stereological morphometric study with electron microscopy. The number of parenchymal cells increased in mice from 20 to 30 days of age, and did not change around puberty, after which the number increased to the adult level. The number of somatotrophs increased with age in almost the same manner as the parenchymal cells. The number of lactotrophs increased with age and were significantly different each time they were measured. The number of non-granulated cells did not increase in mice from 20 days of age to adulthood; at 20 days of age, the number was at the same level as in the adult mice. The other types of cells increased by only a small number. The sizes of all types of cells increased during postnatal life. Somatotrophs and lactotrophs became the same size as in adults by the onset of puberty. Non-granulated cells and other types of cells reached adult size at 5 days after puberty. Lactotrophs and somatotrophs had adult ultrastructural features on the day of puberty. Sizes and ultrastructural features of anterior pituitary cells reached adult levels on the day of puberty, but their numbers were still fewer than in adult mice. The increase in the volume of the anterior pituitary with age arose mostly from an increase in the number and the size of somatotrophs and lactotrophs before puberty, increases in the size of somatotrophs and the number of lactotrophs around puberty, and an increase in the number of both types of cells after puberty.

Fine-structural and immunohistochemical study of anterior pituitary cells of Snell dwarf mice

Snell dwarf mice display remarkable retardation of growth after birth and are known to lack prolactin (PRL), thyroid stimulating hormone (TSH) and growth hormone (GH). The aim of this study was to determine the reason for these hormonal deficiencies. We examined the fine structure of the gland and its immunohistochemical staining pattern with respect to antisera raised against PRL, TSH, GH, adrenocorticotrophic hormone (ACTH) and luteinizing hormone (LH). The gland of control mice reacted immunohistochemically against all antisera used, whereas only ACTH-producing cells (ACTH cells) and LH-producing cells (LH cells) were distinguished in the dwarf mice. ACTH cells in dwarf mice varied in cell shape, although they were similar in size to those of controls. The distribution of secretory granules in the cytoplasm varied from cell to cell. LH cells in the dwarf mice showed immature features, having poorly developed rough endoplasmic reticulum and Golgi apparatus. The cells were about half the size of controls, and secretory granules were smaller. In dwarf mice, non-granulated cells were encountered in addition to granulated ACTH and LH cells. Some of them formed small clusters, characteristic cell junctions being found between the cells; they thus appeared to be follicular cells. The above results suggest that hormone deficiency in Snell dwarf mice is a result of a defect in the hormone-producing cells in the gland.

Immunohistochemical study of Rathke's cleft cyst.

An immunohistochemical study was made of ten cases of asymptomatic and three cases of symptomatic Rathke's cleft cyst. The cysts in the asymptomatic cases had monolayer columnar or cuboidal epithelium. Within the epithelium, cells which were positive for at least one of the pituitary hormones were found. The rate of positive reactions for these six pituitary hormones was between 70% and 100%. In contrast, the cysts in the symptomatic cases had an epithelium which was partly stratified squamous and partly squamous epithelium, and none of the pituitary hormones were found in them. Therefore, when a Rathke's cleft cyst enlarges to the extent that clinical symptoms are produced, we consider that changes have already occurred in structure and function of the cyst epithelium. In addition, we believe there is a tendency for monolayer epithelia to undergo squamous metaplasia and for cells which are positive for pituitary hormones to change into non-granulated cells.

Marginal and Folliculo-Stellate Cells of the Pituitary Gland of the Rat

We examined non-granulated pituitary cells (folliculo-stellate cells and anterior and posterior marginal cells of the pituitary cleft) in lactating and virgin female rats by means of electron microscopy and morphometry. Ultrastructure and morphometric parameters of the cells lining the pituitary cleft were similar in the two groups of animals. On the contrary, folliculo-stellate cells showed a marked nucleocytoplasmic activation in lactating rats in the electron microscope, which was confirmed by morphometric measurements. These results are not consistent with the hypothesis that the three cell types share the same reactivity during pituitary hyperfunction and that they have a common function, as suggested by purely morphological studies in various endocrine conditions. We believe that quantitation of cell response during such stimuli could be useful to further elucidate this matter.

An Ultrastructural Morphometric Analysis of Elastase-tieated Hamster Bronchi Shows Discharge Followed by Progressive Accumulation of Secretory Granule

An ultrastructural morphometric analysis of bronchial secretory cells was carried out on hamsters treated intratracheally with 300 micrograms of human neutrophil elastase (HNE) in 0.5 ml saline, saline alone, or left untreated. Five to 6 animals were killed at 2 h and at 3, 8, and 16 days after treatment. Electron micrographs were prepared from the hilar region of the left main intrapulmonary airway; 125 +/- 16 (mean +/- SE) granulated secretory cells extending from basement membrane to lumen were analyzed for each group. The number (Ng) and area (Ag) of granular profiles per cell, the area of cell profiles (Ac), and the volume density of secretory granules per cell (Vv) were determined using an electronic image analyzer. There were significant decreases in Ng, mean Ag, and Vv in the 2-h HNE group when compared with the saline group. Values of Ng, mean Ag, and Vv were similar for HNE and saline groups at 3 days, but were significantly increased at 8 and 16 days. The Ac of HNE-treated groups was similar to their saline control groups at all time points except at 16 days when the HNE-treated group enlarged to double that of its saline control group. An ultrastructural differential cell count showed a decrease in frequency of granulated secretory cells at 2 h and an increase at 8 and 16 days; there was an inverse change in the frequency of nongranulated secretory cells at these times. The proportion of ciliated, preciliated, and indeterminate cells remained constant over time in all treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Roles of the arcuate nucleus and ovary in the maturation of growth hormone, prolactin, and nongranulated cells in the mouse adenohypophysis during postnatal development: a stereological morphometric study by electron microscopy.

With the use of monosodium L-glutamate (MSG) treatment and ovariectomy, we studied how the arcuate nucleus and ovary are involved in the maturation of GH, PRL, and nongranulated cells in the anterior pituitary lobe during postnatal development. From day 1 to day 7 female pups were injected sc daily with MSG (2 mg/g BW) (MSG mice) or distilled water (control mice). Just before puberty (at 30 days of age) some control and MSG mice were ovariectomized (OVX) (control + OVX mice and MSG + OVX mice, respectively). All animals were killed at 60-70 days of age by decapitation. Populations of GH, PRL, and nongranulated cells in the anterior pituitary lobe were calculated by applying morphometric procedures. Percentages of GH, PRL, and nongranulated cells in MSG mice were not different from those of control mice. In control + OVX mice the percentage of GH cells was greater than that of control mice, while PRL cells were markedly decreased. On the other hand, in MSG + OVX mice the percentage of PRL cells was less than that of MSG mice, while GH cells showed no difference; nongranulated cells were increased. The number of parenchymal cells in the anterior pituitary lobe in MSG mice was approximately two thirds that of control mice. The parenchymal cells in control + OVX mice were the same in number as those in control mice, while those in MSG + OVX mice were approximately half the number of the parenchymal cells in MSG mice. Numbers of GH and PRL cells in MSG mice were half those of control mice. There was no difference in the number of nongranulated cells between MSG and control mice. GH cells in control + OVX mice were more abundant than those of control mice, while PRL cells were fewer. On the other hand, both GH and PRL cells in MSG + OVX mice were fewer when compared with those of MSG mice. The numbers of nongranulated cells in most groups showed no significant differences among each other. All of these data taken from control mice were the same as those taken from normal mice. These results suggest that the arcuate nucleus in the hypothalamus stimulates the differentiation of GH and PRL cells during postnatal life, and that the ovary stimulates the differentiation of PRL cells through a direct effect on the anterior pituitary lobe but inhibits that of GH cells via the arcuate nucleus in the hypothalamus, while the differentiation of nongranulated cells is little affected.

Frequency of mast cell precursors in normal tissues determined by an in vitro assay: antigen induces parallel increases in the frequency of P cell precursors and mast cells.

Persisting (P) cells, homogeneous populations of cells that grow in vitro for prolonged periods provided a specific growth factor is present, resemble mast cells in many respects. An in vitro assay based on limit dilution was used to determine the frequency of precursors capable of giving rise to P cells. The incidences of P cell precursors per 10(6) cells in tissues of CBA mice in representative experiments were as follows: bone marrow, 291; spleen, 30; mononuclear blood cells, 11; popliteal lymph node, 0.5; and mesenteric lymph node, 18. P cell precursors appeared to be relatively undifferentiated, non-granulated cells; no cells with metachromatically staining granules were detected in the bone marrow or peripheral blood. Furthermore, mice of the Wf/Wf genotype that were grossly deficient in mast cells had the same frequencies of P cell precursors in bone marrow and spleen as their normal +/+ littermates. In many tissues in which we found P cell precursors, pluripotential hemopoietic stem cells are present. Among nonepithelial cells from the gut mucosa, however, in which there was a 10-fold higher frequency of P cell precursors than in bone marrow cells, pluripotential hemopoietic stem cells were undetectable, indicating the existence of committed P cell precursors distinct from pluripotential hemopoietic stem cells. The frequency of P cell precursors in mesenteric lymph nodes was more than 30-fold higher than in the popliteal lymph nodes, suggesting that antigenic stimulation influences their numbers. This latter notion is supported by the observation that after immunization in the footpad, the number of P cell precursors in ipsilateral popliteal lymph nodes rose about 35-fold. Immunization was also accompanied by a rise in mast cell numbers in draining popliteal nodes. This correlation between P cell precursors and the local production of mast cells was strengthened by the observation that the frequency of P cell precursors in cells from the gut mucosa of mice of Wf/Wf genotype, which are unable to mount an intestinal mastocytosis, was more than 1000-fold lower than in wild type mice. Thus, the precursors of P cells and probably of at least the T cell-dependent subset of mast cells appear to be generated in the bone marrow and seed as non-granulated cells via the blood to peripheral tissues such as spleen, lymph node, and mucosal surfaces. P cells appear to be in vitro counterparts of the mucosal subset of mast cells.

Ultrastructure of the adenohypophysis in the larval anadromous sea lamprey, Petromyzon marinus L.

The ultrastructure of the adenohypophysis (AH) in the larval anadromous sea lamprey, Petromyzon marinus L., was examined. The AH is subdivided into three regions, the pro-, meso-, and meta-AH. Cells of the nasopharyngeal stalk extend directly beneath the pro- and meso-AH to form the ventral surface of the gland. Some cells in the pro- and meso-AH are arranged into small follicles. Each region of the AH is characterized by a single granulated (secretory) cell type. Granulated cells constitute 80-90% of the pro-AH and contain secretory granules that range from 800 to 2400 A in diameter. Only 10-20% of the cells in the meso-AH are granulated and they contain much smaller secretory granules (400 to 1250 A diameter) than those in the pro-AH. Granulated cells constitute 80-90% of the meta-AH and contain only a few secretory granules, ranging from 1000 to 2500 A in diameter, and many vesicles containing either a loose flocculent or dense granular material. Non-granulated (stellate) cells are found in all regions. They are characterized by their long cell processes, abundant cytoplasmic filaments, and variable electron density. The appearance of organelles in these cells suggests they are nonsecretory. They may play a role in maintaining the structural integrity of the gland and the regulation of granule release in the pro-AH. Two types of nongranulated cells make up 80-90% of the meso-AH. Type I are stellate cells, type II may be undifferentiated cells. The functional significance of the secretory cells in the larval AH is discussed.

Stellate cells as phagocytes of the anuran pars distalis.

When the stellate cells (nongranulated cells) from dissociated-cell preparations of the anuran pars distalis were examined, they were seen to contain debris within phagocytic vacuoles (phagosomes). These phagosomes were variable; some contained granules from secretory cells while others were similar to lipid-like bodies and myelin figures. In situ partes distales from frogs were examined at the breeding season. The tissues were divided into lobules that were bounded by processes of stellate cells located between the secretory cells. Processes of stellate cells in the interior of a lobule interdigitate with processes extending inward from the stellate cells forming the border of the lobule. When these processes come together, a small cavity is formed. In many of the intact frogs the spaces between the stellate and secretory cells were greatly enlarged. At this particular time the processes of the stellate cells were attenuated and enclosed secretory granules that were also present as debris in these dilated, intercellular spaces. Within the cytoplasm of these stellate cells were not only phagosomes containing secretory granules but also organelles that appeared to be lipid bodies and lysosomes. Thus, the stellate cells of the pars distalis function in vivo, as well as in vitro, as phagocytes. In addition, macrophage-like cells moving from the blood may form another component of this system of phagocytes.