Age-related changes of the somatotrophs of the domestic fowl Gallus gallus

Summary The ultrastructure of the somatotrophs of the caudal pituitary of the domestic fowl was studied quantitatively. Two age groups of male chickens were compared: 4–6 weeks and 24–30 weeks post-hatching. With age, somatotrophs decreased from about 40% to about 30% of the pituitary cell population. Their volume density decreased similarly. Mean volume of a somatotroph was the same in young and adult animals. Because the granule volume density of the somatotrophs was unchanged, but the somatotroph volume density of the gland declined, the granule volume density of the caudal pituitary gland dropped in parallel with that of the somatotrophs. Thus the volume of the gland comprised of somatotroph granules fell about 32%: from 6.57% to 4.45%. This lowered pool of stored hormone may be linked to the lowered circulating levels of growth hormone found in older animals by other investigators.The granule volume density of the somatotrophs was unchanged but the numerical density approximately doubled; thus the mean granule size decreased by 47% with age. The relationship of the size reduction of the granules to the lowered plasma growth hormone levels is not understood at present.Supported in part by Hatch and State funds from the New Jersey State Agricultural Experimental Station and NSF grants PCM 8,0227,27 and PCM 8,302197.     

Immunocytochemical studies of chicken somatotrophs and somatotroph granules before and after hatching

Immunocytochemical methods were used to gain information about the embryonic development of chicken somatotrophs before and after hatching. To localize growth hormone, anterior pituitary sections were incubated with growth-hormone antibody, and then an indirect peroxidase method was used for light microscopy and an immunogold method for electron microscopy. The earliest evidence of embryonic somatotrophs was seen at 12 days. At this stage somatotrophs were sparse (0.2% of parenchymal cells) and their granules were pleomorphic with elongated ovoid and lozenge shapes predominating. Few of the immunogold-labeled somatotroph granules of the embryo were spherical until 15 days after fertilization. At 18 days, most of the granules were spherical (their shape in the adult chicken). During the six days between the 15-day-old embryo and the 1-day-old chick, the number of gold particles per granule section approximately doubled suggesting an increase in growth hormone content of the granules. This rise was the result of increases in the size of the granule sections and in the concentration of gold particles in the sections. During the embryonic period of 12–20 days, somatotrophs were not more than 3.6% of the anterior pituitary cell population. During the following two days, between the 20-day-old embryo and the 1-day-old chick, the percentage of somatotrophs in the pituitary parenchymal cell population rose rapidly from 3.6% to 20.7% and then increased slowly to 24.6% during the period of 1–5 days after hatching. Both the sharp percentage rise in somatotrophs (20-day-old embryo to 1-day-old chick) and the rise in growth hormone content of the granules (15-day-old embryo to 1-day-old chick) suggested by gold-particle counts occur close to the time of hatching. These morphological changes may reflect an increased synthesis of growth hormone that is responsible for the rise in plasma growth-hormone concentration that begins about the same time and is especially abrupt two days later (1–3 days after hatching).     

Suppression of somatotrophs in growth hormone injected rats

Summary The response of somatotrophs to large doses of purified bovine growth hormone was examined in uninephrectomized rats drinking 1% sodium chloride in an attempt to prolong the action of the hormone. Growth hormone produced a triphasic response in the ultrastructure of somatotrophs. Uninephrectomy and salt alone induced no changes in somatotrophs as compared to unoperated animals consuming tap water. In the first phase (3 days) the cross sectional area of the cell increased, the size of the largest granule per cell as well as the number of granules per cell increased significantly. All these parameters then reached control level after 7 days of injections during a second phase. Suppression of somatotrophs by exogenous growth hormone was evident by 16 days (third phase) when the area of the cell, the size of the largest secretory granule as well as the number of granules per cell all decreased significantly. Somatotrophs apparently did not react to suppression by a lysosomal mechanism inasmuch as no increase in the number of lysosome-derived structures was seen even at 16 days.Supported by research grant HL 06975 from the National Heart and Lung Institute. Dr. Nakayama is the recipient of a Bertha P. and Henry C. Buswell fellowship. The authors are grateful to Mrs. Neonile Fylipiw, Mrs. Elisabeth Lawson, Mr. Luther Joseph, Mrs. Geneva Joseph and Mr. Robert Linsmair for skilled technical assistance.     

Immuno-electron-microscopic study of the prolactin cells in the pituitary gland of male Wistar rats during aging

Summary Prolactin cells were identified by means of immunocytochemistry with protein-A gold as a marker on ultrathin sections of the pituitary gland of young (3–4 months), middle-aged (16–19 months), and aged (26–30 months) male Wistar rats. Point-counting volumetry revealed that the prolactin (PRL) cell-volume density in middle-aged rats was significantly increased in comparison to the volume densities in young and aged rats. Within the PRL-cell population, four types of PRL cells were distinguished on the basis of the shape and size of their secretory granules. During aging, dramatic changes occurred in the relative volumes of the four cell types. The volume percentage of cells with round granules (type I, granule diameter 150–250 nm, and type IIA, granule diameter 250–350 nm) increased from ±30% in young rats to ±90% in old rats. The volume percentage of cells with round and polymorphic granules (type IIB; granule diameter 350–400 nm and type III; granule diameter 500–600 nm) decreased from ±70% in young rats to ±7% in old rats. Age-related changes in serum PRL levels were not found. It is concluded that although during the life span of the male Wistar rat considerable changes in PRL-cell volume densities and in the ratios of PRL-cell types occur serum, PRL levels remain more or less constant.     

Immunocytochemical distribution of somatotrophs in porcine anterior pituitary

The objective of this immunohistochemical study was to identify the spatial distribution patterns of growth hormone (GH) secreting cells (somatotrophs) in the newborn and prepubertal porcine pituitary. No differences were observed among the total somatotrophs per unit area across the three ages. There were, however, changes in spatial distribution of somatotrophs in porcine pituitary with developmental age. Distinctive characteristics of the pattern included a high population of somatotrophs (44±1.2; mean ± standard error of the mean per 30,495 m²) in regions 1 and 5 and a low population (22±1.4) in regions 2 and 4 at each level (P<0.05). Somatotrophs increased 55% in region 3 from proximal to distal levels at all ages. With increasing age, however, somatotrophs in region 3 at the proximal level decreased 33%. From these results, we suggest that there may be regional specificity of cellular differentiation and transformation to facilitate GH secretion to meet the need for endocrine regulation as the animal ages.     

Dynamics of secretory granules in somatotrophs of rats after stimulation with growth hormone-releasing factor: a stereological analysis

The anterior pituitary tissue of male rats injected with growth hormone-releasing factor (GRF) was either processed for stereology at the light-and electron-microscopic levels, or homogenized for growth hormone (GH) assay 2–60 min after GRF injection. Secretory granules of somatotrophs became smaller but increased in numerical density 2 min after GRF injection. Their volume density began to increase at 5 min. The frequency of exocytosis of the granules was most prominent as early as 2 min after GRF injection and reduced thereafter. GH levels in the tissue were lowest at 2–5 min, and returned to the control value by 60 min. Serum GH levels were highest at 15 min; even at 60 min, this value was higher than in the controls. These findings suggest that secretory granules in somatotrophs are stimulated to divide by GRF, resulting in a decrease in size and an increase in number. The discrepancy between the earlier formation of new secretory granules and the later restoration of intracellular GH levels implies that GRF first stimulates the synthesis of constituents of granules other than GH, and only later the synthesis of GH, and that newly formed small secretory granules contain less GH. From the clearance rate of serum GH and the frequency of granule exocytosis, it can be estimated that about a half million granules are released to maintain 1 ng/ml of serum GH in rats.     

Ultrastructure of pituitary somatotrophs of male rats during liver carcinogenesis by 3'-methyl-4-dimethylaminoazobenzene

Somatotrophs of male Wistar rats fed a carcinogenic dose of the potent hepatocarcinogen 3'methyl-4-dimethylaminoazobenzene-(3'MeDAB) in a complete diet for up to 21 weeks, showed ultrastructural changes within one week of the start of the experiment. Morphometric analysis showed that the area of the somatotrophs occupied by hormone storage granules was reduced significantly at all stages up to 21 weeks, when the experiment was terminated. The cell area occupied by rough endoplasmic reticulum (RER) was significantly increased after three weeks and RER surface density increased after eight weeks, whilst granule size was reduced after four weeks. Pituitary content of radioimmunoassayable growth hormone (RIA-GH) was also lower in treated rats than in controls, from one week after the start of the experiment, but only significantly so at eight and twelve weeks, owing to large standard errors. Serum levels of RIA-GH were similarly lower in 3'MeDAB treated rats than in controls. Withdrawal of 3'MeDAB diet after twelve weeks treatment permitted partial recovery of some cell organelles, in that the content of hormone storage granules increased and the area of the RER decreased compared to rats fed 3'MeDAB continuously. There was also a partial, but non-significant, increase in pituitary RIA-GH. The pituitaries of rats pair-fed 3'MeDAB diet for 15 weeks contained significantly less RIA-GH than matched controls. These results suggest that 3'MeDAB, or its metabolites, may react with pituitary somatotrophs and impair their ability to synthesise and secrete hormone.     

Postnatal development of somatotrophs and mammotrophs in the pars distalis of the C57BL mouse

Summary The pars distalis of the pituitary gland of the C57BL mouse was studied by means of electron microscopy during postnatal stages of males ranging in age from newborn through 24 days, with particular emphasis on the somatotrophs and mammotrophs. During this period, growth curves were plotted in order to correlate postnatal growth patterns with the state of differentiation of the somatotrophs in the pars distalis. In the newborn, the somatotrophs show well developed organelles, including rough endoplasmic reticulum and Golgi complexes. These cells are not as densely packed with granules as the adult somatotrophs; however, from days 5 through 24, they show a progressive accumulation of granules. Although mammotrophs are scarce in the newborn, they are readily distinguishable in the pars distalis at 5 days. Male mice in small litters show a progressive increase in body weight between birth and 11–12 days, at which time the rate slackens until 18–19 days when the rate again increases. Growth curves for mice from large litters are similar to those from smaller litters, except that the transitory decrease in rate is more prominent.Supported by NIH grant no. HD12308 from the National Institute of Child Health and Human Development, USPHSThe authors wish to thank Pratima Ganguly for technical assistance     

Silicon-containing granules of rat liver,kidney and spleen mitochondria

Summary Isolated rat liver mitochondria containing granule aggregates (25–75 nm in diameter) and small (5–10 nm) electron opaque granules were examined by electron probe X-ray microanalysis. The granule aggregates gave an intense Si signal, while the small granules gave both Si and P signals. Isolated mitochondria of rat liver, spleen and kidney, subjected to detergent solubilization and differential centrifugation, produced two granule fractions: (1) a 10,000g fraction consisting predominantly of granule aggregates (25–75 nm) composed of smaller granules (5–10 nm in diameter), and (2) a 10,000–30,000 g fraction of non-aggregated small granules (5–10 nm). Thin sections of isolated granule aggregates gave Si X-ray signals similar to those obtained from in situ granules. In addition S, Cl, Mg, Cr and Fe X-ray signals were observed. Cr occurred only in the large kidney granules, while Fe occurred in both fractions of the spleen and kidney granules. The presence of Si in the granules was confirmed by chemical analysis of the isolated granules and in vivo radiolabeling of the granules with ³¹Si and ⁶⁸Ge. Contamination within the electron microscope was eliminated by a liquid nitrogen anticontamination device.Supported by Grant GM-08229-13-15 from the National Institutes of Health, USPHSWe are grateful to the Perkin-Elmer Corporation and to their Western Branch Manager, Mr. Michael E. Mullen and Microscopist, Mr. Minoru Saito, for use of and assistance with the Hitachi H 500 transmission electron microscope with the scanning attachment, and to the Kevex Corporation for the use of the Kevex X-ray Spectrometer. We also wish to acknowledge Mary Louise Chiappino for her technical assistance in preparing the thin sections, the final micrographs and X-ray spectra photographs and Darlene Lum for technical assistance in the laboratory     

Electron microscopic demonstration of calcitonin in human medullary carcinoma of thyroid by the immuno gold staining method

Summary In a human medullary carcinoma of thyroid gland containing calcitonin in light microscopic demonstration by the avidin biotin complex (ABC) method characteristic secretory granules were found electron microscopically in the cytoplasm of the tumour cells. They consisted in so-called type I granules (270±25 nm) and type II granules (135±17 nm). By the immuno gold staining (IGS) method the content of many secretory granules measuring 85–270 nm (152±18 nm) in diameter could be identified as calcitonin. These granules seemed to be predominantly of type II because of their nearly corresponding size and feature. The type I grnaules were less frequent in number and they showed no or little immunoreactivity. The results indicate that the IGS-method is practicable to demonstrate the ultrastructural localization of calcitonin and to identify clearly the nature of intracytoplasmic granules in electron microscopy.     

In cow anterior pituitary, growth hormone and prolactin can be packed in separate granules of the same cell

The ultrastructural localization of growth hormone and prolactin in cow anterior pituitary was studied by double immunocytochemical labeling using specific antibodies and protein A-gold particles of different sizes. The two hormones were found in specific somatotrophs and mammotrophs as well as in somatomammotropic cells which were multinucleated and predominantly arranged in clusters in the central area of the lobules. In these mixed cells the two hormones were packaged (a) in different granules of the same cell, (b) in the same granules where they were segregated in different portions of the granule content, or (c) in the same granules but evenly intermixed. The relative proportion of these three types of granules varied in somatomammotrophs of different animals. A single large Golgi complex was generally present in somatomammotrophs. Small, immature granules containing either growth hormone or prolactin or both hormones were found randomly distributed along Golgi stacks. This suggests that in these cells the two hormones are processed in the same Golgi cisternae and that mechanism(s) exist(s) to sort out the two hormones from each other.     

Electron-microscopic study on the anterior pituitary gland of spontaneous dwarf rats

Summary The spontaneous dwarf rat is a novel experimental model animal on the study of pituitary dwarfism. The fine structure of the anterior pituitary cells was studied in the immature and mature dwarf rats. Pituitary glands were removed from 5-, 10-, 20-day-old immature dwarfs, adult (45 days-16 weeks) dwarfs and normal 3-month-old rats and processed for electron-microscopic observation. In the control animals, growth hormone cells were readily identified by their ultrastructural characteristics, such as the presence of numerous electron-dense secretory granules, 300–350 nm in diameter, well developed rough endoplasmic reticulum and a prominent Golgi complex. In contrast, growth hormone cells were not found in the anterior pituitary gland of the spontaneous dwarf rat at any age examined. Other pituitary cell types, i.e., luteinizing hormone/ follicle stimulating hormone, thyroid stimulating hormone, adrenocorticotropic hormone and prolactin cells, appeared similar in their fine structure to those found in the control rats. In the pituitary gland of dwarf rats, a number of polygonal cells were observed either with no or relatively few secretory granules. The rough endoplasmic reticulum was arranged in parallel cisternae and the Golgi complex was generally prominent in these cells. In addition, many were found to have abundant lysosomes. A few minute secretory granules were occasionally observed; however, the immunogold technique failed to localize growth hormone or prolactin in the granules. The nature of these cells remained obscure in this study. Since their incidence and fine structural features, other than the secretory granules, were quite similar to those of the growth hormone cells in normal rats, we postulate that these cells are dysfunctional growth hormone cells. These results suggest that the cause of the growth impairment in the spontaneous dwarf rat is due to a defect in the functional growth hormone cells in the pituitary gland, and since other pituitary cell types appeared normal, the disorder seems to be analogous to the isolated growth hormone deficiency in the human.     

Proliferating cells in the rat anterior pituitary during the postnatal period: immunoelectron microscopic observations using monoclonal anti-bromodeoxyuridine antibody

Proliferating cells in the male rat anterior pituitary at 1, 3, 5, and 8 weeks of age were labeled with bromodeoxyuridine (BrdU) and studied by light and electron microscopic immunocytochemistry using anti-BrdU. They decreased in number from 402±31/mm² at 1 week to 50±1.5/mm² at 8 weeks, while their cell area increased by about twofold during this period. They had a slightly higher nucleus/whole cell (N/C) ratio than non-proliferating cells. According to their ultrastructure we classified them into granular and agranular cells. The percentage of granular cells ranged from 73% to 82% of all the proliferating cells during the period studied. They had many granules of various sizes and shapes, and some contained growth hormone and prolactin. Agranular cells, constituting 18–27% of proliferating cells, were small and had a high N/C ratio, indicating their immaturity. Moreover, they showed several features of folliculo-stellate (FS) cells: they showed no secretory granules in the cytoplasm, extended thin cytoplasmic processes, and sometimes they constructed a follicle among them. These results suggest: (1) the majority of proliferating cells were mature cells producing anterior pituitary hormone(s) and (2) most of the agranular proliferating cells maybe FS cells. The possibility of the latter is discussed.     

Die Feinstruktur gefriergebrochener Hormongranula

Zusammenfassung Gefriergebrochene Präparate (mit und ohne Ätzung) der Adenohypophyse und der C-Zellen der Schilddrüse von Ratte und Meerschweinchen wurden elektronenmikroskopisch untersucht. Bei den Hormongranula dieser Zellen verlaufen die Bruchflächen im allgemeinen zwischen den beiden Membranflächen oder zwischen Granulum-Inhalt und Membran. Nur relativ selten werden Granula quergebrochen. Auf beiden Hälften der gespaltenen Membranen der C-Zellgranula werden etwa mit gleicher Häufigkeit Proteinpartikel (100–200 Å) gefunden. Bei den Granula der somatotropen Zellen treten auf der dem Plasma anliegenden Membranhälfte deutlich mehr Proteinpartikel auf als auf der dem Granuluminhalt anliegenden Hälfte. Der Inhalt der somatotropen und C-Zellgranula erscheint bei dieser Präparationsmethode aus einer dichten Packung von 80–100 Å großen Partikeln zu bestehen.Eine besonders strukturierte Zone zwischen Membran und Granuluminhalt konnte bei den bisherigen Untersuchungen nicht festgestellt werden. Durch Ätzung der Gefrierbrüche ließen sich keine zusätzlichen strukturellen Details der Granula darstellen.Eine durch Auswertung von stereoskopischen Aufnahmen gewonnene Größenverteilungskurve für die C-Zellgranula wird vorgelegt.

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The fine structure of freeze-fractured hormone granules

Summary Freeze-fractured preparations (with and without etching procedures) of guineapig and rat thyroid (C-) cells and anterior pituitary (somatotropic-) cells have been investigated with the electron microscope. The hormone granules of these cells in general split either between the two lamellae of their unit membrane or between the granule contents and the unit membrane. Only rarely cross-broken granules have been observed. Inner and outer lamella of the unit membrane of the C-cell granules contain in more or less similar frequency moderate amounts of protein particles of 100–200 Å diameter. In case of the somatotrophs the outer lamella contains higher numbers of these particles than the inner one. The contents of the C-cell and somatotroph granules seems to consist of densely packed 80–100 Å particles. A particular zone between contents and membrane (as observed on micrographs with conventional electron microscopy) could not be detected on freeze-fractured preparations. The etching procedure does not reveal additional details of the granule structure.A size distribution curve of the C-cell granules as determined from stereo-pairs, is given.

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Stability of microsclerotial inoculum of Colletotrichum truncatum encapsulated in wheat flour–kaolin granules

Maintaining adequate viability of microorganisms in products for biocontrol is critically important for commercial reasons. Microsclerotia (MS) of the mycoherbicide agent, Colletotrichum truncatum, are its hardy, over-wintering fungal structures. Microsclerotial inoculum at 2, 7, and 23 MS/granule were matrix-encapsulated in wheat flour–kaolin granules (Pesta), in which the flour provided gluten for the matrix and a food base for the fungus. Pesta granules were dried to a water activity of 0.18–0.29. After storage for 52 weeks at 25°C, granules containing 7 and 23 MS were 100% viable and granules with 2 MS were 95% viable. Granules with 7 MS were 50% viable after 36 weeks at 35°C. Pesta granules (440 granules/g) with conidial inoculum at 3.3×105 c.f.u./g were less storage-stable than granules at the 2 MS/granule level. At all MS inoculum levels, granules stored for up to 2years produced 108 c.f.u./g in vitro when incubated on water agar. High water activity was detrimental to long-term viability. In the greenhouse, 7 MS/granule samples controlled 94% of hemp sesbania when incorporated into the soil pre-planting. The strategy of encapsulation of the naturally stable C. truncatum MS and drying to a favourable water activity led to excellent shelf-life for a live biocontrol agent.     

A comparative study of primary and secondary granules in monocytopoiesis and myelopoiesis of mouse bone marrow

Summary The differentiation and maturation of monocytes and neutrophil granulocytes were studied in bone marrow of normal mice by electron microscopy and cytochemical assessment of peroxidatic activity. The granule populations of the mature cells of bone marrow were identified and investigated to obtain a basis for the analysis of the earlier stages of maturation. Mature monocytes and neutrophils showed primary and secondary granules, and mature neutrophils had more of both kinds. The size, shape, and number of primary granules proved to offer the most reliable criteria for distinguishing promonocytes and promyelocytes. The primary granules of monocytes were smaller than those of mature neutrophils and were either spherical (smallest diameter 50–200 nm) or elongate (100×400 nm). Both granules had a homogeneous matrix. The granules of the granulocytes were either spherical (smallest diameter 200–300 nm) or elongate (150–200×300–500 nm), and some of them had a crystalline inclusion.     

Qualitative and quantitative analysis of granules in atrial appendage cardiocytes in different physiological states

Summary Atrial appendage cardiocytes of mammals, including man, contain multiple cytoplasmic granules that vary in number in different physiological states. Using morphologic and comprehensive morphometric techniques, these granules were assessed in Sprague-Dawley rats following dehydration for 5 days, volume-loading by substituting 1% NaCl as drinking water for 7 days, unilateral nephrectomy plus volume-loading for 7 days, and in late term pregnant animals (18–20 days; term 21 days). Although principally located in the paranuclear region, granules were observed throughout the sarcoplasm. Cytological features indicative of synthetic activity and granule formation were readily apparent in all groups with the exception of pregnant rats where they were infrequently observed. Granule contents were released by exocytosis and observed in the right appendage of control, dehydrated and nephrectomy/volume-loaded groups and left appendage of volumeloaded animals. Exocytosis was not observed in pregnant animals. By point counting, the proportional volume of cardiocytes occupied by granules (V _v ) in controls was significantly greater for right than for left appendage (2.12±0.22% vs 1.29±0.16%; mean±SEM;p<0.05). A significantly similar difference was found for nephrectomy/volume-loaded animals. There was no significant difference inV _v for right appendage between the control and experimental groups; for left appendage there was a significant increase inV _v to 2.42±0.09% (p<0.05) for volume-loaded animals only. Estimation of the maximum diameter of granule profiles in control animals was 238±9 nm and 230±6 nm for right and left appendages, respectively. The profile diameters in the left appendages of dehydrated (202±9 nm) and pregnant (200±7 nm) animals were significantly (p<0.05) less than those of the control animals. The morphometric findings did not correlate with predictions based upon published biochemical data. In the course of this study, a previously unreported bimembranous, circular to ovoid structure was observed in the cardiocyte sarcoplasm of all animals; the nature and function of this structure is unknown.     

Formation of crystalloid inclusions in the small intestine of neonatal pigs: an immunocytochemical study using colloidal gold

Summary Enterocytes of the small intestine in 1-day-old suckling piglets contain numerous vesicles in the apical cytoplasm and a large granule located beneath the nucleus. Within the next 3 days, these granules transform into electron-dense crystalloid inclusions. These membrane-bound inclusions are up to 10 m in length and 1–2 m in diameter, and they are composed of electron-dense lamellae 3.9 nm apart. Postembedding immunocytochemistry, using rabbit anti-porcine IgG and goat anti-rabbit IgG conjugated to 10 nm colloidal gold, revealed that both the granules and the crystalloid inclusions contained a high concentration of maternal IgG. Although the IgG content of the crystalloid inclusions was detected on epoxy-embedded sections, the use of LR White resin resulted in a much higher density of labelling. Quantification of the labelling density showed that the concentration of IgG in the crystalloid inclusions was approximately ten times higher than that in the lumenal colostrum and approximately three times higher than that in the granules. These observations showed that there are at least three compartments involved in the accretion of IgG in the small intestine of neonatal piglets: smaller apical endocytotic vesicles, large subnuclear granules and crystalloid inclusions. The role of these compartments in maternal immunoglobulin absorption and in the acquisition of passive immunity has yet to be explored.     

Correlative immunocytochemical and electron microscopic studies: Identification of (Entero)glucagon-somatostatin- and pancreatic polypeptide-like-containing cells in the human colon

Summary Correlative immunocytochemical and electron microscopic studies, using the semi thin-thin technic, were performed to identify the (entero) glucagon, somatostatin and pancreatic polypeptide-like immunoreactive cells of the human colonic mucosa. Mean granule diameter for each cell type was estimated according to two methods and histograms showing the granule size distribution were constructed. A total of 139 immunostained cells identified at the ultrastructural level were analyzed. Mean granule diameter for (entero)glucagon-containing cells was 318±11 nm but a reduction of granule size with age was noteworthy: granules were larger in the fetus (mean diameter 350±15) than in adults (mean diameter 310±10 nm). Somatostatin-containing cells, very rare in adults, were present in the fetal distal colon. Their general mean granule diameter was 354±18 nm but many cells had a mean granule diameter of more than 400 nm. A pancreatic polypeptidelike immunoreactivity was found only in (entero)glucagon-containing cells, pointing out the possible occurrence of both peptides (or of similar sequences) in the same cells. Previous ultrastructural studies dealing with a tentative classification of the human colonic endocrine cells were compared with the present data.This work was supported by the Institut National de la Santé et de la Recherche Médicale (INSERM).     

Pituitary growth hormone and somatotrophs in rats bearing chemically induced hepatomas.

Measurement of pituitary growth hormone (GH) content by polyacrylamide gel electrophoresis (PAGE) showed that rats bearing diethylnitrosamine (DENA) induced hepatomas contained significantly less GH than age-related controls. Light microscopy of these pituitaries revealed many apparently dying somatotrophs in tumour bearing rats. By electron microscopy the somatotrophs of DENA hepatoma rats were relatively inactive and some clearly dying, which could account for the reduced hormone content. However, electron microscopy of somatotrophs of rats with 2-acetyl-aminofluorene (2-AAF) hepatic tumour nodules revealed many very active cells with greatly increased content of free ribosomes and many granules poised for imminent release. Other somatotrophs of these rats showed lysosomal activity indicating reduced secretion after an active phase.