Takao Setoguti

Electron-microscopic studies on the relationship between the frequency of parathyroid storage granules and serum calcium levels in the rat

SummaryIn the rat parathyroid, the mean number of storage granules (NSG) per chief cell has been electron-microscopically studied and correlated with the mean serum calcium level (SCL). In animals given 4% CaCl2 plus vitamin D2 for 3 days, SCL is significantly elevated and NSG is increased. When these animals are injected with 2% EDTA, SCL is lowered to 8 mg/dl, but NSG is not affected; in those injected with 4% EDTA, however, SCL declines to a minimum (5.8 mg/dl) after 30 min, and NSG is also decreased. Control SCL are 8.9 mg/dl. These results indicate that storage granules may not be released until SCL is depressed to a certain level.In rats 3 weeks after castration, the chief cells show hyperplastic changes and SCL is at a low concentration (8.0 mg/dl). NSG, however, remains almost within control limits. Castrated animals injected with 4% EDTA show a hypocalcemia and a decrease in NSG, but NSG gradually recovers over a period of 6h. These data suggest that storage granules can be produced even under lower calcium concentrations. It is concluded that storage granules may be constantly produced and stored, and are released only as an emergency supply of hormone.

Effects of short-term treatment with calcium on the parathyroid gland of the rat, under particular consideration of the alteration of storage granules

SummaryShort-term effects of CaCl2-treatment on parathyroid cells of the rat, especially on their storage granules, were studied at the ultrastructural level. After an injection of 4% CaCl2, serum calcium levels (SCL) rapidly increased from 9.1 mg/dl (controls) to a maximum of 14.9 mg/dl at 20 min. At 5 min after the injection, the number of type-I storage granules (large core) [NSG-I] and that of type-II storage granules (small core) [NSG-II] remained unchanged, in spite of elevated SCL (12.4 mg/dl). As soon as SCL rose to 13.2 mg/dl at 7.5 min, NSG-I gradually decreased to a minimum at 30 min; in contrast, NSG-II gradually increased to a maximum at 30 min. Vacuolar bodies also increased together with the augmentation of type-II storage granules. The average diameter of the core of the storage granules decreased significantly after the injection. Protein A-gold method for immunocytochemistry showed that the cores of these granules contain parathormone. Acid-phosphatase activity was occasionally found in storage granules of both types, especially in those of type II. It is concluded (i) that type-I storage granules may be transformed into vacuolar bodies via type-II granules as a result of hydrolysis, and (ii) that these processes may be accelerated during hypercalcemia.

Specific organization of intracytoplasmic filaments in the dog testicular interstitial cell.

SummaryIn the dog testicular interstitial cells the cytoplasmic filaments are occasionally arranged in large bundles piled closely in an extensive area adjacent to the Golgi region in the cytoplasm. Some of the large bundles show conspicuous circular or spiral configurations which are composed of elaborate arrangements of both circular and longitudinal filaments and accompany tubules of agranular endoplasmic reticulum running parallel to the longitudinal filaments.

Immunocytochemical localization of parathormone in the mammalian parathyroid gland using the protein A-gold technique

SummaryElectron-microscopic immunocytochemistry for the demonstration of parathormone in parathyroid chief cells was performed in adult male rats, gerbils, mice, and dogs, using the protein A-gold technique. Protein A-gold particles were detected over both large and small secretory granules in all the animals examined. In the former, they were concentrated not only over type-I granules with a large core, but also over type-II granules with a small core. They were also located over atypical granules, including heterogeneously dense granules, granules having vesicles in a finely particulate core, and distorted granules. All labelled secretory granules were characterized by the presence of a clear halo of varying width around the core. Occasionally, Golgi cisternae as well as Golgi vacuoles with a finely particular content were also labelled. The labelling of the secretory granules was strong in dogs, moderate in rats and gerbils, and weak in mice. In addition, it was more intense in the non-osmicated preparations than in the osmicated preparations. The frequency of both types of large granules showed species differences. The possible factors involved in these differences are discussed.

Freeze-Fracture Replica Studies of Tight Junctions in Normal Human Bronchial Epithelium

Using freeze-fracture techniques, tight junctional networks were observed in the human normal bronchial epithelium. They were morphologically classified into three types: type I was a loosely interconnected, most complicated network consisting of 7-11 roughly parallel wavy strands and situated between ciliated cells; type II was a randomly anastomosing, simple network made up of 2-4 strands and present between goblet cells; type III was an irregularly anastomosing network composed of 4-7 strands and located between a ciliated cell and a goblet cell. Type III junctions, when a goblet cell was strongly bulged, were located on the swollen ridge, the upper surface of which was separated by a deep groove from the bulged apical surface, around the lateral surface of the cell at the level of the luminal surface. The possible relation between the orientation of strands of these networks and extra- or intracellular stress was discussed.

Freeze-fracture study of the rat parathyroid gland under hypo- and hypercalcemic conditions, with special reference to secretory granules

SummaryFreeze-fracture images of the parenchymal cells in the parathyroid gland of rats were observed after vitamin D2 plus calcium chloride-suppression and EGTA-activation of secretion. In cells of the suppressed glands, large bulges protruded from the Golgi cisternae, and large granules with a stalk, which are identified as storage granules, suggest that, during maturation, some storage granules may be connected by long tubules with the Golgi cisternae and supplied with secretory products from the Golgi cisternae via these tubules.In the activated glands, presumptive exocytotic and endocytotic specializations of intramembranous particles of the parenchymal cell plasma membrane were frequently observed. In addition, elevations and complementary shallow depressions of various shape and extent were occasionally encountered in the intercellular space. From their morphological characteristics it was concluded that these originated from secretory granule cores, which are discharged from the parenchymal cells into the intercellular space by exocytosis, and it was suggested that discharged granule cores may retain their spherical shape until they fuse to form a flat conglomerate.

Electron-microscopic studies on the threshold value of calcium concentration for the release of storage granules and the acceleration of their degradation in the rat parathyroid gland

SummaryTo determine both a threshold value of calcium concentration (CC) for the release of storage granules and that for the acceleration of degradation of these granules, the rat parathyroid glands were perfused in situ with HEPES-Ringer solutions containing different concentration of Ca2+ for 10 min. With perfusates containing 0.83–1.21 mM Ca2+ (equivalent to 8–11 mg/dl serum calcium), the number of type-I storage granules (large core) [NSG-I] and that of type-II storage granules (small core) [NSG-II] remained unchanged. With perfusates containing 0.83 mM Ca2+ (7.5 mg/dl) or less, however, both NSG-I and NSG-II decreased remarkably and the former was larger than the latter. On the contrary, with perfusates containing 1.27 mM Ca2+ (11.5 mg/dl) or more, NSG-II increased and the ratio of NSG-I to NSG-II was changed reversely. We concluded that a thereshold value of CC required for the release of storage granules may be present between 0.88 and 0.83 mM Ca2+ (8 and 7.5 mg/dl) and that a threshold value of CC for accelerating the transformation of type-I granules into type-II, the degradation of storage granules, may be situated at about 1.27 mM Ca2+ (11.5 mg/dl). Additionally, it was suggested that both prosecretory and storage granules are not only formed at the innermost Golgi cisterna but also at the trans-Golgi network.

Effects of Pilocarpine Treatment and of Electrical Stimulation of the Vagus Nerve on the Rat Parathyroid Gland, with Special Reference to the Alteration of Storage Granules

Effects of pilocarpine treatment and of electrical vagal stimulation on the rat parathyroid were studied ultrastructurally. The number of type I storage granules with a narrow halo (NSG-I) and that of type II storage granules having a wide halo (NSG-II) were calculated. After pilocarpine treatment, NSG-I gradually decreased and reached a minimum at 30 min; in contrast, NSG-II gradually increased and reached a maximum at 20 min, but thereafter it slightly decreased and instead vacuolar bodies increased. Excluding these alterations, the ultrastructure of parenchymal cells showed no remarkable changes. Electrical vagal stimulation furthermore confirmed these results. Acid phosphatase activity was occasionally found in storage granules of both types in control and experimental rats. It was concluded that storage granules normally may be transformed from type I into type II and finally into vacuolar bodies as a result of hydrolysis, and that these processes may be accelerated by parasympathetic stimulation.

Effects of Epinephrine Treatment on the Rat Parathyroid Gland, with Special Reference to the Frequency of Storage Granules

Effects of epinephrine treatment on the rat parathyroid gland were studied morphologically. The mean number of storage granules per cell section (NSG) was rapidly decreased as early as 5 min after an injection of epinephrine and seemed to reach a minimum between 5 and 30 min. During this period, serum calcium levels (SCL) gradually rose and reached a maximum at 30 min. The ultrastructure of chief cells in these epinephrine-injected rats showed no marked difference as compared with that in control rats. In slightly hypocalcemic rats, induced previously by 2% EDTA-treatment, NSG was more rapidly decreased. It was suggested that storage granules may be released promptly by epinephrine treatments in spite of high SCL and that they are more promptly released under hypocalcemia.

Freeze-Fracture Replica Studies of the Human Adrenal Cortex, with Special References to Microvillous Projections

Freeze-fracture replicas of human adrenocortical cells revealed en face fractures (both P and E faces) of the cell surface to be classified into three patterns: (1) the fracture face with no microvillous projections; (2) the fracture face mainly with short microvillous projections which arise horizontally from the edges of the small cavity on the cell surface to form a flower bud-like configuration on the P face; (3) the fracture face mainly with varying-sized, longer microvillous projections which arise almost vertically from the cell surface and show no specific organization. The former two fracture faces were supposed to correspond to the plasma membranes facing the interstitium including pericapillary spaces, since they had no gap junctions and frequently appeared along sinusoidal capillaries. On the other hand, the latter fracture face seemed to be equivalent to the plasma membranes facing intercellular spaces, since it frequently showed gap junctional specialization of intramembranous particles and usually both the P and E fracture faces of plasma membrane on the same surface. En face fractures of these gap junctions were various in shape and size. Tight junctions and their complex form with gap junctions were not encountered. Cell organelles and the nucleus had more numerous intramembranous particles on the P fracture face than on the E fracture face.