The corpuscles of Stannius in arawana (Osteoglossum bicirrhosum), an ancient teleost

The corpuscles of Stannius of arawana (Osteoglossum bicirrhosum), an ancient teleost, were examined by routine light and electron microscopy and following their immunoreactivity to salmon and trout stanniocalcin antisera. Periodic acid-Schiff positive cells of the corpuscles of Stannius had a follicular arrangement and demonstrated a strong immunohistochemical reaction with both stanniocalcin antisera. Fine structural analysis of the paired, posteriorly located, and perirenal ovoid glands revealed two morphologically distinct cell types the basal laminae of which were ramified by nerve terminals. Immunocytochemistry demonstrated that osmiophilic secretory granules in both cell types were immunoreactive to the stanniocalcin antisera. When extracts of arawana corpuscles of Stannius were subjected to sodium dodecyl sulphate electrophoresis and Western blot analysis a diffuse molecular weight band was evident ( approximately 68 kDa) in the non-reduced condition. In all cases, immunoreactivity was abolished by preabsorption of the antisera with salmon stanniocalcin or with a crude extract of arawana corpuscles of Stannius. The corpuscles of Stannius of arawana are similar to those in more recent teleosts with respect to cell structure and their anatomical distribution but their stanniocalcin is more similar in molecular weight to that present in at least one other non-teleost actinopterygian (the gar) which has an ancient lineage.     

Stanniocalcin-like immunoreactivity in the corpuscles of Stannius of the bowfin Amia calva L.

Summary We used an antiserum against salmon stanniocalcin in an immunohistochemical, immunocytochemical, and Western blot analysis of bowfin (Amia calva) corpuscles of stannius. The bowfin is one of two extant holostean species with ancient ancestral links to modern-day bony fishes. The corpuscles of Stannius (white corpuscles) of the bowfin were scattered throughout much of the kidney among the adrenocortical homolog (yellow corpuscles) but could be distinguished from the adrenocortical homolog by their positive staining with both the periodic acid-Schiff reaction and a salmon stanniocalcin antiserum. Immunoreactivity was confined to cytoplamic granules and was absent when the antiserum was blocked with salmon stanniocalcin or with a crude extract of bowfin corpuscles of Stannius. When bowfin corpuscle-of-Stannius extracts were subjected to sodium dodecyl sulphate electrophoresis and Western blot analysis, two closely spaced bands were evident (43–45 kDa). Staining of both bands was abolished by pre-absorption of the antiserum with salmon stanniocalcin. In comparison to salmon stanniocalcin, the reputed bowfin hormone migrated faster in gels, suggesting a smaller apparent size. The purification of bowfin stanniocalcin should yield important new information regarding the evolution of this unique calcium-regulating hormone.     

Immunoreactivity of the corpuscles of Stannius of the garpike,Lepisosteus osseus,to antisera against salmon and trout stanniocalcins

Stanniocalcin-immunoreactive cells were localized in the corpuscles of Stannius of a holostean fish, the garpike (Lepisosteus osseus), using antisera against salmon and trout stanniocalcins and the peroxidase-antiperoxidase and protein A-gold immunohistochemical methods. The stanniocalcin-immunoreactive cells were periodic acid-Schiff-positive, and antibody staining was abolished if the antiserum was preabsorbed with corpuscle homogenate. Immunocytochemistry revealed two reactive cell types in the glandular parenchyma, and immunoreactivity was confined to the secretory granules. Staining of the granules was also abolished when the antisera were blocked with crude corpuscle homogenate. When corpuscle extracts from garpike were subjected to sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blot analysis, a single dense band was evident with a molecular weight of 68 kDa under non-reducing conditions, whereas three bands were observed (29, 31, and 34 kDa) under reducing conditions. Staining of all bands disappeared following preabsorption of the antiserum with salmon stanniocalcin, trout stanniocalcin, or garpike corpuscle extract. The results are compared with stanniocalcins from another extant holostean, the bowfin (Amia calva), and from more modern bony fishes, the teleosts.     

Fine Structure of the Adrenocortical Homolog and the Corpuscles of Stannius of Amia calva L.

A comparison is made between the fine structure of yellow corpuscles and white corpuscles located within the kidneys of the holostean fish, Amia calva L. The yellow corpuscles are composed of epithelial cells possessing all the features of steroid-producing tissues, namely an abundance of vacuoles, tubular smooth endoplasmic reticulum, and mitochondria with tubular cristae. The Golgi apparatus is also a conspicuous component of their cytoplasm. These cells are homologous to adrenocortical cells of higher vertebrates and they have cytoplasmic projections which extend into the lumina of surrounding sinusoids. The white corpuscles possess epithelial cells of variable appearance but all cells contain secretory granules and an extensive rough endoplasmic reticulum. The secretory granules appear to originate at the Golgi apparatus and occasionally are observed intact in the intercellular space. However the method of release of these granules was not clearly defined. These corpuscles are similar to the corpuscles of Stannius which have been described in modern bony fish. The presence of multivesicular bodies and smooth endoplasmic reticulum in some cells may reflect the origin of the corpuscles of Stannius from the tubular nephron. A. calva appears to be a suitable organism for comparative studies into the function of the adrenocortical homolog and corpuscles of Stannius in “primitive” fish.     

鱼类斯钙素的研究进展

本文概述了近年来有关硬骨鱼类斯坦尼氏小体（ｃｏｒｐｕｓｃｌｅｓｏｆＳｔａｎｎｉｕｓ,ＣＳ）分泌激素———斯钙素（ｓｔａｎｎｉｏｃａｌｃｉｎ,ＳＴＣ）的研究进展。ＳＴＣ是糖蛋白类激素,为同型二聚体,其表观分子量在天然状态下从４６（大麻哈鱼）～５６（虹鳟）ｋＤａ,还原状态下则为２３～２８ｋＤａ。ＳＴＣ单体的氨基酸序列分析表明,大麻哈鱼、银大麻哈鱼和澳大利亚鳗鲡的氨基酸残基数分别为１７９、２２３和２３１个。研究还表明,ＳＴＣ的分泌受血钙浓度的调节,并且胆碱能神经参与ＳＴＣ的释放。     

Chum salmon (Oncorhynchus keta) stanniocalcin inhibitis in vitro intestinal calcium uptake in Atlantic cod (Gadus morhua)

Summary Chum salmon (Oncorhynchus keta) stanniocalcin was purified, partially identified and tested for bioactivity in an assay on the intestinal calcium uptake in a marine teleost (Gadus morhua). Basic ethanol extraction, ion exchange chromatography, gel filtration and reverse-phase high-performance liquid chromatography resulted in the isolation of a homogenous glycoprotein that appears as a 46-kDa product under non-reducing conditions and as a 23-kDa product under reducing conditions after sodium dodecylsulphate-polyacrylamide gel electrophoresis. The glycoprotein is likely to be a homodimer composed of two subunits of 23 kDa each. Further characterization indicates homology to Australian eel, sockeye salmon, coho salmon and rainbow trout stanniocalcin, and the glycoprotein is thus concluded to be stanniocalcin. Stanniocalcin-like immunoreactivity was demonstrated in the corpuscles of Stannius of the Atlantic cod, with a specific antiserum raised against purified chum salmon stanniocalcin. The physiological importance and the biological activity of chum salmon stanniocalcin was tested by evaluating its effect on intestinal calcium uptake by the Atlantic cod in vitro. The intestine was perfused, both vascularly and through the intestinal lumen, and the calcium mucosa-to-serosa flux was measured using ⁴⁵Ca²⁺ as a tracer. Stanniocalcin decreased the intestinal calcium uptake in a dose-related manner by 13.5% and 22.4% at doses of 2.2 and 10.9 nM stanniocalcin, respectively. The results establish the intestine as a target organ for stanniocalcin in marine teleosts.Abbreviations BIS balanced intestinal solution - CS corpuscles of Stannius - dpm disintegrations per minute - FW freshwater - J _in ^Ca influx of calcium across the intestinal mucosa - MW molecular weight - NRS normal rabbit serum - PBS phosphate buffered saline - PBST phosphate buffered saline containing 0.05% Tween-20 - PITC phenyl isothiocyanate - rp-HPLC reverse phase - SW seawater - STC phenyl isothiocyanate - rp-HPLC reverse phase - SW seawater - STC stanniocalcin - TFA Trifluoroacetic acid - Tris Tris(hydroxymethyl) aminomethan - V volume per fraction     

Endocrine Control of Calcium Metabolism in Teleosts

It is evident that fishes regulate their serum calcium efficientlybut that endocrine systems involved may be different from thosein tetrapods. A functional parathyroid gland has not yet beendemonstrated in fishes. The majority of evidence indicates thatcalcitonin has little or no effect on fish calcium regulation.Instead, the corpuscles of Stannius and the pituitary glandare necessary for maintaining fish serum calcium levels. Inthe killifish, Fundulus heteroclitus, the removal of the corpusclesproduces hypercalcemia in sea water but not in artificial seawater deficient in calcium. Transplants of the corpuscles orthe administration of corpuscle homogenate corrects the increasein calcium. On the other hand, hypophysectomy elicits hypocalcemiaunder calcium deficient conditions but not in calcium rich seawater. Replacement therapy with pituitary homogenate or hypophysialtransplant prevents the fall in calcium. It is postulated thatthe hypocalcemic corpuscles of Stannius and the hypercalcemicpituitary gland enable the euryhaline killifish to regulateits serum calcium levels in high calcium sea water and low calciumfresh water, respectively.     

The kidney,adrenocortical homolog,and corpuscles of Stannius in the cockscomb pricklebackAnoplarchus purpurescens

The morphology of the kidney, adrenocortical homolog, and the corpuscles of Stannius was examined in the cockscomb prickleback,Anoplarchus purpurescens, a marine teleost which inhabits the intertidal zone. The paired kidneys of this fish are fused throughout most of their length, there is essentially a single posterior cardinal vein on the right side, they possess renal corpuscles, and there is no distal segment of the tubule. The tubule is specialized, in descending order, into ciliated neck and two proximal segments before entering the system of collecting tubules and ducts. The cells of the latter system are specialized for mucous secretion, as are cells of the main excretory ducts, the paired archinephric ducts. Tubulogenesis occurs in the kidneys in close apposition to the archinephric ducts. The presumptive adrenocortical homolog is located around the posterior cardinal veins in the head kidney while paired corpuscles of Stannius are confined to the posterior end of the kidney. All of the above features are consistent with those found in the kidneys of many other marine teleosts.     

Hormones and Calcium Regulation in Fundulus heteroclitus

For the past 20 years, our laboratory has been involved in studyingthe endocrine control of calcium balance in the killifish, Fundulusheteroclitus. We have surveyed almost all endocrine systemsand discovered two main ones directly involved in plasma calciumregulation. They are the pituitary gland and the corpusclesof Stannius. When fish adapted to low-calcium seawater werehypophysectomized, hypocalcemia and tetany were observed. Whenfish adapted to high-calcium seawater were Stanniectomized,hypercalcemia was seen. In both cases, other electrolytes wereunaffected and replacement therapy corrected the plasma calciumchanges. We have tried to characterize the active principlesin both glands. We discovered that prolactin is hyperacalcemic.The pituitary gland also seems to contain a second hypercalcemicfactor which may be located in the PAS-positive pars intermediacells. For the Stannius corpuscle factors, we developed a bioassayand named the active substance(s) hypocalcin. In collaborationwith Dr. Hirofumi Sokabe at Jichi Medical School, Japan, weshowed that the Stannius corpuscles also contain a renin-likesubstance capable of generating a hypocalcemic angiotensin-likesubstance. The exact chemical nature of the hypocalcemic substanceis being investigated. Calcium balance in the whole fish wasalso studied with 47-calcium. These studies were carried outin collaboration with Dr. Nicole Mayer-Gostan at Villefranche-sur-Mer,France. We discovered that killifish depend on the environmentrather than bone as a calcium reservoir. Hormones may be involvedin the exchanges with the environment.     

On the fine structure of corpuscles of Stannius of the eel,Anguilla japonica

Summary The Stannius corpuscle of the eel (Anguilla japonica) consists of numerous ovoid or polymorphic lobules separated by loose connective tissue containing blood capillaries. Each lobule is composed of a number of columnar secretory cells, containing numerous secretory granules, and arranged more or less radially. Each secretory granule, spherical, 0.5–1.0 in diameter, and osmiophilic, is surrounded by a limiting membrane derived from the Golgi membrane. The well developed rough-surfaced endoplasmic reticulum is closely associated with the Golgi field. Some Golgi elements might also be intimately associated with the outer nuclear membrane. Numerous glycogen particles are distributed throughout the cytoplasm. The secretory granules which tend to accumulate near the basal part of the cell might be released into the loose connective tissue. From these facts, the corpuscles of Stannius are considered to be protein-secretory endocrine glands without any similarity to the adrenal gland.     

Detection of FMRFamide-like immunoreactivities in the sea scallopPlacopecten magellanicus by immunohistochemistry and Western blot analysis

FMRFamide-like immunoreactivity was detected histochemically in the sea scallopPlacopecten magellanicus. Most immunoreactivity was concentrated in the cerebral, pedal, and parietovisceral ganglia, particularly in the cortical cell bodies and in their fibers which extend into the central neuropile. Whole-mount immunofluorescence studies were used to localize concentrations of immunoreactive cells on the dorsal and ventral surfaces of each ganglion. Immunoreactivity was also detected in nerves emanating from the ganglia. Strong immunoreactivity was localized in peripheral organs, including the gut and gills of juvenile and adult scallops. Weak immunoreactivity was detected in the gonads, heart, and adductor muscle of the adults. A broad FMRFamide-like immunoreactive band of 2.5–8.2 kDa was detected by Western blotting of acetone extracts of the parietovisceral ganglia. In the presence of protease inhibitors, two FMRFamide-like immunoreactive bands (7.2–8.2 kDa and >17 kDa) were obtained. Neither of these bands comigrated with the FMRFamide standard. It is concluded that peptides of the FMRFamide family are probably regulators of numerous central and peripheral functions inP. magellanicus.     

Development of the corpuscles of Stannius in the anabantid teleost, Colisa lalia

The development of Stannius corpuscles in the teleost Colisa lalia is described. Certain cells of the mesonephric tubules differentiate and proliferate to form buds which evaginate and finally become separated from the structure of origin. The separated cellular masses undergo further histologic differentiation to attain the adult structures. The corpuscles in C. laha, in contrast to some other teleosts, develop only from mesonephric tubules. A possible explanation for the different types of distribution of Stannius corpuscles in teleosts is suggested, and the possible homology of these structures with part of the adrenal cortex or higher vertebrates is discussed.     

家鸽Herbst小体对振动的反应特征

鸟类的Herbst小体是一种形态特殊的感觉性神经末梢器官.本文利用电生理学方法,研究了家鸽腿部胫骨-腓骨之间的Herbst小体对振动刺激的反应特征.这种小体对振动刺激非常敏感,当振动频率在600—800赫时,它们有反应的最低阈值约为0.3微米.不同的Herbst小体的反应阈值与频率的关系曲线表明:这种小体具有明显的带通滤波的特征,对振动反应的最佳频率范围为400—1000赫.在适宜频率、超阈值强度的振动刺激下,Herbst小体能以1:1的方式作出反应,即相对于每次正弦波振动刺激都有一个锁相的神经脉冲产生.在背根脊神经节内的细胞外记录表明:对振动敏感的神经节细胞具有和Herbst小体完全相似的反应特征.     

Involvement of Ca/calmodulin-dependent protein kinases in mycelial growth of the basidiomycetous mushroom, Coprinus cinereus

Although multifunctional Ca²⁺/calmodulin-dependent protein kinases (CaM-kinases) are widely distributed in animal cells, the occurrence of CaM-kinases in the basidiomycetous mushroom has not previously been documented. When the extracts from various developmental stages from mycelia to the mature fruiting body of Coprinus cinereus were analyzed by Western blotting using Multi-PK antibodies, which had been generated to detect a wide variety of protein serine/threonine kinases (Ser/Thr kinases), a variety of stage-specific Ser/Thr kinases was detected. Calmodulin (CaM) overlay assay using digoxigenin-labeled CaM detected protein bands of 65 kDa, 58 kDa, 46 kDa, 42 kDa, and 38 kDa only in the presence of CaCl₂, suggesting that these bands were CaM-binding proteins. When the CaM-binding fraction was prepared from mycelial extract of C. cinereus by CaM-Sepharose and analyzed with Multi-PK antibodies, two major immunoreactive bands corresponding to 65 kDa and 46 kDa were detected. CaM-binding fraction, thus obtained, exhibited Ca²⁺/CaM-dependent protein kinase activity toward protein substrates such as histones. These CaM-kinases were found to be highly expressed in the actively growing mycelia, but not in the resting mycelial cells. Mycelial growth was enhanced by the addition of CaCl₂ in the culture media, but inhibited by the addition of EGTA or trifluoperazine, a potent CaM inhibitor. This suggested that CaM-dependent enzymes including CaM-kinases play crucial roles in mycelial growth of basidiomycete C. cinereus.     

Characterization of trypsinogens 1 and 2 in two human pancreatic adenocarcinoma cell lines; CFPAC-1 and CAPAN-1.

Proteins with trypsin-like immunoreactivity (first detected by a specific immunoenzymatic assay) were isolated from CAPAN-1 and CFPAC-1 cell culture-conditioned media by chromatography on an immunoadsorbent prepared with a polyclonal antibody directed against trypsin 1. The adsorbed proteins were devoid of free trypsin activity but trypsin activity was present after enterokinase activation demonstrating that the immunoreactive trypsin present in cell supernatants corresponds to trypsinogens. When characterised by Western blotting using a monoclonal antibody directed against human trypsin 1 two protein bands corresponding to trypsinogen 1 (23 kDa) and trypsinogen 2 (25 kDa) gave a positive reaction. These results demonstrate the presence of trypsinogens 1 and 2 in CAPAN-1 and CFPAC-1 cells and in their culture-conditioned media.     

Differential Activity of Stanniocalcin in Male and Female Fresh Water Teleost Mastacembelus armatus (Lacepede) during Gonadal Maturation

The present study was carried out to analyze the differences in the activity of hormone stanniocalcin (STC) between male and female fishes of Mastacembelus armatus during their gonadal cycle. A large variation in nuclear diameter of cells of corpuscles of Stannius (CS) were recorded in relation to testicular cycle as well as ovarian cycle which indicates that the cellular activity varied with different phases of reproductive cycle in both male and female fish. Similar changes in nuclear diameter of CS cells were also observed after 17alpha-methyltestosterone administration in males and 17 β-estradiol administrations in females. A positive correlation was observed between plasma STC levels, gonadosomatic index (GSI) and the sex steroids in both sexes, suggesting that STC has a role in the processes involved in gonadal development. In addition females showed remarkable changes in plasma calcium level during gonadal cycle while no such change for males were observed. In females the plasma calcium level estimated during different phases of reproductive cycle indicates positive correlation between plasma level of calcium and gonad growth. Thus hyperactivity of CS cells was noted in both male and female fishes during gonadal cycle along with the differences in the activity of STC as well. In female it may act as hypocalcemic factor and bring the level of calcium to normal which increases during preparatory and pre spawning phases to fulfill the increased demand of calcium for vitellogenesis. However data of male fishes indicated that plasma STC concentration varied widely during gonadal cycle but showed no consistent relationship to plasma calcium level.     

The murine stanniocalcin 1 gene is not essential for growth and development

The stanniocalcin 1 (STC1) gene is expressed in a wide variety of tissues, including the kidney, prostate, thyroid, bone, and ovary. STC1 protein is considered to have roles in many physiological processes, including bone development, reproduction, wound healing, angiogenesis, and modulation of inflammatory response. In fish, STC1 is a hormone that is secreted by the corpuscles of Stannius and is involved in calcium and phosphate homeostasis. To determine the role of STC1 in mammals, we generated Stc1-null mice by gene targeting. The number of Stc1-/- mice obtained was in accordance with Mendelian ratios, and both males and females produced offspring normally. No anatomical or histological abnormalities were detected in any tissues. Our results demonstrated that Stc1 function is not essential for growth or reproduction in the mouse.     

Origin of 33 kDa protein of vitelline membrane of quail egg: Immunological studies

The inner layer of vitelline membrane is an investment of avian ovum at the time of ovulation, but its formation is poorly understood. In order to elucidate the origin of the inner layer of vitelline membrane, a 33 kDa protein, one of the components of the inner layer, was purified from quail eggs and polyclonal antibody was raised against this protein. The tissue distribution of protein interacted with the antibody was studied by Western blotting technique. No immunoreactive component could be observed in extracts of liver, kidney, heart, lung, small intestine, brain, infundibulum, albumen-secreting region of oviduct, uterus, and wall of small white follicles. The intensive band was detected in the granulosa layer, which was isolated from the large preovulatory follicles as a monolayer of granulosa cells sandwiched between the inner layer of vitelline membrane and the basal lamina. The granulosa cells isolated from the granulosa layer also reacted with this antibody. Theca layer had no immunoreactive components. The position of the band of the 33 kDa protein on SDS-PAGE was sifted to higher molecular weight in follicular tissues as compared with that in the laid eggs, indicating that the structural change of the protein occurs after ovulation. These studies indicate that the material reactive to the antibody raised against a 33 kDa protein of quail vitelline membrane is synthesized by the granulosa cells.     

Evidence of hypocalcemic factor from corpuscles of Stannius of the teleost Heteropneustes fossilis (Bloch)

The administration of acid extract of corpuscles of Stannius has no effect on the plasma calcium level, in the normal fish, whereas the same dose shows typical hypocalcemic response in hypercalcemic fish, H. fossilis. It is concluded that acid extract of corpuscles of Stannius is effective only in hypercalcemic condition.