Hypocalcemic effects of bovine parathyroid hormone (1-34) and Stannius corpuscle homogenates in teleost fish adapted to low-calcium water

Injections of bovine parathyroid hormone (PTH 1-34) and homogenates of corpuscles of Stannius produce hypocalcemia in male killifish and tilapia adapted to calcium-deficient seawater or fresh water, respectively. In fish from water with normal calcium concentrations no effects are noticeable. These results suggest similarity in bioactivity between PTH, the hypercalcemic hormone of terrestrial vertebrates, and the hypocalcemic factor of the corpuscules of Stannius in teleost fish.     

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.     

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.     

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.     

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.     

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.     

Fine structure of the corpuscles of stannius in the toadfish.

The micro-anatomy of the corpuscles of Stannius of the toadfish, Opsanus tau, an aglomerular marine teleost, has been studied by light and electron microscopy. The corpuscles are composed of extensively anastomosed cords of epithelial cells which maintain intimate contact with blood capillaries. Most of the epithelial cells contain acidophilic granules which also show a positive reaction with the periodic acid-Schiff technique and aldehyde fuchsin. On the basis of fine structural criteria, three cell types can be recognized. The granular cells contain abundant quantities of granular endoplasmic reticulum, ribosomes, Golgi apparatus with prosecretory granules, coated vesicles, polymorphic mitochondria with lamellar cristae, filaments, microtubules, a cilium, a variety of lysosome-like dense bodies, glycogen particles, lipid droplets, secretory granules and intranuclear lipid-like inclusions. One variety of agranular cell (type I) is characterized by the total absence of secretory granules, but it contains large amounts of granular endoplasmic reticulum and ribosomes, conspicuous profiles of Golgi apparatus, coated vesicles and sometimes an abundance of glycogen. Another variety of agranular cell (type II) has poorly developed cytoplasmic organelles. The perivascular space between the capillary and parenchyma contains connective tissue cells and abundant nerve fibers. The different types of epithelial cells observed in the corpuscles of Stannius of this fish may represent functional stages of the secretory cycle in a single cell type.     

Seasonal changes in the corpuscles of Stannius and the gonads of the catfish, Heteropneustes fossilis (Bloch).

The seasonal changes in the corpuscles of Stannius (CS) and the gonads of the catfish, Heteropneustes fossilis were studied. The annual sex cycle of the fish has been divided into 4 phases on the basis of the variations in the gonosomatic index and histocytological features displayed by the testes and ovaries. There is a rise in the percentage of aldehyde fuchsin (AF)-positive cells in the CS and an increase in the nuclear diameter, at the beginning of preparatory period (February). In the prespawning period (May--June) the AF-positive cells undergo degranulation. A slight regranulation and rise in the percentage of AF-positive cells occurs during early spawing period (July). During the postspawning phase (September--January) the corpuscles remain predominated by AF-negative cells and show histolytical changes; the nuclear indices are reduced. In view of the concomitant changes occurring in the CS and the gonads the possibility of some direct or indirect relationship between the two has been discussed, although it is difficult to ascertain whether the changes in the corpuscles are cause or consequence of the gonadal cycle.     

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.     

Corpuscles of Stannius and stanniocalcin-like immunoreactivity in the white sucker (Catostomus commersoni): evidence for a new cell-type

The distribution of stanniocalcin immunoreactivity was examined in the corpuscles of Stannius of the white sucker (Catostomus commersoni) by using a chum salmon stanniocalcin antiserum, Western blotting, and light and electron microscopy. The white sucker possesses at least two stanniocalcin-immunoreactive corpuscles in the most posterior portions of the kidneys. Immunocytochemistry and ultrastructure revealed two cell-types in the corpuscle parenchyma, only one of which was immunoreactive. The nonimmunoreactive cells contained dense-cored vesicles and long processes that extended between the immunoreactive cells and terminated at perivascular spaces. When corpuscle extracts were subjected to electrophoresis and Western blotting, three nonreduced stanniocalcin-like immunoreactive bands (approximately 56, 61, and 64 kDa) were observed. However, in the presence of a reductant, a diffuse band migrating in the range of 28 to 32 kDa was noted. The results of this study on the white sucker demonstrate the presence of a dimeric stanniocalcin-like molecule and present evidence of a previously uncharacterized cell-type in the corpuscles of Stannius.     

Evolution of Calcium Regulation in Lower Vertebrates

Calcium regulation in lower vertebrates appears to be a continuum.The predominant hypercalcemic hormone in reptiles, birds andmammals is parathyroid hormone, while the major hypercalcemiccontrol in fishes is a pituitary factor, probably prolactin.In the amphibians dual controlling mechanisms are at work, sothat both the pituitary and parathyroids exert their influence.Prolactin may still retain some hypercalcemic potency in thehigher vertebrate groups, either directly or indirectly by influencingthe secretion of other hypercalcemic hormones. On the otherhand, parathyroid hormone does not occur in, nor does it elevateblood calcium in fishes. It thus seems to be a new inventionof tetrapods, or possibly to have evolved from a pituitary factorof fishes. The ability to lower blood calcium seems to be veryimportant in seawater fishes, in which the corpuscles of Stanniusexert major control. In terrestrial forms, the corpuscles ofStannius are not present, and hypocalcemic factors assume aminor role in overall calcium regulation     

Immunocytochemical localization of hypocalcin in the endocrine cells of the corpuscles of Stannius in three teleost species (trout,flounder and goldfish)

Summary In order to identify the cell-type responsible for the production of hypocalcin (the recently isolated hypocalcemic hormone of teleost fish), the corpuscles of Stannius (CS) of trout, flounder and goldfish, were immunocytochemically stained with antisera raised against trout hypocalcin. The secretory granules of the type-1 cells of the CS, considered to be the hypocalcin-producing cells, showed intense immunoreactivity in all species examined. However, in trout and flounder, the secretory granules produced by the type-2 cells, which have been suggested to represent a functionally different cell-type, also showed an intense immunoreactivity. In goldfish, no type-2 cells were observed. We tentatively conclude that type-1 and type-2 cells represent structurally different forms of the same functional cell-type.     

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。     

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.     

The Calcium-sensitive Cells of the Pars Intermedia in the Eel

In the pituitary, the PAS-positive calcium-sensitive (Ca-s) cells of the pars intermedia appear less active in seawater (SW)- than in freshwater (FW)-adapted eels. The kinetics of their response during adaptation to SW or readaptation to FW was investigated. Morphometric studies show that transfer to SW induces a rapid nuclear atrophy which accentuates in eels kept for several weeks in SW. Readaptation to FW stimulated the Ca-s cells after 2–10 days; after 1 or 2 months, the cells tend to be similar to those of eels kept in FW. Plasma calcium decreases slightly but significantly in SW eels. The response of the Ca-s cells is not modified by an ovine prolactin treatment inducing hypercalcemia, hypernatremia and stimulation of the corpuscles of Stannius. Minor changes occurring in the MSH cells remain difficult to interpret; the short stimulation during readaptation to FW may be related to a stress effect and/or to release of other peptides present in the MSH cells of fish.     

Vascularization,innervation, and ultrastructure of the endocrine cell types of stannius corpuscles in the teleost Gasterosteus aculeatus

The blood circulation of the Stannius corpuscles, like that of the kidneys to which the corpuscles are attached, represents a portal system. The corpuscles receive blood from the dorsal caudal vein and from a vein coming from the hypaxial musculature. They are drained by veins which enter the caudal parts of the kidneys and therefore endocrine substances released by the corpuscles pass through the kidneys before they enter the general body circulation. The corpuscles are penetrated by sympathetic nerves coming from a small subvertebral ganglion. It is likely that these nerves innervate the muscular coat around the blood vessels. The muscular coat surrounding the renal blood vessels, the collecting tubules and part of the ureters, is innervated by nerves from the same ganglion. The secretory activity of the gland cells appears to be controlled by blood borne factors, because neither synaptic contacts with these cells, nor gap junctions among the cells, have been found in thin sections and freeze-etch replicas of the corpuscles. The corpuscles contain two cell types, both presumed to have endocrine function. Histochemical and ultrastructural data indicate that the gland cells produce glycoproteins. It is likely that the contents of the secretory granules are released by exocytosis. One cell type is structurally similar to the cells described in many other teleosts and thought to be engaged in the synthesis of a hypocalcemic hormone. The ultrastructure of the second cell type resembles cells described only in other migratory species: salmonids and eels. It may be involved in the control of monovalent ions.     

A second cell type in stannius bodies of two euryhaline teleost species

In the corpuscles of Stannius of sticklebacks and eels two cell types are described of presumably endocrine nature. The predominating type, comparable to the cells observed in other species, responds to variation in calcium content of the medium and possibly produces a hypocalcemic hormone. The second cell type is unreactive to calcium. Since it is more active in freshwater than in seawater specimens, this cell type is possibly involved in ionic regulation. It was not found in two seawater teleost species.     

Studies on a hill-stream fish, Pseudecheneis sulcatus (McClelland). II. Histocytology of the corpuscles of Stannius.

There are 1...2 corpuscles of Stannius in the Indian hill-stream teleost, Pseudecheneis sulcatus. They are encapsulated bodies, measuring about 400 micrometer in length and 200 micrometer in width. The secretory cells of CS have fine granules in their cytoplasm which are AF, PTAH, CAH and silverpositive. The cells are arranged along blood sinusoids in which their secretions are probably discharged. Individual neurons and other neural structures are found scattered among the secretory cells.     

Effects of changes in environmental calcium on prolactin secretion in Japanese eel,Anguilla japonica

Effects of changes in environmental Ca²⁺ on the secretion of prolactin, a possible hypercalcemic hormone, were examined both in vivo and in vitro in the Japanese ecl, Anguilla japonica. Transfer of seawater- or freshwater-adapted fish to fresh water, fresh water containing 10 mmol Ca²⁺ · 1^-1 sea water, Ca²⁺-free sea water, or deionized water was accompanied by significant changes in plasma Ca²⁺ levels after 7 days, except for the fish transferred from fresh water to fresh water and from sea water to sea water. Changes in external Ca²⁺ concentrations did not affect plasma prolactin levels, although plasma prolactin levels as well as pituitary prolactin contents were significantly greater in fish in a hypotonic environment than those in a hypertonic environment, regardless of the external Ca²⁺ concentration. Hypercalcemia, induced by removal of the corpuscles of Stannius, did not alter plasma prolactin levles. Incubation of the pituitary in the medium with different Ca²⁺ concentrations (up to 2.9 mmol·l^-1) did not affect the basal release of prolactin, except at an extremely low Ca²⁺ concentration (less than 0.1 mmol·l^-1) where prolactin release was inhibited. Addition of Ca²⁺ ionophore (A23187) to the medium led to a marked and significant increase in prolactin release, indicating that an increase in intracellular Ca²⁺ stimulates prolactin release. However, the effect was not specific to prolactin cells; a similar increase was seen in growth hormone release. These results indicate that changes in environmental Ca²⁺ concentration may not be the primary factor influencing prolactin secretion in the eel; changes in environmental osmolality or Na⁺ levels seem to be more critical for the regulation of prolactin secretion.Abbreviations CSX stanniectomy - DMSO dimethylsulphoxide - DW deionized water - FW fresh water - GH growth hormone - PRL prolactin - SW sea water