Ultimobranchial gland respond in a different way in male and female fresh water teleost Mastacembelus armatus (Lacepede) during reproductive cycle.

The present study was carried out to analyze the differences in the activity of ultimobranchial gland (UBG) between male and female fresh water teleost Mastacembelus armatus during reproductive cycle. Considerable variations in the nuclear diameter of UBG cells and plasma calcitonin (CT) levels during different reproductive phases of testicular and ovarian cycle suggested that the activity of the UBG depends upon the sexual maturity of fishes. A positive correlation was observed between plasma CT and sex steroid levels and the gonadosomatic index in both sexes which further confirmed the involvement of UBG in the processes related to gonadal development in fishes irrespective of the sex. Sudden increase in the level of plasma CT and nuclear diameter of UBG cells after administration of 17 α-methyltestosterone in males and 17 ß-estradiol in females during resting phase of the reproductive cycle clearly showed that UBG becomes hyperactive with increases in the level of sex steroids. Plasma calcium level was also found to be positively correlated with gonadal maturation in females. However no such change in plasma calcium level in relation to testicular cycle was observed. Thus it can be concluded that UBG becomes hyperactive during gonadal maturation but its role differs between male and female fishes. In females it may involved in both gonadal maturation and plasma calcium regulation while in males its involvement in calcium regulation was not justified. Variations in the level of CT during various phases of testicular cycle evidenced its involvement in gonadal maturation only.

Histological alterations in the ultimobranchial gland of teleost Heteropneustes fossilis in response to chlorpyrifos treatment.

In this study, an experiment was performed on Heteropneustes fossilis for short-term (1.76 mg/L chlorpyrifos, i.e., 0.8 of 96-h LC50) and long-term (0.44 mg/L chlorpyrifos, i.e., 0.2 of 96-h LC50) exposure. The fish were sacrificed after 24, 48, 72 and 96 h in the short-term experiment and after 7, 14, 21 and 28 days in the long-term experiment. On these intervals, blood was collected and analysis of serum calcium was done. Ultimobranchial glands were also fixed for histological study. The serum calcium levels of H. fossilis exhibit a decline after 24 h following exposure to chlorpyrifos. This decrease continues until the end of the experiment (96 h). The serum calcium levels of chronically exposed fish exhibit a decrease on day 7. Thereafter, the levels continue to fall progressively until the end of the experiment (28 days). The ultimobranchial gland of chlorpyrifos treated fish exhibits no histological change up to 48 h. After 72 h, there is a decrease in the staining response of cytoplasm of the ultimobranchial cells. The nuclear volume of these cells is slightly decreased. After 96 h following chlorpyrifos exposure, these changes become exaggerated. In chlorpyrifos-treated fish there is no change in the histological structure of the ultimobranchial gland up to 14 days. After 21 days, the cytoplasm of ultimobranchial cells stain feebly and the nuclear volume of these cells exhibits a decrease. Following 28 days treatment, the nuclear volume of these cells records a further decrease and the gland depicts vacuolization and degeneration at certain areas.

Influence of calcitonin administration on ultimobranchial and parathyroid glands of pigeon, Columba livia.

Pigeons were divided into two numerically equal groups (A and B) containing 30 specimens each. Birds from group A were intraperitoneally injected daily with 0.1 mL/100 g body wt of vehicle (0.1 M acetate buffer, pH 4.3 containing 0.1% gelatin). Specimens from group B were intraperitoneally injected daily with 1 microg/100 g body wt of salmon calcitonin. Six birds were sacrificed from each group 2 h after the last injection on 1st, 3rd, 5th, 10th, and 15th day of the experiment. After collection of blood samples, ultimobranchial and parathyroid glands were fixed for histological studies. In calcitonin-treated C. livia, the plasma calcium levels exhibit a progressive decline from day 1 till day 5. On day 15, the levels become more or less similar to the control value. No change has been noticed on day 1 in the plasma phosphate levels of calcitonin-treated C. livia. The levels decrease progressively from day 3 to day 5; thereafter, it exhibits an elevation so that on day 15, normal plasma phosphate levels is achieved. The ultimobranchial gland of C. livia exhibits no change up to day 5 following calcitonin treatment. The nuclear volume of ultimobranchial cells exhibits a decrease on day 10. This response progresses up to day 15. Few degenerating cells are also discerned following 15 days calcitonin treatment. The parathyroid gland of calcitonin-treated C. livia exhibits no histological alteration up to day 3. The nuclear volume of parathyroidal cells exhibits a progressive increase from day 3 till the close of the experiment (day 15). Moreover, the gland exhibits more compactness on day 10 and day 15. Few degenerating cells are encountered after day 15 following calcitonin treatment.

Ultimobranchial gland of a freshwater teleost, Heteropneustes fossilis, in response to cadmium treatment.

Heteropneustes fossilis were subjected to 288 mg/L (0.8 of 96 h LC(50)) and 72 mg/L (0.2 of 96 h LC(50)) of cadmium chloride for short-term and long-term experiments, respectively. After sacrificing the fish, the blood was collected on 24, 48, 72, and 96 h in short-term and after 7, 14, 21, and 28 days in long-term experiment and analyzed for plasma calcium levels. Also, ultimobranchial glands were fixed on these intervals. The plasma calcium levels of short-term cadmium-exposed fish remain unchanged after 24 h. The levels exhibit a progressive decrease from 48 h onwards. The fish exposed to cadmium for 7 days exhibit a decrease in the plasma calcium level. Thereafter, the levels progressively decrease till the end of the experiment (28 days). Up to 72 h exposure of the fish to cadmium, the ultimobranchial gland exhibits no histological change. After 96 h, a decrease in the staining response of the cytoplasm of ultimobranchial cells has been noticed. The nuclear volume of these cells records a slight decrease. Up to 14 days of cadmium exposure, there is no change in the histological structure of ultimobranchial gland. After 21 days following the exposure, the ultimobranchial cells exhibit a slight decrease in the staining response of the cytoplasm and the nuclear volume of these cells records a decrease. Following 28 days cadmium exposure the nuclear volume exhibits a further decrease, and degeneration and vacuolization sets in.

Ultimobranchial and parathyroid glands of the pigeon Columba livia in response to 1,25OH2D3 administration.

Columba livia were given daily intraperitoneal injections of 100 pmol of 1,25(OH)(2)D(3)/100 g body wt. for 15 days. Ultimobranchial and parathyroid glands were fixed on 1st, 3rd, 5th, 10th and 15th day of the experiment. Following 1,25(OH)(2)D(3) treatment, the plasma calcium levels of pigeon remain unchanged on day 1. The levels increase significantly after day 3 which progresses up to day 10. The plasma calcium level becomes normal at day 15. The plasma inorganic phosphate levels of Columba livia injected with 1,25(OH)(2)D(3) remain unaffected up to day 3. The levels exhibit a progressive increase from day 5 to day 10. The levels become normophosphatemic at day 15. Up to day 3 following 1,25(OH)(2)D(3) treatment, there is no change in the ultimobranchial gland of Columba livia. The gland exhibits an increased activity after 5 days 1,25(OH)(2)D(3) treatment which is evident by the increased nuclear volume and weak staining response of the cytoplasm of ultimobranchial cells. After day 10, the nuclear volume depicts a further increase and a few completely exhausted cells are discerned. Following 15-day 1,25(OH)(2)D(3) treatment the nuclear volume records an increase and degenerating cells have been observed at certain places. The parathyroid glands of 1,25(OH)(2)D(3)-treated Columba livia remain unaffected up to day 5. After day 10 and day 15, there is a progressive decrease in the nuclear volume of parathyroidal cells and reduced chromaticity of nuclei has been noticed. Moreover, after 15 days few degenerating cells are observed.

Calcitonin is expressed in the chicken pituitary gland: influence of gonadal steroids and sexual maturation.

Calcitonin (CT) is primarily produced by the thyroid C cells in mammals or by the ultimobranchial gland in chickens. CT is also expressed by the pituitary gland in rats in which it functions as a paracrine factor causing decreased lactotroph proliferation and prolactin (PRL) secretion. Gonadal steroids influence CT expression in the rat pituitary gland. However, the expression of the CT gene in the pituitary gland of chickens or of any other avian species has not previously been reported. We have tested the hypotheses that CT is expressed in the chicken pituitary gland, and that its expression is influenced by sexual maturation or in response to ovarian steroid administration. We have detected robust expression of CT cDNA in the chicken pituitary gland by reverse transcription/polymerase chain reaction (PCR). The sequence of the pituitary-derived CT cDNA is identical to that of the ultimobranchial gland. CT-immunoreactive (ir) cells have been observed throughout the anterior pituitary gland by confocal microscopy. Many of the PRL-ir cells show co-localization with CT-ir cells. Quantitative real-time PCR analysis has revealed an inverse relationship between the quantities of PRL mRNA and CT mRNA in the pituitary gland: sexually mature hens contain lower amounts of CT mRNA but larger quantities of PRL mRNA compared with sexually immature chickens. Estradiol and/or progesterone treatment of sexually immature chickens leads to a significant decrease in the quantity of pituitary CT mRNA relative to that in the vehicle-treated chickens. We conclude that pituitary CT plays an important paracrine/autocrine role in the control of lactotroph function and PRL secretion in the chicken.

Dynamics of plasma Ca and calcitonin levels in stonefish (Inimicus japonicus) administered a high-Ca solution into the stomach.

In stonefish, changes in plasma total Ca and calcitonin levels were examined after administration of a high-Ca solution into the stomach. Blood was taken successively at 0, 1, 3, 9, 33, and 81 hr from a fine tube cannulated into the aortic bulb. Plasma Ca levels increased acutely at 1 hr and attained the peak after 3-9 hr of the administration. Although plasma calcitonin levels did not exhibit conspicuous changes for 1-3 hr, they began to rise significantly at 33 hr. The plasma Ca level began to decline significantly at 33 hr, although the level was still significantly higher than the initial level. At 33 hr, however, the plasma calcitonin level still continued to increase. At 81 hr, the plasma Ca level had returned to the initial level. At that time, the plasma calcitonin level was also significantly lower than that at 33 hr. These results suggest that, in stonefish, the ultimobranchial gland has the ability to respond physiologically to rises in plasma Ca levels, to secrete calcitonin, and to cease the secretion when the plasma Ca levels return to the initial level.

Responses of the ultimobranchial gland to vitamin D3 treatment in freshwater mud eel, Amphipnous cuchia, kept in different calcium environments.

Freshwater mud eel, Amphipnous cuchia, were injected intraperitoneally daily with 100 ng of vitamin D3/100 g body weight and maintained in media containing either no calcium or different calcium concentrations. The eels were killed after 1, 3, 5, 10 and 15 days following the treatment and their serum calcium levels were measured. The ultimobranchial glands were fixed and processed using the routine paraffin method for histological studies. The results of the present study indicate that vitamin D3 can induce hypercalcaemia in eels kept in different calcium environments. Also, the ultimobranchial glands became hyperactive following vitamin D3 treatment. It is concluded that in mud eels, the gland has a calcium-regulating function.

Direct raises in blood Ca levels by infusing a high-Ca solution into the blood stream accelerate the secretion of calcitonin from the ultimobranchial gland in eels.

In eels, a CaCl(2) solution was infused into the pneumatic duct vein. Plasma Ca levels were significantly increased during 3 hr and were followed by significant raises in plasma calcitonin levels. These results strongly suggest that, in eels, direct raises in blood Ca levels by infusion of a high-Ca solution via blood vessels can accelerate the secretion of calcitonin from the ultimobranchial gland.

Influence of prolactin administration on the ultimobranchial body and parathyroid glands of the freshwater snake, Natrix piscator.

The effects of ovine prolactin (10 I.U./100 g body wt) on the ultimobranchial body and parathyroid glands were investigated in Natrix piscator for 15 days. The ultimobranchial body of the prolactin treated specimens depicts signs of increased activity whereas the parathyroid glands exhibit reduced activity.

Laminin-induced process outgrowth from isolated fetal rat C-cells.

A method for isolation of C-cells from rat fetuses was developed, and the morphological plasticity of the cells in primary culture systems was tested. Thyroid-parathyroid-ultimobranchial body (UB) complexes from 16-day rat fetuses were treated with 0.1% collagenase and 1000 PU/ml Dispase at 37 degrees C for 1 h. After dissociation by pipetting, UBs were obtained as remaining cell aggregates with diameters of 150-200 microns. The isolated UBs were cultured on untreated, fibronectin-coated, or laminin-coated substratum in Dulbecco's modified Eagle's medium/Ham's nutrient mixture F-12 (1:1) supplemented with 5% fetal calf serum. In some experiments, the medium was changed to serum-free medium after 24 h of incubation, until the UBs had formed cell sheets. At Day 4 in vitro, the cultures were subjected to immunostaining using anti-calcitonin antiserum. On untreated or fibronectin-coated substratum, most of the C-cells exhibited polygonal or ovoid shapes, and 5-8% of them were found to project processes. On laminin-coated substratum, the ratio of process-bearing C-cells to total C-cells was 23% in serum-supplemented medium and 51% in serum-free medium. The longest processes reached 150 microns in length. The processes were intensely reactive with anti-alpha-tubulin antibody and were completely disintegrated by colcemid, suggesting that the microtubule cytoskeleton participated in the maintenance of the processes. Thus it was demonstrated that fetal rat C-cells are still responsive to environmental signals, such as laminin, and extend neuritic processes.

Dose-dependent vitamin D3, 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3-induced hypercalcemia and hyperphosphatemia, and the correlative changes in the activity of ultimobranchial and parathyroid glands of the toad, Bufo andersoni Boulenger.

Vitamin D3, 25(OH) vitamin D3 and 1,25 (OH)2D3 were administered daily to unfed male Bufo andersoni for 15 days which resulted in a significant hypercalcemia and hyperphosphatemia in a dose-dependent fashion. 1,25(OH)2D3 is more potent than the other metabolites. The treatment activated ultimobranchial gland but induced degenerative changes in parathyroid of the toads.

Ultimobranchial body and parathyroid gland of the frog, Rana tigrina in response to calcitonin administration.

The effects of salmon calcitonin (0.25 MRC mU/g body wt) on the serum calcium and phosphate levels as well as on the activity of ultimobranchial body and parathyroid glands was investigated in the frog, Rana tigrina for 15 days. The hormone evokes hypocalcemia (on day 1 and day 3) which is followed by a significant hypercalcemia on day 10. Thereafter, the level of calcium decreases again on day 15. Calcitonin induces hypophosphatemia (on day 3 and day 5). Thereafter, hyperphosphatemia is recorded on day 10. By day 15 normal serum phosphate value is achieved. After treatment with calcitonin, the ultimobranchial body becomes inactive and the parathyroid glands get activated.

Ultimobranchial body and parathyroid gland of the parrot Psittacula psittacula in response to experimental hypercalcemia.

Psittacula psittacula when subjected to long term hypercalcemia by intramuscular injections of vitamin D2 (20,000 I.U.) on alternate days and by increasing dietary calcium, exhibit a rise in the serum calcium level after 10, 20 and 30 days of treatment as compared to their corresponding controls. The ultimobranchial cells show progressive hypertrophy up to 20th day of the treatment. From 20th day till the end of the experiment (30 days) these cells show feeble staining response. The parathyroid glands suffer from degenerative changes due to its inactivity under chronic hypercalcemia.

Structure and behaviour of ultimobranchial gland in response to vitamin D3--induced hypercalcemia in male Clarias batrachus.

Hypercalcemia was induced in Clarias batrachus by treating them with vitamin D3 (5,000 I.U./100 g body wt.) and/or 0.5% solution of CaCl2. The animals were killed on 1st, 3rd, 5th, 9th, 13th and 17th days after the initiation of the experiment. Histological preparations of the ultimobranchial gland (UBG) were made. The gland exhibits nuclear hypertrophy, hyperplasia and loss of staining response corresponding to the rise in serum calcium levels. At later intervals, the UBG shows exhaustion and degeneration which is evident from vacuolization and nuclear shrinkage of the ultimobranchial cells after day 13 in groups B and C and day 9 in group D.

Ultimobranchial body of Notopterus notopterus in relation to calcium and sodium rich environments.

The UBB of Notopterus notopterus is a paired structure situated between the oesophagus and sinus venosus. Both right and left lobes of the gland are enveloped by a common thick connective tissue which gets constricted between the lobes and separates them. Numerous follicles of varying sizes are encountered in each gland. In N. notopterus the effects of hypercalcaemia (caused by keeping the specimens in 0.5% of CaCl2 solution and by injecting 4000 I.U. of vitamin D2 on alternate days) on UBB has been observed. The effects of NaCl rich environment (created by keeping the fish in 0.5% NaCl solution) on this gland has also been studied. In the UBB of N. notopterus the activity of the gland is observed in terms of: 1. increase in the blood supply of the gland and the dilation of the blood vessel, 2. increase in the height of the follicular epithelium, 3. cytoplasmic hypertrophy resulting in the increase in secretory processes, 4. appearance of pseudostratified epithelium in place of single layered cuboidal follicular epithelium and 5. nuclear and cellular hypertrophy. According to these characteristics it is evident that the gland from group II shows gradual activity from the 2nd day onwards and is maximum on the 6th day. From 8th day to the close of the experiment gradual inactivity of the gland is discerned--follicles get atrophied and the cells appear in clumps. The gland from group III shows a good response to its environment and is more hypertrophied as compared to that of group II. The activity of the gland closely parallels serum sodium levels which increase up to the 8th day when UBB shows the maximum activity. The serum sodium level rises from a normal of 110 m eq/l to a peak of 180 m eq/l on 8th day. After 10 days onwards the gland shows gradual inactivity and degeneration. The serum sodium level is 130 m eq/l on 12th day. These observations support the view that the main role of UBB in N. notopterus lies in sodium metabolism and it is only partially responsible for calcium regulation.