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Biomedical subjects

B Borg

Publications and source records attributed to B Borg.

At least 19 recordsLinked to original sources

Activation of polyphosphoinositide metabolism at artificial maturation of Patella vulgata oocytes.

The metabolism of polyphosphoinositides (PPI) has been investigated during the meiosis reinitiation of the oocytes of a prosobranch mollusk, the limpet Patella vulgata. Meiosis reinitiation which leads to germinal vesicle breakdown (GVBD) and metaphase-1 spindle formation was artificially induced by treating the prophase-blocked oocytes with 10 mM NH4Cl, pH 8.2. This treatment, which results in a rise in intracellular pH, triggered a general increase in polyphosphoinositide synthesis. Determinations of phosphorus content showed that maturation induced a 30 to 50% increase in both phosphatidylinositol (PI) and phosphatidylinositol-1 monophosphate (PIP) concentrations. Incorporations of 32PO4 and [3H]inositol have been measured in three classes of polyphosphoinositides: PI, PIP, and phosphatidylinositol 4,5-bisphosphate (PIP2). By comparing incorporation rates of the radiolabeled precursors into PPI before and after meiosis reinitiation, we found that artificial maturation by ammonia induced a 50-fold increase in the turnover of these lipids. No significant burst of inositol 1,4,5-trisphosphate (IP3) was observed after maturation. We suggest that modifications in PPI metabolism occurring at maturation of Patella oocytes might ensure the formation of an important stock of PPI that would be available for the profuse production of IP3, the messenger responsible for the Ca2+ signal at fertilization.

Ammonia

Metabolism of androstenedione and 11-ketotestosterone in the kidney of the three-spined stickleback, Gasterosteus aculeatus.

Stickleback kidneys were incubated with tritiated androstenedione (A4) or 11-ketotestosterone (OT). After the A4 incubations the following steroids were found, testosterone (T), 5 beta-androstane-3,17-dione (5 beta Ad), etiocholanolone (Et), 5 beta-dihydrotestosterone (5 beta DHT), 5 beta-androstane-3 alpha,17 beta-diol (5 beta A3 alpha 17 beta diol), androsterone, 5 alpha-androstane-3,17-dione, 5 alpha-androstane-3 alpha,17 beta-diol (5 alpha A3 alpha 17 beta diol), as well as glucuronides of T, Et, 5 beta DHT, 5 beta A3 alpha 17 beta diol, and 5 alpha A3 alpha 17 beta diol. The metabolites found in the largest amounts were usually T, 5 beta Ad, Et, and the glucuronides of T, Et, and especially of 5 beta A3 alpha 17 beta diol. These results suggest the main pathway to be A4-5 beta Ad-Et-5 beta A3 alpha 17 beta diol-5 beta A3 alpha 17 beta diol-glucuronide or Et-Et-glucuronide-5 beta A3 alpha 17 beta diol-glucuronide. The formation of 5 beta-reduced compounds was larger in postbreeding males and females than in breeding males. The opposite was the case for 5 alpha-reduced compounds. The total formation of glucuronides was lower in the breeding males than in the other groups. After the OT incubations 11-ketoandrostenedione, 17 beta-hydroxy-5 beta-androstane-3,11-dione (tentatively identified), and OT-glucuronide were found. 17 beta-Hydroxy-5 alpha-androstane-3,11-dione was not present.

Androstane-3,17-diol

Temperature, but not photoperiod, influences gonadotropin-releasing hormone binding in the pituitary of the three-spined stickleback, Gasterosteus aculeatus.

Gonadotropin-releasing hormone (GnRH) binding characteristics in pituitaries of stickle-backs under different physiological conditions were studied using D-Arg6-Pro9-salmonGnRH-NEt as labeled ligand. Both males and females displayed marked seasonal changes in the capacity of high-affinity GnRH binding sites; there was a high content in the breeding season (summer) (800-1500 pmol/pituitary) and no detectable high affinity (< 150 pmol) binding in late winter-early spring. The binding capacity was lower in postbreeding fish (ca. 400 pmol/pituitary in females, ca. 900 pmol in males) than in breeding fish (females: ca. 1850, males ca. 1400 pmol). GnRH binding sites were also studied in fish exposed to long and short photoperiod in combination with high and low temperature in winter. Only long photoperiod in combination with high temperature stimulated sexual maturation. The capacity of the GnRH binding sites was similar in fish exposed to long (females 1550 pmol, males 1000 pmol) and short (females 1800, males 900) photoperiod in combination with high temperature. In fish exposed to low temperature, binding was nondetectable irrespective of the photoperiod.

Amino Acid Sequence

Polyphosphoinositide metabolism during the fertilization wave in sea urchin eggs.

A transient increase in intracellular free calcium is believed to be the signal responsible for the stimulation of the egg metabolism at fertilization and the resumption of the cell cycle. We have studied how the polyphosphoinositides (PPI) turn over at fertilization in sea urchin eggs, in order to determine the relationship between the metabolism of these lipids and the calcium signal. We compare the patterns of PPI turnover that occur during the first minute following fertilization in eggs in which PPI are labelled to steady state with [3H]inositol or [3H]arachidonate with that in which PPI are labelled for a shorter period with [3H]inositol. When eggs are labelled to apparent isotopic equilibrium with either [3H]inositol or [3H]arachidonate, no early increase in [3H]PtdInsP2 occurs while PtdIns decreases slightly. On the contrary, when not labelled to isotopic equilibrium, all [3H]PPI increase during the first 15 seconds following fertilization. We find that, within seconds, fertilization triggers a 600-fold increase in the turnover of PPI, producing an amount of InsP3 apparently sufficient to trigger calcium release. We suggest that phosphoinositidase C and PtdInsP kinase, responsible respectively for the hydrolysis and synthesis of PtdInsP2, are both stimulated to a comparable degree in the first 30 seconds following fertilization and that net changes in the amount of PtdInsP2 at fertilization are very sensitive to the relative levels of activation of the two enzymes. Activating the eggs with the calcium ionophore A23187 showed that both these enzymes are sensitive to calcium, suggesting that calcium-dependent InsP3 production might play a role in the initiation and/or the propagation of the fertilization calcium wave.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Phosphatidylinositol 4-Kinase

Effects of castration and androgen treatment on aromatase activity in the brain of mature male Atlantic salmon (Salmo salar L.) parr.

Brain homogenates from male Atlantic salmon parr aromatized tritiated androstenedione to estrogens. The aromatase activity in homogenates of whole brains from castrated male parr was lower than that in homogenates from sham-operated male parr in autumn. This was also found in homogenates of the telencephalon and diencephalon, but not in homogenates of the tectum opticum. Treatment of castrated males with testosterone (T) and 11-ketoandrostenedione (OA) increased the aromatase activity in whole brains. T, but not OA, also significantly increased aromatase activity in all brain parts. In spring, half a year after the peak breeding period, mature parr males had a lower aromatase activity in telencephalon, diencephalon and tectum than that found in castrated males in autumn, whereas immature males displayed an even lower level of activity.

Androgens

Seasonal changes in and effect of castration/androgen replacement on the plasma levels of five androgens in the male three-spined stickleback, Gasterosteus aculeatus L.

The seasonal changes in plasma levels of the androgens 11-ketotestosterone (OT), testosterone (T), 11 beta-hydroxytestosterone (OHT), 11-ketoandrostenedione (OA), and 11 beta-hydroxyandrostenedione (OHA) were measured in the male three-spined stickleback (Gasterosteus aculeatus L). OT was the dominant plasma androgen in the breeding season in summer and was the only androgen that peaked during this period. The levels of OT correlated closely with the development of male secondary sexual characters and reproductive behavior. T and OHT were low in all seasons, whereas OHA and OA displayed the highest levels in early winter. During the postbreeding period, the time of active spermatogenesis, all measured steroids were low. Castration resulted in an almost complete loss of plasma OT and reduced T, whereas OHT, OHA, and OA were not reliably influenced. Androstenedione implants in castrated fish increased plasma T and OA implants increased plasma OT, suggesting a nontesticular site of conversion.

Androgens

Plasma growth hormone levels increase during seawater exposure of sexually mature Atlantic salmon parr (Salmo salar L.).

At the time of smoltification in May, smolts and sexually mature male parr were transferred to seawater (25% salinity) and sampled after 6 and 24 hr. Plasma levels of growth hormone (GH) were measured by radioimmunoassay. There was no difference in GH levels between smolts and mature parr in fresh water. GH levels did not change during exposure of smolts to seawater. In the mature male parr, plasma GH levels increased after 24 hr, when the levels were almost five times those of the freshwater controls. In the mature male parr, there was an increase in plasma osmolality, sodium, and magnesium after 24 hr in seawater; magnesium also increased after 6 hr. The levels of potassium and calcium did not change in either immature parr or mature male parr. The increase in plasma GH levels in the mature parr in seawater may be part of a mechanism to increase hypoosmoregulatory ability in fish not ready for seawater entry.

Animals

Conversion of 11-ketoandrostenedione to 11-ketotestosterone by blood cells of six fish species.

Blood cells from Baltic salmon, Salmo salar, three-spined stickleback, Gasterosteus aculeatus, eel pout, Zoarces viviparus, crucian carp, Carassius carassius, African catfish, Clarias gariepinus, and reedfish, Calamoichthys calabaricus, were incubated with tritiated 11 beta-hydroxyandrostenedione (OHA) or 11-ketoandrostenedione (OA). In all fish there was conversion of OA to 11-ketotestosterone (OT), indicating that 17 beta-hydroxysteroid dehydrogenase activity was present in the blood cells. On the other hand, OHA was not converted to 11 beta-hydroxytestosterone in any fish. The addition of serum to the incubates largely prevented the OA-OT conversion by salmon blood cells.

Androstenes

Effects of castration on pituitary gonadotropic cells of the male three-spined stickleback, Gasterosteus aculeatus L., under long photoperiod in winter: indications for a positive feedback.

Male three-spined sticklebacks caught in winter were castrated or sham-operated and subsequently kept under long photoperiod at about 20 degrees for a month. With this treatment the sham-operated fish attained breeding condition. The gonadotropic cells of the sham-operated fish contained significantly more dilated endoplasmic reticulum and fewer granules than those of the castrated fish, indicating a higher secretory activity of the gonadotrops in the sham-operated fish. These findings suggest the existence of a physiological positive feedback within the gonadal-pituitary axis of the male stickleback when stimulated into its breeding condition by long photoperiod.

Animals

Characterization of gonadotropin-releasing hormone binding sites in the pituitary of the three-spined stickleback, Gasterosteus aculeatus.

Binding sites for gonadotropin-releasing hormone (GnRH) in stickleback pituitary homogenates were characterized using an iodinated, superactive analog of salmon GnRH (sGnRH), D-Arg6-Pro9-sGnRH-NEt (sGnRHa). Binding of 125I-sGnRHa reached equilibrium after 60 min incubation at 4 degrees and was a function of tissue concentration. The specificity of 125I-sGnRHa binding was demonstrated by displacement with sGnRHa, sGnRH, and Buserelin [D-Ser(t-Bu)6-Pro9-GnRH-NEt]. Both Scatchard analyses of saturation data and displacement curves revealed a single class of high-affinity binding sites (Ka = 0.71 +/- 0.03 X 10(9) M-1, Bmax = 1087 +/- 165 fmol/mg protein).

Animals

Steroid metabolism in the testes of the breeding and nonbreeding three-spined stickleback, Gasterosteus aculeatus.

Steroid metabolism in the testes of sticklebacks was studied in vitro by tissue incubations with [3H]pregnenolone or [3H]androstenedione as precursors. In males in full reproductive condition (nesting), [3H]pregnenolone was mainly converted via progesterone and 17 alpha-hydroxyprogesterone into androstenedione, 11 beta-hydroxyandrostenedione, and 11-ketoandrostenedione. The latter was the largest product formed. The main products from the [3H]androstenedione incubation, 11 beta-hydroxyandrostenedione and 11-ketoandrostenedione, confirm these findings. The rate of androgen synthesis, especially of 11-ketoandrostenedione, was much lower after the end of the breeding season.

Androstenes

Aromatase activity in the brain of the three-spined stickleback, Gasterosteus aculeatus. III. Effects of castration under different conditions and of replacement with different androgens.

Non-breeding stickleback males were gonadectomized or sham-operated in winter. They were then exposed to a high temperature and either a short photoperiod which did not induce breeding or to a longer, stimulatory, photoperiod. In the latter experiment empty Silastic capsules or capsules filled with the aromatizable androgen, androstenedione or the non-aromatizable androgen, 11-ketoandrostenedione were implanted intraperitoneally in the operated fish. Furthermore, males that were already in breeding condition were gonadectomized or sham-operated in late spring. Brains were freeze-sectioned and punches from the nucleus preopticus-nucleus anterioris periventricularis region were incubated with [19-3H]-androstenedione. After extraction of steroids with CH2CI2, the tritium activity remaining in the water phase gives a measure of aromatization as tritiated water and formic acid are formed when [19-3H]-androstenedione is aromatized to oestrogens. The aromatase activity was higher in sham-operated than in castrated fish in all three experiments. Both androstenedione and 11-ketoandrostenedione abolished the effect of gonadectomy on aromatization.

Androgens

Aromatase activity in brain and pituitary of immature and mature Atlantic salmon (Salmo salar L.) parr.

Brain homogenates from Atlantic salmon parr converted tritiated androstenedione to testosterone, estrone, and 17 beta-estradiol. The formation of estrogens was markedly higher in homogenates of whole brains from mature parr males than from immature parr males. The highest estrogen synthesis was found in the telencephalon and diencephalon. In both of these parts the estrogen production was higher in mature males than in immature males. There was no difference in estrogen formation by pituitaries from immature female and immature male parr, whereas the formation of estrogens and testosterone was higher in the mature male parr.

Age Factors

Aromatase activity in the brain of the three-spined stickleback, Gasterosteus aculeatus. I. Distribution and effects of season and photoperiod.

Tritiated androstenedione was in vitro aromatized to estrone and estradiol by the stickleback brain. Highest aromatase activity was found in the diencephalon, particularly in the periventricular hypothalamic region containing the nucleus preopticus (NPO), in the area containing the nucleus lateralis tuberis, and in the pituitary. This localization suggests a role of aromatase in controlling the secretion of gonadotropic hormone. The activity in the area containing the NPO was higher in May than in December. The possible role of seasonal changes in aromatase activity in the control of the yearly reproductive cycle is discussed.

Androstenedione

Aromatase activity in the brain of the three-spined stickleback, Gasterosteus aculeatus. II. Effects of castration in winter.

Non-breeding stickleback males were gonadectomized or sham-operated in winter. Treatment with long photoperiod at high temperature stimulated sexual maturation in control fish. Brains were freeze-sectioned and punches were taken with hollow needles from different diencephalic regions and the punches incubated with [19-3H]-androstenedione. After extraction of steroids with CH2Cl2 tritium activity remaining in the water phase was measured. This gives a measure of aromatization as tritiated water and formic acid is formed when [19-3H]-androstenedione is aromatized to estrogens. Aromatase activity in the area containing the nucleus preopticus and the nucleus anterioris periventricularis was dramatically higher in sham-operated males than in gonadectomized ones or in the initial controls. There was no effect of gonadectomy in the other studied regions, including the nucleus lateralis tuberis, lobi inferioris, nucleus posterioris periventricularis and thalamic regions.

Androstenedione

Stimulatory effects of methyltestosterone on pituitary gonadotropic cells and testes leydig cells of the three-spined stickleback, Gasterosteus aculeatus L., in winter.

Male three-spined sticklebacks were kept for a month in short photoperiod at ca. 20 degrees in winter. During this period methyltestosterone (MT), dissolved in the water (35 micrograms/liter), stimulated the appearance of breeding colors and kidney hypertrophy, characters which were both absent in the controls. MT furthermore prevented the onset of spermatogenesis. The gonadotropic cells of the MT-treated fish contained significantly more dilated endoplasmic reticulum and less granules than those of the controls. The nuclei of the testes Leydig cells were enlarged after treatment with MT. These findings suggest the existence of a positive feedback of androgen on the gonadal-pituitary axis of the male stickleback in winter.

Animals