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Biomedical subjects

I P Callard

Publications and source records attributed to I P Callard.

At least 55 records · Page 3Linked to original sources

Plasma estradiol, testosterone, and progesterone levels during the ovulatory cycle of the skate (Raja erinacea).

Amounts of estradiol, testosterone, and progesterone in plasma were measured during the reproductive cycle of female Raja erinacea. Estradiol titers correlated directly with follicle size in females undergoing ovarian recrudescence, while highest concentrations were found in females with preovulatory follicles. These data indicate that as follicles grow, their steroidogenic capacity increases. In mature, nonspawning females, titers of estradiol and testosterone varied markedly. Progesterone was not detected in peripheral plasma of skates that did not produce eggs during the observation period. In females producing eggs, estradiol and testosterone predominated during the follicular phase of each spawning cycle. While estradiol and testosterone were elevated, progesterone was not detectable in the peripheral circulation. As ovulation and formation of capsules approached, plasma estradiol and testosterone declined to near baseline levels. Circulating progesterone rose sharply two days before encapsulation of ovulated eggs and remained elevated for only two days. On the day of encapsulation, concentrations of plasma progesterone had fallen to nearly baseline levels. Progesterone titers remained low throughout egg retention and oviposition. These measurements demonstrate that progesterone titers are elevated at specific times during the reproductive cycle of the skate and clearly suggest that progesterone is critically involved in events occurring at ovulation, encapsulation, and possibly oviposition.

Animals↗

Characterization of the testicular binding site for iodinated rat FSH in the turtle, Chrysemys picta.

1. To correlate the morphological observations with the known gonadotropic activity of FSH in the turtle testis, studies of the binding of iodinated FSH were conducted. 2. These demonstrated the presence of gonadotropin-binding sites of high affinity (apparent Kd = 10(-10) M) for [125I]rFSH in turtle testicular membrane preparations. 3. Although these sites did not bind iodinated human LH or avian LH, these hormones, as well as PMSG and FSH, were effective competitive inhibitors of the binding of the radioligand. 4. Binding of the radioligand to the testis was influenced by duration of incubation and temperature. 5. Binding activity was lost after incubation with proteolytic enzymes (trypsin, pronase) but not with DNAase, lipase, collagenase and neuraminidase. 6. The binding exhibited target organ specificity (no binding observed in brain, epididymis, lung, muscle and pancreas). 7. In addition, the number of binding sites varied according to the stage spermatogenesis, being highest when the tubules contained spermatocytes and spermatids, intermediate when the tubules consisted to Sertoli cells and spermatogonia and lowest st spermiation.

Animals↗

Validation of radioimmunoassay systems for the measurement of 11-keto- and 11 beta-hydroxytestosterone in teleost blood.

Highly specific antisera for 11-keto- and 11 beta-hydroxytestosterone have been raised in sheep. Assay systems for the simultaneous measurement of 11-ketotestosterone, 11 beta-hydroxytestosterone, testosterone, progesterone and estradiol-17 beta were validated for Ictalurus nebulosus plasma and Carassius auratus serum. In males of both species 11-ketotestosterone and testosterone were the major steroids detected. In females, testosterone and estradiol-17 beta were the predominant steroids measured. Data from samples taken at different stages of the annual cycle suggest that seasonal fluctuations in gonadal steroid secretion occur in I. nebulosus and C. auratus.

Aging↗

Effects of relaxin and insulin on reproductive tract size and early fetal loss in Squalus acanthias.

These experiments demonstrate that both porcine relaxin and bovine insulin can increase the cervical cross-sectional area in Squalus acanthias. When given after estradiol priming, the magnitude of this response is greater. This effect is limited to Stage C females in which pregnancy is over 75% complete. The overall result is premature loss of developing fetuses. The effect of relaxin or insulin on cervical cross-sectional area and fetal loss is unrelated to an effect on blood sugar. Cervical weights are affected little by hormone treatment. Relaxin and insulin do not increase the cross-sectional area of the anterior uterine constriction. These results suggest the involvement of a relaxin-like molecule during normal parturition in the spiny dogfish.

Animals↗

High affinity binding of [3H]R5020 and [3H]progesterone by putative progesterone receptors in cytosol and nuclear extract of turtle oviduct.

The purpose of this investigation was to identify and characterize putative cytosolic and nuclear forms of progesterone receptor in the female reproductive tract of a turtle, Chrysemys picta. A dextran-coated charcoal adsorption assay and DNA-cellulose affinity chromatography were used as the primary methodologies with [3H]R5020 [3H-labeled 17 alpha-dimethyl-19-norpregna-4,9-diene-3,20-dione) and [3H]progesterone (P4) as the ligands. The receptor was of high affinity (Kd = 4.7 X 10(-10) M for [3H]P4; 2.2 X 10(-10) M for [3H]R5020) and limited capacity (500-6000 fmol/g tissue wet wt). Association was rapid (apparent equilibrium being reached in 30-40 min), as was dissociation (t1/2 = 45 min for [3H]P4 and 180 min for [3H] R5020). The putative receptor demonstrated strict steroid specificity, binding progestins but not estrogens, androgens, or glucocorticoids. Heterogeneity of the cytosolic receptor was demonstrated as two forms eluting off DNA-cellulose columns at 0.2- and 0.3-M salt concentrations. Binding of cytosolic receptor to DNA-cellulose was not increased by preexposure of cytosol to 25 C for 30 min. Some variations in cytosolic, but not nuclear, receptor were associated with different stages of the reproductive cycle and were positively correlated with body weight. Preliminary studies using an explant culture system suggest that the progesterone receptor in turtle oviduct may be maintained by estrogen and translocated from the cytosol to the nucleus by P4. In summary, we have partially characterized a putative P4 receptor in the oviduct of the turtle that is similar to mammalian and avian P4 receptors in specificity, affinity, and other physicochemical properties, supporting the idea that steroid receptor proteins have been highly conserved in vertebrate evolution. However, temperature sensitivity of activation and DNA affinity are different in the turtle and suggest modifications that may be related to physiological adaptation in such a poikilothermic species.

Animals↗

Estradiol binding activity in epididymal cytosol of the turtle, Chrysemys picta.

Estrogen binding activity was investigated in the epididymis of the turtle, Chrysemys picta using DNA-cellulose affinity chromatography. A component binding estradiol-17 beta specifically with high affinity (Kd:8.0 X 10(-10) M) and limited capacity (20 fmol/mg protein) was demonstrated in the epididymal cytosol. In addition, binding of estradiol-17 beta was sensitive to excess (100-fold) diethylstilbestrol or natural estrogens (estradiol-17 beta, estrone, and estriol) but not to progesterone or androgens (testosterone and 5 alpha-dihydrotestosterone). The specific estrogen binding macromolecules eluted from DNA-cellulose columns sedimented at 4-5 S in linear 5-20% sucrose gradients. These characteristics suggest the presence of an estrogen receptor in this androgen target organ.

Animals↗

Characterization of an estrogen receptor in the turtle testis.

Using DNA-cellulose affinity chromatography, an estrogen-binding component having the properties of a classical estrogen receptor was characterized from testicular cytosol of the freshwater turtle, Chrysemys picta. This putative cytoplasmic receptor exhibited high affinity (Kd = 7.0 X 10(-10) M), low capacity (1-4 fmol/mg protein), and steroid binding that was specific for estrogens. It was not present in plasma, muscle, kidney, or lung. A temperature-dependent conversion of turtle testicular estrogen receptor from 4 to 5 S occurred on DNA-cellulose columns, and resembled that in mammalian testis and other target tissues. After a single injection of estrogen at 3 hr, cytoplasmic receptors were depleted with a concomitant increase of nuclear receptors. Identification in turtle testis of an estrogen-binding macromolecule having the physicochemical properties of mammalian estrogen receptors is further evidence that receptors have been widely conserved in many tissue types through vertebrate phylogeny and supports the idea that the testis is an important target of estrogen action.

Animals↗

Estrogen receptors in the turtle brain.

Estrogen binding activity in the CNS of the freshwater turtle, Chrysemys picta, was investigated using DNA-cellulose affinity chromatography. An estrogen binding component (EBC) with the characteristics of an estrogen receptor species was demonstrated in the brain cytosol extracts from both sexes. This EBC exhibited high affinity (K d = 10 -10 M), low capacity (n = 0.8 to 6.0 fmol/mg cytosol protein) and binding specificity. The bound estradiol-17 beta which adhered to DNA-cellulose was sensitive to excess synthetic and natural estrogens (DES, E2, E1 and E3) but not to progesterone and androgens (5 alpha- DHT and T). Specific estradiol-17 beta binding was not detected in plasma or non-target tissues such as lung, kidney and muscle. The topographic distribution of cytoplasmic EBC was similar in males and females, with binding highest in the hypothalamus-preoptic areas (HPOA) followed by the remaining forebrain (RFB). The mid/hindbrain (HB), consisting of the optic lobes, cerebellum and underlying brain stem had significantly lower concentrations of EBC. Monthly data (from May to October) suggest that variations in EBC concentrations occur during the year.

Animals↗

Radioimmunoassay of plasma progesterone, testosterone, total estrogen and immunoreactive gonadotropin in the nesting and non-nesting green sea turtle, Chelonia mydas (L.).

Plasma progesterone, testosterone, total estrogens and immunoreactive gonadotropin were measured in nesting and non-nesting sea turtles, Chelonia mydas. Progesterone and gonadotropin concentrations were significantly higher in nesting than in non-nesting turtles, testosterone was not significantly different in either group and total estrogens appeared to be slightly in the nesting group.

Animals↗

Sex steroid binding proteins in non-mammalian vertebrates.

High affinity cytosolic and nuclear steroid binding macromolecules have been identified in steroid target tissues of representative amphibians, reptiles and birds. In addition to these traditional "receptors" evidence exists which indicates the presence of other steroid binding proteins, varying in affinity and specificity, in the cytosol of steroid targets (liver, oviduct) of these same species. The possible relationships between cytosolic binding proteins of differing affinity and specificity and their relevance to hormone action in non-mammalian species is discussed.

Amphibians↗