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D R Idler

Publications and source records attributed to D R Idler.

At least 19 recordsLinked to original sources
PubMed1992

Steroids involved with final oocyte maturation in the winter flounder.

A number of androgens and progestogens including 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20-P) were examined in female winter flounder as possible maturation inducing steroids (MIS). During final oocyte maturation serum levels of testosterone (T) and 17 beta-hydroxy-5 beta-androsten-3-one (5 beta-T) peaking at over 200 ng/ml and pregnenolone (PE) at 40 ng/ml were the predominant steroids found from each major group. High levels of T and 5 beta-T were correlated with oocyte stages characterized by germinal vesicle migration. Of the PEs measured, maximum serum levels of PE, 3 beta,17 alpha-hydroxy-5-pregnen-20-one (17-PE) and 3 beta,17 alpha, 20 beta-dihydroxy-5-pregnene (17,20-PE) were found during later oocytes stages associated with germinal vesicle breakdown. Levels of 17,20-P, an established MIS in most fish, were almost non-detectable (less than 0.1 ng/ml serum) in females throughout all stages of final oocyte maturation. Incubations of ovarian follicles in vitro with physiological concentrations of T and 5 beta-T indicated that these steroids could induce all stages of final oocyte maturation. Similar in vitro incubations showed that 17-PE and 17,20-PE were only effective on germinal vesicle breakdown. The principal conclusions are that T, 5 beta-T and the PEs can be considered as MISs in winter flounder and the PE pathway predominates during the final stages of oocyte maturation in winter flounder in contrast to progesterones which predominate in other fish species, mostly salmonids, studies to date.

Androgens↗
PubMed1992

Growth enhancement in transgenic Atlantic salmon by the use of an "all fish" chimeric growth hormone gene construct.

We have developed an "all fish" growth hormone (GH) chimeric gene construct by using an antifreeze protein gene (AFP) promoter from ocean pout linked to a chinook salmon GH cDNA clone. After microinjection into fertilized, nonactivated Atlantic salmon eggs via the micropyle, transgenic Atlantic salmon were generated. The presence of the transgene was detected by polymerase chain reaction (PCR) using specific oligonucleotide primers. A number of these transgenic fish showed dramatic increases in their growth rate. At one year old, the average increase of the transgenic fish was 2 to 6 fold and the largest transgenic fish was 13 times that of the average non-transgenic control.

Animals↗
PubMed1991

Growth hormone heterogeneity in American plaice pituitaries: isolation, characterization, and partial amino acid sequence.

Growth hormones (GHs) have been isolated from pituitary glands of American plaice (Hippoglossoides platessoides), a marine flatfish, using affinity and gel filtration chromatography, followed by preparative polyacrylamide gel electrophoresis (PAGE). A bioassay based on serum triiodothyronine elevation in immature rainbow trout was used to monitor biological activity. These GHs originate from two molecular mass regions, 42K and less than 33K relative molecular mass (Mr), in their native state. The 42K Mr region yielded two forms of GH, which differ in terms of quantity and net charge as evidenced by native PAGE, a major variant with a relative mobility of (Rf) 0.22 and a lesser variant with Rf 0.28. The less than 33 Mr region has a single GH species with Rf 0.22. Upon sodium dodecyl sulfate-PAGE, without reduction, both GH variants from the 42K Mr region gave Mrs of 21K, while the GH from the less than 33K Mr region was 20K Mr, typical of monomeric vertebrate GHs. The proteins composing the 42K Mr region are proposed as GH dimers since they yield 21K Mr peptides. The less than 33K Mr region contains a GH monomer (20K Mr) in its native state. An amino-terminal amino acid sequence, identical for both the 42K and the 20K Mr Rf 0.22 forms, has good homology with other complete fish GH sequences near their carboxyl-terminal regions (between amino acids 130 and 196). The GH dimers (42K Mr) predominate in the plaice pituitary, contributing 93% of the total, of which 86% gives rise to the Rf 0.22 variant.

Amino Acid Sequence↗
PubMed1991

Isolation of sockeye salmon growth hormone utilizing serum triiodothyronine enhancement in rainbow trout to monitor biological activity.

Growth hormone (GH) was isolated from sockeye salmon (Oncorhynchus nerka) pituitary glands using established techniques of affinity and gel filtration chromatography, and preparative polyacrylamide gel electrophoresis. The GH activity was followed throughout the fractionation procedure with a bioassay based on the increase of serum triiodothyronine (T3) in rainbow trout (Oncorhynchus mykiss), measured by radioimmunoassay (RIA). Amino-terminal amino acid sequence analysis and subsequent comparison with established GH sequences from other Oncorhynchus sp. were used to confirm the isolation of sockeye salmon GH (ssGH). The bioassay was sensitive to a dose of 55 ng of purified ssGH/g fish. Monomeric GHs, located in the carbohydrate-poor protein fraction, were the only pituitary components that elevated serum T3. Twenty-four hours after GH injection was an appropriate and practical time to blood sample, allowing completion of the bioassay, including RIA, in 3 days. The generic homology, between the source of pituitaries and the bioassay animals used in this study, should permit the bioassay to be useful during GH isolation from pituitaries of all Oncorhynchus sp.

Analysis of Variance↗
PubMed1991

Stimulation of in vitro ovarian estradiol-17 beta synthesis in the rainbow trout by the carbohydrate-poor protein fraction from sockeye salmon pituitary glands.

The role of carbohydrate-poor (Con A I) and carbohydrate-rich (Con A II) pituitary protein fractions, isolated from sockeye salmon (Oncorhynchus nerka), were investigated pertaining to in vitro estradiol-17 beta (E2) production by rainbow trout (Oncorhynchus mykiss) ovarian follicles. During the early vitellogenic phase of the reproductive cycle, using defolliculated ovarian follicle preparations (outer epithelium-thecal layer absent), it was demonstrated that the Con A I fraction was capable of increasing E2 production, in the presence of exogenous testosterone (T) as the substrate. Under similar conditions the Con A II fraction (containing the maturational gonadotropin) was inactive. However the Con A II fraction or T, separately, increased E2 production by intact ovarian follicles, whereas the Con A I fraction did not. A mechanism proposed to explain the regulation of ovarian E2 synthesis involves the Con A I fraction enhancing aromatase activity in granulosa cells permitting an increased conversion of T to E2.

Animals↗
PubMed1989

Gonadotropin stimulation of K+ secretion and Na+ absorption by brook trout (Salvelinus fontinalis) sperm duct epithelium.

The sperm duct epithelium from mature spermiating brook trout (Salvelinus fontinalis) was mounted in vitro to examine control of Na+ absorptive and K+ secretory transport. Na+ absorption (measured as the short-circuit current) and K+ secretion (measured using 86Rb+ as tracer) were stimulated by 3-isobutyl-1-methylxanthine and cyclic AMP while unstimulated tissues had no net ion transport. Purified chum salmon (Oncorhynchus nerka) Con AII carbohydrate-rich gonadotropin produced a rapid, sustained rise in Rb+ secretion and Na+ uptake in a log linear dose-dependent manner. Addition of gonadotropin to either apical (mucosal) or basolateral (serosal) sides evoked the response, but addition to the apical side produced the more rapid effect, indicating that receptors for the hormone are present on both sides of the transporting cells and suggesting that subepithelial tissue may slow the response to serosally added hormone. This is the first indication that gonadotropin may directly regulate ion transport functions of the blood-testis barrier of vertebrates and in this way regulate seminal plasma ionic composition.

Absorption↗
PubMed1989

Regulation of antifreeze protein production in winter flounder: a unique function for growth hormone.

Salmon pituitary extract and the protein fraction unabsorbed on concanavalin A-Sepharose, the carbohydrate-poor fraction, depressed plasma levels of antifreeze proteins (AFP) when the pituitary fractions were administered to flounder in late fall or winter. The active pituitary protein occurred in the fraction with a mean molecular weight of 25,000. The two major isohormones of growth hormone (GH) were the only biologically active proteins identified from the pituitary. Hypophysectomized flounder synthesize AFP in the spring and the two isohormones of GH suppress the synthesis. The fraction of flounder pituitaries containing putative GH depressed flounder plasma levels of AFP in late fall.

Acclimatization↗
PubMed1988

Molecular cloning and expression of salmon prolactin cDNA.

Prolactin was purified from chum salmon pituitaries. It was resolved into two variants by reverse-phase high-performance liquid chromatography. A cDNA library was prepared from Pacific chinook salmon pituitaries. Salmon prolactin gene was screened using a synthetic oligonucleotide based on partial protein sequence. A positive clone (PRL-10) was identified and sequenced. It is a full-size clone containing 1.1 kb and coding for a preprolactin of 211 amino acids. A modified prolactin plasmid (PRL-10A), in which the 5' untranslated sequence and the nucleotide sequence coding for the signal peptide of prolactin were deleted, was reconstructed into an expression vector using the heat-inducible lambda pL promotor. Mature prolactin, a single polypeptide of 22 kDa, was efficiently expressed in the bacteria at an elevated temperature.

Amino Acid Sequence↗
PubMed1987

Adrenocorticotropin- and opiate-like hormones from pituitaries of the sockeye salmon Oncorhynchus nerka.

The pituitaries of vitellogenic sockeye salmon (Oncorhynchus nerka) were extracted with a mixture of acetone, water, and hydrochloric acid. The precipitate which formed upon the addition of a copious volume of acetone to the extract, designated acid acetone powder, was subjected to salt fractionation and desalting, followed by ion-exchange chromatography on CM-cellulose. An unadsorbed fraction (S-1) and four adsorbed fractions (S-2, S-3, S-4 and S-5) were obtained. Adrenocorticotropic activity was detected in the fractions by their ability to stimulate isolated rat adrenal decapsular cells to produce corticosterone and by their immunoreactivities in an adrenocorticotropin-specific radioimmunoassay. The steroidogenic activities of all fractions, except S-4, were blocked by corticotropin inhibiting peptide. Opiate activity was detected in the fractions by their ability to inhibit the binding of either [3H]naloxone or (D-ala2, D-leu5)-[3H]enkephalin to rat brain membranes. There was a discrepancy in the potencies of the five fractions in the two opiate radioreceptor assays, indicating the presence of opiate peptides with different affinities of binding to the micron- and delta-opiate receptors of the rat brain. There was a separation between adrenocorticotropic and opiate receptor binding activities, suggesting that the activities were due to separate molecular entities.

Adrenal Cortex↗
PubMed1986

The cellular location of 1 alpha-hydroxycorticosterone binding protein in skate.

Antiserum to 1 alpha-hydroxycorticosterone binding protein was used to investigate its location in selected tissues of the skate Raja ocellata. Immunofluorescence, using an indirect technique, showed 1 alpha-hydroxycorticosterone binding sites in potential target tissue: gill. chloride cells, rectal gland parenchyma, and a portion of the kidney tubule. The binding protein was not detectable in the ventricle or the intestinal spiral valve but was associated with liver parenchyma and interrenal cells. The intracellular location of the binding protein is apparently tissue specific.

Animals↗
PubMed1986

Maturational steroids and gonadotropin in upstream migratory sockeye salmon.

The circulating serum concentrations of various steroid hormones in mature sockeye salmon were measured at four different developmental stages in their upstream migration to spawn at Adams River in British Columbia, Canada. In females, a high level of estradiol-17 beta was found in fish at the first location, and it persisted until immediately before reaching the spawning grounds, 485 km upstream, where it decreased to a minimal level. Free testosterone was extremely high throughout the migration but decreased significantly after spawning while its glucuronide changed reciprocally. Free and conjugated 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one (17 alpha, 20 beta-P) peaked at the last location before spawning with the glucuronide only 20% of the free steroid in concentration. After spawning, the concentration of free steroid declined but the glucuronide remained constant. 17 alpha-hydroxyprogesterone increased significantly in serum before and after the fish had spawned. During the migration 11-deoxycortisol was present in the serum at all stages but maximal levels were found in postspawned fish. Throughout the migration, the males had high serum levels of 11-ketotestosterone. Lesser amounts of free testosterone were also consistently present but the ratio of free: conjugated decreased from 1.7 at the beginning of the migration to 0.15 on the arrival at the Adams River and 0.10 in postspawned fish. Only low levels of 11 beta-hydroxytestosterone were found except in the postspawned males where the value was equal to one-half that of free testosterone. As in the females, there was a substantial increase in the levels of 17 alpha, 20 beta-P and its glucuronide in the males captured at the spawning grounds. In both sexes gonadotropin levels were low during the migration and high in the postspawned fish.

Animals↗
PubMed1985

Epidermal cellular proliferation and differentiation in sexually mature male Salmo salar with androgen levels depressed by oil.

Sexually mature male Salmo salar exhibit epidermal thickening and an increase in goblet cell concentration during the spawning season. The ventral skin, which is likely to experience most abrasive contact during the spawning period, has the thickest epidermis and the greatest goblet cell concentration. Following exposure to crude oil there is inhibition of cellular proliferation and elongation associated with epidermal thickening, and also inhibition of mucigenesis. Data on the androgen levels in these fish, and data from earlier studies involving treatment with hormones, indicate that oil-related epidermal effects during the spawning period are most likely systemic in origin, probably arising from reduced plasmatic androgen levels.

Androgens↗
PubMed1985

Progestogens, androgens and their glucuronides in the terminal stages of oocyte maturation in landlocked Atlantic salmon.

Peripheral serum levels of free and conjugated steroids were correlated with seven terminal stages of oocyte maturation in female landlocked Atlantic salmon. Pregnenolone levels were maximal at stage 1, and there was a surge in testosterone after initiation of germinal vesicle migration. Testosterone remained high, above 25 ng/ml until ovulation, and its glucuronide was always lower than the corresponding free form. 17 alpha,20 beta-Dihydroxy-4-pregnen-3-one increased progressively from 0.71 to 68 ng/ml from oocyte stage 1 to 6, with the glucuronide predominating in stages 1 to 4 and the free steroid in stages 5 to 7. Thus, very low levels of free 17 alpha,20 beta-dihydroxy-4-pregnen-3-one were present when migration of the germinal vesicle was initiated but considerable free steroid was present at and after germinal vesicle breakdown. Negligible amounts of progesterone, 17 alpha-hydroxypregnenolone, 5-pregnene-3 beta,17 alpha,20 beta-triol were found in the serum during final maturation. In male salmon with expressible milt, 17 alpha,20 beta-dihydroxy-4-pregnen-3-one was present with a gradual increase of both free and conjugated form at the advent of spawning, suggesting that this steroid may also play a role in reproduction in male fish. The action of steroids in vitro on oocytes at various stages was consistent with conclusions based on blood levels.

Androgens↗
PubMed1985

Plasma vitellogenin in landlocked Atlantic salmon (Salmo salar Ouananiche): isolation, homologous radioimmunoassay and immunological cross-reactivity with vitellogenin from other teleosts.

Vitellogenin was isolated by affinity chromatography and gel filtration from landlocked Atlantic salmon plasma. Vitellogenin was labelled with iodine-131 using iodogen and an homologous radioimmunoassay was developed. There was poor immunological cross-reactivity with vitellogenin or plasma from other teleosts. Parallelism of the vitellogenin standard to the displacement by plasma of vitellogenic salmon allowed the assay to be used to evaluate the seasonal concentration profile of vitellogenin in female adult salmon. Extracts of liver or ovary from female Atlantic salmon also yielded displacements parallel to the vitellogenin standard in the assay.

Animals↗
PubMed1984

Failure of antibody to carbohydrate-rich gonadotropin to inhibit rapid ovarian growth in landlocked Atlantic salmon.

The discovery of a carbohydrate-poor gonadotropin has resulted in a reassessment of the role of the classical, carbohydrate-rich gonadotropin (Con A II GtH) in teleost ovarian development. Prolonged treatment of landlocked Atlantic salmon with antibody to Con A II GtH failed to inhibit rapid ovarian growth or accumulation of triglyceride or vitellogenin derivative. Sera from antibody-treated fish showed substantial capacity to bind Con A II GtH indicating that antibody treatment successfully removed endogenous Con A II GtH from the circulation. These results are consistent with previous reports that a factor other than Con A II GtH is responsible for stimulating yolk accumulation.

Animals↗