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Biomedical subjects

G M Weber

Publications and source records attributed to G M Weber.

12 recordsLinked to original sources
PubMed2001

In vitro hormone induction of final oocyte maturation in striped bass (Morone saxatilis) follicles is inhibited by blockers of phosphatidylinositol 3-kinase activity.

Oocyte germinal vesicle breakdown (GVBD) was induced in striped bass ovarian fragments when tissues were incubated with 100-nM recombinant human insulin-like growth factor-I (rhIGF-I), 25-IU human chorionic gonadotropin (hCG) ml(-1), or 290 nM of the maturation-inducing steroid (MIS), 17,20beta,21-trihydroxy-4-pregnen-3-one (20beta-S). Inhibitors of phosphatidylinositol 3-kinase (PI 3-K), wortmannin (100 nM) and LY 294002 (50 microM), inhibited GVBD induced by these hormones. Furthermore, the inhibitors attenuated hCG-induced steroid hormone synthesis. Previous studies report that gap junction uncouplers inhibit GVBD induced by hCG, but not by rhIGF-I, in striped bass. We show that 20beta-S-induced GVBD is also attenuated by 1 mM 1-heptanol or 1-octanol without being affected by incubation with 3 mM ethanol. Thus, the effects of inhibiting PI 3-K activity on GtH and MIS actions are similar to effects of uncoupling gap junctions. These data suggest that PI 3-K activity is required for GtH- MIS- and IGF-I induction of GVBD in striped bass. Our data are also consistent with the notion that a ligand that regulates PI 3-K activity, possibly an IGF, participates in maintenance of gap junctional communication required for maximal GtH and MIS action.

Androstadienes↗
PubMed2001

Sequence conservation among fish myostatin orthologues and the characterization of two additional cDNA clones from Morone saxatilis and Morone americana.

Myostatin (MSTN) negatively regulates mammalian skeletal muscle growth and development by inhibiting myoblast proliferation. Mice and cattle possessing mutant MSTN alleles display a 'double muscling' phenotype characterized by extreme skeletal muscle hypertrophy and/or hyperplasia. MSTN orthologues have been previously characterized in 12 vertebrate species, including the white bass Morone chrysops. Presented herein is the identification and characterization of novel cDNA clones from two additional Morone species: saxatilis (striped bass) and americana (white perch), which were obtained by PCR amplification and subsequent TA-cloning. The predicted amino acid sequence of each cDNA clone contains a putative signal sequence, conserved cysteine residues and a RXXR proteolytic processing site. The different Morone proteins were 97-99% identical to each other and approximately 91, 81, 68 and 67% identical to the tilapia, zebrafish, mammalian and avian proteins, respectively. However, the bioactive domains, which lie downstream of each processing site, were considerably more conserved. They were 99-100% identical within the genus and were approximately 99, 95, 88 and 88% identical to the tilapia, zebrafish, mammalian and avian domains, respectively. This high level of sequence conservation among all known MSTN orthologues suggests that the structure/function relationship of each is equally well conserved among vertebrates.

Amino Acid Sequence↗
PubMed2001

Argus--a new database system for Web-based analysis of multiple microarray data sets.

The ongoing revolution in microarray technology allows biologists studying gene expression to routinely collect >10(5) data points in a given experiment. Widely accessible and versatile database software is required to process this large amount of raw data into a format that facilitates the development of new biological insights. Here, we present a novel microarray database software system, named Argus, designed to process, analyze, manage, and publish microarray data. Argus imports the intensities and images of externally quantified microarray spots, performs normalization, and calculates ratios of gene expression between conditions. The database can be queried locally or over the Web, providing a convenient format for Web-publishing entire microarray data sets. Searches for regulated genes can be conducted across multiple experiments, and the integrated results incorporate images of the actual hybridization spots for artifact screening. Query results are presented in a clone- or gene-oriented fashion to rapidly identify highly regulated genes, and scatterplots of expression ratios allow an individual ratio to be interpreted in the context of all data points in the experiment. Algorithms were developed to optimize response times for queries of regulated genes. Supporting databases are updated easily to maintain current gene identity information, and hyperlinks to the Web provide access to descriptions of gene function. Query results also can be exported for higher-order analyses of expression patterns. This combination of features currently is not available in similar software. Argus is available at http://vessels.bwh.harvard.edu/software/Argus.

Databases, Factual↗
PubMed2001

Isolation and characterization of myostatin complementary deoxyribonucleic acid clones from two commercially important fish: Oreochromis mossambicus and Morone chrysops.

In mammals, skeletal muscle mass is negatively regulated by a muscle-derived growth/differentiating factor named myostatin (MSTN) that belongs to the transforming growth factor-beta superfamily. Although putative MSTN homologs have been identified from several vertebrates, nonmammalian orthologs remained poorly defined. Thus, we isolated and characterized MSTN complementary DNA clones from the skeletal muscle of the tilapia Oreochromis mossambicus and the white bass Morone chrysops. The nucleic and amino acid sequences from both fish species are highly homologous to the previously identified mammalian and avian orthologs, and both possess conserved cysteine residues and putative RXXR proteolytic processing sites that are common to all transforming growth factor-beta family members. Western blotting of conditioned medium from human embryonal kidney (HEK293) cells overexpressing a His-tagged tilapia MSTN indicates that the secreted fish protein is processed in a manner similar to mouse MSTN. However, in contrast to mice, MSTN expression in tilapia is not limited to skeletal muscle as it occurs in many tissues. Furthermore, the timing of MSTN expression in developing tilapia larvae coincides with myogenesis. These results suggest that the biological actions of MSTN in the tilapia and possibly in other fishes may not be limited to myocyte growth repression, but may additionally influence different cell types and organ systems.

Aging↗
PubMed2000

Effects of insulin-like growth factor-I on in vitro final oocyte maturation and ovarian steroidogenesis in striped bass, Morone saxatilis.

Recombinant human (rh) insulin-like growth factor-I (IGF-I) was more potent than rhIGF-II at inducing in vitro germinal vesicle breakdown (GVBD), a marker for resumption of meiosis, in oocytes of striped bass. Treatment of ovarian fragments containing oocytes in intact follicles with rhIGF-I increased concentrations of estradiol-17beta and maturation-inducing steroid (MIS) 17,20beta, 21-trihydoxy-4-pregnen-3-one (20beta-S) in the culture medium and decreased testosterone levels. The follicles were too immature for oocytes to complete GVBD in response to 20beta-S (MIS incompetent) or hCG. Addition of 20beta-S to cultures did not increase the percentage of oocytes completing GVBD in response to rhIGF-I or rhIGF-II. Bovine insulin was without effect on GVBD or steroid production. Incubation of MIS-competent follicles with actinomycin D, cyanoketone, trilostane, 1-heptanol, or 1-octanol had no effect on rhIGF-I-induced GVBD, but attenuated hCG-induced GVBD and 20beta-S production. Cycloheximide inhibited rhIGF-I-induced GVBD. Collectively, these observations indicate that IGF-I can induce GVBD via MIS- and transcription-independent pathways without coupled gap junctions between oocytes and granulosa cells or among granulosa cells, but requires protein synthesis to do so. An rhIGF-I analogue that does not bind IGF-binding proteins, des(1,3)IGF-I, was more potent than rhIGF-I in inducing GVBD, suggesting ovarian IGF-binding proteins may inhibit IGF-I action.

1-Octanol↗
PubMed1999

Changes in serum concentrations and pituitary content of the two prolactins and growth hormone during the reproductive cycle in female tilapia, Oreochromis mossambicus, compared with changes during fasting.

Patterns of change in serum concentrations and pituitary content of GH and two tilapia prolactins (PRL177 and PRL188) were examined during the reproductive cycle of female tilapia, Oreochromis mossambicus, adapted to fresh water and to seawater. Changes in these hormones during fasting were examined to elucidate whether changes observed during brooding could be attributed to a reduction in feeding during brooding. Serum concentrations of GH increased prior to pituitary content during the brooding phase of the reproductive cycle. In contrast, pituitary content of GH increased prior to serum concentrations during fasting. There was no consistent pattern of change in serum or pituitary PRL levels during the reproductive cycle, among experiments. Serum concentrations of PRL177 were elevated in all fasted fish, whereas PRL188 was elevated during fasting in males but not females. The increases in the serum concentration of PRLs and GH, and in the pituitary content of GH in response to fasting support the notion that these hormones are involved in the regulation of the use of metabolic substrates in tilapia. We conclude that reduced food intake during brooding may contribute to changes in serum and pituitary levels of the PRLs and GH observed during the reproductive cycle. Nevertheless, differences between changes in serum and pituitary GH during brooding and fasting suggest GH has actions in reproduction, and changes in GH during brooding are not only in response to fasting.

Analysis of Variance↗
PubMed1998

3,5,3'-Triiodothyronine (T3) clearance and T3-glucuronide (T3G) appearance kinetics in plasma of freshwater-reared male tilapia, Oreochromis mossambicus.

Distribution and metabolism of the thyroid hormone 3,5, 3'-l-triiodothyronine (T3) were studied in several ways to gain insights into these processes in the warm water fish tilapia Oreochromis mossambicus. Trace doses of 125I-labeled T3 (T*3)1 were injected intraarterially, extraarterially, or intraperitoneally in freshwater-reared male tilapia to explore plasma clearance kinetic responses to these different input modalities. Multicompartmental analysis of the plasma clearance data indicated a kinetic distribution of T*3 much like that reported for the rat and human, with about 2% of total body T*3 in plasma, 5% in rapidly exchanging tissues such as kidney and liver, and 93% in slowly exchanging tissues such as muscle. However, plasma clearance rates (PCR, 5.37 mL/h . 100 g body wt) and plasma appearance rates (PAR3 = PCR x [T3] plasma = 36.3 ng/h . 100 g body wt) were quite different than these indices in rat and human and 5 to 50 times larger than values reported for rainbow trout. On a whole-body basis, normalized for body weight, the tilapia we studied produced and accumulated much more T3 than rat, human, or rainbow trout. Enzymatic and chromatographic analyses of the plasma clearance data samples indicated substantial production of labeled glucuronide, but not sulfate, conjugates of iodothyronines (TiG) of unknown origin appearing in plasma. The TiG appeared beginning a few hours postinjection, peaked at 6 hours, and yielded a predicted steady-state TiG level of 8.3% of the T3 level in plasma. In contrast, in published studies, no conjugates were detected in rainbow trout plasma from 2 to 24 h after iv injection of T*3, T*4, or reverse-T*3, although conjugates of all were present in bile. To our knowledge, although T3 and T4 sulfate conjugates are present in the sera of several mammals, this is the first quantification of iodothyronine glucuronides reported in blood of any species under normal conditions. This might have physiological significance for the tilapia, with T3G providing a reversible storage form of T3 in blood, as has been suggested for sulfate conjugates of T3 and T4 in blood of several mammals.

Animals↗
PubMed1997

Evidence that gonadotropin-releasing hormone (GnRH) functions as a prolactin-releasing factor in a teleost fish (Oreochromis mossambicus) and primary structures for three native GnRH molecules.

Three forms of gonadotropin-releasing hormone (GnRH) are isolated and identified here by chemical sequence analysis for one species of tilapia, Oreochromis niloticus, and by HPLC elution position for a second species of tilapia, O. mossambicus. Of the three GnRH forms in O. mossambicus, chicken GnRH-II (cGnRH-II) and sea bream GnRH (sbGnRH) are present in greater abundance in the brain and pituitary than salmon GnRH (sGnRH). These three native forms of GnRH are shown to stimulate the release of prolactin (PRL) from the rostral pars distalis (RPD) of the pituitary of O. mossambicus in vitro with the following order of potency: cGnRH-II > sGnRH > sbGnRH. In addition, a mammalian GnRH analog stimulated the release of PRL from the pituitary RPD incubated in either iso-osmotic (320 mosmol/l) or hyperosmotic (355 mosmol/l) medium, the latter normally inhibiting PRL release. The response of the pituitary RPD to GnRH was augmented by co-incubation with testosterone or 17 beta-estradiol. The effects of GnRH on PRL release appear to be direct effects on PRL cells because the RPD of tilapia contains a nearly homogeneous mass of PRL cells without intermixing of gonadotrophs. Our data suggest that GnRH plays a broad role in fish, depending on the species, by affecting not only gonadotropins and growth hormone, but also PRL.

Amino Acid Sequence↗
PubMed1995

Consumer concerns about modern technology in agriculture: considerations for undergraduate and graduate teaching.

A number of events over the last several decades have sensitized society to the possible negative impacts of technology. The majority of our population is one or more generations removed from direct experience on farms and ranches. Thus, few individuals are aware of the dramatic changes that have occurred in agriculture. Since the establishment of the land-grant university system, agriculture in the United States has changed from a system employing 10 million farmers and farm workers each feeding five people to a system in which 2.1 million farmers feed more than 100 people each and support exports of more than $35 billion annually. There are perceptions, as well as survey data, indicating society is concerned about the use and impacts of technology in the food and agriculture system. Survey data indicate 87% of people disagreed that "economic growth is more important than environmental protection." In addition, the survey data indicate society is concerned about how technology is used and who decides which applications are to be pursued. More than 8 out of 10 people (85%) surveyed believed "citizens deserve a greater role in decisions about science and technology." There seems to be concern regarding the use of technology and resultant impacts on the environment, food safety, animal well-being, and the size and numbers of farms in the United States. Education, both formal and informal, is the key to helping the public make informed decisions regarding the role of science and technology in the food and agriculture system. The colleges of agriculture need to carefully evaluate their research, teaching, and extension portfolios to "rediscover" their mission.(ABSTRACT TRUNCATED AT 250 WORDS)

Agriculture↗
PubMed1994

Changes in whole-body thyroxine and triiodothyronine concentrations and total content during early development and metamorphosis of the toad Bufo marinus.

This study describes changes in the thyroid hormones thyroxine (T4) and triiodothyronine (T3) in the toad Bufo marinus during development from early embryonic stages through metamorphosis. Both the total content and concentrations of T4 and T3 were calculated to assess whether changes in these hormones might derive from changes in body mass or water content rather than from changes in hormone production or metabolism. Three clutches of eggs were collected from ponds during the day following fertilization and raised through metamorphosis. Samples of five or more individuals were collected daily for thyroid hormone measurement and body mass determination. Hormones were measured by radioimmunoassay utilizing miniature Sephadex columns after whole-body extraction. Both T4 and T3 were elevated at gastrulation (1 day after fertilization), but declined to low levels within the first week. This is the first report that thyroid hormones occur in anuran embryos prior to thyroid differentiation. This suggests that thyroid hormones are deposited in the anuran egg during oogenesis in a manner similar to that described for teleost fishes and the domestic hen. Levels of T3 and T4 began to rise at about 2 weeks after fertilization (stage 31; staged according to Limbaugh and Volpe, 1957) and peaked during early metamorphic climax (stage 43). This pattern coincides well with the notion that thyroid hormones are central regulators of metamorphosis in B. marinus as has been shown in other amphibians examined to date. Generally, both total content and concentration of the two hormones varied in parallel. Overall, whereas individual mass and water changes may affect the magnitude of hormone changes, they appear to have little input into their direction.

Animals↗
PubMed1992

Patterns of thyroxine and triiodothyronine in serum and follicle-bound oocytes of the tilapia, Oreochromis mossambicus, during oogenesis.

This study describes simultaneous measurements of thyroid hormones, thyroxine (T4) and triiodothyronine (T3), in the oocytes and serum of a female teleost fish over a complete reproductive cycle. We have identified patterns in circulating T4 and T3 levels as well as their accumulation into oocytes during the reproductive cycle of the tilapia (Oreochromis mossambicus). This is the first description of the patterns with which thyroid hormones accumulate in teleost oocytes. The sampling strategy used in the study eliminated the possible influences of covarying environmental factors that may affect thyroid hormone levels independently of reproductive events. Hormones in serum and oocytes were measured by radioimmunoassay utilizing miniature Sephadex columns. The total content of both thyroid hormones in the oocytes increased throughout most of the ovarian cycle as the oocytes increased in size from less than 2 mg to approximately 6.5 mg by ovulation. By contrast, concentrations of thyroid hormones in the oocytes rose only during the first third of post-spawning oocyte growth (up to approximately 2 mg) before attaining plateaus at approximately 6 ng/g for T4 and 13 ng/g for T3. Serum concentrations of T4 and T3 varied in cyclical patterns during oogenesis, dropping to lows of 3.4 ng/ml (T4) and 2.7 ng/ml (T3) when the oocytes were 1.5 and 2 mg, respectively, and then increasing to 6.5 ng/ml (T4) and 4.8 ng/ml (T3) when the oocytes reach approximately 6 mg. The concentrations of both hormones decreased shortly before spawning. Maximum concentrations of thyroid hormones in the oocytes were reached approximately 10 days prior to those in the serum. Although the serum levels of T4 were greater than those of T3, the reverse was found in the oocytes. Triiodothyronine appears to be accumulated selectively over T4 and the patterns with which both thyroid hormones accumulate in the oocytes of the tilapia do not appear to be tied to serum levels.

Analysis of Variance↗