308 research outputs found
Luteinizing hormone induces ovulation via tumor necrosis factor α-dependent increases in prostaglandin F2α in a nonmammalian vertebrate
Ovulation is induced by the preovulatory surge of luteinizing hormone (LH) that acts on the ovary and triggers the rupture of the preovulatory ovarian follicle by stimulating proteolysis and apoptosis in the follicle wall, causing the release of the mature oocyte. The pro-inflammatory cytokine tumor necrosis factor α (TNFα) and prostaglandin (PG) F2α (PGF2α) are involved in the control of ovulation but their role mediating the pro-ovulatory actions of LH is not well established. Here we show that Lh induces PGF2α synthesis through its stimulation of Tnfα production in trout, a primitive teleost fish. Recombinant trout Tnfα (rTnfα) and PGF2α recapitulate the stimulatory in vitro effects of salmon Lh (sLh) on contraction, proteolysis and loss of cell viability in the preovulatory follicle wall and, finally, ovulation. Furthermore, all pro-ovulatory actions of sLh are blocked by inhibition of Tnfα secretion or PG synthesis and all actions of rTnfα are blocked by PG synthesis inhibitors. Therefore, we provide evidence that the Tnfα-dependent increase in PGF2α production is necessary for the pro-ovulatory actions of Lh. The results from this study shed light onto the mechanisms underlying the pro-ovulatory actions of LH in vertebrates and may prove important in clinical assessments of female infertility
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Distribution and juvenile ecology of bull trout (Salvelinus confluentus) in the Cascade Mountains
Distribution and juvenile habitat use of bull trout (Salvelinus
confluentus) were surveyed in selected areas of the Upper Willamette,
Deschutes, upper Yakima, and upper Cedar River basins in Oregon and
Washington from 1989 to 1991, using day snorkeling, night snorkeling,
and electrofishing. These methods were selected after a preliminary
diel streamside study of juvenile bull trout showed fry (age 0) counts
were significantly higher (P<0.00l) during the day, while counts of
juvenile fish (age 1 and 2) were significantly higher (P<0.00l) at
night. The highest counts of juveniles occurred during a "quiet period"
immediately after dusk, during which time fish were inactive, out of
cover, and easily counted with underwater flashlights.
In a comparison of four sampling methods on Jack Creek,
electrofishing was significantly correlated (P<0.05) with day (r=0.8l)
and night (r=0.89) snorkeling counts, but not with streambank counts.
In a comparison of day and night snorkeling in 10 streams, total density
estimates were significantly greater (P<0.0l) for night snorkeling than
day. The diel study and sampling methods comparison suggested surveys
of distribution and habitat use of bull trout should include night
surveys as well as day. Distribution surveys found that, except for one stream, juvenile
bull trout were found only in, or near, spring-fed areas created by
recent lava flows. Presence of bull trout in a stream was related to
cold groundwater temperatures, as they were not found in streams with
temperatures above 14°C. Distribution of bull trout in Oregon and
Washington followed a pattern of decreasing elevation with increasing
latitude and longitude(Ra=-0.9l6) Presence at lower than expected
elevations were explained when groundwater temperatures were predicted
from mean annual air temperatures. Actual water temperatures for these
spring-fed streams were significantly lower (p<0.00l) than predicted
based on elevation, latitude, and longitude.
Comparison of historical distribution of bull trout showed extant
bull trout were found in the Willamette and Deschutes River Basins of
Oregon, respectively, in only 26.2 and 56.2% of their former ranges.
Factors associated with bull trout demise in Oregon Cascade streams were
(1) isolation and inundation of spring-fed stream habitat by water
control structures, (2) introduction of brook trout and brown trout, and
(3) large flood events and habitat degradation.
Juvenile habitat use was analyzed at the macrohabitat (habitat
unit) level in five river basins. Diel and seasonal microhabitat use
was also documented for one spring-fed stream, Jack Creek. There was a
clear difference in habitat use day and night, associated with the low
water temperature of the spring-fed streams (mean=8.0°C). All habitat
unit types (pool, riffle, glide, side channel) were used day and night,
but bull trout only elected to use side channels in both time periods.
Bull trout were found at night in (1) a higher percentage of habitat
units, (2) in higher densities by unit, and (3) in increasing numbers with increasing habitat unit area. In Jack Creek, bull trout elected to
use shallow water depths, low mean velocities, instream woody debris,
and small substrates
Divergent responses to peptidoglycans derived from different E. coli serotypes influence inflammatory outcome in trout, Oncorhynchus mykiss, macrophages
Background: Pathogen-associated molecular patterns (PAMPs) are structural components of pathogens such as lipopolysaccharide (LPS) and peptidoglycan (PGN) from bacterial cell walls. PAMP-recognition by the host results in an induction of defence-related genes and often the generation of an inflammatory response. We evaluated both the transcriptomic and inflammatory response in trout (O. mykiss) macrophages in primary cell culture stimulated with DAP-PGN (DAP; meso-diaminopimelic acid, PGN; peptidoglycan) from two strains of Escherichia coli (PGN-K12 and PGN-O111:B4) over time. Results: Transcript profiling was assessed using function-targeted cDNA microarray hybridisation (n = 36) and results show differential responses to both PGNs that are both time and treatment dependent. Wild type E. coli (K12) generated an increase in transcript number/diversity over time whereas PGN-O111:B4 stimulation resulted in a more specific and intense response. In line with this, Gene Ontology analysis (GO) highlights a specific transcriptomic remodelling for PGN-O111:B4 whereas results obtained for PGN-K12 show a high similarity to a generalised inflammatory priming response where multiple functional classes are related to ribosome biogenesis or cellular metabolism. Prostaglandin release was induced by both PGNs and macrophages were significantly more sensitive to PGN-O111:B4 as suggested from microarray data. Conclusion: Responses at the level of the transcriptome and the inflammatory outcome (prostaglandin synthesis) highlight the different sensitivity of the macrophage to slight differences (serotype) in peptidoglycan structure. Such divergent responses are likely to involve differential receptor sensitivity to ligands or indeed different receptor types. Such changes in biological response will likely reflect upon pathogenicity of certain serotypes and the development of disease
Extending immunological profiling in the gilthead sea bream, sparus aurata, by enriched cDNA library analysis, microarray design and initial studies upon the inflammatory response to PAMPs
This study describes the development and validation of an enriched oligonucleotide-microarray platform for Sparus aurata (SAQ) to provide a platform for transcriptomic studies in this species. A transcriptome database was constructed by assembly of gilthead sea bream sequences derived from public repositories of mRNA together with reads from a large collection of expressed sequence tags (EST) from two extensive targeted cDNA libraries characterizing mRNA transcripts regulated by both bacterial and viral challenge. The developed microarray was further validated by analysing monocyte/macrophage activation profiles after challenge with two Gram-negative bacterial pathogen-associated molecular patterns (PAMPs; lipopolysaccharide (LPS) and peptidoglycan (PGN)). Of the approximately 10,000 EST sequenced, we obtained a total of 6837 EST longer than 100 nt, with 3778 and 3059 EST obtained from the bacterial-primed and from the viral-primed cDNA libraries, respectively. Functional classification of contigs from the bacterial- and viral-primed cDNA libraries by Gene Ontology (GO) showed that the top five represented categories were equally represented in the two libraries: metabolism (approximately 24% of the total number of contigs), carrier proteins/membrane transport (approximately 15%), effectors/modulators and cell communication (approximately 11%), nucleoside, nucleotide and nucleic acid metabolism (approximately 7.5%) and intracellular transducers/signal transduction (approximately 5%). Transcriptome analyses using this enriched oligonucleotide platform identified differential shifts in the response to PGN and LPS in macrophage-like cells, highlighting responsive gene-cassettes tightly related to PAMP host recognition. As observed in other fish species, PGN is a powerful activator of the inflammatory response in S. aurata macrophage-like cells. We have developed and validated an oligonucleotide microarray (SAQ) that provides a platform enriched for the study of gene expression in S. aurata with an emphasis upon immunity and the immune response
Bacterial lipopolysaccharide induces apoptosis in the trout ovary
BACKGROUND: In mammals it is well known that infections can lead to alterations in reproductive function. As part of the innate immune response, a number of cytokines and other immune factors is produced during bacterial infection or after treatment with lipopolysaccharide (LPS) and acts on the reproductive system. In fish, LPS can also induce an innate immune response but little is known about the activation of the immune system by LPS on reproduction in fish. Therefore, we conducted studies to examine the in vivo and in vitro effects of lipopolysaccharide (LPS) on the reproductive function of sexually mature female trout. METHODS: In saline- and LPS -injected brook trout, we measured the concentration of plasma steroids as well as the in vitro steroidogenic response (testosterone and 17alpha-hydroxyprogesterone) of ovarian follicles to luteinizing hormone (LH), the ability of 17alpha,20beta-dihydroxy-4-pregnen-3-one to induce germinal vesicle breakdown (GVBD) in vitro, and that of epinephrine to stimulate follicular contraction in vitro. We also examined the direct effects of LPS in vitro on steroid production, GVBD and contraction in brook trout ovarian follicles. The incidence of apoptosis was evaluated by TUNEL analysis. Furthermore, we examined the gene expression pattern in the ovary of saline- and LPS-injected rainbow trout by microarray analysis. RESULTS: LPS treatment in vivo did not affect plasma testosterone concentration or the basal in vitro production of steroids, although a small but significant potentiation of the effects of LH on testosterone production in vitro was observed in ovarian follicles from LPS-treated fish. In addition, LPS increased the plasma concentration of cortisol. LPS treatment in vitro did not affect the basal or LH-stimulated steroid production in brook trout ovarian follicles. In addition, we did not observe any effects of LPS in vivo or in vitro on GVBD or follicular contraction. Therefore, LPS did not appear to impair ovarian steroid production, oocyte final maturation or follicular contraction under the present experimental conditions. Interestingly, LPS administration in vivo induced apoptosis in follicular cells, an observation that correlated with changes in the expression of genes involved in apoptosis, as evidenced by microarray analysis. CONCLUSION: These results indicate that female trout are particularly resistant to an acute administration of LPS in terms of ovarian hormone responsiveness. However, LPS caused a marked increase in apoptosis in follicular cells, suggesting that the trout ovary could be sensitive to the pro-apoptotic effects of LPS-induced inflammatory cytokines
Early marine survival of steelhead smolts in Puget Sound
Smolt-to adult survival rates for Puget Sound steelhead populations have declined substantially over the last 25 years and remain at or near historic lows. From 2006-2009, nearly 1,400 steelhead smolts from 9 watersheds within Puget Sound were tracked from river mouth to the Pacific Ocean using acoustic telemetry to: (1) estimate early marine survival through Puget Sound, (2) identify common areas of abnormally high mortality along the migration route, and (3) to identify factors that may influence survival. Cormac-Jolly-Seber mark-recapture models were used to jointly estimate survival and detection rate at telemetry arrays. Estimated survival rates from river mouths to near the Pacific Ocean ranged from 1.5% (Skokomish River hatchery smolts in 2009) to 34.0% (Big Beef Creek wild smolts in 2006), and averaged 14.9% for all populations. Factors influencing survival included population, migration segment, migration year, and rearing type (i.e., hatchery or wild), while geographic region, body length, and tag type (i.e., 7mm or 9mm) showed lesser effects. Comparison of survival rates between migration segments implicated central Puget Sound and Admiralty Inlet as potential areas of heightened mortality. Early marine survival rates estimated here are very low considering that steelhead smolts spend only about two to three weeks in Puget Sound before entering the Pacific Ocean. Mortality in Puget Sound may be a major driver behind low observed smolt-to adult survival rates. This study addresses a major gap in steelhead marine life history knowledge and can help to inform future Puget Sound steelhead recovery planning efforts
Cellular and molecular evidence for a role of tumor necrosis factor alpha in the ovulatory mechanism of trout
Background: The relevance of immune-endocrine interactions to the regulation of ovarian function in teleosts is virtually unexplored. As part of the innate immune response during infection, a number of cytokines such as tumor necrosis factor alpha (TNF alpha) and other immune factors, are produced and act on the reproductive system. However, TNF alpha is also an important physiological player in the ovulatory process in mammals. In the present study, we have examined for the first time the effects of TNF alpha in vitro in preovulatory ovarian follicles of a teleost fish, the brown trout (Salmo trutta). Methods: To determine the in vivo regulation of TNF alpha expression in the ovary, preovulatory brook trout (Salvelinus fontinalis) were injected intraperitoneally with either saline or bacterial lipopolysaccharide (LPS). In control and recombinant trout TNF alpha (rtTNF alpha)-treated brown trout granulosa cells, we examined the percentage of apoptosis by flow cytometry analysis and cell viability by propidium iodide (PI) staining. Furthermore, we determined the in vitro effects of rtTNF alpha on follicle contraction and testosterone production in preovulatory brown trout ovarian follicles. In addition, we analyzed the gene expression profiles of control and rtTNF alpha-treated ovarian tissue by microarray and real-time PCR (qPCR) analyses. Results: LPS administration in vivo causes a significant induction of the ovarian expression of TNF alpha. Treatment with rtTNF alpha induces granulosa cell apoptosis, decreases granulosa cell viability and stimulates the expression of genes known to be involved in the normal ovulatory process in trout. In addition, rtTNF alpha causes a significant increase in follicle contraction and testosterone production. Also, using a salmonid-specific microarray platform (SFA2.0 immunochip) we observed that rtTNF alpha induces the expression of genes known to be involved in inflammation, proteolysis and tissue remodeling. Furthermore, the expression of kallikrein, TOP-2, serine protease 23 and ADAM 22, genes that have been postulated to be involved in proteolytic and tissue remodeling processes during ovulation in trout, increases in follicles incubated in the presence of rtTNF alpha. Conclusions In view of these results, we propose that TNF alpha could have an important role in the biomechanics of follicle weakening, ovarian rupture and oocyte expulsion during ovulation in trout, primarily through its stimulation of follicular cell apoptosis and the expression of genes involved in follicle wall proteolysis and contraction
Lack of growth enhancement by exogenous growth hormone treatment in yellow perch (Perca flavescens) in four separate experiments
Author Posting. © The Authors, 2005. This is the author's version of the work. It is
posted here by permission of Elsevier B. V. for personal use, not for redistribution. The
definitive version was published in Aquaculture 250 (2005): 471-479, doi:10.1016/j.aquaculture.2005.03.019.The effect of exogenous growth hormone (GH) treatment on the growth of juvenile yellow perch (Perca flavescens) was investigated in four experiments. In the first two experiments, juvenile yellow perch were reared at either 13°C or 21°C, and injected weekly with bovine GH (bGH) at 0.1, 1.0 or 10.0 μg/g body weight for 84 days. No significant growth enhancement in GH-treated fish was measured in fish in either of the experiments. In the third experiment, juvenile yellow perch were treated with estradiol-17β (E2, 15 μg/g of diet), bGH (1.0 μg/g body weight) injected weekly or both hormones for 70 days at 21°C. E2 alone stimulated growth, but no further growth stimulation occurred in the E2 + bGH-treated fish. In addition, no growth enhancement was found in fish treated with bGH alone. We measured no difference in serum insulin-like growth factor-I (IGF-I) levels between the treatment groups at 12 and 24 h after the final injection of GH; however, a drop in IGF-I levels after 24 h was observed. In a fourth study, the effect of recombinant yellow perch GH (rypGH, 0.2 or 1.0 μg/g body weight) injected weekly was evaluated in yellow perch juveniles. The fish were reared for 42 days at 18°C. Neither GH dosages improved growth compared to control-injected and non-injected fish. Taken together, the lack of effect of mammalian GH or rypGH in our experiments suggests (1) low binding affinity between these hormones and the GH receptor in yellow perch, (2) that the endogenous GH levels were already at biologically maximal levels or (3) that other endocrine factors are needed in order for GH to promote yellow perch growth. The reduction in IGF-I levels 24 h after handling suggests a negative effect of handling stress on the GH-IGF-I axis in yellow perch.This work was supported by the University of Wisconsin-Madison College of Agricultural and Life Sciences and School of Natural Resources; the Wisconsin Department of Natural Resources; the University of Wisconsin Sea Grant College Program, National Oceanic and Atmospheric Administration, US Department of Commerce; the State of Wisconsin (Federal Grant NA46RG0481, Project No. R/AQ-38); and the USDA NOAA Project R/A-05-99, grant #NA86RG0048 to FG and SR. This study was also funded by the Norwegian Research Council (NFR)
Using global genome approaches to address problems in cod mariculture
Author Posting. © The Authors, 2005. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in ICES Journal of Marine Science 63 (2006): 393-399, doi:10.1016/j.icesjms.2005.10.006.A number of techniques including expressed sequence tag (EST) analysis, serial analysis
of gene expression, and microarrays are available to study the global expression and regulation of genes. Many of these techniques are being used for intensively reared fish such as trout, salmon and catfish to study genes involved in growth, reproduction and health. In contrast, relatively little is known about the composition and regulation of transcriptomes in gadids.
However, several bottlenecks in cod mariculture might benefit from the discovery and analysis of genes involved in reproduction, growth and disease. As a result, we have begun EST analysis
of genes in the cod ovary. Complimentary DNA (cDNA) libraries of cod ovaries taken from
females at oocyte final maturation and ovulation have been constructed, and 1,361 ESTs have been analyzed. As expected, several oocyte-related genes were observed including various zona pellucida egg membrane proteins. However, pivotal cell cycle regulators such as cyclins, genes involved in the regulation of apoptosis such as the Bcl-2-related ovarian killer protein, and
hormone receptor components were also observed. Finally, a cDNA for a potential novel cod antifreeze protein was observed 12 times, suggesting the existence of a cod egg-specific antifreeze protein.This work was supported in part by grant #139630/140 from the Research Council of Norway
to BN and USDA grant #2004-35204-14232 to FWG
Behavioural fever is a synergic signal amplifying the innate immune response
Behavioural fever, defined as an acute change in thermal preference driven by pathogen recognition, has been reported in a variety of invertebrates and ectothermic vertebrates. It has been suggested, but so far not confirmed, that such changes in thermal regime favour the immune response and thus promote survival. Here, we show that zebrafish display behavioural fever that acts to promote extensive and highly specific temperature-dependent changes in the brain transcriptome. The observed coupling of the immune response to fever acts at the gene-environment level to promote a robust, highly specific time-dependent anti-viral response that, under viral infection, increases survival. Fish that are not offered a choice of temperatures and that therefore cannot express behavioural fever show decreased survival under viral challenge. This phenomenon provides an underlying explanation for the varied functional responses observed during systemic fever. Given the effects of behavioural fever on survival and the fact that it exists across considerable phylogenetic space, such immunity-environment interactions are likely to be under strong positive selection
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