-
[show abstract]
[hide abstract]
ABSTRACT: A stable supply of viable eggs and embryos is crucial for successful farming of Atlantic cod. Stress during broodstock rearing can have negative effects on offspring, but little is known about the molecular mechanisms that cause abnormal development. Maternally transferred mRNAs have been shown to be essential for normal development, and stress may therefore influence their expression and the subsequent embryonic development. We investigated if mimicked stress in cod females affects mRNA concentrations in eggs/embryos, and if this can be linked to viability of embryos. Three weeks before peak spawning, 20 fish were intraperitoneally implanted with either cortisol-containing or cortisol-free (sham) osmotic pumps. At peak spawning all individuals were stripped and eggs were fertilized and incubated until hatching. Samples were collected from unfertilized eggs and embryos for analysis of gene expression (microarray), viability, steroids and vitellogenin. Plasma concentration of cortisol (ng/ml) in treated females was significantly higher at spawning (127.1±20.9) than that of sham control (11.3±6.7). This difference was also reflected in eggs and embryos. Percent fertilization, asymmetric cell division and hatching were not affected. However, numerous genes were differentially expressed in eggs and embryos in response to elevated cortisol, especially in maternal (oocyte and blastula) stages. Among these differentially expressed genes, some were found to be linked to cytogenesis (stxbp6, fbxw2, capn12, thbs4, sytl2, coro1c, sel1l3), induction of mesodermal fate (fgfrl1) and import of the glucocorticoid receptor to the cell nucleus (ipo7). Gene ontology overrepresentation analysis on the whole set of differentially expressed genes at maternal stages (539 genes) revealed enriched activity in membrane associated regions, which largely corresponds to cytogenesis related processes. These results suggest that despite no visible phenotypic effects in early embryos, broodstock stress affects the egg/embryonic transcriptome, especially in relation to cytogenesis. Furthermore, effects related to egg/embryo phenotypes are difficult to measure at early stages of development, and instead might become apparent at later life stages.
General and Comparative Endocrinology 05/2013; · 3.27 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To better understand the role(s) of progesterone in fish spermatogenesis, we cloned the nuclear progesterone receptor (Pgr) of Atlantic cod. The open-reading frame of the cod pgr consists of 2076bp, coding for a 691-amino acids-long protein that shows the highest similarity with other piscine Pgr proteins. Functional characterization of the receptor expressed in mammalian cells revealed that the cod Pgr exhibited progesterone-specific, dose-dependent induction of reporter gene expression, with 17α,20β-dihydroxy-4-pregnen-3-one (DHP), a typical piscine progesterone, showing the highest potency in activating the receptor. During ontogenesis, the pgr mRNA was undetectable in embryo's 24h after fertilization, but became detectable 4days after fertilization. During the larval stage, the expression levels increased steadily with the development of the larvae. In adult fish, pgr was predominantly expressed in gonads of both sexes. During the onset of puberty, testicular pgr transcript levels started to increase during rapid spermatogonial proliferation, and peaked when spermiation started. In situ hybridization studies using testis tissue during the rapid growth phase containing all germ cell stages indicated that in cod, pgr mRNA is predominantly located in Sertoli cells that are in contact with proliferating spermatogonia. Taken together, our data suggests that the Pgr is involved in mediating progestagen stimulation of the mitotic expansion of spermatogonia, and in processes associated with the spermiation/spawning period in Atlantic cod.
General and Comparative Endocrinology 08/2012; 179(1):71-7. · 3.27 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: In differentiated gonochoristic species, a bipotential gonad develops into an ovary or testis during sex differentiation. Knowledge about this process is necessary to improve methods for masculinizing genetically female Atlantic cod for the subsequent purpose of producing all-female populations.
Gonads were examined histologically in juveniles from 14 to 39 mm total body length (TL). Number and size of germ cells were determined in a subset of the samples. Relevant genes were cloned, and mRNA levels determined by qPCR of amh, cyp19a1a; dax1 (nr0b2); shp (nr0b2a) and sox9b in a mixed-sex and an all-female population ranging from 12-49 mm TL.
Individuals between 14-20 mm TL could be separated in two subgroups based on gonad size and germ cell number. Ovarian cavity formation was observed in some individuals from 18-20 mm TL. The mixed sex population displayed bimodal expression patterns as regards cyp19a1a (starting at 12 mm TL) and amh (starting at 20 mm TL) mRNA levels. After approximately 30 mm TL, cyp19a1a and amh displayed a gradual increase in both sexes. No apparent, sex-dependent expression patterns were found for dax1, shp or sox9b transcripts. However, shp levels were high until the larvae reached around 35 mm TL and then dropped to low levels, while dax1 remained low until 35 mm TL, and then increased sharply.
The morphological sex differentiation in females commenced between 14-20 mm TL, and ovarian cavities were evident by 18-20 mm TL. Testis development occurred later, and was morphologically evident after 30 mm TL. This pattern was corroborated with sexually dimorphic expression patterns of cyp19a1a from 12-13 mm TL, and a male-specific increase in amh from 20 mm TL.
Reproductive Biology and Endocrinology 06/2012; 10:47. · 2.05 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Produced water (PW), a by-product of the oil-production process, contains large amount of alkylphenols (APs) and other harmful oil compounds. In the last 20 years, there have been increasing concerns regarding the environmental impact of large increases in the amounts of PW released into the North Sea. We have previously shown that low levels of APs can induce disruption of the endocrine and reproductive systems of Atlantic cod (Gadus morhua). The aims of this follow-up study were to: (i) identify the lowest observable effect concentration of APs; (ii) study the effects of exposure to real PW, obtained from a North Sea oil-production platform; and (iii) study the biological mechanism of endocrine disruption in female cod. Fish were fed with feed paste containing several concentrations of four different APs (4-tert-butylphenol, 4-n-pentylphenol, 4-n-hexylphenol and 4-n-heptylphenol) or real PW for 20 weeks throughout the normal period of vitellogenesis in Atlantic cod from October to January. Male and female cod, exposed to AP and PW, were compared to unexposed fish and to fish fed paste containing 17β-oestradiol (E(2)). Approximately 60% of the females and 96% of the males in the unexposed groups were mature at the end of the experiment. Our results show that exposure to APs and E(2) have different effects depending on the developmental stage of the fish. We observed that juvenile females are advanced into puberty and maturation, while gonad development was delayed in both maturing females and males. The AP-exposed groups contained increased numbers of mature females, and significant differences between the untreated group and the AP-treated groups were seen down to a dose of 4 μg AP/kg body weight. In the high-dose AP and the E(2) exposed groups, all females matured and no juveniles were seen. These results suggest that AP-exposure can affect the timing of the onset of puberty in fish even at extremely low concentrations. Importantly, similar effects were not seen in the fish that were exposed to real PW.
Aquatic toxicology (Amsterdam, Netherlands) 06/2011; 105(1-2):136-50. · 3.12 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Pituitary mRNA levels of gonadotropin β-subunits and of their cognate receptors in the testis were studied during puberty in Atlantic cod under normal and experimental photoperiod conditions that suppressed, delayed or accelerated testis maturation. Results are discussed in context with changes in testicular histology and plasma androgen levels, considered as end points of gonadotropic regulation. Up-regulation of fshb was closely associated with the onset of puberty, decreased when spermatogenesis was completed and reached minimum levels after spawning. These results demonstrate, for the first time using an experimental approach, that activation of Fsh-dependent signaling is associated with spermatogonial proliferation and formation of spermatogenic cysts. Changes in fshr expression were less prominent and could be explained by changes in the cellular composition and RNA content of cod testis tissue. At more advanced stages of development (spermiogenesis, spermiation and spawning), lhb and, one month later, lhcgr transcript levels increased and reached peak values in spawning fish, in a positive feedback loop involving plasma androgens and Lh/Lhcgr-dependent signaling. This loop was broken by a loss of lhb expression at the end of the spawning season. Continuous light (LL) from summer solstice, ~8 months prior to spawning, suppressed the start of testis maturation and the changes in gonadotropin and receptor mRNA levels, while LL from winter solstice initially up-regulated lhb and lhcgr expression, before resulting in a precocious termination of the spawning season and low expression of all four genes. Our studies provide experimental evidence for a clear functional discrimination of cod gonadotropins.
General and Comparative Endocrinology 05/2011; 173(1):111-9. · 3.27 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To better understand the role(s) of progestogens during early stages of spermatogenesis, we carried out studies on the nuclear progesterone receptor (Pgr) of the Atlantic salmon. Its open-reading frame shows the highest similarity with other piscine Pgr proteins. When expressed in mammalian cells, salmon Pgr exhibited progestogen-specific, dose-dependent induction of reporter gene expression, with 17α,20β-dihydroxy-4-pregnen-3-one (DHP) showing the highest potency. We then analyzed testicular pgr mRNA and DHP plasma levels in animals during the onset of spermatogenesis, which were exposed to natural light or to constant light, to induce significant differences in testis growth. Grouping of the animals according to their progress through spermatogenesis showed that testicular pgr mRNA levels as well as DHP plasma levels first increased when germ cells had reached the stage of late type B spermatogonia and further increased when entered meiosis, i.e. when spermatocytes were present. However, in situ hybridization studies revealed that pgr mRNA expression was restricted to Sertoli cells, with a strong signal in Sertoli cells contacting type A/early type B spermatogonia, while Sertoli cells contacting larger germ cell clones with further differentiated stages (e.g. late type B spermatogonia) were less intensely/not stained. We conclude that the increase in pgr mRNA levels per pair of testis reflects, at least in part, the increased number of Sertoli cells enveloping type A and early type B spermatogonia. We propose that Sertoli cell-expressed Pgr may mediate DHP-stimulated early steps in spermatogenesis in Atlantic salmon, such as an increase in the number of new spermatogonial cysts.
Reproduction 01/2011; 141(4):491-500. · 2.58 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Growth hormone in fish regulates many important physiological processes including growth, metabolism and potentially reproduction. In salmonid fish, GH secretion is episodic with irregularly spaced GH peaks. Plasma GH reflects secretion episodes as well as the clearance rate of the hormone, and plasma levels may thus not always reflect the level of activation of the GH axis. This study measured the production dynamics of GH over a 17-month period in sexually maturing female Atlantic salmon which included final maturation and spawning. For the first time, the level of pituitary GH mRNA, pituitary GH protein and plasma GH protein were analyzed concurrently in the same individuals. mRNA and protein were extracted in parallel from the same samples with subsequent real time quantitative PCR to measure mRNA transcripts and radioimmunoassay to measure pituitary and plasma GH protein. Further, the effects of photoperiod manipulation on these parameters were studied. The results show no correlation between mRNA and protein levels except at some time points, and indicate that it is inappropriate to correlate pooled temporal data and averages in time series unless the relationship among the variables is stable over time. The results indicate complex and shifting relationships between pituitary GH mRNA expression, pituitary GH content and plasma GH levels, which could result from changes in clearance rather than secretion rate at different times and its episodic secretion. The study also suggests that there is a functionally important activation of the GH system during spring leading up to maturation and spawning.
General and Comparative Endocrinology 02/2010; 167(1):77-85. · 3.27 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The gonadotropins Fsh and Lh interact with their receptors (Fshr and Lhr, respectively) in a highly specific manner in mammals with little overlap in biological activities. In fish, the biological activities seem less clearly separated considering, for example, the steroidogenic potency of both Fsh and Lh. Important determinants of the biological activity are the specificity of hormone-receptor interaction and the cellular site of receptor expression. Here, we report the pharmacological characterization of Atlantic salmon Fshr and Lhr, identify receptor-expressing cells in the ovary, and validate receptor mRNA quantification systems. For the pharmacological studies, we used highly purified coho salmon gonadotropins and found that the Fshr preferentially responded to Fsh, but was also activated by approximately 6-fold higher levels of Lh. The Lhr was specific for Lh and did not respond to Fsh. Photoperiod manipulation was used to generate ovarian tissue samples with largely differing stages of maturation. Specific real-time, quantitative (rtq) PCR assays revealed up to 40-fold (fshr) and up to 350-fold (lhr) changes in ovarian expression levels, which correlated well with the differences in ovarian weight, histology, and circulating oestrogen levels recorded in January and June, respectively. Vitellogenic ovaries were used to localise receptor-expressing cells by in situ hybridization. Granulosa cells of small and large vitellogenic follicles were positive for both receptors. Also theca cells of small and large vitellogenic follicles expressed fshr mRNA, while only in large vitellogenic follicles theca cells were (weakly) positive for lhr mRNA. While only ovulatory Lh levels seem high enough to cross-activate the Fshr, expression by both receptors by granulosa and theca cells suggests that homologous ligand receptor interaction will prevail.
General and Comparative Endocrinology 06/2009; 163(3):329-39. · 3.27 Impact Factor
-
Geir Lasse Taranger,
Manuel Carrillo,
Rüdiger W Schulz,
Pascal Fontaine,
Silvia Zanuy,
Alicia Felip,
Finn-Arne Weltzien,
Sylvie Dufour,
Orjan Karlsen,
Birgitta Norberg, Eva Andersson,
Tom Hansen
[show abstract]
[hide abstract]
ABSTRACT: Puberty comprises the transition from an immature juvenile to a mature adult state of the reproductive system, i.e. the individual becomes capable of reproducing sexually for the first time, which implies functional competence of the brain-pituitary-gonad (BPG) axis. Early puberty is a major problem in many farmed fish species due to negative effects on growth performance, flesh composition, external appearance, behaviour, health, welfare and survival, as well as possible genetic impact on wild populations. Late puberty can also be a problem for broodstock management in some species, while some species completely fail to enter puberty under farming conditions. Age and size at puberty varies between and within species and strains, and are modulated by genetic and environmental factors. Puberty onset is controlled by activation of the BPG axis, and a range of internal and external factors are hypothesised to stimulate and/or modulate this activation such as growth, adiposity, feed intake, photoperiod, temperature and social factors. For example, there is a positive correlation between rapid growth and early puberty in fish. Age at puberty can be controlled by selective breeding or control of photoperiod, feeding or temperature. Monosex stocks can exploit sex dimorphic growth patterns and sterility can be achieved by triploidisation. However, all these techniques have limitations under commercial farming conditions. Further knowledge is needed on both basic and applied aspects of puberty control to refine existing methods and to develop new methods that are efficient in terms of production and acceptable in terms of fish welfare and sustainability.
General and Comparative Endocrinology 06/2009; 165(3):483-515. · 3.27 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To elucidate the role of the gonadotropins in Atlantic cod (Gadus morhua), complete coding sequences with partially or fully un-translated regions for the three subunits GPalpha, FSHbeta, and LHbeta were determined. The sequences of the corresponding genomic loci were also determined, allowing the design of mRNA-targeting quantitative PCR assays. Relative expression was analyzed during a complete seasonal sexual maturation cycle in Atlantic cod females. Increasing levels of lhbeta mRNA were observed during gonadal growth, peaking at spawning in February-March which corresponds to maximum gonadosomatic index. In contrast, both gpalpha and fshbeta gradually increased to a peak in December, two months before spawning started, and decreased in January just prior to spawning. Both mRNAs increased again and remained high during the spawning season, with a decline at the end of the spawning period, a further decrease in spent females, followed by a new gradual increase concurrent with the start of the next reproductive cycle. In addition to its role in vitellogenesis prior to spawning, FSH seems to have additional functions during the spawning period, possibly related to vitellogenesis that runs in parallel with final oocyte maturation and ovulation of the multiple batch spawner Atlantic cod.
Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology 04/2009; 153(3):288-95. · 1.61 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Atlantic salmon (Salmo salar L.) females (2 SW), maturing for the first time, were reared under one of three temperature regimes (high: 14.3 +/- 0.5 degrees C; natural: 10.6 +/- 1.0 degrees C; and cold: 6.9 +/- 1.0 degrees C) in combination with one of two experimental treatments; an injection of GnRH analogue (GnRHa) contained in biodegradable microspheres, or a sham injection (microspheres only). The six experimental groups were then reared under simulated natural photoperiod for 4 weeks. Blood samples were drawn for analysis of plasma steroid levels and the fish were inspected for ovulation weekly. Batches of stripped eggs were incubated in triplicate incubators in raceways until the eyed stage. Treatment with GnRHa resulted in a substantial advancement and synchronization of ovulation at all temperatures, while exposure to cold water also appeared to advance ovulation slightly. While 75% (warm and cold) to 90% (natural) of GnRHa fish ovulated during the 4-week trial, only 30% of sham-treated females exposed to cold water, and none of the sham-treated fish held at higher temperatures, ovulated during this period. Survival rates of embryos to the eyed-stage were significantly higher for broodstock exposed to cold water. Plasma levels of testosterone (T), 17beta-oestradiol (E2), and 17alpha,20beta-dihydroxy-4-pregnen-3-one (17,20betaP) were all significantly affected by treatment with GnRHa and, to a lesser extent, temperature. The efficiency of GnRHa in counteracting the negative effects of high temperature on ovulation and the associated changes in circulating sex steroids suggest that temperature inhibition operates at least in part at the brain or pituitary.
Fish Physiology and Biochemistry 10/2008; 34(3):289-98. · 1.53 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Somatolactin (Sl) is a fish specific adenohypophyseal peptide hormone related to growth hormone (Gh). Some species, including salmonids, possess two forms: Sl alpha and Sl beta. The somatolactin receptor (slr) is closely related to the growth hormone receptor (ghr). Sl has been ascribed many physiological functions, including a role in sexual maturation. In order to clarify the role of Sl in the sexual maturation of female Atlantic salmon (Salmo salar), the full length cDNAs of slr, Sl alpha and Sl beta were cloned and their expression was studied throughout a seasonal reproductive cycle using real-time quantitative PCR (RTqPCR).
Atlantic salmon Sl alpha, Sl beta and slr cDNAs were cloned using a PCR approach. Gene expression of Sl alpha, SL beta and slr was studied using RTqPCR over a 17 month period encompassing pre-vitellogenesis, vitellogenesis, ovulation and post ovulation in salmon females. Histological examination of ovarian samples allowed for the classification according to the degree of follicle maturation into oil drop, primary, secondary or tertiary yolk stage.
The mature peptide sequences of Sl alpha, Sl beta and slr are highly similar to previously cloned salmonid forms and contained the typical motifs. Phylogenetic analysis of Atlantic salmon Sl alpha and Sl beta shows that these peptides group into the two Sl clades present in some fish species. The Atlantic salmon slr grouped with salmonid slr amongst so-called type I ghr. An increase in pituitary Sl alpha and Sl beta transcripts before and during spawning, with a decrease post-ovulation, and a constant expression level of ovarian slr were observed. There was also a transient increase in Sl alpha and Sl beta in May prior to transfer from seawater to fresh water and ensuing fasting.
The up-regulation of Sl alpha and Sl beta during vitellogenesis and spawning, with a subsequent decrease post-ovulation, supports a role for Sl during gonadal growth and spawning. Sl could also be involved in calcium/phosphate mobilization associated with vitellogenesis or have a role in energy homeostasis associated with lipolysis during fasting. The up-regulation of both Sl alpha and Sl beta prior to fasting and freshwater transfer, suggests a role for Sl linked to reproduction that may be independent of the maturation induced fasting.
Reproductive Biology and Endocrinology 10/2008; 6:42. · 2.05 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Abstract
Background
Somatolactin (Sl) is a fish specific adenohypophyseal peptide hormone related to growth hormone (Gh). Some species, including salmonids, possess two forms: Sl alpha and Sl beta. The somatolactin receptor (slr) is closely related to the growth hormone receptor (ghr). Sl has been ascribed many physiological functions, including a role in sexual maturation. In order to clarify the role of Sl in the sexual maturation of female Atlantic salmon (Salmo salar), the full length cDNAs of slr, Sl alpha and Sl beta were cloned and their expression was studied throughout a seasonal reproductive cycle using real-time quantitative PCR (RTqPCR).
Methods
Atlantic salmon Sl alpha, Sl beta and slr cDNAs were cloned using a PCR approach. Gene expression of Sl alpha, SL beta and slr was studied using RTqPCR over a 17 month period encompassing pre-vitellogenesis, vitellogenesis, ovulation and post ovulation in salmon females. Histological examination of ovarian samples allowed for the classification according to the degree of follicle maturation into oil drop, primary, secondary or tertiary yolk stage.
Results
The mature peptide sequences of Sl alpha, Sl beta and slr are highly similar to previously cloned salmonid forms and contained the typical motifs. Phylogenetic analysis of Atlantic salmon Sl alpha and Sl beta shows that these peptides group into the two Sl clades present in some fish species. The Atlantic salmon slr grouped with salmonid slr amongst so-called type I ghr. An increase in pituitary Sl alpha and Sl beta transcripts before and during spawning, with a decrease post-ovulation, and a constant expression level of ovarian slr were observed. There was also a transient increase in Sl alpha and Sl beta in May prior to transfer from seawater to fresh water and ensuing fasting.
Conclusion
The up-regulation of Sl alpha and Sl beta during vitellogenesis and spawning, with a subsequent decrease post-ovulation, supports a role for Sl during gonadal growth and spawning. Sl could also be involved in calcium/phosphate mobilization associated with vitellogenesis or have a role in energy homeostasis associated with lipolysis during fasting. The up-regulation of both Sl alpha and Sl beta prior to fasting and freshwater transfer, suggests a role for Sl linked to reproduction that may be independent of the maturation induced fasting.
Reproductive Biology and Endocrinology. 01/2008;
-
[show abstract]
[hide abstract]
ABSTRACT: The key component regulating vertebrate puberty and sexual maturation is the endocrine system primarily effectuated along the brain-pituitary-gonad (BPG) axis. By far most investigations on the teleost BPG axis have been performed on salmonids, carps, catfish and eels. Accordingly, earlier reviews on the BPG axis in teleosts have focused on these species, and mainly on females (e.g. 'Fish Physiology, vol. IXA. Reproduction (1983) pp. 97'; 'Proceedings of the Fourth International Symposium on the Reproductive Physiology of Fish. FishSymp91, Sheffield, UK, 1991, pp. 2'; 'Curr. Top. Dev. Biol. 30 (1995) pp. 103'; 'Rev. Fish Biol. Fish. 7 (1997) pp. 173'; 'Proceedings of the Sixth International Symposium on the Reproductive Physiology of Fish. John Grieg A/S, Bergen, Norway, 2000, pp. 211'). However, in recent years new data have emerged on the BPG axis in flatfish, especially at the level of the brain and pituitary. The evolutionarily advanced flatfishes are important model species both from an evolutionary point of view and also because many are candidates for aquaculture. The scope of this paper is to review the present status on the male teleost BPG axis, with an emphasis on flatfish. In doing so, we will first discuss the present understanding of the individual constituents of the axis in the best studied teleost models, and thereafter discuss available data on flatfish. Of the three constituents of the BPG axis, we will focus especially on the pituitary and gonadotropins. In addition to reviewing recent information on flatfish, we present some entirely new information on the phylogeny and molecular structure of teleost gonadotropins.
Comparative Biochemistry and Physiology - Part A Molecular & Integrative Physiology 04/2004; 137(3):447-77. · 2.23 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To elucidate the role of the gonadotropins in the multiple spawner Atlantic halibut (Hippoglossus hippoglossus) full length cDNAs encoding FSHbeta, LHbeta, and the common alpha-subunit were cloned from pituitary glands by RACE-PCR. The three cDNAs consisted of 614, 595, and 666 nucleotides encoding peptides of 131, 146, and 124 amino acids, respectively. Halibut FSHbeta and LHbeta showed unique structural features among the vertebrate glycoprotein hormones. First, in contrast to all known FSHbeta, which contain either one or two conserved N-glycosylation sites, no potential binding site was found in Atlantic halibut FSHbeta. Second, the conserved glycosylation site in the N-terminus of all vertebrate LHbeta has been substituted with a unique C-terminal binding site in Atlantic halibut LHbeta. Furthermore, a specific cysteine residue of importance for the folding and heterodimerization of mammalian FSH is lacking in the FSHbeta from Atlantic halibut as well as many other teleosts. However, teleost FSHbeta is characterized by an additional N-terminal cysteine, which has likely replaced the missing residue, implicating a modified folding pattern of this subunit. In situ hybridization of mature male pituitaries revealed that FSHbeta and LHbeta mRNA were expressed in distinct cell types throughout the proximal pars distalis of the adenohypophysis, while alpha-subunit mRNA was identified in all parts of the proximal pars distalis, and also along the periphery of pars intermedia. Consistently, Northern blot analysis of pituitary RNA from mature males showed that FSHbeta, LHbeta, and alpha-subunit mRNAs were highly expressed. In juvenile male pituitaries very few cells containing FSHbeta, LHbeta, and alpha-subunit mRNA were identified by in situ hybridization. Low mRNA levels encoding LHbeta and the alpha-subunit were also demonstrated by Northern blot analysis of the juvenile pituitaries, while no FSHbeta mRNA was detected using this less sensitive technique. The results suggest that both FSH and LH play a role during both the very early and the final reproductive stages in Atlantic halibut males.
General and Comparative Endocrinology 05/2003; 131(2):87-96. · 3.27 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The eight distinct hormone-producing cell types in the adenohypophysis of male Atlantic halibut (Hippoglossus hippoglossus L.) were identified and localized using immunohistochemistry and in situ hybridization. Lactotropes either occupied most of the rostral pars distalis (RPD) or they were arranged in follicular structures located along the periphery of the RPD. Corticotropes were confined to a thin layer of RPD cells bordering the pars nervosa (PN). The somatotropes were arranged in multicellular layers bordering the highly convoluted PN penetrating the proximal pars distalis (PPD), while thyrotropes, scattered in small islets in between the somatotropes, were located in the centro-dorsal part of the PPD. Gonadotropes were found throughout the PPD. Immunoreactivity to glycoprotein-alpha and luteinizing hormone beta-subunit was also observed along the periphery of the pars intermedia (PI), indicating that a thin extension of the PPD surrounded the PI. In situ hybridization showed that follicle-stimulating hormone and luteinizing hormone were produced in distinct cells of the PPD. PI contained somatolactotropes bordering the highly convoluted PN, and melanotropes that showed positive immunostaining against both anti-alpha-melanocyte-stimulating hormone and anti-beta-endorphin. The general cellular organization was similar to that of other teleost fish. These results lay the basis for future investigations on Atlantic halibut pituitary physiology.
Comparative Biochemistry and Physiology - Part A Molecular & Integrative Physiology 03/2003; 134(2):315-27. · 2.23 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Photoperiod and temperature modulated the seasonal pattern of ovarian gene expression of P450 aromatase, Follicle-Stimulating Hormone receptor and Luteinizing Hormone receptor in Atlantic salmon broodstock.
Fish Physiology and Biochemistry 02/2003; 28(1):411-411. · 1.53 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Three forms of GnRH, chicken (c) GnRH-II, salmon (s) and seabream (sb) GnRH, were immunologically characterized in the brain and pituitary of turbot by ELISA. cGnRH-II and sGnRH were detected in the brain, while sbGnRH and sGnRH (but not cGnRH-II) were detected in the pituitary. In females, the levels of cGnRH-II in the turbot brain extracts increased from May to July, concomitant with an increase in oocyte diameter. In the pituitary, sbGnRH was found to be the dominant form, with levels 100–600-fold those of sGnRH. Both sGnRH and sbGnRH in the pituitary showed variation during the spawning season; sbGnRH increased from May to July and correlated with the increase in oocyte diameter, while sGnRH decreased. The overall patterns were the same for male turbot, although levels were generally lower. These findings suggest that sbGnRH could be controlling reproduction in the turbot. However, the seasonal variation in sGnRH indicates a potential physiological role in turbot reproduction. This study gives the first immunological indications that sbGnRH is present in the pituitary of a pleuronectiform fish, and will provide the basis for further studies on the endocrine regulation of reproduction in flatfish.
Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology 07/2001; 129:551-558. · 1.92 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Radioimmunoassay (RIA) studies on highperformance liquid chromatography (HPLC) fractions of brain extracts of the three-spined stickleback, Gasterosteus aculeatus, provided evidence for at least two forms of gonadotropin-releasing hormone (GnRH). One form showed chromatographic and immunological properties similar to that of synthetic salmon GnRH (sGnRH). A second, unidentified form of GnRH eluted in the same position as chicken GnRH I (cGnRH-I); however, it did not cross-react in a cGnRH-I RIA. Furthermore, it cannot be excluded that chicken GnRH II (cGnRH-II) and maybe one other unidentified form are present in the stickleback. The distribution of GnRH in the brain of breeding adult male sticklebacks was studied by use of immunohistochemistry. Two antisera against sGnRH and antisera against mGnRH and cGnRH-II were applied on cryosections and visualized using the peroxidase-antiperoxidase method. Staining patterns were similar after incubations with all four antisera. Immunoreactive fibers were found in most parts of the brain. Three distinct groups of GnRH-immunoreactive perikarya were found in the nucleus olfactoretinalis, in the nucleus anterior periventricularis, and in the nucleus lateralis tuberis. Moreover, weakly stained cells occurred in a periventricular position in the midbrain. The proximal pars distalis of the pituitary, housing the gonadotropic cells, was richly innervated by GnRH-positive fibers. In the pars intermedia and in the rostral pars distalis, immunoreactive fibers were absent.
Cell and Tissue Research 02/1995; 279(3):485-493. · 3.11 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The cDNAs encoding the catfish prepro-gonadotropin-releasing hormone and the chicken pre-pro-gonadotropin-releasing hormone II of the African catfish (Clarias gariepinus) have been isolated and sequenced. The catfish gonadotropin-releasing-hormone precursor and the chicken gona-dotropin-releasing-hormone-II precursor have the same overall architecture as other gonadotropin-releasing-hormone precursors identified so far; each is composed of a signal peptide, gonadotropin-releasing hormone and a gonadotropin-releasing-hormone-associated peptide which is connected to gonadotropin-releasing hormone by a Gly-Lys-Arg sequence. The amino acid sequences of catfish gonadotropin-releasing hormone and chicken gonadotropin-releasing hormone II, in combination with the Gly-Lys-Arg sequence, are highly conserved during evolution when compared with the corresponding regions of mammalian, avian (chicken gonadotropin-releasing hormone I) and other fish gonadotropin-releasing-hormone precursors. However, the gonadotropin-releasing-hormone-as-sociated peptide regions are markedly divergent. Northern-blot analysis revealed the presence of a single catfish gonadotropin-releasing-hormone mRNA species of about 470 bases, and the presence of a single chicken gonadotropin-releasing-hormone-II mRNA species of about 650 bases in the African catfish brain. In situ hybridization revealed catfish gonadotropin-releasing-hormone cell bodies rostro-caudally scattered in the olfactory nerve, along both sides of the midline of the telencephalon, in the preoptic area of the ventral hypothalamus, and in the infundibular stalk close to the pituitary. Chicken gonadotropin-releasing-hormone-II cell bodies, however, were exclusively found in the midbrain tegmentum.
European Journal of Biochemistry. 05/1994; 222(2):541 - 549.
