Physiological concentrations of ouabain rapidly inhibit prolactin release from the tilapia pituitary

Ouabain, a cardiac glycoside and inhibitor of Na(+), K(+)-ATPase, is now believed to be a steroid hormone in mammals. We have recently identified ouabain immunoreactivity in the plasma of the tilapia, a euryhaline teleost. Changes in plasma concentrations of immunoreactive ouabain (20-40 pM) in response to salinity change were well correlated with the changes in plasma osmolality and cortisol. Our previous studies have shown that cortisol rapidly inhibits prolactin (PRL) release from the tilapia pituitary by suppressing intracellular Ca(2+) ([Ca(2+)]i) and cAMP. In the present study, low doses of ouabain (10-1000 pM) inhibited PRL release dose-dependently during 2-24 h of incubation. There was no effect on growth hormone (GH) release, except for a significant increase at 1000 pM during 8-24 h of incubation. Significant dose-related increases in PRL release were observed at higher doses of ouabain (100-1000 nM), whereas significant inhibition was seen in GH release at 1000 nM during 2-24h of incubation. Ouabain at 1-100 pM had no effect on Na(+), K(+)-ATPase activity of the pituitary homogenate. The enzyme activity was inhibited by higher concentrations of ouabain, 10% at 1 nM, 15% at 10 nM, 28% at 100 nM, and 45% at 1000 nM. Ouabain also attenuated stimulation of PRL release by the Ca(2+) ionophore, A23187, and by a combination of dibutyryl cAMP and a phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthin. Intracellular Ca(2+) concentrations were monitored in the dispersed PRL cells with the Ca(2+)-sensitive dye, fura-2. Ouabain at 1 nM reversibly reduced [Ca(2+)]i within seconds, whereas 1 microM ouabain increased [Ca(2+)]i. A rapid reduction in [Ca(2+)]i was also observed when PRL cells were exposed to 1 microM cortisol, whereas there was no consistent effect at 1 nM. These results suggest that ouabain at physiological concentrations rapidly inhibits PRL release from the tilapia pituitary by suppressing intracellular Ca(2+) and cAMP metabolism. The stimulation of PRL release by high concentrations of ouabain (100-1000 nM) may result from an increase in [Ca(2+)]i, and subsequent depolarization due to the inhibition of Na(+), K(+)-ATPase activity.     

Isolation and characterization of a homologue of mammalian prolactin-releasing peptide from the tilapia brain and its effect on prolactin release from the tilapia pituitary

In the tilapia (Oreochromis mossambicus), as in many teleosts, prolactin (PRL) plays a major role in osmoregulation in freshwater. Recently, PRL-releasing peptides (PrRPs) have been characterized in mammals. Independently, a novel C-terminal RF (arginine-phenylalanine) amide peptide (Carrasius RF amide; C-RFa), which is structurally related to mammalian PrRPs, has been isolated from the brain of the Japanese crucian carp. The putative PrRP was purified from an acid extract of tilapia brain by affinity chromatography with antibody against synthetic C-RFa and HPLC on a reverse-phase ODS-120 column. The tilapia PrRP cDNA was subsequently cloned by polymerase chain reaction. The cDNA consists of 619 bp encoding a preprohormone of 117 amino acids. Sequence comparison of the isolated peptide and the preprohormone revealed that tilapia PrRP contains 20 amino acids and is identical to C-RFa. Incubation of the tilapia pituitary with synthetic C-RFa (100 nM) significantly stimulated the release of two forms of tilapia PRL (PRL188 and PRL177). However, the effect of C-RFa was less pronounced than the marked increase in PRL release in response to hyposmotic medium. The ability of C-RFa to stimulate PRL release appears to be specific, since C-RFa failed to stimulate growth hormone release from the pituitary in organ culture. In contrast, rat and human PrRPs had no effect on PRL release. C-RFa was equipotent with chicken GnRH in stimulating PRL release in the pituitary preincubated with estradiol 17beta. Circulating levels of PRL were significantly increased 1 h after intraperitoneal injection of 0.1 microg/g of C-RFa in female tilapia in freshwater but not in males. These results suggest that C-RFa is physiologically involved in the control of PRL secretion in tilapia.     

Alpha-MSH, the melanocortin-1 receptor and background adaptation in the Mozambique tilapia, Oreochromis mossambicus

The regulation of skin darkness in vertebrates is mediated by alpha-melanophore-stimulating-hormone (alphaMSH). For this action, alphaMSH binds to the melanocortin (MC)-1 receptor, a 7-transmembrane receptor located in melanophore cell membranes. The Mozambique tilapia, Oreochromis mossambicus, can change the hue of its body in response to a change in background, a process that may involve alphaMSH and the MC1R. Scale melanophores were isolated from tilapia that were acclimatised for 25 days to a black, control grey or white background and then tested for their sensitivity to des-, mono-, and di-acetylated alphaMSH. On all backgrounds, mono-acetylated alphaMSH was the dominant isoform present in pituitary homogenates. Mono-acetylated alphaMSH also had the highest potency to disperse melanosomes. Black background adapted fish showed the highest dispersing response to alphaMSH, independent of the isoform applied. We elucidated the nucleotide and amino acid sequence of the tilapia MC1R. We show that its expression in skin does not change when tilapia are acclimatised for 25 days to a black, grey or white background, while a clear change in hue is visible. This finding, combined with the absence of differential MC1R gene expression following background acclimation indicates that the increased sensitivity to alphaMSH is most likely a result of changes in the intracellular signalling system in melanophores of black background adapted fish, rather than up-regulation of the MC1R.     

Hormone release is tied to changes in cell size in the osmoreceptive prolactin cell of a euryhaline teleost fish, the tilapia, Oreochromis mossambicus

Prolactin (PRL) cells from a teleost fish, the tilapia, Oreochromis mossambicus, facilitate the direct study of osmoreception. The release of two prolactins, PRL(188) and PRL(177), which act in freshwater osmoregulation in teleost fish, rises in vitro within 5 min after extracellular osmolality falls. An increase in cell size accompanied this rise. Cell size and PRL release also increased, albeit more slowly, following the partial replacement of medium NaCl (55 mOsmolal) with an equivalent concentration of urea, a membrane-permeant molecule. Similar replacement using mannitol, which is membrane-impermeant, elicits no response. These findings suggest that osmoreception is linked to changes in cell volume rather than to extracellular osmolality per se.     

Effects of fasting on growth hormone, growth hormone receptor, and insulin-like growth factor-I axis in seawater-acclimated tilapia, Oreochromis mossambicus

Effects of fasting on the growth hormone (GH)--growth hormone receptor (GHR)-insulin-like growth factor-I (IGF-I) axis were characterized in seawater-acclimated tilapia (Oreochromis mossambicus). Fasting for 4 weeks resulted in significant reductions in body weight and specific growth rate. Plasma GH and pituitary GH mRNA levels were significantly elevated in fasted fish, whereas significant reductions were observed in plasma IGF-I and hepatic IGF-I mRNA levels. There was a significant negative correlation between plasma levels of GH and IGF-I in the fasted fish. No effect of fasting was observed on hepatic GHR mRNA levels. Plasma glucose levels were reduced significantly in fasted fish. The fact that fasting elicited increases in GH and decreases in IGF-I production without affecting GHR expression indicates a possible development of GH resistance.     

Disparate release of prolactin and growth hormone from the tilapia pituitary in response to osmotic stimulation

In most teleost fishes, prolactin (PRL) plays a key role in freshwater (FW) adaptation, whereas growth hormone (GH) is involved in seawater (SW) adaptation in salmonids and certain euryhaline species including the tilapia, Oreochromis mossambicus. Consistent with its osmoregulatory activity, PRL release increases in response to physiologically relevant reductions in extracellular osmolality. When dispersed PRL and GH cells from FW-acclimatized fish were incubated in media of varying osmolalities, PRL release increased significantly in response to a 12% reduction in medium osmolality during 1 and 4h of exposure. By contrast, cells from SW-acclimatized fish responded only to a 24% reduction in osmolality. Growth hormone release on the other hand increased whether medium osmolality was reduced or raised. Cell volume increased together with PRL release during the perifusion of dispersed PRL cells in direct proportion to the reduction in medium osmolality. Growth hormone release increased whether GH cell volume increased or decreased. In in vivo studies, circulating PRL levels increased as early as 1h after the transfer of fish from SW to FW, whereas GH levels remained unchanged during 24h of acclimatization. These results indicate that while PRL and GH cells are osmosensitive, the PRL cells respond to reductions in extracellular osmolality in a manner that is consistent with PRL's physiological role in the tilapia. While the rise in GH release following the reduction in osmolality is of uncertain physiological significance, the rise in GH release with the elevation of medium osmolality may be connected to its role in SW adaptation.     

Background adaptation and water acidification affect pigmentation and stress physiology of tilapia, Oreochromis mossambicus

The ability to adjust skin darkness to the background is a common phenomenon in fish. The hormone alpha-melanophore-stimulating hormone (alphaMSH) enhances skin darkening. In Mozambique tilapia, Oreochromis mossambicus L., alphaMSH acts as a corticotropic hormone during adaptation to water with a low pH, in addition to its role in skin colouration. In the current study, we investigated the responses of this fish to these two environmental challenges when it is exposed to both simultaneously. The skin darkening of tilapia on a black background and the lightening on grey and white backgrounds are compromised in water with a low pH, indicating that the two vastly different processes both rely on alphaMSH-regulatory mechanisms. If the water is acidified after 25 days of undisturbed background adaptation, fish showed a transient pigmentation change but recovered after two days and continued the adaptation of their skin darkness to match the background. Black backgrounds are experienced by tilapia as more stressful than grey or white backgrounds both in neutral and in low pH water. A decrease of water pH from 7.8 to 4.5 applied over a two-day period was not experienced as stressful when combined with background adaptation, based on unchanged plasma pH and plasma alphaMSH, and Na levels. However, when water pH was lowered after 25 days of undisturbed background adaptation, particularly alphaMSH levels increased chronically. In these fish, plasma pH and Na levels had decreased, indicating a reduced capacity to maintain ion-homeostasis, implicating that the fish indeed experience stress. We conclude that simultaneous exposure to these two types of stressor has a lower impact on the physiology of tilapia than subsequent exposure to the stressors.     

Activation of the growth hormone/insulin-like growth factor axis by treatment with 17 alpha-methyltestosterone and seawater rearing in the tilapia,Oreochromis mossambicus

Effects of 17 alpha-methyltestosterone (MT) treatment and environmental salinity on the growth hormone (GH)/insulin-like growth factor (IGF) axis were examined in the euryhaline tilapia, Oreochromis mossambicus. Yolk-sac fry were collected from brood stock in fresh water (FW). After yolk-sac absorption, they were assigned randomly to 1 of 4 groups: FW, MT treatment in FW, SW, and MT treatment in seawater (SW). After 147 days, FW controls had the lowest levels of GH mRNA followed by FW fish treated with MT and SW control fish. Seawater fish fed with a diet containing MT, which grew the fastest, had significantly higher levels of GH mRNA than all the other groups. A significant correlation was observed between GH mRNA and the size of the individual fish. By contrast, plasma GH levels did not vary significantly among the groups. Pituitary GH mRNA levels, plasma IGF-I levels, and fish size varied in a correlated pattern, i.e., SW+MT>FW+MT=SW control>FW control. The tilapia pituitary produces two prolactins (PRLs), PRL(177) and PRL(188). Prolactin(177), but not PRL(188), exhibits growth-promoting actions in FW tilapia. Pituitary mRNA levels of both PRLs were significantly higher in fish reared in FW than those reared in SW. Treatment with MT significantly increased mRNA levels of both PRLs in FW, but had no effect on SW fish. No correlation was seen between plasma PRL levels and growth or between PRL mRNA levels and growth. These results indicate that SW rearing and MT treatment stimulate the GH/IGF-I axis, and suggest that pituitary GH mRNA at this stage of development is a better indicator of growth than plasma levels of GH and IGF-I.     

Changes in plasma concentrations of immunoreactive ouabain in the tilapia in response to changing salinity: is ouabain a hormone in fish?

Ouabain, a cardiac glycoside and inhibitor of Na(+), K(+)-ATPase, is now believed to be a steroid hormone in mammals, involved in blood pressure and volume regulation and possibly acting as a natriuretic hormone. We have identified ouabain-like immunoreactivity in the plasma and tissues of a euryhaline teleost, the tilapia (Oreochromis mossambicus), by means of solid-phase extraction followed by a specific radioimmunoassay. Plasma concentrations of immunoreactive ouabain were 5-20pg/ml. Ouabain immunoreactivity was detected in all the tissues examined, with highest concentrations in the head kidney followed by intestine and body kidney. When the fish in fresh water were transferred to seawater, plasma osmolality increased significantly after 2, 4, 8, and 24h. Significant increases were observed in plasma ouabain immunoreactivity after 4 and 24h, and a significant correlation was seen between ouabain immunoreactivity and plasma osmolality. There was also a significant correlation between the plasma osmolality and cortisol concentrations. Upon transfer from seawater to fresh water, significant increases were seen in plasma cortisol after 4 and 8h and in immunoreactive ouabain after 4h. When the correlation was analyzed using all the data obtained during the two transfer experiments, plasma ouabain immunoreactivity and cortisol were significantly correlated with plasma osmolality, whereas there was a significant negative correlation between plasma prolactin and osmolality. A significant positive correlation was also seen between plasma cortisol and ouabain immunoreactivity. These results suggest that immunoreactive ouabain may be involved, together with cortisol, in the maintenance of hydromineral balance in the tilapia.     

Tissue-specific regulation of the growth hormone/insulin-like growth factor axis during fasting and re-feeding: Importance of muscle expression of IGF-I and IGF-II mRNA in the tilapia

The effects of prolonged nutrient restriction (fasting) and subsequent restoration (re-feeding) on the growth hormone (GH)/insulin-like growth factor (IGF) axis were investigated in the tilapia (Oreochromis mossambicus). Mean weight and specific growth rate declined within 1 week in fasted fish, and remained lower than controls throughout 4 weeks of fasting. Plasma levels of IGF-I were lower than fed controls during 4 weeks of fasting, suggesting a significant catabolic state. Following re-feeding, fasted fish gained weight continuously, but did not attain the weight of fed controls at 8 weeks after re-feeding. Specific growth rate increased above the continuously-fed controls during the first 6 weeks of re-feeding, clearly indicating a compensatory response. Plasma IGF-I levels increased after 1 week of re-feeding and levels were not otherwise different from fed controls. Plasma GH levels were unaffected by either fasting or re-feeding. No consistent effect of fasting or re-feeding was observed on liver expression of GH receptor (GH-R), somatolactin (SL) receptor (SL-R), IGF-I or IGF-II. In contrast, muscle expression of GH-R increased markedly during 4 weeks of fasting, and then declined below control levels upon re-feeding for weeks 1 and 2. Similarly, muscle expression of SL-R increased after 4 weeks of fasting, and reduced below control levels after 1 and 2 weeks of re-feeding. On the other hand, muscle expression of IGF-I was strongly reduced throughout the fasting period, and levels recovered 2 weeks after re-feeding. Muscle expression of IGF-II was not affected by fasting, but was reduced after 1 and 2 weeks of re-feeding. These results indicate that GH/IGF axis, particularly muscle expression of GH-R, SL-R and IGF-I and -II, is sensitive to nutritional status in the tilapia.     

Effects of insulin-like growth factors (IGF-I and -II) on growth hormone and prolactin release and gene expression in euryhaline tilapia,Oreochromis mossambicus

We investigated in vitro effects of insulin-like growth factors (IGF-I and -II) on growth hormone (GH) and prolactin (PRL) release and gene expression in euryhaline tilapia, Oreochromis mossambicus. Pituitaries were removed from freshwater-acclimated adult males and incubated for 2-24h in the presence of human IGF-I or -II at doses ranging from 1-1000 ng/ml (0.13-130 nM). IGF-I at concentrations higher than 10 ng/ml and IGF-II higher than 100 ng/ml significantly inhibited GH release after 8, 16, and 24h. No effect of IGFs was seen during the first 4h of incubation. IGFs at the same concentrations also significantly attenuated GH gene expression after 24h, although no effect was seen at 2h. By contrast, PRL(188) release was stimulated significantly and in a dose-related manner by IGF-I at concentrations higher than 10 ng/ml and by IGF-II at concentrations higher than 100 ng/ml within 2h. No stimulation was observed after 4h. Similarly, both IGFs at concentrations higher than 10 ng/ml increased PRL(177) release within 2h. However, no significant effect of IGF-I or -II was observed on mRNA levels of both PRLs after 2 and 24h at all concentrations examined. These results clearly indicate differential regulation of GH and PRL release and synthesis by IGFs in the tilapia pituitary, i.e., rapid-acting, stimulatory effects of IGFs on PRL release and slow-acting, inhibitory effects on GH release and synthesis.     

体外培养妊娠早期人胎肝细胞对乙型肝炎病毒易感性的变化

目的观察妊娠早期(孕10周)人胎肝细胞在不同的体外培养条件下对乙型肝炎病毒(HBV)易感性的变化，为HBV受体的发现和鉴定打下基础。方法设计无血清培养基，分别添加不同的细胞分化诱导物，在不同时相以HBV感染原代培养10周孕龄人胎肝细胞，检测细胞形态、功能及HBV感染标志物。结果培养基中添加2．5mmol／L苯巴比妥钠，培养6d后细胞逐渐出现成熟肝细胞标志，并可被HBV感染；其他培养条件下则不能观察到HBV的成功感染。结论苯巴比妥钠可在体外培养时诱导早期人胎肝细胞分化，并出现HBV易感性。     

Effects of angiotensin II and natriuretic peptides of the eel on prolactin and growth hormone release in the tilapia,Oreochromis mossambicus

The effects of angiotensin II (ANG II) and natriuretic peptides (NPs) of the eel (ANP, atrial natriuretic peptide; CNP, C-type natriuretic peptide; and VNP, ventricular natriuretic peptide) on prolactin (PRL(188) and PRL(177)) and growth hormone (GH) release from the organ-cultured tilapia pituitary were examined. Eel ANG II at concentrations greater than 1 nM stimulated the release of PRL(188) and PRL(177) in a dose-related manner during the first hour of incubation. Significant stimulation by 100 nM ANG II on PRL(177) release was observed until 4h of incubation, and on PRL(188) release until 12 h. No effect of ANG II was seen on GH release. None of the NPs altered the release of PRLs at any time point. On the other hand, eel VNP at concentrations greater than 1 nM stimulated GH release in a dose-related manner after 4 h, and significant stimulation was observed until 48 h. Eel CNP was less effective than eel VNP; significant stimulation of GH release was observed at 1 and 10 nM during 24-48 h of incubation. No significant effect of eel ANP on GH release was seen at any concentration. ANG II had no effect on GH release at any time point. There was no change in mRNA levels of PRLs or GH in the pituitaries incubated with ANG II for 8 h or those incubated with the NPs for 48 h. These results indicate rapid and short-lasting stimulation by ANG II on PRL release and slow and long-lasting stimulation by VNP and CNP on GH release from the tilapia pituitary.     

Changes in gene expression levels of somatolactin in the pituitary and morphology of gill mitochondria-rich cells in Mozambique tilapia after transfer to acidic freshwater (pH 3.5)

Mozambique tilapia, Oreochromis mossambicus, is easily acclimated to highly acidic water, and thus presents a useful model to unravel endocrine regulation of adaptation to acidic water in fish. We analyzed gene expression of somatolactin (sl), growth hormone (gh) and prolactin (prl), in the pituitary gland and size distribution of mitochondria-rich (MR) cells in the gills after transfer from normal freshwater (FW, pH 7.2) to acidified freshwater (AW, pH 3.5). Plasma osmolality drastically decreased until 2 days after transfer to AW, but had restored to normal after 1 week of acclimation, and this confirmed the excellent acid tolerance of tilapia. Expression levels of sl, gh and prl were all up-regulated during short-term exposure to AW. The expression of sl remained elevated up to 7 days after transfer; the expression of gh and prl was back to initial levels at that time. These findings point to an important and specific role of SL in adaptation to acid water in this tilapia, although temporal contribution of GH and PRL cannot be ruled out. The size distribution of branchial MR cells changed drastically during acclimation to AW. The mean MR cell size was 1.5-fold larger in the fish exposed to AW for 7 days compared to controls in FW. The gills and their MR cells are a likely site of important acid-base regulation, and SL may change ion-transport functions of MR cells to correct plasma osmotic balance disturbed by acid exposure.     

An analysis of the androgens of musth in the Asian bull elephant (Elephas maximus)

During musth in bull elephants, the androgens testosterone (T), dihydrotestosterone (DHT), and androstenedione all increase significantly. Given the unusual endocrine physiology that has been discovered in female elephants, it is also possible that bull elephants produce some unusual androgens. A cell-based androgen receptor assay was used to explore this possibility using two different methods. The first method compared the level of T measured by radioimmunoassay (RIA) with the level of androgen receptor (AR) activity measured in the serum of eight bull elephants during musth and non-musth periods. A ratio was calculated for T/AR activity for non-musth and musth, to determine if there was a change in the ratio between these two states. The second method used HPLC to separate two pooled serum samples (one non-musth and one musth) into fractions using a protocol which separates known androgens into specific, previously identified fractions. Each fraction was then tested with the AR assay to determine the androgenicity of any compounds present. This was done to determine if there were any fractions which had androgenic activity but did not contain any previously identified androgens. Results from the first analysis indicated no change in the T/AR ratio between non-musth and musth states. Clearly whatever active androgens are present during musth, they increase proportionately with T. Findings from the second analysis suggested that the only bioactive androgen present in the serum of non-musth Asian bulls is a low level of T. During musth, the only bioactive androgens detected were T and DHT; of these, T was by far the predominant active androgen present. Taken together, these two analyses suggest that T is by far the predominant active androgen present during musth in Asian bull elephants, and that no previously unidentified bioactive androgen is present.     

Masculinization of the European sea bass (Dicentrarchus labrax) by treatment with an androgen or aromatase inhibitor involves different gene expression and has distinct lasting effects on maturation

The objective of this study was to contribute to our understanding of the role of sex steroids in fish sex differentiation and male maturation. Sexually undifferentiated sea bass were administered 17α-methyldihydrotestosterone (MDHT), estradiol-17β (E₂), fadrozole (Fz), cyproterone acetate (CPA) or tamoxifen (Tx). MDHT produced 100% males whereas E₂ and Tx resulted in 100% females. Fz produced 95% males while CPA did not alter sex ratios. E₂ treatment did not affect gonadal aromatase (cyp19a) expression levels, supporting the possibility that the feminizing effect of exogenous E₂ are not directly related to cyp19a regulation. Both MDHT and Fz decreased cyp19a expression. Moreover, androgen receptor (ar) expression levels increased during development in all but the MDHT group, suggesting that early exposure to an androgen down-regulates subsequent ar expression in males and that Fz does not interact with the androgen receptor. Together, these observations indicate that although MDHT and Fz result in a similar phenotype, the molecular pathways involved are likely different, and show that Fz masculinization is the consequence of inhibited ovarian differentiation rather than of a direct androgenic effect. Further, since CPA did not alter sex ratios when administered during the period of highest androgen sensitivity, we suggest that androgens are not required for initial testicular differentiation in the sea bass. MDHT and Fz did not alter the number of precocious males but reduced and increased, respectively, their gonadosomatic index (GSI). In addition, Fz had lasting effects on the GSI of precocious and non-precocious males, probably due to alterations of estrogen function in the testis.     

Vitellogenin, steroid plasma levels and spawning performance of cultured female Senegalese sole (Solea senegalensis)

The Senegalese sole (Solea senegalensis) is a high value market flatfish, which aquaculture is compromised by severe reproductive problems; these are mostly found in soles hatched and raised in captivity (F1 generation). To gain knowledge on the reproductive dysfunctions observed in cultured (F1) Senegalese sole, this work aimed at developing a specific vitellogenin (VTG) ELISA, for the measurement of plasma VTG levels in this species. Profiles of VTG were correlated with those of sexual steroids and spawning performance of an F1 broodstock, during three consecutive years. The Senegalese sole VTG (ssVTG) was purified by precipitation with MgCl(2)-EDTA and anion-exchange chromatography and showed a molecular mass of 172 kDa, by SDS-PAGE. Specific antibodies were obtained and used to develop a competitive ELISA, which had a sensitivity of 3.6 ng ml(-1), and inter- and intra-assay coefficients of variation of 9.5% (n=29) and 6.7% (n=12), respectively. Annual profiles of plasma VTG showed a major peak at pre-spawning, and a second minor rise around autumn, which mirrored plasma profiles of both estradiol (E(2)) and testosterone (T) levels. Spontaneous spawning occurred every year in the spring season, but no fertilized eggs were obtained. In conclusion, this study described, for the first time, the purification and development of a sensitive and specific ELISA for Senegalese sole VTG. The endocrine and spawning data suggested that F1 female broodstock showed normal VTG and steroid releasing profiles in captivity with occurrence of spontaneous spawning, but no fertilization of the eggs was recorded.     

Androgen rather than estrogen up-regulates brain-type cytochrome P450 aromatase (cyp19a1b) gene via tissue-specific promoters in the hermaphrodite teleost ricefield eel Monopterus albus

CYP19A1 in the brain and pituitary of vertebrates is important for reproductive and non-reproductive processes. In teleosts, it is broadly accepted that estradiol (E₂) up-regulates cyp19a1b gene via a positive autoregulatory loop. Our present study, however, showed that E₂ did not up-regulate ricefield eel cyp19a1b in the hypothalamus and pituitary, whereas dihydrotestosterone (DHT) or testosterone (T) stimulated cyp19a1b expression only in the pituitary. Two tissue-specific promoters, namely promoter I and II directing the expression in the brain and pituitary respectively, were identified. Promoter I contained a non-consensus estrogen response element (ERE), and consequently did not respond to E₂. Promoter II contained an androgen response element (ARE) and consequently responded to DHT. Taken together, these results demonstrated a novel steroidal regulation of cyp19a1b gene expression and an alternative usage of tissue-specific cyp19a1b promoters in the brain and pituitary of a teleost species, the ricefield eel.