Somatolactin Research Articles

Effects of Water Temperature on the Body Color and Expression of the Genes Related to Body Color Regulation in the Goldfish.

Melanin-concentrating hormone (MCH), melanocyte-stimulating hormone (MSH), and somatolactin (SL) in the hypothalamus-pituitary axis are associated with body color regulation in teleost fish. Although these hormones' production and secretion respond well to light environments, such as background color, little is known about the effects of different water temperatures. We investigated the effects of water temperature, 10°C, 20°C, and 30°C, on body color and the expression of these genes and corresponding receptor genes in goldfish. The body color in white background (WBG) becomes paler at the higher water temperature, although no difference was observed in black background (BBG). Brain mRNA contents of proMCH genes (pmch1 and pmch2) increased at 30°C and 20°C compared to 10°C in WBG, respectively. Apparent effects of background color and temperature on the pituitary mRNA contents of a POMC gene (pomc) were not observed. The pituitary mRNA contents of the SLα gene were almost double those on a WBG at any temperature, while those of the SLβ gene (slb) at 30°C tended to be higher than those at 10°C and 20°C on WBG and BBG. The scale mRNA contents of the MCH receptor gene (mchr2) in WBG were higher than those in BBG at 30°C. The highest scale mRNA contents of MSH receptor (mc1r and mc5r) on BBG were observed at 20°C, while the lowest respective mRNA levels were observed at 30°C on WBG. These results highlight the importance of temperature for the endocrinological regulation of body color, and darker background color may stabilize those endocrine functions.

Source diversity of Artemia enrichment boosts goldfish (Carassius auratus) performance, β-carotene content, pigmentation, immune-physiological and transcriptomic responses

This study aimed to assess the impact of spirulina and/or canthaxanthin-enriched Artemia on the goldfish (Carassius auratus) growth, pigmentation, blood analysis, immunity, intestine and liver histomorphology, and expression of somatolactin (SL) and growth hormone (GH) genes. Artemia was enriched with spirulina and/or canthaxanthin for 24 h. Goldfish (N = 225, 1.10 ± 0.02 g) were tested in five experimental treatments, three replicates each: (T1) fish fed a commercial diet; (T2) fish fed a commercial diet and un-enriched Artemia (UEA); (T3) fish fed a commercial diet and spirulina-enriched Artemia (SEA); (T4) fish fed a commercial diet and canthaxanthin-enriched Artemia (CEA); and (T5) fish fed a commercial diet and spirulina and canthaxanthin-enriched Artemia (SCA) for 90 days. The results showed that the use of spirulina and/or canthaxanthin increased performance, β-carotene content and polyunsaturated fatty acids of Artemia. For goldfish, T5 showed the highest growth performance, β-carotene concentration and the lowest chromatic deformity. T5 also showed improved hematology profile, serum biochemical, and immunological parameters. Histomorphology of the intestine revealed an increase in villi length and goblet cells number in the anterior and middle intestine, with normal liver structure in T5. SL and GH gene expression in the liver and brain differed significantly among treatments with a significant increase in enriched Artemia treatments compared to T1 and T2. In conclusion, the use of spirulina and/or canthaxanthin improved performance of Artemia. Feeding goldfish spirulina and/or canthaxanthin-enriched Artemia improved performance, β-carotene content, pigmentation, health status and immune-physiological response.

Neuropeptides and hormones in hypothalamus-pituitary axis of Chinese sturgeon (Acipenser sinensis)

The hypothalamus and pituitary serve as important neuroendocrine center, which is able to secrete a variety of neuropeptides and hormones to participate in the regulation of reproduction, growth, stress and feeding in fish. Chinese sturgeon is a basal vertebrate lineage fish with a special evolutionary status, but the information on its neuroendocrine system is relatively scarce. Using the transcriptome data on the hypothalamus-pituitary axis of Chinese sturgeon as reference, we found out 46 hypothalamus neuropeptide genes, which were involved in regulation of reproduction, growth, stress and feeding. The results of sequence alignment showed that the neuroendocrine system of Chinese sturgeon evolves slowly, which confirms that Chinese sturgeon is a species with a slow phenotypic evolution rate. In addition, we also isolated six pituitary hormones genes from Chinese sturgeon, including reproductive hormones: follicle-stimulating homone (FSH) and luteinizing hormone (LH), growth-related hormones: growth hormone (GH)/prolactin (PRL)/somatolactin (SL), and stress-related hormone gene: proopiomelanocortin (POMC). Similar to teleost, immunostaining localization analysis in Chinese sturgeon pituitary showed that LH and FSH were located in the pituitary proximal pars distalis, SL was located in the pituitary rostral pars distalis, and POMC was located in the pituitary pars intermedia and pituitary rostral pars distalis. This study will give a contribution to enrich our information on the neuroendocrine system in Chinese sturgeon.

Immunohistochemical description and dispensation of the hypophyseal cell types in Liza ramada

ABSTRACTIntroduction: The localization and characterization of the hypophyseal cell types in L. ramada is so far lacking. Aim of the work: The present study aimed to describe and determine the hypophyseal cells in L. ramada. Material and methods: Immunohistochemical localization of the hormone-secreting cells in the hypophysis of L. ramada was completed by antisera against both mammalian and piscine hormones.Results: The hypophysis of L. ramada was comprised two main parts, the glandular part (adenohypophysis) and the nervous part (neurohypophysis). The adenohypophysis was divided into three subdivisions; rostral pars distalis (RPD), proximal pars distalis (PPD) and pars intermedia (PI). Seven of hormone-secreting cells were distinguished in the glandular part according to their grouping, distribution, and immunoreactivity. The acidophilic prolactin (PRL) cells occupied the bulk of the RPD and showed positive and specific immunoreaction with salmon PRL antiserum. Lead hematoxylin-positive (PbH+) cells, which were bordered the neurohypophysis in the RPD, gave positive immunoreactivity with human adrenocorticotrophic (ACTH) antiserum. The acidophilic cells of the PPD immunoreacted positively and strongly with antiserum against salmon growth hormone (GH). In addition, the central region of PPD comprised basophilic cells which were showed specific and strong immunoreaction with antisera against salmon gonadotropin (GTH) subunits; Iβ and IIβ. The presumptive thyrotropin (TSH) secreting cells were distinguished in the neurohypophysis between the RPD and the PPD, and they were negative immunoreacted with antiserum against rat thyrotropin (rBetaTSH). Two cell types were found in the PI; the Periodic Acid-Schiff-positive (PAS+) cells which had positive immunoreaction with anti-salmon somatolactin (SL) and the Lead hematoxylin-positive (PbH+) cells which immunostained with anti-alpha-melanin-stimulating hormone (α-MSH). Conclusion: The localization of hypophyseal cells of L. ramada, consider as basis for understanding and tracking hormonal changes during its reproductive cycle in captivity.

Effects of background color and feeding status on the expression of genes associated with body color regulation in the goldfish Carassius auratus

Alpha-melanocyte-stimulating hormone (α-MSH), a peptide derived from proopiomelanocortin (POMC), and melanin-concentrating hormone (MCH), act as neuromodulators and regulate food intake in vertebrates. In teleosts, these peptides are also involved competitively in body color regulation; α-MSH induces a dark body color, while MCH induces a pale body color. Similarly, members of the growth hormone (GH) family, somatolactin (SL) and prolactin (PRL), which are involved in the regulation of energy metabolism, are also associated with body color regulation in teleosts. Since these hormones are involved in both body color regulation and energy metabolism, it is possible that feeding status can affect body color. Here, we examined the effects of fasting on the response of goldfish body coloration to changes in background color. Goldfish were acclimated for one week in tanks with a white or black background under conditions of periodic feeding or fasting. The results showed that body color and expression levels of pmch1 and pomc were affected by background color, irrespective of feeding status. Expression levels of sla were higher in fish maintained in tanks with a black background than in tanks with a white background, and higher in the fasted fish compared to the fed fish. However, the pattern of slb expression was almost the opposite of that observed in sla expression. The expression levels of gh and prl in the pituitary, and pmch2a and pmch2b in the brain, were not affected by background color. These results suggest that MCH, α-MSH, SLα, and SLβ might be involved in body color regulation and that they are affected by background color in goldfish. The results also suggest that feeding status may affect body color regulation via SLα and SLβ, although these effects might be limited compared to the effect of background color.

Immunoreactivity of somatolactin (SL)-producing cells during larval development of Nile tilapia

ABSTRACTIntroduction: A growing body of evidence confirmed the involvement of somatolactin (SL) in various physiological processes including the acid-base as well as calcium regulation, fat metabolism and osmoregulation. However, less is known about the immunoreactivity of SL- producing cells during developmental stages of Nile tilapia, Oreochromis niloticus. Aim of the work: The aim of the present work was to examine the immunoreactivity of SL-producing cells during different developmental larval stages of Nile tilapia, O. niloticus. Material and methods: We performed immunohistochemical technique using specific antibody against chum salmon SL to identify the anatomical distribution and change in the activity of SL-producing cells accompanied the different larval stages of Oreochromis niloticus, and study the possible involvement of this hormone in the early endocrine integration. Results: The present immunocytochemical assessments revealed a few and dispersed numbers of SL-positive cells at the first day after hatching. Moreover, we observed a gradual increase in the activity of SL-positive cells in the pituitary gland as reflected with the increased cell number and size as well as SL immunoreactivity concomitant with the continued larval development from the first day post-hatching onwards and reached the highest levels on day 35 post- hatching. In parallel to the advanced development of larvae, the number of SL-positive cells and the intensity of immunoreaction were clearly and gradually increased especially after the onset of exogenous feeding at 21-28 days post-hatching. Consequently, there is an obvious increase in both the larval body length and weight during the experimental period of rearing.Conclusion: The early appearance of SL-producing cells and the gradual changes in their immunoreactivity during development suggest crucial effects of this hormone in different physiological processes during larval rearing of O. niloticus especially in the period after yolk sac resorption.

Gonadal remodeling and hormonal regulation during sex change of juvenile dusky grouper Epinephelus marginatus (Teleostei, Serranidae), an endangered protogynous hermaphrodite fish.

Sex change was induced in Epinephelus marginatus juveniles using a nonsteroidal aromatase inhibitor (AI), a synthetic androgen (17α-methyltestosterone; MT), and a combination of both (MT + AI) in a 90-day experiment. A detailed remodeling of the gonads, the plasma level of gonadal steroids, and immunostaining of pituitary follicle-stimulating hormone (FSH), luteinizing hormone (LH), and somatolactin (SL) cells were analyzed. Sex inversion reached the final spermatogenesis stages using MT, while AI triggered spermatogenesis, but reaching only the spermatid stage. Estradiol (E2) levels did not change in fish treated with AI but decreased throughout the experimental period in animals treated with MT and MT + AI. Testosterone (T) levels increased in animals treated with MT during the first 60 days (and combined with AI in the first 30 days), decreasing in all experimental groups at 90 days, while AI-treated animals had increased plasma 11-ketotestosterone (11-KT) levels after 90 days. In control fish, FSH- and SL-producing cells (ir-FSH and ir-SL) were restricted to pars intermedia (PI) of the adenohypophysis. Pituitary ir-FSH cells were decreased at the end of the experimental period in all treatments compared with the CT animals. LH-producing cells (ir-LH) were present in proximal pars distalis (PPD) and pars intermedia (PI) of adenohypophysis and did not change after the experimental period. The decreased number of ir-FSH cells at the end of the experiment in all treatments could be related to the negative feedback loop triggered by the increase in natural and/or synthetic androgens.

Brain Control Reproduction by the Endocrine System of Female Blue Gourami (Trichogaster trichopterus).

Blue gourami belongs to the Labyrinithici fish and the Anabantiform order. It is characterized by a specific organ located above its gills for the respiration of atmospheric oxygen. This specific adaptation to low oxygen levels affects reproduction that is controlled by the brain, which integrates different effects on reproduction mainly through two axes—the gonadotropic brain pituitary gonad axis (BPG) and the hypothalamic-pituitary-somatotropic axis (HPS axis), including the interactions between them. This brain control reproduction of the Anabantoidei suborder summarizes information that has been published on the hormones involved in controlling the reproduction system of a model female blue gourami fish (Trichogaster trichopterus), including unpublished data. In the whole-brain transcriptome of blue gourami, 17 transcription genes change during vitellogenesis in the brain. The hormones involved in reproduction in blue gourami described in the present paper include: Kisspeptin 2 (Kiss 2) and its receptors 1 and 2 (KissR 1 and 2); gonadotropin-releasing hormone 1, 2 and 3 (GnRH1, 2 and 3); GnRH receptor; pituitary adenylate cyclase-activating polypeptide (PACAP) and its related peptide (PRP); somatolactin (SL); follicle-stimulating hormone (FSH); luteinizing hormone (LH); growth hormone (GH); prolactin (PRL), 17β-estradiol (E2); testosterone (T); vitellogenesis (VTL); and 17α,20β- dihydroxy-4-pregnen-3-one (17,20P). A proposed quality model is presented regarding the brain control oogenesis in blue gourami that has a Labyrinth organ about which relatively little information has been published. This paper summarizes the complex various factors involved in the interactions between external and internal elements affecting the brain of fish reproduction in the Anabantiform order. It is suggested to study in the future the involvement of receptors of hormones, pheromones, and genome changes in various organs belonging to the reproduction system during the reproduction cycles about which little is known.

Evolution of the growth hormone, prolactin, prolactin 2 and somatolactin family

Growth hormone (GH), prolactin (PRL), prolactin 2 (PRL2) and somatolactin (SL) belong to the same hormone family and have a wide repertoire of effects including development, osmoregulation, metabolism and stimulation of growth. Both the hormone and the receptor family have been proposed to have expanded by gene duplications in early vertebrate evolution. A key question is how hormone-receptor preferences have arisen among the duplicates. The first step to address this is to determine the time window for these duplications. Specifically, we aimed to see if duplications resulted from the two basal vertebrate tetraploidizations (1R and 2R). GH family genes from a broad range of vertebrate genomes were investigated using a combination of sequence-based phylogenetic analyses and comparisons of synteny. We conclude that the PRL and PRL2 genes arose from a common ancestor in 1R/2R, as shown by neighboring gene families. No other gene duplicates were preserved from these tetraploidization events. The ancestral genes that would give rise to GH and PRL/PRL2 arose from an earlier duplication; most likely a local gene duplication as they are syntenic in several species. Likewise, some evidence suggests that SL arose from a local duplication of an ancestral GH/SL gene in the same time window, explaining the lack of similarity in chromosomal neighbors to GH, PRL or PRL2. Thus, the basic triplet of ancestral GH, PRL/PRL2 and SL genes appear to be unexpectedly ancient. Following 1R/2R, only SL was duplicated in the teleost-specific tetraploidization 3R, resulting in SLa and SLb. These time windows contrast with our recent report that the corresponding receptor genes GHR and PRLR arose through a local duplication in jawed vertebrates and that both receptor genes duplicated further in 3R, which reveals a surprising asynchrony in hormone and receptor gene duplications.

Immunohistochemical study of gonadotropin-releasing hormone and somatolactin during induced spawning of Liza ramada

Introduction: Gonadotropin releasing hormone (GnRH) is a crucial regulator of gonadal development and reproduction in fish. In addition, it is a possible secretagogue of somatolactin (SL) in teleosts. Little is known about the possible association of GnRH and SL during reproduction of Liza ramada.Aim of the work: The present study was designed to investigate the GnRH and SL immunoreactivities during inducing the final stages of maturation, ovulation and spawning of Liza ramada; to assess the possible actions of these hormones on sexual maturation and spawning of teleosts.Materials and Methods: Histological and immunohistochemical techniques were generally performed to describe the immunoreactivity of both GnRH and SL during ovarian maturation and spawning of Liza ramada.Results: The mammalian GnRH (mGnRH) cell bodies were observed in the medulla oblongata. However, SL-immunoreactive (-IR) cells were detected in the pars intermedia (PI) of the pituitary gland. The GnRH-IR nerve terminals are in close contact with SL-IR pituitary cells. Importantly, the activity of both mGnRH-IR neurons and SL-IR pituitary cells were increased as reflected with the increased cell number and size during sexual maturation and spawning. The activity of these cells was more pronounced as indicated by the increased cells number and size with more immunoreactivity in pre-spawning female. During final maturation and spawning, mGnRH-IR neurons and SL cells showed an increase in the secretory activity as reflected by their small sizes, vacuolated appearance and weak immunoreactivity.Conclusions: Taken together, these findings suggest that the close contact of mGnRH-IR terminals with SL-IR cells in the pituitary, together with the concomitant changes of GnRH and SL immunoreactivity in response to progress in the reproductive events suggest that GnRH may regulate SL release and the regulation of reproductive activity in L. ramada.

Evolution of the receptors for growth hormone, prolactin, erythropoietin and thrombopoietin in relation to the vertebrate tetraploidizations

The receptors for the pituitary hormones growth hormone (GH), prolactin (PRL) and somatolactin (SL), and the hematopoietic hormones erythropoietin (EPO) and thrombopoietin (TPO), comprise a structurally related family in the superfamily of cytokine class-I receptors. GH, PRL and SL receptors have a wide variety of effects in development, osmoregulation, metabolism and stimulation of growth, while EPO and TPO receptors guide the production and differentiation of erythrocytes and thrombocytes, respectively. The evolution of the receptors for GH, PRL and SL has been partially investigated by previous reports suggesting different time points for the hormone and receptor gene duplications. This raises questions about how hormone-receptor partnerships have emerged and evolved. Therefore, we have investigated in detail the expansion of this receptor family, especially in relation to the basal vertebrate (1R, 2R) and teleost (3R) tetraploidizations. Receptor family genes were identified in a broad range of vertebrate genomes and investigated using a combination of sequence-based phylogenetic analyses and comparative genomic analyses of synteny. We found that 1R most likely generated EPOR/TPOR and GHR/PRLR ancestors; following this, 2R resulted in EPOR and TPOR genes. No GHR/PRLR duplicate seems to have survived after 2R. Instead the single GHR/PRLR underwent a local duplication sometime after 2R, generating separate syntenic genes for GHR and PRLR. Subsequently, 3R duplicated the gene pair in teleosts, resulting in two GHR and two PRLR genes, but no EPOR or TPOR duplicates. These analyses help illuminate the evolution of the regulatory mechanisms for somatic growth, metabolism, osmoregulation and hematopoiesis in vertebrates.

Dopamine inhibits somatolactin gene expression in tilapia pituitary cells through the dopamine D2 receptors

Dopamine (DA) is an important neurotransmitter in the central nervous system of vertebrates and possesses key hypophysiotropic functions. Early studies have shown that DA has a potent inhibitory effect on somatolactin (SL) release in fish. However, the mechanisms responsible for DA inhibition of SL gene expression are largely unknown. To this end, tilapia DA type-1 (D1) and type-2 (D2) receptor transcripts were examined in the neurointermediate lobe (NIL) of the tilapia pituitary by real-time PCR. In tilapia, DA not only was effective in inhibiting SL mRNA levels in vivo and in vitro, but also could abolish pituitary adenylate cyclase-activating polypeptide (PACAP)- and salmon gonadotropin-releasing hormone (sGnRH)-stimulated SL gene expression at the pituitary level. In parallel studies, the specific D2 receptor agonists quinpirole and bromocriptine could mimic the DA-inhibited SL gene expression. Furthermore, the D2 receptor antagonists domperidone and (−)-sulpiride could abolish the SL response to DA or the D2 agonist quinpirole, whereas D1 receptor antagonists SCH23390 and SKF83566 were not effective in this respect. In primary cultures of tilapia NIL cells, D2 agonist quinpirole-inhibited cAMP production could be blocked by co-treatment with the D2 antagonist domperidone and the ability of forskolin to increase cAMP production was also inhibited by quinpirole. Using a pharmacological approach, the AC/cAMP pathway was shown to be involved in quinpirole-inhibited SL mRNA expression. These results provide evidence that DA can directly inhibit SL gene expression at the tilapia pituitary level via D2 receptor through the AC/cAMP-dependent mechanism.

Production and Purification of Recombinant Somatolactin and its Effects on Insulin-like Growth Factors Gene Expression in Tilapia Hepatocytes

Somatolactin (SL), the member of the growth hormone (GH)/prolactin (PRL) family, is fish-specific pituitary hormone with diverse functions. However, little is known about its biological function in fish hepatocytes. Using tilapia as a model, SLtranscripts were shown to be widely expressed in various extrapituitary tissues with the relatively high expression level in liver. To explore the biological action of SL in hepatocytes, we produced and purified recombinant tilapia SL protein which could induce pigment aggregation in tilapia melanophores. Further, the antiserum for the SL was produced and its specificity was confirmed by antiserum preabsorption. During 4 week starvation, hepatic SL transcripts in starved fish were significantly higher than the control fish starting on the 1st week of starvation until 4th week. After re-feeding, the SL transcripts level returned to normal. Using primary cultures of tilapia hepatocytes, insulin-like growth factors (IGF1 and IGF2) gene expression were elevated by static incubation with recombinant tilapia SL. In contrast, removal endogenous SL by immunoneutralization using SL antiserum was shown to inhibit IGF1 and IGFgene expression. These findings, taken together, provide evidence for the first time that SL may serve as a novel regulator in fish stimulating IGF1 and IGF gene expression in hepatocytes.

Gonadotropins and Growth Hormone Family Characterization in an Endangered Siluriform Species, Steindachneridion parahybae (Pimelodidae): Relationship With Annual Reproductive Cycle and Induced Spawning in Captivity.

The aim of this study was to identify and characterize pituitary cells of Steindachneridion parahybae females in captivity, highlighting the possible relationship with reproductive disorders at this level, since this species shows oocyte final maturation, ovulation and spawning dysfunction in captivity. The localization and distribution of growth hormone (GH), prolactin (PRL), somatolactin (SL), β-luteinizing hormone (β-LH), and β-follicle stimulating hormone (β-FSH) immunoreactive (-ir) cells in the adenohypophysis was studied by immunohistochemical and Western blot methods. In addition, cellular morphometric analyses and semi-quantification of ir-cells optical density (OD) during the annual reproductive cycle and after artificial induced spawning (AIS) were performed. Results showed that the distribution and general localization of pituitary cell types were similar to that of other teleost species. However, the morphometrical study of adenohypophysial cells showed differences along the reproductive cycle and following AIS. In general, females at the vitellogenic stage presented greater OD values for GH, PRL and SL than at other maturation stages (previtellogenic and regression stages), probably indicating an increased cellular activity during this stage. Conversely, β-LH OD did not vary during the annual reproductive cycle. After AIS, β-LH, SL and GH ir-cells showed an increase in OD values suggesting a possible involvement on oocyte final maturation, ovulation and spawning or a feedback control on the brain-pituitary-gonads axis. Reproductive dysfunction in S. parahybae females in captivity may be due to alteration of the synthesis pathways of β-LH. In addition, GH family of hormones could modulate associated mechanisms that influence the reproductive status in this species.

Adrenergic Regulation of Somatolactin Gene Expression in Tilapia Pituitary Cells

Epinephrine is an important neuroendocrine regulator to control growth hormone (GH) secretion in vertebrates. Somatolactin (SL), the latest member of the GH family, is a novel pituitary hormone with diverse functions in fish. In a previous report it was shown that epinephrine had a potent inhibitory effect on SL release in fish. However, very little is known about the mechanisms responsible for epinephrine inhibition of SL gene expression. In primary cultures of tilapia neurointermediate lobe (NIL) cells, epinephrine not only reduced SL mRNA levels, but could also abolish pituitary adenylate cyclase-activating polypeptide (PACAP)-stimulated SL gene expression. The inhibitory effects of epinephrine on SL gene expression were mimicked by additions of α2-adrenergic agonists clonidine and UK14304, whereas similar treatments with the α1-agonist cirazoline or the β-agonist isoproterenol had no effects in this regard. In parallel experiments, the SL response to epinephrine was significantly abolished by co-incubations with the α2- antagonist yohimbine, but the α1- or β-antagonist was not effective in this regard. In tilapia NIL cells, the α2-adrenergic agonist clonidine suppressed cAMP production and blocked forskolin and PACAP induction of total cAMP production. By using a pharmacological approach, the adenylate cyclase (AC) activator- and cAMP analog-stimulated SL responses were blocked treatment with clonidine. Furthermore, adrenergic inhibition of SL gene expression was also mimicked by inhibiting AC and blocking protein kinase A (PKA). These results, as a whole, suggest that α2-adrenergic stimulation can downregulate SL gene transcription by inhibiting the AC/cAMP-dependent mechanism at the tilapia pituitary level.

Somatolactin Transcription during Oogenesis in Female Blue Gourami (<i>Trichogaster trichopterus</i>)

Somatolactin (SL), a specific pituitary hormone belonging to the prolactin (PRL) super family, is involved in background adaptation, osmoregulation, reproduction and fatty acid metabolism. The goal of this study was to examine the gene transcription of SL changes in the ovary of blue gourami females (Trichogaster trichopterus) during oogenesis by quantitative real-time polymerase chain reaction (Real-Time PCR). Somatolactin in the pituitary was higher in females at low vitellogenesis compared to females with oocytes in maturation, and the difference was significant (p < 0.05). No significant differences were found in mRNA levels between low and high vitellogenesis, and high vitellogenesis and maturation. The findings of this and previous studies demonstrate that SL, growth hormone (GH) and PRL are involved in oogenesis in blue gourami; however, considerably more studies are required in order to separate the functions of these hormones.

Hypoxia acclimation protects against oxidative damage and changes in prolactin and somatolactin expression in silver catfish (Rhamdia quelen) exposed to manganese

The aim of this study was to assess the Mn toxicity to silver catfish considering Mn accumulation and oxidative status in different tissues, as well as pituitary hormone expression after acclimation to hypoxia. Silver catfish acclimated to hypoxia for 10 days and successively exposed to Mn (9.8mgL−1) for an additional 10 days exhibited lower Mn accumulation in plasma, liver, kidneys and brain and prevented the hematocrit decrease observed in the normoxia group. Hypoxia acclimation also modified Mn-induced oxidative damage, which was observed by lower reactive species (RS) generation in gills and kidneys, decreased lipid peroxidation (LP) levels in gills, liver and kidneys and decreased protein carbonyl (PC) levels in liver, kidneys and brain. Manganese accumulation showed positive correlations with LP levels in gills and kidneys, as well as with PC levels in gills, liver and brain. In addition, hypoxia acclimation and Mn exposure increased catalase (CAT) activity in gills and kidneys and Na+/K+-ATPase activity in gills, liver and brain. Silver catfish that were acclimated under normoxia and exposed to Mn displayed increased pituitary prolactin (PRL) and decreased somatolactin (SL) expression. Interestingly, hypoxia acclimation prevented hormonal fluctuation of PRL and SL in fish exposed to Mn. These findings indicate that while the exposure of silver catfish to Mn under normoxia was related to metal accumulation and oxidative damage in tissues together with endocrine axis disruption, as represented by PRL and SL, hypoxia acclimation reduced waterborne Mn uptake, thereby minimizing oxidative damage and changes in hormonal profile. We hypothesized that moderate hypoxia is able to generate adaptive responses, which may be related to hormesis, thereby ameliorating Mn toxicity to silver catfish.

The influence of stocking density and food deprivation in silver catfish (Rhamdia quelen): A metabolic and endocrine approach

The influence of stocking density and food deprivation on energy metabolism, stress processes and the pituitary endocrine system of silver catfish ( Rhamdia quelen ) was investigated after a period of 14 days, in which plasmatic and hepatic parameters and the mRNA expression of prolactin (PRL), growth hormone (GH) and somatolactin (SL) were assessed. The fish were subjected to four experimental conditions: (1) fed under high stocking density (32 kg/m 3 , HSD); (2) fed under mean stocking density (16 kg/m 3 , MSD); (3) fed under low stocking density (8 kg/m 3 , LSD); and (4) food-deprived under low stocking density (8 kg/m 3 , LSD-FD). After 14 days, plasma and liver samples were obtained to analyze the metabolite levels and enzymatic activities related to metabolism, and pituitary glands were obtained to analyze hormone expression (PRL, GH and SL). Liver weight and the hepato-somatic index (HSI) revealed that specimens maintained at HSD and/or MSD had higher hepatic stores, which were observed in the triglyceride and glycogen levels in this tissue, than animals submitted to the LSD and LSD-FD groups. Triglyceride levels in the plasma and liver revealed the consumption of fatty acid reserves in the fasting group. Enzymatic activities, such as glutamate dehydrogenase (GDH), phosphorylase (GPase), pyruvate kinase (PK), aspartate transaminase (AST) and glycerol-3-phosphate dehydrogenase (G3PDH), indicated an increase in gluconeogenic pathways in the HSD group and an increase in glycolitic metabolism in the LSD groups. The expression of PRL was not affected by stocking density and/or food deprivation and GH decreased with increased density and increased in fasting conditions. A negative effect of density and fasting was observed on the expression of SL. Overall, the data suggested that juvenile silver catfish reared at stocking densities of 16 to 32 kg/m 3 were better maintained than those maintained at the lowest density. • Stocking density is an important factor for the cultivation of fish. • Silver catfish were submitted to 32, 16 and 8 kg/m 3 for 14 days with food or fasting. • Better stocking conditions are 16 to 32 kg/m 3 . • Lowest stocking density increased glycolytic and glycogenolytic pathways. • An increase in gluconeogenic pathways in the group of the highest density occurred.

Differential response of two somatolactin genes to zinc or estrogen in pituitary of Cyprinus carpio

Environmental changes affect gene expression that we addressed in the pituitary, a central regulatory organ at the interface between the central nervous system and the endocrine system. With the aim to reveal effects of changes in the aquatic environment on the expression of hypothalamo-hypophyseal factors, we characterized somatolactin (SL) in Cyprinus carpio. SL, a fish specific pituitary hormone belonging to the prolactin (PRL) superfamily, is involved in background adaptation, osmoregulation, reproduction and fatty acid metabolism. Two sl genes, α and β, were discovered in carp and transcripts of both were detected in pituitaries. Clearly, expression of slα and slβ was modulated significantly in pituitary of male adult carp in response to treatment with ZnCl2 (Zn), but only slβ responded to 17β-estrogen (E2), relative to control carp as shown by RT-qPCR analyses. Furthermore, the amount of mRNA of related factors was assessed revealing variable effects on prl, growth hormone (gh), and factors involved in sl regulation: the pituitary transcription factor pit1 and hypothalamic pituitary adenylase cyclase activating peptide (pacap). In parallel, the physiological response of the experimental animals to Zn or E2 was confirmed by showing a significant increase of metallothionein (mt) or vitellogenin (vg) gene expression in liver, classical sentinels for exposure to heavy metal or estrogens.These data suggest that the sl genes seem to be involved in the response to Zn, as well as to estrogen, and could contribute to evaluate biological relevant changes in the aquatic environment.

Kisspeptin regulates the somatic growth-related factors of the cinnamon clownfish Amphiprion melanopus

This study aimed to test the effects of kisspeptin (Kiss) on somatic growth in the cinnamon clownfish Amphiprion melanopus. We investigated the effects of Kiss treatment on the growth by measuring the mRNA expressions of the growth hormone (GH), insulin-like growth hormone factor (IGF-I), somatolactin (SL), and melatonin receptor (MT). The expression levels of GH and SL of the pituitary gland and IGF-I of the liver increased after Kiss treatment (in vivo and in vitro). In addition, the MT mRNA expression increased in the pituitary gland and brain after Kiss treatment (in vivo and in vitro). These results support the hypothesis that Kiss directly regulates the somatic growth-related factors, such as GH, SL, and MT, and IGF-I in the cinnamon clownfish. Further, injection of Kiss resulted in significantly higher levels of plasma melatonin than that in the control. We, therefore, conclude that Kiss plays a role in modulating growth and artificially induced rapid growth in cinnamon clownfish.