Pituitary gland volumes and stress: Results of a population-based adult sample.

Early and chronic stress was reported to alter the hypothalamic-pituitary-adrenal axis functioning which regulates the secretion of cortisol. Nevertheless, few studies mainly focused on specific study populations (e.g. adolescents, pregnant women, and psychiatric patients), and researched interactive associations of pituitary volumes and single stress markers. The present study used pituitary volumes of two adult general-population cohorts of the Study of Health in Pomerania (SHIP-START-2: N = 1026, 54% Men, 30-90 years; SHIP-TREND-0: N = 1868, 53% Men, 21-82 years). In linear regression models, main effects of the pituitary volumes as well as interaction effects with childhood abuse and neglect (Childhood Trauma Questionnaire) were estimated using depressive symptoms (Beck Depression Inventory-II), and serum cortisol concentrations as outcome variables. The results of both cohorts were integrated via meta-analyses. No main effect between pituitary volumes and depressive symptoms was observed (START-2: ß = -0.004 [-0.082; 0.075], p = .929; TREND-0: ß = 0.020 [-0.033; 0.073], p = .466; Meta-analysis: ß = 0.012 [-0.031; 0.056], p = .580). However, larger pituitary volumes were associated with more depressive symptoms in participants with more severe childhood neglect (START-2: ß = 0.051 [-0.024; 0.126], p = .183; TREND-0: ß = 0.083 [0.006; 0.159], p = .034; Meta-analysis: ß = 0.066 [0.013; 0.120], p = .015). Further, larger pituitary volumes were associated with lower serum cortisol concentrations in participants with more severe depressive symptoms (START-2: ß = -0.087 [-0.145; -0.030], p = .003; TREND-0: ß = -0.053 [-0.091; -0.015], p = .006; Meta-analysis: ß = -0.063 [-0.095; -0.032], p = 8.39e-05). Summarizing, larger pituitary volumes were associated with more severe psychopathological symptoms, particularly in participants reporting early life stress. This was supported by stronger associations between pituitary volumes and cortisol concentrations in participants with more severe depressive symptoms. Future studies are needed to transfer these results into developmental stages of high hormonal changes and patient samples.

Vasoactive Intestinal Peptide Indirectly Elicits Pituitary LH Secretion Independent of GnRH in Female Zebrafish.

Vasoactive intestinal peptide (Vip) regulates luteinizing hormone (LH) release through the direct regulation of gonadotropin-releasing hormone (GnRH) neurons at the level of the brain in female rodents. However, little is known regarding the roles of Vip in teleost reproduction. Although GnRH is critical for fertility through the regulation of LH secretion in vertebrates, the exact role of the hypophysiotropic GnRH (GnRH3) in zebrafish is unclear since GnRH3 null fish are reproductively fertile. This phenomenon raises the possibility of a redundant regulatory pathway(s) for LH secretion in zebrafish. Here, we demonstrate that VipA (homologues of mammalian Vip) both inhibits and induces LH secretion in zebrafish. Despite the observation that VipA axons may reach the pituitary proximal pars distalis including LH cells, pituitary incubation with VipA in vitro, and intraperitoneal injection of VipA, did not induce LH secretion and lhß mRNA expression in sexually mature females, respectively. On the other hand, intracerebroventricular administration of VipA augmented plasma LH levels in both wild-type and gnrh3-/- females at 1 hour posttreatment, with no observed changes in pituitary GnRH2 and GnRH3 contents and gnrh3 mRNA levels in the brains. While VipA's manner of inhibition of LH secretion has yet to be explored, the stimulation seems to occur via a different pathway than GnRH3, dopamine, and 17ß-estradiol in regulating LH secretion. The results indicate that VipA induces LH release possibly by acting with or through a non-GnRH factor(s), providing proof for the existence of functional redundancy of LH release in sexually mature female zebrafish.

FOXO Transcription Factors Are Required for Normal Somatotrope Function and Growth.

Understanding the molecular mechanisms underlying pituitary organogenesis and function is essential for improving therapeutics and molecular diagnoses for hypopituitarism. We previously found that deletion of the forkhead factor, Foxo1, in the pituitary gland early in development delays somatotrope differentiation. While these mice grow normally, they have reduced growth hormone expression and free serum insulin-like growth factor-1 (IGF1) levels, suggesting a defect in somatotrope function. FOXO factors show functional redundancy in other tissues, so we deleted both Foxo1 and its closely related family member, Foxo3, from the primordial pituitary. We find that this results in a significant reduction in growth. Consistent with this, male and female mice in which both genes have been deleted in the pituitary gland (dKO) exhibit reduced pituitary growth hormone expression and serum IGF1 levels. Expression of the somatotrope differentiation factor, Neurod4, is reduced in these mice. This suggests a mechanism underlying proper somatotrope function is the regulation of Neurod4 expression by FOXO factors. Additionally, dKO mice have reduced Lhb expression and females also have reduced Fshb and Prl expression. These studies reveal FOXO transcription factors as important regulators of pituitary gland function.

Successive seminal collections from Leiarius marmoratus during the reproductive period.

This study aim was to verify whether milt quality of male Leiarius marmoratus is maintained among successive samples collected during the same reproductive period. Ten reproductively mature males were used to evaluate four successive sperm samples collected at 10-day intervals. For these collections, seven males were injected with carp pituitary homogenate (CPH) at a dosage of 3.0 mg/kg body weight, in two applications (30% and 70%), at an interval of 10 h. The other three males were administered only saline (control). Injection with CPH or saline occurred prior to each of the four collections. Only one male from the control group released a small volume of milt (0.33 mL), and only during the first collection period. Of the seven males treated with CPH, five released milt during all four collections. Milt volume of the first sample collected (0.63 mL) did not differ from that of other samples (0.59-1.38 mL; P > 0.05). Sperm concentration was greater in the first samples collected (1.98 × 109 spermatozoa/mL) compared to the other samples (0.35 × 109 at 0.92 × 109 spermatozoa/mL; P < 0.05). Sperm motility, curvilinear velocity, straightness, and morphological normality did not differ among the consecutive samples (P > 0.05). Average path velocity, straight-line velocity, oscillation, linearity, progression, and membrane integrity decreased slightly in the samples collected subsequent to the first sample (P > 0.05). In conclusion, milt quality decreased among successive collections; however, quality of all samples from all collections was sufficient for use for fertilization of oocytes.

Dynamic Expression of Imprinted Genes in the Developing and Postnatal Pituitary Gland.

In mammals, imprinted genes regulate many critical endocrine processes such as growth, the onset of puberty and maternal reproductive behaviour. Human imprinting disorders (IDs) are caused by genetic and epigenetic mechanisms that alter the expression dosage of imprinted genes. Due to improvements in diagnosis, increasing numbers of patients with IDs are now identified and monitored across their lifetimes. Seminal work has revealed that IDs have a strong endocrine component, yet the contribution of imprinted gene products in the development and function of the hypothalamo-pituitary axis are not well defined. Postnatal endocrine processes are dependent upon the production of hormones from the pituitary gland. While the actions of a few imprinted genes in pituitary development and function have been described, to date there has been no attempt to link the expression of these genes as a class to the formation and function of this essential organ. This is important because IDs show considerable overlap, and imprinted genes are known to define a transcriptional network related to organ growth. This knowledge deficit is partly due to technical difficulties in obtaining useful transcriptomic data from the pituitary gland, namely, its small size during development and cellular complexity in maturity. Here we utilise high-sensitivity RNA sequencing at the embryonic stages, and single-cell RNA sequencing data to describe the imprinted transcriptome of the pituitary gland. In concert, we provide a comprehensive literature review of the current knowledge of the role of imprinted genes in pituitary hormonal pathways and how these relate to IDs. We present new data that implicate imprinted gene networks in the development of the gland and in the stem cell compartment. Furthermore, we suggest novel roles for individual imprinted genes in the aetiology of IDs. Finally, we describe the dynamic regulation of imprinted genes in the pituitary gland of the pregnant mother, with implications for the regulation of maternal metabolic adaptations to pregnancy.

Di-n-butyl phthalate diminishes testicular steroidogenesis by blocking the hypothalamic-pituitary-testicular axis: relationship with germ cell apoptosis in Japanese quail.

Although di-n-butyl phthalate (DBP) induces germ cell apoptosis, the underlying mechanism is not yet clear in quail. In this study, prepubertal quails were given a single dose of 500mg kg-1 DBP by gavage and were then killed 3, 6 and 24h after treatment. There was a significant reduction in intratesticular testosterone (ITT) concentrations and testicular steroidogenic enzyme mRNA expression and a significant increase in germ cell apoptosis in DBP-treated compared with control quails at all time points. Maximum apoptosis was detected 6h after treatment and the maximum reduction in testosterone concentrations was at 3h. To investigate whether DBP suppressed testicular steroidogenesis by affecting the hypothalamic-pituitary-testicular axis, we analysed pituitary LH subunit ß (Lhb) mRNA expression and serum LH concentrations. At all time points, pituitary Lhb expression and serum LH concentrations were significantly decreased following DBP treatment. The present observations suggest the possibility that DBP blocked LH secretion from the hypothalamus and/or pituitary, thereby decreasing LH stimulation of Leydig cells and reducing ITT concentrations. DBP-induced decreases in ITT concentrations may cause changes to the physical structure of Sertoli cells, which, in turn, may induce germ cell apoptosis.

Alpha-melanocyte stimulating hormone immunoreactivity in the brain of the cichlid fish Oreochromis mossambicus.

This study reports the distribution of a pro-opiomelanocortin-derived neuropeptide α-MSH in the brain of the cichlid fish Oreochromis mossambicus. α-MSH-ir fibres were found in the granule cell layer of the olfactory bulb, the medial olfactory tract, the pallium and the subpallium, whereas in the preoptic area of the telencephalon, few large α-MSH-ir perikarya along with extensively labeled fibres were observed close to the ventricular border. Dense network of α-MSH-ir fibres were seen in the hypothalamic areas such as the nucleus preopticus pars magnocellularis, the nucleus preopticus pars parvocellularis, the suprachiasmatic nucleus, the nucleus anterior tuberis, the paraventricular organ, the subdivisions of the nucleus recessus lateralis and the nucleus recessus posterioris. In the nucleus lateralis pars medialis, some α-MSH-ir perikarya and fibres were found along the ventricular margin. In the diencephalon, numerous α-MSH-ir fibres were detected in the nucleus posterior tuberis, the nucleus of the fasciculus longitudinalis medialis and the nucleus preglomerulosus medialis, whereas in the mesencephalon, α-MSH-ir fibres were located in the optic tectum, the torus semicircularis and the tegmentum. In the rhombencephalon, α-MSH-ir fibres were confined to the medial octavolateralis nucleus and the descending octaval nucleus. In the pituitary gland, densely packed α-MSH-ir cells were observed in the pars intermedia region. The widespread distribution of α-MSH-immunoreactivity throughout the brain and the pituitary gland suggests a role for α-MSH peptide in regulation of several neuroendocrine and sensorimotor functions as well as darkening of pigmentation in the tilapia.

Effect of hormonal stimulation on captive broodstock maturation, induced breeding and spawning performance of striped snakehead, Channa striata (Bloch, 1793).

In the present study, there was assessment of effects of gonadotropin treatments on broodstock maturation, induced breeding, and spawning outcomes of striped snakehead in captivity. The striped snakehead (n = 128) were equally distributed in four concrete tanks (15 m2) and hormone implants (500 IU human chorionic gonadotropin (hCG)/kg body weight) were inserted intramuscularly and striped snakehead broodstock administered this treatment were confined in two tanks and striped snakehead of a non-implanted group were confined in two tanks. The hormone implanted striped snakehead had a greater (P < 0.05) gonadosomatic index (GSI) and oocyte diameter in comparison to non-implanted striped snakehead. In a subsequent experiment, hCG and carp pituitary homogenate (CPH) were evaluated for inducing breeding. Dosages of hCG used were, 2,000 (TH1), 3000 (TH2), and 4000 (TH3) IU hCG/kg body weight of females. Dosages of CPH were, 20 (TP1), 30 (TP2), and 40 (TP3) mg CPH/kg body weight of females. Males were administered 0.75 of the dosage administered to females. The values for reproductive variables were estimated. Fertilization (89.0 ±â€¯3.0 %) and hatching (92.0 ±â€¯1.0 %) rates were greater (P < 0.05) in the TH1 group of implanted striped snakehead. Relative fecundity (19,023 ±â€¯2211), as well as fertilization (96.2 ±â€¯2.4 %), and hatching (96.6 ±â€¯1.7 %) rates were greater in the TP2 group of the implanted striped snakehead. The results from the present study indicate broodstock treated with gonadotropins had greater spawning outcomes which might facilitate mass scale breeding and fertilized egg as well as juvenile production of striped snakehead in captivity.

Pharmacokinetics and Tissue Distribution of Loratadine, Desloratadine and Their Active Metabolites in Rat based on a Newly Developed LC-MS/MS Analytical Method.

Loratadine (LOR) and its major metabolite, desloratadine (DL) are new-generation antihistamines. The hydroxylated metabolites of them, 6-OH-DL, 5-OH-DL and 3-OH-DL are also active because of their ability to inhibit binding of pyrilamine to brain H1 receptors and a tendency for distributing to specific immune-regulatory tissues. In this study, a new validated LC-MS/MS method to simultaneously quantify LOR, DL, 6-OH-DL, 5-OH-DL and 3-OH-DL in plasma and tissues was established and applied to an investigation of their pharmacokinetics and target-tissue distribution tendency for the first time. Pharmacokinetics parameters in rat were measured and the results suggest that the body's exposure to active metabolites were much higher than to the prodrug with LOR, but much lower with DL. The tissue distribution study shows that LOR, DL and their active metabolites were widely distributed in the liver, spleen, thymus, heart, adrenal glands and pituitary gland. For immune-regulatory tissues, the concentrations of LOR, DL and their active metabolites in the spleen were much higher than in the thymus, which is related to the spleen, one of the sites where immune responses occur. LOR and its metabolites might inhibit immune-mediated allergic inflammation through the hypothalamic-pituitary-adrenal (HPA) axis. It was also found that the concentration of LOR in the heart was highest after liver and adrenal glands while those of DL, 6-OH-DL and 5-OH-DL in the liver, adrenal glands and spleen were all higher than those in the heart, which suggests that LOR may have a greater tendency to distribute in the heart than its metabolites.

Differential impacts of carp and salmon pituitary extracts on induced oogenesis, egg quality, molecular ontogeny and embryonic developmental competence in European eel.

Low egg quality and embryonic survival are critical challenges in aquaculture, where assisted reproduction procedures and other factors may impact egg quality. This includes European eel (Anguilla anguilla), where pituitary extract from carp (CPE) or salmon (SPE) is applied to override a dopaminergic inhibition of the neuroendocrine system, preventing gonadotropin secretion and gonadal development. The present study used either CPE or SPE to induce vitellogenesis in female European eel and compared impacts on egg quality and offspring developmental competence with emphasis on the maternal-to-zygotic transition (MZT). Females treated with SPE produced significantly higher proportions of floating eggs with fewer cleavage abnormalities and higher embryonic survival. These findings related successful embryogenesis to higher abundance of mRNA transcripts of genes involved in cell adhesion, activation of MZT, and immune response (dcbld1, epcam, oct4, igm) throughout embryonic development. The abundance of mRNA transcripts of cldnd, foxr1, cea, ccna1, ccnb1, ccnb2, zar1, oct4, and npm2 was relatively stable during the first eight hours, followed by a drop during MZT and low levels thereafter, indicating transfer and subsequent clearance of maternal mRNA. mRNA abundance of zar1, epcam, and dicer1 was associated with cleavage abnormalities, while mRNA abundance of zar1, sox2, foxr1, cldnd, phb2, neurod4, and neurog1 (before MZT) was associated with subsequent embryonic survival. In a second pattern, low initial mRNA abundance with an increase during MZT and higher levels persisting thereafter indicating the activation of zygotic transcription. mRNA abundance of ccna1, npm2, oct4, neurod4, and neurog1 during later embryonic development was associated with hatch success. A deviating pattern was observed for dcbld1, which mRNA levels followed the maternal-effect gene pattern but only for embryos from SPE treated females. Together, the differences in offspring production and performance reported in this study show that PE composition impacts egg quality and embryogenesis and in particular, the transition from initial maternal transcripts to zygotic transcription.

Cloning, characterisation and expression profile of kisspeptin1 and the kisspeptin1 receptor in the hypothalamic-pituitary-ovarian axis of Chinese alligator Alligator sinensis during the reproductive cycle.

Kisspeptin1 (Kiss1), a product of the Kiss1 gene, plays an important role in the regulation of reproduction in vertebrates by activating the Kiss1 receptor (Kiss1R) and its coexpression with gonadotrophin-releasing hormone (GnRH) in GnRH neurons. The purpose of this study was to clone the Kiss1 and Kiss1R genes found in the brain of Alligator sinensis and to explore their relationship with reproduction. The full-length cDNA of Kiss1 is 816bp, the open reading frame (ORF) is 417bp and the gene encodes a 138-amino acid precursor protein. The full-length cDNA of Kiss1R is 2348bp, the ORF is 1086bp and the gene encodes a 361-amino acid protein. Quantitative polymerase chain reaction showed that, except for Kiss1R expression in the hypothalamus, the expression of Kiss1 and Kiss1Rduring the reproductive period of A. sinensis was higher than that in the hypothalamus, pituitary gland and ovary during the hibernation period. The changes in GnRH2 mRNA in the hypothalamus were similar to those of GnRH1 and peaked during the reproductive period. This study confirms the existence of Kiss1 and Kiss1R in A. sinensis and the findings strongly suggest that Kiss1 and Kiss1R may participate in the regulation of GnRH secretion in the hypothalamus of alligators during the reproductive period. Furthermore, this is the first report of the full-length cDNA sequences of Kiss1 and Kiss1R in reptiles.

Adaptive response of reproduction to high-altitude hypoxic stress by altering mRNA expression of hypoxia-inducible factors in female yaks (Bos grunniens).

Hypoxia-inducible factors (HIFs) are oxygen-dependent transcriptional activators, but there is little information about their role in yak (Bos grunniens) reproduction. The present study, for the first time, investigated the adaptive mechanism of yak reproduction to high-altitude hypoxic stress by comparing the expression of HIF mRNAs between female yaks at high-altitude and cattle at low-altitude. Hypothalamus, anterior pituitary, oviduct, ovary and uterus tissue samples were collected from five adult female yaks and cattle. mRNA expression was determined by the quantitative real-time polymerase chain reaction. Both HIF-1α and HIF-2α were expressed in all five tissues examined from both species, albeit at different levels. In yaks, the highest mRNA levels of HIF-1α and HIF-2α occurred in the oviduct and anterior pituitary, respectively. Both HIF-1α and HIF-2α mRNA levels were higher in yaks than in cattle (p < 0.01). These data provide evidence that adaptation of reproduction to hypoxic conditions is associated with a greater expression of HIF-1α and HIF-2α in the reproductive axis of female yaks than cattle.

Effect of different treatment agents and post-treatment latency times on spermiation stimulation of northern pike (Esox lucius) under controlled conditions.

The effect of different treatment agents, namely, carp pituitary homogenate (CPH), Ovaprim ([D-Arg6, Pro9NEt]-sGnRH + domperidone) and a dopamine-receptor antagonist (metoclopramide), on the stimulation of northern pike (Esox lucius) spermiation was tested under controlled conditions. To carry out the experiment, males (n = 84) were divided into four groups: control (n = 21); CPH (n = 21); Ovaprim (n = 21); metoclopramide (n = 21). The control group was given 0.9% NaCl but no hormonal treatment. After 24 h, sperm was collected from seven males belonging to control (n = 7), CPH (n = 7), Ovaprim (n = 7) and metoclopramide (n = 7). This procedure was repeated after 48 and 72 h post-treatment. At each time, sperm was collected from seven males from each group only once. After collection, the quantity and quality of sperm were determined. It was confirmed that the treatment agent and latency time (the time between treatment and sperm collection) are two factors affecting the quantity and quality of northern pike sperm collected under controlled conditions. The highest total sperm volume and total sperm production (TSP) were noted in the CPH group compared to the Ovaprim, metoclopramide and control groups. In contrast, the time of sperm collection affected the sperm concentration (SC), TSP and sperm pH. With increasing time, SC and TSP decreased, which indicated the occurrence of sperm hydration being part of the final sperm maturation process. Sperm maturation is in turn a consequence of increases in sperm pH and seminal plasma osmotic pressure between 48 and 72 h post-treatment. Sperm motility and sperm kinetic parameters were affected by treatment agent and the time of sperm collection. This indicates that the sperm's ability to move that is achieved in the optimal environment (in spermatic ducts) is dependent on both factors which determine sperm maturation in northern pike under controlled condition.

Hypothalamic and pituitary transcriptome profiling using RNA-sequencing in high-yielding and low-yielding laying hens.

The reproductive physiology and laying performance of laying hens are regulated by the hypothalamus and pituitary. To understand the mechanism of egg laying regulation, we sequenced and analysed the hypothalamus and pituitary expression profiles in high- and low-yielding laying Chinese Dagu Chickens (CDC) using RNA-seq. More than 46 million clean reads and 24,873 tentative genes were obtained using the Gallus gallus genome as a reference. Transcriptome analysis in hypothalamus and pituitary revealed seven and 39 differentially expressed genes (DEGs) between high- and low-yielding CDC hens, respectively. A total of 24 and 22 DEGs were up-regulated and down-regulated, respectively, and 13 novel genes were identified. Functional annotation and pathway enrichment analysis showed that DEGs in the hypothalamus were mainly enriched in glycosaminoglycan biosynthesis. DEGs significantly enriched in the pituitary primarily affected the extracellular matrix, the protein extracellular matrix, and the extracellular space. Pathways involving phenylalanine metabolism, 2-oxocarboxylic acid metabolism, the glycosphingolipid biosynthesis-ganglion series, and local adhesion were significantly enriched in the pituitary. Eight DEGs, PRDX6, TRIB2, OVCH2, CFD, Peptidase M20, SLC7A10, and two other amino acid transporters, are involved in the metabolism and transport of amino acids. To our knowledge, this is the first study comparing the hypothalamus and pituitary transcriptomes of high- and low-yielding laying hens. Our findings suggest that putative differences in gene expression can provide a base for further research in this field. Moreover, we identified increased expression of genes involved in amino acid metabolism, glycosaminoglycan biosynthesis, and oestrogen negative feedback systems in low-yielding laying hens, highlighting their potential as biomarkers of egg production.

In-depth characterization of the pituitary transcriptome in Simmental and Chinese native cattle.

The pituitary gland is a key endocrine organ responsible for growth and development. To get closer to understanding important molecular mechanisms at work in the bovine pituitary and identifying the core genes controlling growth, in the present study we have analyzed the transcriptome profiling of the pituitary glands of two cattle breeds (Wenshan and Simmental cattle) with extreme growth differences using high-throughput RNA sequencing. Our study revealed that the overall gene expression levels are quite similar between the two breeds. A total of 18,065 annotated genes were identified, which accounted for 85% of the annotated genes in cattle. The five most abundant hormone genes (GH, PRL, POMC, CGA, and LHB) were relatively stable in both breeds, indicating their pivotal roles in maintaining the basic functions of the pituitary. In addition, 105 genes were identified as differentially expressed between Wenshan and Simmental pituitary glands, including 83 known and 22 novel genes. Functional association analyses of the differentially expressed genes between the breeds revealed 60 enriched gene ontology terms and 3 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Interestingly, the most enriched KEGG pathway, cell adhesion molecules, can modulate pituitary hormone secretion by cell-cell contact. Our findings demonstrated that SYTL2, SLC38A4, and NCAM2 are new candidates for crucial functions in the secretory pathways of the pituitary gland. These results will further understanding of the important molecular mechanisms at work in the bovine pituitary.

Ovopel® and carp pituitary extract for induction of reproduction in Colossoma macropomum females.

The aim of this study was to evaluate the efficiency of the hormonal inducers Ovopel® and carp pituitary extract (CPE) for induction of reproduction in Colossoma macropomum females. The treatments were CPE at the dose of 5.5 mg/kg divided into two applications (10%; and 90% after 12 h) and Ovopel® at doses of 0.2 and 0.4 pellet/kg body weight in a single application. Eight replicates were used in each of the three treatments, totaling 24 experimental units. The females spawned when treated with the 0.2 pellet of Ovopel® (100.0%), 0.4 pellet of Ovopel® (62.5%), and CPE (87.5%), but there were no significant differences among the treatment groups in spawning rate. When there was treatment with Ovopel® spawning occurred with greater (P < 0.05) degree-hours (average water temperature × number of hours until spawning; 0.2 pellet: 417.7; 0.4 pellet: 412.3) in relation to the CPE treatment (268.9). The total oocyte weight was similar when there was treatment with Ovopel® (0.2 pellet: 832.3 g; 0.4 pellet: 798.9 g) and CPE (688.3 g). By contrast, the production index was greater (P < 0.05) with the Ovopel® treatments (0.2 pellet: 8.8%; 0.4 pellet: 9.0%) as compared with CPE (6.7%). Fertility and hatching rates were similar among the treatment groups. Ovopel® and CPE are efficient in induction of reproduction in C. macropomum females. Of the two Ovopel® treatments assessed in this study, the dose of 0.2 pellet/kg body weight is sufficient for effective induction of reproductive processes.

Effect of carp pituitary homogenate (CPH) and sGnRHa (Ovaprim) on northern pike (Esox lucius) spermiation stimulation and its effect on quantity and quality of sperm.

The effect of carp pituitary homogenate (CPH, n = 7) at a dose of 2.0 mg kg-1 and (D-Arg6, Pro9NET)-sGnRH + domperidone (Ovaprim, n = 7) at a dose of 0.5 ml kg-1 in northern pike (Esox lucius) sperm maturation under controlled conditions was examined. On the control group, 0.9% NaCl at a dose of 1.0 ml kg-1 (n = 7) was used. Sperm was collected 48 h following injection. Sperm quantity (total sperm volume, total sperm production and sperm concentration), sperm motility using the Computer-assisted sperm analysis (CASA) system and sperm and seminal plasma quality (sperm pH, seminal plasma osmotic pressure and seminal plasma pH) were determined for each male separately in each group. The results of the present study demonstrate that hormonal treatment had a positive effect on sperm maturation in northern pike, regardless of the hormonal preparation used. However, even though no differences were found in total sperm volume and total sperm production between fish injected with either CPH or Ovaprim, it should be highlighted that the highest progressive motile sperm (PRG), straight-linear velocity (VSL) and movement linearity (LIN) was noted in fish treated with Ovaprim. It was also found that it is possible to collect sperm from non-hormonally manipulated fish. However, in such a case, only a small sperm volume (around 0.1 ml) characterised by lowered PRG (below 40%) was noted. Considering the fact that only after Ovaprim application sperm motility with progressive movement of sperm above 50% was observed, the hatchery practice of collection of sperm 48 h after its application (at a dose of 0.5 ml kg-1) may be recommended.

Neuropeptidome of the Hypothalamus and Pituitary Gland of Indicine × Taurine Heifers: Evidence of Differential Neuropeptide Processing in the Pituitary Gland before and after Puberty.

Puberty in cattle is regulated by an endocrine axis, which includes a complex milieu of neuropeptides in the hypothalamus and pituitary gland. The neuropeptidome of hypothalamic-pituitary gland tissue of pre- (PRE) and postpubertal (POST) Bos indicus-influenced heifers was characterized, followed by quantitative analysis of 51 fertility-related neuropeptides in these tissues. Comparison of peptide abundances with gene expression levels allowed assessment of post-transcriptional peptide processing. On the basis of classical cleavage, 124 mature neuropeptides from 35 precursor proteins were detected in hypothalamus and pituitary gland tissues of three PRE and three POST Brangus heifers. An additional 19 peptides (cerebellins, PEN peptides) previously reported as neuropeptides that did not follow classical cleavage were also identified. In the pre-pubertal hypothalamus, a greater diversity of neuropeptides (25.8%) was identified relative to post-pubertal heifers, while in the pituitary gland, 38.6% more neuropeptides were detected in the post-pubertal heifers. Neuro-tissues of PRE and POST heifers revealed abundance differences ( p < 0.05) in peptides from protein precursors involved in packaging and processing (e.g., the granin family and ProSAAS) or neuron stimulation (PENK, CART, POMC, cerebellins). On their own, the transcriptome data of the precursor genes could not predict the neuropeptide profile in the exact same tissues in several cases. This provides further evidence of the importance of differential processing of the neuropeptide precursors in the pituitary before and after puberty.

Production of live larvae following in vitro maturation of zebrafish oocytes.

This study aimed to assess the effects of carp pituitary extract (CPE), follicle stimulating hormone (FSH) and luteinizing hormone (LH) on zebrafish oocyte maturation and the ability of these mature oocytes to be fertilized and developed until hatching. Stage III follicles were matured in eight treatments: five concentrations of CPE (16, 32, 48, 64 and 80 µg/mL), one of FSH (0.5 µg/mL), one of LH (0.5 µg/mL), or one combination of FSH (0.5 µg/mL) and LH (0.5 µg/mL). Maturation rates in CPE treatments were 12.8% (16 µg/mL), 24.8% (32 µg/mL), 27.0% (48 µg/mL), 22.7% (64 µg/mL) and 9.6% (80 µg/mL); in FSH was 15.7% (0.5 µg/mL), in LH was 31.8% (0.5 µg/mL) and in FSH (0.5 µg/mL) combined with LH (0.5 µg/mL) it was 50.4%. In vitro fertilization was performed in all treatments; however, only the treatment combining FSH and LH resulted in fertilized oocytes. After maturation using FSH combined with LH, the cleavage rate was 33.3% and hatching rate of live larvae was 20.0%. These results showed that FSH combined with LH was effective in IVM of zebrafish oocyte.

Sequencing and Identification of Endogenous Neuropeptides with Matrix-Enhanced Secondary Ion Mass Spectrometry Tandem Mass Spectrometry.

Matrix-enhanced secondary ion mass spectrometry (ME-SIMS) has overcome one of the biggest disadvantages of SIMS analysis by providing the ability to detect intact biomolecules at high spatial resolution. By increasing ionization efficiency and minimizing primary ion beam-induced fragmentation of analytes, ME-SIMS has proven useful for detection of numerous biorelevant species, now including peptides. We report here the first demonstration of tandem ME-SIMS for de novo sequencing of endogenous neuropeptides from tissue in situ (i.e., rat pituitary gland). The peptide ions were isolated for tandem MS analysis using a 1 Da mass isolation window, followed by collision-induced dissociation (CID) at 1.5 keV in a collision cell filled with argon gas, for confident identification of the detected peptide. Using this method, neuropeptides up to m/z 2000 were detected and sequenced from the posterior lobe of the rat pituitary gland. These results demonstrate the potential for ME-SIMS tandem MS development in bottom-up proteomics imaging at high-spatial resolution.