Changes in corticotropin releasing factor system transcript levels in relation to feeding condition in Acipenser dabryanus.

CRF system, structural conservation, has an association with feeding regulation in mammals. However, mammals and fish have different physiological mechanisms, the potential role of CRF system for feeding regulation in teleost fish are most unknown. To better explore possible feeding mechanisms of CRF system in Acipenser dabryanus, the gene expression patterns of CRF system have been investigated after different energy status. CRF and two receptors have been studied in Acipenser dabryanus in previous study, thus, four components of CRF system (UI, UCN2, UCN3 and CRF-BP) have been studied in this study. Results showed post-prandial increased UCNs mRNA expressions, and 10 days fasting decreased UCNs mRNA expressions, and the mRNA abundance of CRF-BP has no significant differences. Above, this study confirmed the CRF system has potential role for feeding regulation in Acipenser dabryanus.

The suppression effects of feeding and mechanisms in CRF system of animals.

CRF system is comprised of 4 homologous lineages, 2 main receptors (CRF-R1 and CRF-R2), and a binding protein CRF-BP. The homologous lineages are corticotropin-releasing factor (CRF), urotensin I (UI)/sauvagine (SVG)/urocortin 1 (UCN1), urocortin 2 (UCN2), and urocortin 3 (UCN3), and UI, SVG, UCN1 are orthologous genes. CRF system genes are widely distributed in the brain and gastrointestinal tract, which may relate to feeding regulation. According the research progress about CRF system on mammals and non-mammals, this paper summarized the discovery, structure, tissue distribution, appetite regulation and mechanism of CRF system in animals, which can provide the reference for further research and production of feeding regulation and growth in mammals and fish species.

Progress in understanding the roles of Urocortin3 (UCN3) in the control of appetite from studies using animal models.

Urocortin3 (UCN3), the newest member of corticotrophin releasing hormone (CRH) family polypeptides, is an anorexic factor discovered in 2001, which has a strong inhibitory effect on animal appetite regulation. UCN3 is widely distributed in various tissues of animals and has many biological functions. Based on the research progress of UCN3 on mammals and non-mammals, this paper summarized the discovery, tissue distribution, appetite regulation and mechanism of UCN3 in animals, in order to provide a reference for feeding regulation and growth in mammals and fish in further research and production.

The transcripts of CRF and CRF receptors under fasting stress in Dabry's sturgeon (Acipenser dabryanus Dumeril).

Dabry's sturgeon (Acipenser dabryanus Dumeril, 1868) belongs to Sturgeon and is distributed throughout the mainstream of the upper Yangtze River. While there is little research onphysiological mechanism of Dabry's sturgeon, such as feeding regulation by the CRF system. At present, CRF is thought to regulate feeding via CRF receptors (CRF-Rs) in several mammals, but relatively few studies of CRF and feeding exist in teleosts. Herein, the transcripts of CRF and CRF-Rs under fasting stress in Dabry's sturgeon (Acipenser dabryanus Dumeril) have been explored. A full length Dabry's sturgeon CRF cDNA of 953 bp was identified, which contained a 447 bp open reading frame (ORF). A partial CRF-R1 cDNA of 1053 bp and CRF-R2 cDNA of 906 bp corresponding to the coding sequences (CDS) was obtained. In addition, analysis of the tissue distribution of CRF and CRF-Rs mRNAs revealed they were widely distributed in the central and peripheral nervous systems. Furthermore, periprandial (preprandial and postprandial), fasting, and re-feeding experiments revealed CRF mRNA was significantly increased 1 h and 3 h after feeding and CRF and CRF-Rs transcripts were significantly decreased after 10 days fasting, and significantly increased on re-feeding on day 10. These results suggest that CRF and CRF-Rs might regulate feeding by acting as satiety factors.

UCN3 suppresses food intake in coordination with CCK and the CCK2R in Siberian sturgeon (Acipenser baerii).

Urocortin-3 (UCN3) as a brain-gut peptide inhibits food intake of animal, but the underlying mechanism is not clear. To explore the appetite mechanism about the action of UCN3 in fish, intraperitoneal injection of UCN3 with CCK8, Lorglumide (CCK1R antagonist) or LY225910 (CCK2R antagonist) were conducted. Siberian sturgeon administrated with UCN3 and CCK8 showed a drastic reduction in food intake. The anorectic effect of UCN3 was significantly blocked by LY225910, but not affected by Lorglumide. Furthermore, LY225910 could effectively reverse appetite factor mRNA expressions, including cck, pyy, cart, npy, ucn3, apelin and nucb2 in the whole brain, stomach and intestinum valvula, but Lorglumide could only partially reverse these effects, suggesting the anorectic effect of UCN3 may be primarily mediated CCK2R in Siberian sturgeon. This study indicates for the first time in fish that UCN3 may inhibit food intake in coordination with CCK and CCK2R.

Intraperitoneal injection of nesfatin-1 primarily through the CCK-CCK1R signal pathway affects expression of appetite factors to inhibit the food intake of Siberian sturgeon (Acipenser baerii).

Nesfatin-1 is an 82-amino acid protein derived from nucleobindin 2 (NUCB2), which could inhibit food intake in fish and mammals. However, the neuroendocrine mechanism of nesfatin-1 in animal appetite regulation is unclear. To explore the feeding mechanism of nesfatin-1 in Siberian sturgeon (Acipenser baerii), intraperitoneal injections of nesfatin-1 and sulfated cholecystokinin octapeptide (CCK8), Lorglumide (CCK1R selective antagonist), or LY 225,910 (CCK2R selective antagonist) were performed. Co-injection of nesfatin-1 and CCK8 synergistically significantly decreased the food intake in 1 h. Lorglumide reversed the anorectic effect of nesfatin-1, but LY 225,910 had no effect. Moreover, Lorglumide could also reverse the expressions of appetite factors including nucb2, cck, unc3, cart, apelin, pyy, and npy induced by nesfatin-1 in the brain, stomach, and liver, while LY 225,910 partially reversed these changes. These results indicate that nesfatin-1 inhibits the appetite of Siberian sturgeon mainly through the CCK-CCK1R signaling pathway.

The inhibitory effect of NUCB2/nesfatin-1 on appetite regulation of Siberian sturgeon (Acipenser baerii Brandt).

Since NUCB2 was discovered, the information about NUCB2/nesfatin-1 in appetite regulation in both mammals and teleost has been still limited. The present study aims to determine the effects of nesfatin-1 on food intake and to explore the appetite mechanism in Siberian sturgeon. In this study, nucb2 cDNA sequence of 1571 bp was obtained, and the mRNA expression of nucb2 was abundant in brain and liver. Levels of nucb2 were appreciably increased in brain after feeding 1 and 3 h, while significantly decreased within fasting 15 days. Except for fasting 1 day, the expression pattern of nucb2 in the liver was similar to the brain. Acute intraperitoneal (i.p.) injection of nesfatin-1 inhibited the food intake during 0-1 h in a dose-dependent manner and 50 or 100 ng/g BW nesfatin-1 significantly decreased the cumulative food intake during 3 h. The daily food intake and cumulative food intake were remarkably reduced post chronic (7 days) i.p. injection. Moreover, chronic i.p. injection of nesfatin-1 affected the expression of appetite factors including cart, apelin and pyy in the brain, stomach and liver with the consistent pattern of change, while the levels of cck, ucn3 and nucb2 in these have different patterns. This study demonstrates that nesfatin-1 acts as a satiety factor in reducing the short-term and long-term food intake of Siberian sturgeon. Therefore, the data suggesting nesfatin-1 inhibits the appetite through different signal pathways in the central and peripheral endocrine systems of Siberian sturgeon.

One evidence of cocaine- and amphetamine-regulated transcript (CART) has the bidirectional effects on appetite in Siberian sturgeon (Acipenser baerii).

Cocaine- and amphetamine-regulated transcript (CART), discovered in 1995, with various biological functions, has received much attention recently due to its role in the regulation of appetite in mammals. However, the function of CART on the appetite control in fish species is still not very clear. In this study, Siberian sturgeon (Acipenser baerii Brandt) cart gene was cloned for the first time, and the cart mRNA levels in 11 feeding-related tissues was investigated. The Siberian sturgeon cart gene sequence was 1459 base pairs (bp), including a 3'-terminal untranslated region (3'-UTR) of 39 bp, a 5'-terminal untranslated region (5'-UTR) of 52 bp, and an open reading frame (ORF) of 348 bp encoding 115 amino acids. Siberian sturgeon cart gene has three exons and two introns including 341 bp intron 1 and 679 bp intron 2. The result of tissue distribution showed that cart was widely distributed in 11 tissues with the highest expression in the whole brain. The effects of periprandial (pre- and post-feeding), fasting, and re-feeding on cart mRNA abundance in the whole brain were assessed. Periprandial result showed the expression of cart mRNA in the whole brain significantly elevated after feeding for 3 h. However, fasting experiment showed that the level of cart significantly decreased after 1 day of fasting, but that significantly increased after 3-17 days of food deprivation and returned to the basic level after 3 days of re-feeding in the fishes which were fasted for 15 days. In conclusion, this study suggests that CART has the bidirectional effects on appetite, which acts as a satiety factor in short-term feeding regulation but as a starvation factor in long-term appetite regulation in Siberian sturgeon.

CCK reduces the food intake mainly through CCK1R in Siberian sturgeon (Acipenser baerii Brandt).

To explore the effect of CCK on food intake in Siberian sturgeon, cck cDNA sequence of 1005 bp was obtained, and cck mRNA possessed the highest expression in brain. The expressions of cck were significantly increased after feeding 1 and 3 h, while displaying significant decrease after fasting within 15 days in brain and duodenum. Re-feeding for 3 days induced cck level returned to basic level. Acute i.p. injection experiment showed 100 and 200 ng/g BW CCK8 inhibited the food intake in 0-1 h together with the cumulative food intake within 3 h. 7 days chronic i.p. injection of 100 and 200 ng/g BW CCK8, both daily food intake and cumulative food intake were significantly decreased. In addition, chronic i.p injection of CCK8 induced the expression of feeding related factors changes including cck, ucn3, cart, apelin, pyy and npy in respective organization. Moreover, as revealed by the results, Lorglumide, the CCK1R selective antagonist, effectively reversed the inhibitory effects of CCK8 on food intake and the levels of feeding related factors. On the other hand, LY 225910, the CCK2R selective antagonist, partially reversed these effects. These results indicate CCK is a satiety factor inhibits the feeding of Siberian sturgeon primarily through CCK1R.

The evidence of apelin has the bidirectional effects on feeding regulation in Siberian sturgeon (Acipenser baerii).

Apelin is a peptide, mainly produced in the brain, which participates in several physiologic effects. However, knowledge about the mechanism of appetite regulation in teleosts, including the role of apelin is not well understood. The aim of this study is to explore the effect of feeding status on the expression of apelin mRNA in the whole brain and the effects of injection of apelin on food intake in Siberian sturgeon (Acipenser baerii). In this study, we first cloned the apelin cDNA sequence of the Siberian sturgeon. We obtained a 1046-bp cDNA fragment, including a 237-bp open reading frame (ORF) that encoded 78 amino acids. Apelin was widely distributed in 11 tissues related to feeding regulation, with the highest expression in thewhole brain, followed by the spleen and trunk kidney. In addition, we measured the effects of periprandial (preprandial and postprandial) change, fasting and re-feeding on apelin mRNA expression in whole brain. The level of apelin mRNA was significantly decreased 1h after feeding. The results of the fasting experiment showed that the expression of apelin mRNA in the brain was significantly reduced after 1day of fasting but consistently increased throughout the 15-day food deprivation period. When the 15-day fasted fish were re-fed, apelin mRNA expression in the brain was significantly increased as compared to that of the control. These results suggest that apelin may play a bidirectional role in the regulation of food intake in the Siberian sturgeon. In order to further examine the effect of apelin on feeding regulation in Siberian sturgeons, acute and chronic intraperitoneal (i.p.) injection experiments were performed and food intakes were recorded. Results showed that acute i.p. injection of apelin-13 reduced food intake, however, chronic i.p. injection apelin-13 increased the food intake for 7days in Siberian sturgeons. In conclusion, our results show that apelin has a bidirectional effect on feeding regulation in Siberian sturgeons by acting as a satiety factor in short-term feeding regulation and a starvation factor in long-term feeding regulation.

Intraperitoneal injection urocortin-3 reduces the food intake of Siberian sturgeon (Acipenser baerii).

Urocortin-3 (UCN3), one of the corticotropin releasing factor (CRF) family peptides, which was discovered in 2001, has a variety of biological functions. However, the researches of UCN3 in fish were scarce. In order to understand whether UCN3 play a role in regulating food intake in fish, we first cloned the ucn3 cDNAs sequence of Siberian sturgeon (Acipenser baerii Brandt), and investigated the ucn3 mRNA levels in 11 tissues. The Siberian sturgeon ucn3 cDNA sequence was 1044bp, including an open reading frame (ORF) of 447bp that encoded 148 amino acids with a mature peptide of 40 amino acids, a 5'-terminal untranslated region (5'-UTR) of 162bp and a 3'-terminal untranslated region (3'-UTR) of 435bp. The result of tissue distribution showed that ucn3 widely distributed in 11 tissues with highest expression in brain. We also assessed the effects of periprandial (pre- and post-feeding), fasting and re-feeding on ucn3 mRNAs abundance in brain. The results showed the expression of ucn3 mRNA in brain was significantly elevated after feeding, decreased after fasting 17 days and increased after re-feeding. To further investigate the food intake role of UCN3 in Siberian sturgeon, we performed intraperitoneal (i.p.) injection of Siberian sturgeon UCN3 (SsUCN3) with three doses (60, 120 or 240ng/g) and recorded the food intake. Acute and chronic i.p. injection SsUCN3 reduced the food intake in a dose-dependent pattern. In conclusion, this study indicates that SsUCN3 acts as a satiety factor to inhibit the food intake of Siberian sturgeon.

Bioequivalence and comparison of pharmacokinetic properties of 4-mg tablet formulations of rosiglitazone hydrochloride and rosiglitazone maleate: a single-dose, randomized, open-label, two-period crossover study in healthy adult male Chinese volunteers.

BACKGROUND: Rosiglitazone is an insulin-sensitizing oral thiazolidinedione used for treating patients with type 2 diabetes mellitus. There are 9 oral generic and branded formulations of rosiglitazone available in the People's Republic of China (PRC); however, a literature search did not identify any published data concerning the bioavailability of these formulations in the Chinese population. OBJECTIVES: The aims of this study were to compare the pharmacokinetic properties and determine the bioequivalence of 2 formulations of rosiglitazone 4-mg tablets-rosiglitazone hydrochloride (test) and rosiglitazone maleate (reference)-in healthy adult male Chinese volunteers. METHODS: This single-dose, randomized, open-label, 2-period crossover study was conducted at Zhejiang Provincial People's Hospital, Hangzhou, PRC. Healthy adult male Chinese volunteers were randomly assigned to receive a single 4-mg dose of the test or reference formulation, followed by a 7-day washout period and administration of the alternate formulation. The study drugs were administered after a >or=12-hour (overnight) fast. Plasma was analyzed for rosiglitazone concentration using a validated high-performance liquid chromatography method. For analysis of pharmacokinetic properties, including C(max), AUC(0-t), and AUC(0-infinity), blood samples were drawn before (0 hour; baseline) and at 10, 20, 30, 45, 60, 75, and 90 minutes and 2, 4, 6, 8, 12, and 24 hours after administration. The formulations were to be considered bioequivalent if the logarithm-normal (ln)-transformed ratios of C(max) and AUC were within the predetermined range of 80% to 125%, as established by the US Food and Drug Administration. Tolerability was assessed using physical examination, including monitoring of vital signs, laboratory testing (hematology, blood biochemistry, hepatic function, renal function, and urinalysis), and questioning subjects about adverse events (AEs). RESULTS: Twenty subjects were enrolled and completed the study (mean [SD] age, 21.1 [1.4] years; weight, 62.6 [4.6] kg; height, 171 [5] cm; and body mass index, 21.4 [1.3] kg/m(2)). No period or sequence effects were detected. The 90% CIs for the ln-transformed ratios of C(max), AUC(0-24), and AUC(0-infinity) were 96.79 to 109.73, 92.14 to 102.93, and 91.64 to 102.60, respectively (all, P < 0.001), meeting the predetermined criterion for bioequivalence. No AEs were reported in this study. CONCLUSIONS: In this small study in these healthy adult male Chinese volunteers, a single 4-mg dose of rosiglitazone hydrochloride appeared to be bioequivalent to rosiglitazone maleate, according to the regulatory definition, based on the rate and extent of absorption. Both formulations were well tolerated.