Colocalization of 5HTR_B- and PTTH-ir in the adult BR-SOG of

<p><b><i>A. pernyi</i></b>. <i>Bm</i>PTTH-ir/<i>Ap</i>PTTH-ir in the DL region of protocerebrum of female adult less than one day old was co-localized with <i>Ap</i>5HTR_B-ir (red filled circles). EH-ir in the adult brain of <i>A. pernyi</i> and its colocalization with <i>Ap</i>5HTR_B-ir (blue filled circles) and unique distribution (open circles) of EH-ir in other regions of the brain. (A) The location of detected cells. Lower-case letters correspond to the regions shown in the photographs (e.g., b to B). (B) Two pairs <i>Bm</i>EH-ir cells in the DC region. (C, G) 5HTR_B-ir in the DL region. (D) <i>Bm</i>PTTH-ir in the DL region. (E) Merged image of <i>Bm</i>PTTH-ir and 5HTR_B-ir in the DL region. (F) <i>Ap</i>PTTH-ir in the DL region. (H) Merged image of <i>Ap</i>PTTH-ir and 5HTR_B-ir in the DL region. (I) A 5HTR_B-ir cell in the SOG. (J) One <i>Bm</i>EH-ir cell in the SOG. (K) Merged image of <i>Bm</i>EH-ir and 5HTR_B-ir in the SOG region. Scale bar = 100 µm.</p

Colocalization of 5HTR_A- and PTTH-ir in the early pupal BR-SOG of

<p><b><i>A. pernyi</i></b>. <i>Bm</i>PTTH-ir/<i>Ap</i>PTTH-ir was co-localized with <i>Ap</i>5HTR_A-ir in the BR-SOG of 5-day-old pupa. (A) The location of detected cells. Lower-case letters correspond to the regions shown in the photographs (e.g., b to B). (B) Two large PTTH-ir neurons in the DL region. (C, D, H) Two large 5HTR_A-ir neurons in the DL region. (E) <i>Bm</i>PTTH-ir in the DL region. (F) Merged image of <i>Bm</i>PTTH- and 5HTR_A-ir in the DL region. (G) <i>Ap</i>PTTH-ir in the DL region. (I) Merged image of <i>Ap</i>PTTH-ir and 5HTR_A-ir in the DL region. Scale bar = 100 µm.</p

Pharmacological confirmation of RNAi effect targeting at 5HTR_B.

<p>Effect of injections of 5HT, 5,7-DHT and luzindole on photoperiodism. (A) Diapause pupae were injected either with 5 µl water and 5 µl DMSO (Mock injection) or 5 pmoles Luzindole plus 5 pmoles 5HT in the same volume of solvent and placed under LD: 16:8 at 25°C. Cumulatively 5% adults emerged in 40 days after injection. Cont.: untreated. M: injection with distilled water and DMSO. Luzindole +5HT: luzindole and 5HT co-injected. (B) Diapause pupae were injected with 5,7-DHT at three doses and thereafter the pupae were kept under LD 12:12 at 25°C. Cont.: untreated. M: mock injection with 10 µl distilled water. 5,7-DHT: injected with 5,7-DHT dissolved in the same volume as in the mock. 5 percentage points, 30 percentage points and 100 percentage points adult emergence was observed in 40 days. Asterisks indicate significant difference from control by Kaplan-Meier. ** <i>p</i><0.01.</p

Schematic illustration of 5HTRs role on diapause induction/maintenance in pupal diapause of <i>A. pernyi</i>.

<p>The moth has two 5HTR subtypes, 5HTR_A and 5HTR_B.The former subtype shows no transcription rhythm [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079381#B24" target="_blank">24</a>] and did not respond to photoperiodic activation by long day, while the latter showed rhythmic expression and responded to photoperiodic activation. Therefore, it may be regulated by circadian system. Via one type of arylalkylamine N-acetyltransferase, aaNAT. This engyme is encoded by <i>nat</i> is that a circadian-controlled gene (ccg) since dsRNA^CYC, and dsRNA^CLK suppressed <i>nat</i> transcription and dsRNA^NAT dysfunctioned photoperiodism. Transcription of 5HTR_A was not rhythmic. MT-binding closes the endocrine switch to PTTH release that finally terminates diapause, while 5HTR_B cuts this circuit to enforce or initiate diapause. Diapause of <i>A.penyi</i> is therefore under binary regulation and circadian system regulates at least two points in this system, <i>nat</i> transcription and 5HTR_B expression. aaNAT, arylalkylamine N-acetyltransferase. PER, Period protein, a negative regulator of transcription translation feedback. CYC/CLK, heterodimeric circadian transcription regulator. Mel, melatonin. MT, melatonin receptor. LD, long day. SD, short day. PTTH, prothoracicotropic hormone. PTG, prothoracic gland. E, ecdysone. 20E, 20 hydroxyecdysone.</p

Colocalization of 5HTR_A- and PTTH-ir in the adult BR-SOG of

<p><b><i>A. pernyi</i></b>. <i>Bm</i>PTTH-ir/<i>Ap</i>PTTH-ir in the BR-SOG of female adult with 24hrs after emergence was co-localized with <i>Ap</i>5HTR_A-ir. (A) The locations of detected cells. Lower-case letters correspond to the regions (capital letters) shown in the photographs (e.g., b to B). (B) Two large PTTH-ir neurons in the DL region. (C, D, H) Two large 5HTR_A-ir neurons in the DL region. (E) <i>Bm</i>PTTH-ir in the DL region. (F) Merged image of <i>Bm</i>PTTH- and 5HTR_A-ir in the DL region. (G) <i>Ap</i>PTTH-ir in the DL region. (I) Merged image of <i>Ap</i>PTTH and 5HTR_A in the DL region. Scale bar = 100 µm.</p

Serotonin Receptor B May Lock the Gate of PTTH Release/Synthesis in the Chinese Silk Moth, <i>Antheraea pernyi</i>; A Diapause Initiation/Maintenance Mechanism?

<div><p>The release of prothoracicotropic hormone, PTTH, or its blockade is the major endocrine switch regulating the developmental channel either to metamorphosis or to pupal diapause in the Chinese silk moth, <i>Antheraea pernyi</i>. We have cloned cDNAs encoding two types of serotonin receptors (5HTR_{A and B}). 5HTR_A-, and 5HTR_B-like immunohistochemical reactivities (-ir) were colocalized with PTTH-ir in two pairs of neurosecretory cells at the dorsolateral region of the protocerebrum (DL). Therefore, the causal involvement of these receptors was suspected in PTTH release/synthesis. The level of mRNA^5HTRB responded to 10 cycles of long-day activation, falling to 40% of the original level before activation, while that of 5HTR_A was not affected by long-day activation. Under LD 16:8 and 12:12, the injection of dsRNA^5HTRB resulted in early diapause termination, whereas that of dsRNA^5HTRA did not affect the rate of diapause termination. The injection of dsRNA^5HTRB induced PTTH accumulation, indicating that 5HTR_B binding suppresses PTTH synthesis also. This conclusion was supported pharmacologically; the injection of luzindole, a melatonin receptor antagonist, plus 5th inhibited photoperiodic activation under LD 16:8, while that of 5,7-DHT, induced emergence in a dose dependent fashion under LD 12:12. The results suggest that 5HTR_B may lock the PTTH release/synthesis, maintaining diapause. This could also work as diapause induction mechanism.</p> </div

Relative mRNA levels of 5HTRs upon long-day activation.

<p>Diapause pupae were exposed to LD 16:8 for 0, 5 and 10 cycles at 25°C and mRNA level of 5HTR_A (gray bar) and 5HTR_B (white bar) in the BR-SOG was determined by real time PCR. The results are presented as the mean ± S.E.M. from three independent experiments. Asterisks indicate significant difference from 0-day incubation by one-way ANOVA (Fisher's, LSD). <i>p<</i>0.05.</p

Colocalization of 5HTR_B- and PTTH-ir in the erly pupal BR-SOG of

<p><b><i>A. pernyi</i></b>. <i>Bm</i>PTTH-ir/<i>Ap</i>PTTH-ir in the 5-day-old pupal brain of <i>A. pernyi</i> and its colocalization with <i>Ap</i>5HTR_B-ir (red filled circles). <i>Bm</i>EH-ir in the 5-day-old pupa brain and its colocalization with <i>Ap</i>5HTR_B-ir (blue filled circles) and unique distribution (open circles) of <i>Bm</i>EH-ir in other regions of the brain. (A) The loaction of detected cells. Lower-case letters correspond to the regions shown in the photographs (e.g., b to B). (B) One <i>Bm</i>EH-ir neuron in the PI. (C) <i>Bm</i>EH-ir in the DC region. (D, H) 5HTR_B-ir in the DL region. (E) <i>Bm</i>PTTH-ir in the DL region. (F) Merged image of <i>Bm</i>PTTH-ir and 5HTR_B-ir in the DL region. (G) <i>Ap</i>PTTH-ir in the DL region. (I) Merged image of <i>Ap</i>PTTH-ir and 5HTR_B-ir in the DL region. (J) 5HTR_B-ir in the DC region. (K) One EH-ir neuron in the DC. (L) Merged image of <i>Bm</i>EH-ir and 5HTR_B-ir in the DC region. Scale bar = 100 µm.</p

Green tea polyphenols alter lipid metabolism in the livers of broiler chickens through increased phosphorylation of AMP-activated protein kinase

<div><p>Our previous results showed that green tea polyphenols (GTPs) significantly altered the expression of lipid-metabolizing genes in the liver of chickens. However, the underlying mechanism was not elucidated. In this study, we further characterized how GTPs influence AMP-activated protein kinase (AMPK) in the regulation of hepatic fat metabolism. Thirty-six male chickens were fed GTPs at a daily dose of 0, 80 or 160 mg/kg of body weight for 4 weeks. The results demonstrated that oral administration of GTPs significantly reduced hepatic lipid content and abdominal fat mass, enhanced the phosphorylation levels of AMPKα and ACACA, and altered the mRNA levels and enzymatic activities of lipid-metabolizing enzymes in the liver. These results suggested that the activation of AMPK is a potential mechanism by which GTPs regulate hepatic lipid metabolism in such a way that lipid synthesis is reduced and fat oxidation is stimulated.</p></div

Effects of green tea polyphenols on feed intake, body weight, liver-to-body weight ratio, and abdominal fat mass in broiler chickens.

<p>Effects of green tea polyphenols on feed intake, body weight, liver-to-body weight ratio, and abdominal fat mass in broiler chickens.</p