Functional categories by Gene Ontology analysis of the sex accessory gland sequences.

<p>(A) 133 sequences out of a total of 560 sequences; not all unigenes could be annotated and some received multiple annotations. (B) Comparison of different percentages in each GO sub-category is represented in all categories.</p

MALDI-TOF MS spectrum of β2 peptide recovered from the oviduct gland.

<p>MALDI-TOF MS spectrum of β2 peptide recovered from the oviduct gland.</p

BLASTX searches and analyses for the collection of <i>Sepia officinalis</i> contigs and singletons.

<p>BLASTX searches and analyses for the collection of <i>Sepia officinalis</i> contigs and singletons.</p

General characteristics of <i>Sepia officinalis</i> accessory sex gland ESTs.

a<p>length of sequences used for comparison after editing (<100-bp et N-rich inserts were excluded).</p>b<p>ESTs with minimum 90% identity over 100-bp region were clustered together, forming a cluster.</p>c<p>that did not sufficiently match any sequence in the data set to allow assembly.</p>d<p>number of ESTs assembled in clusters/total ESTs.</p

Description of sex pheromone precursors.

<p>(A) Amino acid alignments of the 3 precursors. Asterisks (*) indicate strict identity between sequences; colons (:) indicate strong similarity; dots (.) indicate weak similarity; and hyphens (-) represent alignment gaps. Predicted signal sequences are underlined. Cysteines are filled in grey; putative amidations are filled in black; and potential basic residue cleavage sites are boxed. (B) Schematic diagrams showing the organization of <i>Sepia officinalis</i> pheromone precursors. Precursors encode a complex cocktail of peptides and polypeptides resulting from dibasic cleavages. Black box, signal peptide; arrow, predicted site of signal sequence cleavage; vertical black line, potential dibasic residue cleavage site; asterisk, predicted N-linked glycosylation site; S, Cys residue.</p

Tissue-specific patterns of expression for candidate pheromones assayed via reverse transcription PCR.

<p>Expression patterns for SPα, SPα′ and SPβ transcripts and actin were assayed from three females and three males. Gels (A) and (B) correspond to SPα-α′ and SPβ, respectively. M, weight marker; O, ovary; MNG, main nidamental gland; OG, oviduct gland; G, gills; NS, nervous system; T, testis; SG, secondary glands; P, Penis; Ctrl−, negative control (no template added to PCR mix); Ctrl+, positive control (the template was a plasmid containing the insert of interest).</p

Peptides resulting from dibasic cleavages of SP precursors.

<p>Peptides resulting from dibasic cleavages of SP precursors.</p

MALDI-TOF detected ions resulting from truncated α3 and α3′ peptides.

<p>MALDI-TOF detected ions resulting from truncated α3 and α3′ peptides.</p

Presentation_1_Tachykinin-3 Genes and Peptides Characterized in a Basal Teleost, the European Eel: Evolutionary Perspective and Pituitary Role.PDF

<p>In mammals, neurokinin B (NKB) is a short peptide encoded by the gene tac3. It is involved in the brain control of reproduction by stimulating gonadotropin-releasing hormone (GnRH) neurons, mainly via kisspeptin. We investigated tac3 genes and peptides in a basal teleost, the European eel, which shows an atypical blockade of the sexual maturation at a prepubertal stage. Two tac3 paralogous genes (tac3a and tac3b) were identified in the eel genome, each encoding two peptides (NKBa or b and NKB-related peptide NKB-RPa or b). Amino acid sequence of eel NKBa is identical to human NKB, and the three others are novel peptide sequences. The four eel peptides present the characteristic C-terminal tachykinin sequence, as well as a similar alpha helix 3D structure. Tac3 genes were identified in silico in 52 species of vertebrates, and a phylogeny analysis was performed on the predicted TAC3 pre-pro-peptide sequences. A synteny analysis was also done to further assess the evolutionary history of tac3 genes. Duplicated tac3 genes in teleosts likely result from the teleost-specific whole genome duplication (3R). Among teleosts, TAC3b precursor sequences are more divergent than TAC3a, and a loss of tac3b gene would have even occurred in some teleost lineages. NKB-RP peptide, encoded beside NKB by tac3 gene in actinopterygians and basal sarcopterygians, would have been lost in ancestral amniotes. Tissue distribution of eel tac3a and tac3b mRNAs showed major expression of both transcripts in the brain especially in the diencephalon, as analyzed by specific qPCRs. Human NKB has been tested in vitro on primary culture of eel pituitary cells. Human NKB dose-dependently inhibited the expression of lhβ, while having no effect on other glycoprotein hormone subunits (fshβ, tshβ, and gpα) nor on gh. Human NKB also dose-dependently inhibited the expression of GnRH receptor (gnrh-r2). The four eel peptides have been synthesized and also tested in vitro. They all inhibited the expression of both lhβ and of gnrh-r2. This reveals a potential dual inhibitory role of the four peptides encoded by the two tac3 genes in eel reproduction, exerted at the pituitary level on both luteinizing hormone and GnRH receptor.</p

<i>urp1</i> mRNA is restricted to the ventral spinal cord and hindbrain at early stages of development in zebrafish.

<p>Expression of <i>urp1</i> revealed by ISH (BM purple, violet) on nacre embryos at 22 <b>(A)</b>, 28 <b>(B)</b> and 48 hpf <b>(C)</b>. At 22 hpf and 28 hpf, <i>urp1</i>⁺ cells occur only in the spinal cord at the level of the lateral floor plate <b>(A, B)</b>, while from 48 hpf, they are mainly visible in the hindbrain <b>(C)</b>. <b>A1</b>, <b>B1</b>, <b>B5</b> and <b>C1</b>, dorsal views; <b>A2</b>, <b>B2, B3</b> and <b>C2</b>, lateral views with dorsal up; <b>B4</b>, coronal section with dorsal up; all embryos oriented anterior left; <b>B3</b> and <b>B5</b> are details at higher magnifications of B2 and B1, respectively. Ch, chord; Cc, central canal; Nt, neural tube; Scale bar: 100 μm.</p