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First published online September 15, 2004
Journal of Experimental Biology 207, 3681-3691 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.01186

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Sequence and expression of the CAPA/CAP2b gene in the tobacco hawkmoth, Manduca sexta

Poh Kheng Loi^* and Nathan J. Tublitz

Institute of Neuroscience, University of Oregon, Eugene, OR 97403, USA

View larger version (40K): [in a new window]   Fig. 1. Complete sequence of the Manduca sexta CAPA (cardioacceleratory peptide 2b; Mas-CAPA-1) gene. Nucleotide and predicted peptide sequences are listed for the coding region, which starts at nucleotide 1. Orange, Mas-CAPA-2 peptide encoding region; purple, Mas-CAPA-1 encoding region; blue, Mas-PK-1 peptide encoding region; green, glycine used for C-terminal amidation; red, dibasic post-translational cleavage sites; STOP, stop codon.

View larger version (17K): [in a new window]   Fig. 2. Summary of all cells in the larval and adult Manduca sexta central nervous system (CNS) labeled with the in situ probe for the CAPA gene. SEG, sub-esophageal ganglion; T1–3, thoracic ganglia 1–3; A1–8, abdominal ganglia 1–8.

View larger version (23K): [in a new window]   Fig. 3. Spatial and temporal expression of CAPA expressing cells in the Manduca sexta CNS. L, larval stages; W, wandering stage of the 5th instar larva; P, pupa; Ph, pharate; SEG, sub-esophageal ganglion; T1–3, thoracic ganglia 1–3; A1–8, abdominal ganglia 1–8.

View larger version (72K): [in a new window]   Fig. 4. CAPA transcript expression in the brain of Manduca sexta. Dorsal (A) and ventral (B) 3rd-instar brain. (C) Anterior view of a different 3rd-instar brain showing the 10 pairs of CAPA-expressing cells. Arrowheads, cluster of five cells near dorsal midline; concave arrowhead, single dorsal cell in the middle of each brain hemisphere; convex arrowhead, single dorsal cell located antero-laterally in each hemisphere; convex arrow, single dorsolateral cell; arrows, cluster of three ventral cells. (D, top) Anterior view of a pharate adult brain showing the CAPA-expressing cell (arrowheads); (D, bottom) anterior view of the pharate adult subesophageal ganglion fused to the brain, showing a pair of CAPA-expressing cells (concave arrowheads). Scale bar, 100 µm.

View larger version (57K): [in a new window]   Fig. 5. CAPA transcript expression in cells in the subesophageal ganglion of a 1st-instar Manduca sexta larva showing the posterioventral (A) and anteriodorsal cells (B,C, arrowheads). Scale bar, 100 µm.

View larger version (80K): [in a new window]   Fig. 6. CAPA transcript expression in cells in the 3rd thoracic ganglion of Manduca sexta. (A) 3rd thoracic ganglion from a 3rd-instar larva. (B) Absence of CAPA expression in the 3rd thoracic ganglion from a slipped-head 4th-instar larva. Scale bars, 100 µm.

View larger version (66K): [in a new window]   Fig. 7. CAPA transcript expression in cells in the abdominal ganglia of Manduca sexta. (A) CAPA gene expression in the 2nd abdominal ganglion (A2) of a day 1 2nd-instar larva. (B) A2 from a different day 1 2nd-instar larva with no labeled cells. (C) CAPA transcript expression in the 4th abdominal ganglion (A4) from a pharate adult. Scale bar, 100 µm.

View larger version (71K): [in a new window]   Fig. 8. CAPA transcript expression in cells in the terminal ganglia of Manduca sexta. (A) CAPA gene expressing cells in the terminal ganglion of day 1 2nd-instar larva (arrowheads). (B) Terminal ganglion of a late 4th-instar larva with no detectable staining. (C) A pharate adult terminal ganglion with two stained cells in the 6th abdominal neuromere (A6) (convex arrowheads). Scale bars, 100 µm.

View larger version (23K): [in a new window]   Fig. 9. Amino acid alignment between the CAPA gene in Manduca sexta and the CAPA gene in Drosophila melanogaster. Alignment derives from the DIALIGN2 protein sequence alignment program. Purple, putative peptide sequence; red, post-translated cleavage sites; blue, C-terminal glycine, presumably converted to amides; *, region of homology.