spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

First published online February 29, 2008
Journal of Experimental Biology 211, 900-910 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.013953
This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplementary Material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Mapara, S.
Right arrow Articles by Goldberg, J. I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mapara, S.
Right arrow Articles by Goldberg, J. I.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Identification, molecular structure and expression of two cloned serotonin receptors from the pond snail, Helisoma trivolvis

Sabeen Mapara1, Shawn Parries1, Caitlin Quarrington1, Kee-Chan Ahn1, Warren J. Gallin1 and Jeffrey I. Goldberg1,2,*

1 Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada, T6G 2E9
2 Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada, T2N 1N4


Figure 1
View larger version (40K):
[in this window]
[in a new window]

  		Fig. 1. Phylogenetic reconstruction illustrating the placement of H. trivolvis 5-HT receptor proteins as orthologs of 5-HT1 and 5-HT7 receptors in other animals. The tree illustrates the relationship between the two H. trivolvis receptors and other receptors of the 5-HT1, 5-HT5 and 5-HT7 families. This figure represents a subtree of a larger phylogenetic tree generated from 768 biogenic amine receptors, aligned and trimmed to yield a matrix of 214 amino acid characters. For simplicity, clades encompassing a large number of vertebrate receptors are represented diagrammatically as triangles. Each sequence is represented by the name of the species from which it was obtained and a GenPept identification number that is uniquely assigned to each sequence. The scale bar represents an amount of evolutionary change corresponding to an expected 0.1 changes per site.

 

Figure 2
View larger version (58K):
[in this window]
[in a new window]

  		Fig. 2. Alignment of the seven predicted transmembrane helices of the two H. trivolvis receptors (Hel1, Hel2) with representative vertebrate (Homo sapiens; Hum1D, Hum7) and invertebrate (Drosophila melanogaster; Dmel1, Dmel7) 5-HT1 and 5-HT7 receptors. The conserved residues that have been shown to be involved in 5-HT binding to the receptor are marked with asterisks. The arrows along transmembrane (TM) helices 3 to 7 indicate the portion of the sequence that forms the vestibule leading from the extracellular space to the 5-HT binding site.

 

Figure 3
View larger version (87K):
[in this window]
[in a new window]

  		Fig. 3. Localization of 5-HT1Hel and 5-HT7Hel transcripts in whole-mounts of H. trivolvis embryos. In situ hybridization of digoxigenin (DIG)-labeled anti-sense RNA probes was immunohistochemically localized with alkaline phosphatase-conjugated anti-DIG antibodies and reactants that form either a purple (A–C; light microscope) or fluorescent red (D and E; confocal microscope) precipitate. Using anti-sense probes in stage E45 embryos (A, B and D), 5-HT1Hel RNA expression was localized mainly to the ciliated epithelium of the foot (open arrows), anterior head (filled arrows), tentacles (*), ciliated mantle (filled arrowheads) and gut (open arrowheads). Using sense probes to 5-HT1Hel, no signal was observed (C). In stage E30 embryos (E), 5-HT7Hel RNA expression was localized to the ciliated foot epithelium (open arrows), developing gut (open arrowheads), and ciliated mantle (filled arrowhead). Using sense probes to 5-HT7Hel, no signal was observed (not shown). Scale bar, 100 µm.

 

Figure 4
View larger version (78K):
[in this window]
[in a new window]

  		Fig. 4. Immunofluorescence expression of 5-HT1Hel and 5-HT7Hel in the CNS of mature H. trivolvis. 5-HT1Hel-like immunoreactivity (A) and 5-HT7Hel-like immunoreactivity (B–F) was observed in select neurites within neuropile (arrows) and neurite tracts (arrowhead) in all ganglia of the CNS. The expression observed in the cerebral ganglia (A and B), pleural ganglia (C and D) and buccal ganglia (E and F) was representative of the pattern seen in all ganglia. Immunoreactivity was not observed in neuronal somata. Scale bar, 30 µm.

 

Figure 5
View larger version (106K):
[in this window]
[in a new window]

  		Fig. 5. In situ and cultured neurons display different cellular localization of 5-HT1Hel and 5-HT7Hel receptors. In situ, 5-HT1Hel-like immunoreactivity (A) and 5-HT7Hel-like immunoreactivity (B) was restricted to neurites in neuropile (filled arrows) and neurite tracts within connectives and nerves (arrowhead; also see Fig. 4). Cell bodies (open arrows) were always unstained as revealed by overlaying the differential interference contrast pattern on the fluorescence signal. In contrast, after isolation and culture of cerebral neuron C1, 5-HT1Hel-like immunoreactivity occurred primarily in the cell body (C and D). Preabsorption of antibody with Keyhole limpet hemocyanin (D) and replacement of antiserum with pre-immune serum (E) provided positive and negative controls for antibody specificity, respectively. Scale bar, 50 µm.

 

Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?



© The Company of Biologists Ltd 2008