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 January 18, 2008
Journal of Experimental Biology 211, 382-390 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.013771
This Article
Right arrow Summary Freely available
Right arrow Full Text
Right arrow Full Text (PDF)
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 Google Scholar
Google Scholar
Right arrow Articles by Brugge, V. A. T.
Right arrow Articles by Orchard, I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Brugge, V. A. T.
Right arrow Articles by Orchard, I.

Amino acid sequence and biological activity of a calcitonin-like diuretic hormone (DH31) from Rhodnius prolixus

Victoria A. Te Brugge1,*, David A. Schooley2 and Ian Orchard1

1 Department of Biology, University of Toronto at Mississauga, 3359 Mississauga Road, Mississauga, Ontario, Canada, L5L 1C6
2 Department of Biochemistry, University of Nevada, Reno, NV 89557, USA


Figure 1
View larger version (12K):
[in this window]
[in a new window]

  		Fig. 1. (A) DH31 ELISA standard curve ({blacksquare}) and the dilutions of the R. prolixus CNS (•). Note the log scale for Rhopr/Dippu-DH31. (B) Quantification of DH31-like immunoreactive material in the CNS of 5th instar R. prolixus (male and female). Abbreviations: CC, corpus cardiacum; SOG, suboesophageal ganglion; PRO, prothoracic ganglion; MTGM, mesothoracic ganglionic mass; and ABN, abdominal nerves.

 

Figure 2
View larger version (13K):
[in this window]
[in a new window]

  		Fig. 2. Purification of DH31-like material from 150 R. prolixus CNS through RPLC system A. The active fraction 28 was then run through system B. Aliquots were tested by a DH31 ELISA to determine DH31-like immunoreactive fractions. The elution times of Drome-DH31 and Dippu-DH31 are indicated by the asterisks.

 

Figure 3
View larger version (11K):
[in this window]
[in a new window]

  		Fig. 3. MALDI-TOF (matrix-assisted laser desorption/ionisation time-of-flight) mass spectrometry of DH31-like material from phenyl column (system B) fraction 40. Peak at 2986.6 matches the MH+ determined for Dippu-DH31. M, mass number; Z, atomic number.

 

Figure 4
View larger version (49K):
[in this window]
[in a new window]

  		Fig. 4. Rhodnius prolixus dorsal cuticle with dorsal diaphragm, dorsal vessel and alary muscles. (A) Line diagram of dorsal abdominal segments. Box indicates the approximate location of images B and C. (B) Phalloidin staining of the heart/dorsal vessel (filled arrow) and alary muscles (open arrow). (C) Dippu-DH31-like staining of the dorsal diaphragm and alary muscles (open arrow).

 

Figure 5
View larger version (2K):
[in this window]
[in a new window]

  		Fig. 5. Trace of a single contraction of the heart alary muscles and the dorsal vessel. Deflections in the trace from the impedance converter indicate the contraction of the alary muscles (a), heart (b) and dorsal vessel (c).

 

Figure 6
View larger version (39K):
[in this window]
[in a new window]

  		Fig. 6. Examples of heart bioassay for R. prolixus. The first 5 min of a recording in saline applied at the open arrow and the second 5 min in peptide applied at the filled arrow. (A) Rhopr/Dippu-DH31 tested at 10–10 mol l–1; (B) Rhopr/Dippu-DH31 tested at 10–8 mol l–1; (C) Rhopr/Dippu-DH31 tested at 10–6 mol l–1. Note the increase in frequency and amplitude of heartbeat.

 

Figure 7
View larger version (6K):
[in this window]
[in a new window]

  		Fig. 7. Heart bioassay dose–response curve as percentage increase in the frequency of contractions over saline controls. Concentrations were tested between 10–12 and 10–6 mol l–1 (N=5–9; note the log scale).

 

Figure 8
View larger version (17K):
[in this window]
[in a new window]

  		Fig. 8. Examples of hindgut muscle bioassay for R. prolixus. The first 5 min of a recording in saline applied at the open arrow, and the second 5 min in the peptide applied at the filled arrow. (A) Rhopr/Dippu-DH31 tested at 10–10 mol l–1; (B) Rhopr/Dippu-DH31 tested at 10–8 mol l–1; (C) Rhopr/Dippu-DH31 tested at 10–6 mol l–1. Note the increase in frequency and the variable change in amplitude.

 

Figure 9
View larger version (6K):
[in this window]
[in a new window]

  		Fig. 9. Hindgut dose–response curve as percentage increase in the frequency of contractions over saline controls. Concentrations were tested between 10–12 and 10–5 mol l–1 (N=5–9; note the log scale).

 




© The Company of Biologists Ltd 2008