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Journal of Molecular Endocrinology (2008) 40 137-150    DOI: 10.1677/JME-07-0161
© 2008 Society for Endocrinology

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Purification and characterization of an insulin-related peptide in the desert locust, Schistocerca gregaria: immunolocalization, cDNA cloning, transcript profiling and interaction with neuroparsin

Liesbeth Badisco*, Ilse Claeys*, Matthias Van Hiel, Elke Clynen, Jurgen Huybrechts, Tim Vandersmissen, Sofie Van Soest, Luc Vanden Bosch, Gert Simonet and Jozef Vanden Broeck

Department of Animal Physiology and Neurobiology, Zoological Institute K U Leuven, Naamsestraat 59, PO Box 02465, B-3000 Leuven, Belgium

(Correspondence should be addressed to J Vanden Broeck Email: jozef.vandenbroeck{at}bio.kuleuven.be)

* *(L Badisco and I Claeys contributed equally to this work)

Members of the insulin superfamily are not restricted to vertebrates, but have also been identified in invertebrate species. In the current report, we present the characterization of Scg-insulin-related peptide (IRP), an insulin-related peptide in the desert locust, Schistocerca gregaria. This peptide was isolated from corpora cardiaca (CC) extracts by means of a high-performance liquid chromatography (HPLC)-based purification strategy. Subsequent cloning and sequencing of the corresponding cDNA revealed that the encoded Scg-IRP precursor displays the structural organization that is typical for members of the insulin superfamily. Moreover, immunocytochemistry on brain tissue sections demonstrated the presence of Scg-IRP in median neurosecretory cells of the pars intercerebralis and their projections towards the storage part of the CC. Quantitative real-time RT-PCR studies revealed the presence of Scg-IRP transcripts in a variety of tissues, including nervous tissue and fat body. Furthermore, these transcripts showed a tissue- and phase-dependent, temporal regulation during the reproductive cycle of adult males and females. Finally, we demonstrated that Scg-IRP interacts in vitro with a recombinant neuroparsin, a locust protein displaying sequence similarity with vertebrate IGF binding proteins.







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