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Differential expression of ITP and ITPL indicate multiple functions in the silkworm Bombyx mori

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Abstract

Ion transport peptide (ITP) and a longer ITP-like (ITPL) are alternatively spliced insect neuropeptides involved in the regulation of development and water homeostasis. Using in situ hybridisation and immunohistochemistry, we determined site- and stage-specific expression of each peptide in Bombyx mori. Each peptide was differentially expressed, except for the prominent overlapping expression of both peptides in six pairs of the brain neurosecretory cells Ia2. After metamorphosis, ITP appeared in the male-specific neurons of the abdominal neuromere 9 (MAN9) that innervate the reproductive organs. ITPL was detected in a pair of dorsolateral interneurons (IN-DL) in each thoracic and abdominal ganglion, and in the thoracic neurosecretory cells (NS-VTL2) which terminate in the vicinity of the prothoracic gland. Feeding larvae showed ITPL expression in the abdominal neurosecretory cells M5. ITPL was also expressed in the peripheral L1 neurons that project axons into the thoracic and abdominal transverse nerves. Our results suggest that ITP and ITPL exhibit different sex- and stage-specific functions that may include regulation of reproduction and steroid production. For future functional studies, we identified an upstream regulatory region controlling ITP/ITPL expression in the brain and L1 neurons, and prepared stable transgenic line pITP-Gal4.2 using the piggyBac system.

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Acknowledgements

We are grateful to C.J.P. Grimmelikhuijzen, Y-J. Kim, A. Mizoguchi and J.A. Veenstra for providing valuable antisera.

Funding

This study was supported by the Slovak grant agency, Agentúra na podporu výskumu a vývoja, (APVV-16–0395, APVV-18–0201, APVV-21–0431).

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  1. Institute of Zoology, Slovak Academy of Sciences, Bratislava, Slovakia

    Vanda Klöcklerová, Zuzana Gáliková, Ladislav Roller & Dušan Žitňan

  2. Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia

    Vanda Klöcklerová

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  1. Vanda Klöcklerová
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441_2023_3752_MOESM1_ESM.tiff

Supplementary file1 Fig. S1 Detection of ITP/ITPL transcripts using ISH probe followed by IH staining with ITP antibody in Ia2 cells of the larval brain. Note very strong ISH labelling in cell bodies that suppressed IH staining (arrows). ITP-IR in axonal projections from these cells are clearly visible (arrowheads) (TIFF 2957 KB)

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Klöcklerová, V., Gáliková, Z., Roller, L. et al. Differential expression of ITP and ITPL indicate multiple functions in the silkworm Bombyx mori. Cell Tissue Res 392, 715–731 (2023). https://doi.org/10.1007/s00441-023-03752-y

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