Analysis of the conserved neurotrophic factor MANF in the Drosophila adult brain

https://doi.org/10.1016/j.gep.2015.04.002Get rights and content

Highlights

  • DmMANF expression is much wider in the adult than in the embryonic and larval CNS.

  • DmMANF is expressed in glia and neurons including dopaminergic neurons.

  • DmMANF overexpression or knockdown results in no loss of dopaminergic neuron somas.

Abstract

Mesencephalic astrocyte-derived neurotrophic factor (MANF) is an evolutionarily conserved neurotrophic factor that supports and protects dopaminergic neurons. The Drosophila MANF (DmMANF) null mutant animals die during early development, and DmMANF is required for the maintenance of dopamine positive neurites. The aim of this study was to investigate the role of DmMANF during later developmental stages. Here we report that DmMANF expression in the adult brain is much wider than in the embryonic and larval stages. It is expressed in both glia and neurons including dopaminergic neurons. Clonal analysis showed that DmMANF is not required cell-autonomously for the differentiation of either glia or dopaminergic neurons. In addition, DmMANF overexpression resulted in no apparent abnormal dopaminergic phenotype while DmMANF silencing in glia resulted in prolonged larval stage.

Section snippets

DmMANF is expressed in glial processes

It has been shown that during embryonic stages, DmMANF expression is confined to some glial subpopulations, such as glia surrounding dopaminergic neurons, in longitudinal and in channel glia (Palgi et al., 2009). Here we used immunohistochemistry to identify the cell populations where DmMANF is expressed. We found that in the adult brain, DmMANF is expressed in all glial subtypes, as categorized by Awasaki et al. (2008) and Doherty et al. (2009) (Table 1), thus having a considerably broader

Fly strains

Flies were kept and grown under standard conditions. w1118 flies were regarded as wild type. For the statistical analysis (Fig. 5, Fig. 6), Canton-S flies were used as wild type. We used the following stocks from Bloomington Drosophila Stock Centre (BDSC): UAS-nGFP (4775), UAS-mCD8::GFP (5137), hsFLP (8862), Kr/CyO; D/Tb Sb (7199), FRT82B Gal80 (5135) and UAS-DmMANFRNAi [from Vienna Drosophila RNAi Centre (VDRC) v12835]. The following stocks are the glial subtype drivers obtained from the

Acknowledgements

Most of the antibodies were provided by the Development Studies Hybridoma Bank, and the fly strains were from the Bloomington Drosophila Stock Centre, the Vienna Drosophila RNAi Centre and the Drosophila Genomics Resource Centre. The authors would like to thank Iris Salecker for providing the lama-Gal4 fly stock and Arja Ikävalko for her assistance with crosses and stock maintenance. This work was supported in parts by the Academy of Finland, Marie Curie Early Stage Training, the Finnish

References (34)

  • C. Chotard et al.

    Glial cell development and function in the Drosophila visual system

    Neuron Glia Biol

    (2007)
  • G. Dietzl et al.

    A genome-wide transgenic RNAi library for conditional gene inactivation in Drosophila

    Nature

    (2007)
  • J. Doherty et al.

    Ensheathing glia function as phagocytes in the adult Drosophila brain

    J. Neurosci

    (2009)
  • K. Fischbach et al.

    Optic Lobe Development. Brain Development in Drosophila Melanogaster

    (2008)
  • F. Friggi-Grelin et al.

    Targeted gene expression in Drosophila dopaminergic cells using regulatory sequences from tyrosine hydroxylase

    J. Neurobiol

    (2003)
  • B. Iyengar et al.

    Silencing synaptic communication between random interneurons during Drosophila larval locomotion

    Genes Brain Behav

    (2011)
  • P. Lindholm et al.

    Novel CDNF/MANF family of neurotrophic factors

    Dev. Neurobiol

    (2010)
  • Cited by (17)

    • MANF regulates splenic macrophage differentiation in mice

      2019, Immunology Letters
      Citation Excerpt :

      Besides protection against neuron apoptosis, MANF is greatly beneficial for animal behavioral and brain functional recovery, as well as differentiation and migration of neural progenitor cells to play a role of neural regeneration [22,23]. Some researches on Drosophila brain have shown that MANF silencing in glial cells prolongs growth and development of Drosophila and induces a new cell type of MANF immunoreactive Cells (MiCs) [35,36]. Moreover, there are a number of researches demonstrating the role of MANF in inflammatory reaction of multiple organs.

    • Unconventional neurotrophic factors CDNF and MANF: Structure, physiological functions and therapeutic potential

      2017, Neurobiology of Disease
      Citation Excerpt :

      Interestingly DmManf is expressed in astrocyte-like glia that surround dopamine neurons but not in neurons (Palgi et al., 2009). However, in adult Drosophila brain, DmManf was expressed in glia but also shown to partly co-localize with pan-neuronal marker and dopaminergic marker in cell somas, suggesting that DmManf was expressed in some dopaminergic cells (Stratoulias and Heino, 2015). Outside the nervous system, during larval stages, DmMANF was predominantly expressed in garland cells, salivary glands, Malpighian tubules, fat body, in the gut and in the ring gland which is the major endocrine organ of Drosophila.

    • UPR Responsive Genes Manf and Xbp1 in Stroke

      2022, Frontiers in Cellular Neuroscience
    View all citing articles on Scopus
    View full text