July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Donor-host material transfer is photoreceptor-specific and comprises mRNA but not mitochondria
Author Affiliations & Notes
  • Oliver Borsch
    Center for Regenerative Therapies Dresden, Dresden, Germany
    TU Dresden, Dresden, Germany
  • Sílvia Llonch
    Center for Regenerative Therapies Dresden, Dresden, Germany
    TU Dresden, Dresden, Germany
  • Sylvia Gasparini
    Center for Regenerative Therapies Dresden, Dresden, Germany
    TU Dresden, Dresden, Germany
  • Marius Ader
    Center for Regenerative Therapies Dresden, Dresden, Germany
    TU Dresden, Dresden, Germany
  • Footnotes
    Commercial Relationships   Oliver Borsch, None; Sílvia Llonch, None; Sylvia Gasparini, None; Marius Ader, None
  • Footnotes
    Support  DFG Grant FZT 111, CRTD Grant EXC 168 Cluster of Excellence (M.A.), DFG Grant AD375/6-1 (M.A.), BMBF Research Grant 01EK1613A (M.A.), ProRetina Stiftung Grant Pro-Re/Prom-stip/Ader-Llonch.1–2016 (S.L.)
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 2881. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Oliver Borsch, Sílvia Llonch, Sylvia Gasparini, Marius Ader; Donor-host material transfer is photoreceptor-specific and comprises mRNA but not mitochondria. Invest. Ophthalmol. Vis. Sci. 2019;60(9):2881.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : Recent studies demonstrated that subretinally transplanted photoreceptors remain at the injection site and, instead of structurally integrating into the host tissue, mainly exchange cytoplasmic material with endogenous host photoreceptors by an as yet unknown mechanism. In order to better understand this process, we aimed to assess whether material transfer is (I) photoreceptor-specific and (II) which cytoplasmic components are transferred.

Methods : In order to evaluate (I) the material transfer potential of non-photoreceptor cells after transplantation, we isolated different neuronal (retinal and cortical neurons), and non-neuronal (bone marrow) donor cell types from a Cre-driven fluorescent (tdTomato) reporter mouse line (Ai9) and subretinally transplanted cell suspensions into photoreceptor-specific Cre-expressing mice (B2-Cre, n=4/group). (IIa) To analyse whether mRNA is exchanged, male donor photoreceptors isolated from postnatal day (P) 4 Nrl-GFP reporter mice were transplanted into adult wild-type female hosts (n=4). Single cell RT-qPCR of FACS-sorted GFP+ cells was conducted to assess the presence of GFP-mRNA within female host cells (n=54). (IIb) Exchange of larger cytoplasmic components such as organelles, i.e. mitochondria, was investigated by transplanting P4 donor photoreceptors isolated from a double reporter mouse line labelling mitochondria and cell membranes independently (mito-Dendra2/membrane tdTomato) into adult wild-type mice (n=4).

Results : While (I) transplanted photoreceptors in the in vivo Cre/loxP assay led to reporter expression in both donor and host photoreceptors (3307±2381), only marginal fluorescence was visible upon transplantation of other retinal cells (452±217; p≤0.05), cortical neurons (10±9; p≤0.01) or bone marrow cells (0; p≤0.01), indicating that material transfer is largely photoreceptor-specific. (IIa) Single cell RT-qPCR of the GFP+ host (female) photoreceptors revealed that mRNA is exchanged between donor and host cells (25%). (IIb) After transplanting donor photoreceptors with fluorescent cell membranes and mitochondria, host photoreceptors only displayed the donor membrane-reporter (245±60) but no donor mitochondria signal, indicating that mitochondria are not transferred.

Conclusions : Our results indicate that material transfer is photoreceptor-specific and comprises the transfer of mRNA but not mitochondria.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×