Stiftung Tierärztliche Hochschule Hannover (TiHo)

Cryopreservation of canine primary dorsal root ganglion neurons and Its impact upon susceptibility to paramyxovirus infection

GND
1036124835
Affiliation
Department of Pathology, University of Veterinary Medicine, 30559 Hannover, Germany. sarah.schwarz@tiho-hannover.de.
Schwarz, Sarah L.;
ORCID
0000-0001-9935-3700
Affiliation
Department of Pathology, University of Veterinary Medicine, 30559 Hannover, Germany. ingo.spitzbarth@tiho-hannover.de.
Spitzbarth, Ingo;
GND
142929565
ORCID
0000-0001-8151-5644
Affiliation
Department of Pathology, University of Veterinary Medicine, 30559 Hannover, Germany. Wolfgang.Baumgaertner@tiho-hannover.de.
Baumgärtner, Wolfgang;
GND
1023386887
ORCID
0000-0002-6970-4327
Affiliation
Department of Pathology, University of Veterinary Medicine, 30559 Hannover, Germany. annika.lehmbecker@tiho-hannover.de.
Lehmbecker, Annika

Canine dorsal root ganglion (DRG) neurons, isolated post mortem from adult dogs, could provide a promising tool to study neuropathogenesis of neurotropic virus infections with a non-rodent host spectrum. However, access to canine DRG is limited due to lack of donor tissue and the cryopreservation of DRG neurons would greatly facilitate experiments. The present study aimed (i) to establish canine DRG neurons as an in vitro model for canine distemper virus (CDV) infection; and (ii) to determine whether DRG neurons are cryopreservable and remain infectable with CDV. Neurons were characterized morphologically and phenotypically by light microscopy, immunofluorescence, and functionally, by studying their neurite outgrowth and infectability with CDV. Cryopreserved canine DRG neurons remained in culture for at least 12 days. Furthermore, both non-cryopreserved and cryopreserved DRG neurons were susceptible to infection with two different strains of CDV, albeit only one of the two strains (CDV R252) provided sufficient absolute numbers of infected neurons. However, cryopreserved DRG neurons showed reduced cell yield, neurite outgrowth, neurite branching, and soma size and reduced susceptibility to CDV infection. In conclusion, canine primary DRG neurons represent a suitable tool for investigations upon the pathogenesis of neuronal CDV infection. Moreover, despite certain limitations, cryopreserved canine DRG neurons generally provide a useful and practicable alternative to address questions regarding virus tropism and neuropathogenesis.

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