Abstract
Neurological disorders in ruminants have an important impact on veterinary health, but very few host-specific in vitro models have been established to study diseases affecting the nervous system. Here we describe a primary neuronal dorsal root ganglia (DRG) culture derived from calves after being conventionally slaughtered for food consumption. The study focuses on the in vitro characterization of bovine DRG cell populations by immunofluorescence analysis. The effects of various growth factors on neuron viability, neurite outgrowth and arborisation were evaluated by morphological analysis. Bovine DRG neurons are able to survive for more than 4 weeks in culture. GF supplementation is not required for neuronal survival and neurite outgrowth. However, exogenously added growth factors promote neurite outgrowth. DRG cultures from regularly slaughtered calves represent a promising and sustainable host specific model for the investigation of pain and neurological diseases in bovines.
Highlights
Dorsal root ganglia (DRG), or spinal ganglia are isolated nodular thickenings of the dorsal spinal nerve roots, which contain somas of afferent sensory neurons carrying nociceptive, mechanoceptive and thermoceptive signals from the periphery to the central nervous system (CNS)
Neurite outgrowth and arborisation were evaluated. These growth factors were supplemented either as a single formula in increasing concentrations (NGF: 10, 50, 150 ng/ml; GDNF: 10, 50, 150 ng/ ml; IGF: 5, 20, 50 ng/ml) or as combined formulas (NGF 50 ng/ml combined with GDNF 50 ng/ml: COMBO2; NGF 50ng/ml combined with GDNF 50 ng/ml and IGF20 ng/ml: COMBO3)
We describe a method for the establishment of a primary postnatal bovine DRG neuron culture from slaughtered calves, which can be maintained for extended periods of time
Summary
Dorsal root ganglia (DRG), or spinal ganglia are isolated nodular thickenings of the dorsal spinal nerve roots, which contain somas of afferent sensory neurons carrying nociceptive, mechanoceptive and thermoceptive signals from the periphery to the central nervous system (CNS). These neurons are of pseudo-unipolar type, and their somas are tightly insulated by satellite cells [1,2,3]. PLOS ONE | DOI:10.1371/journal.pone.0168228 December 9, 2016
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