Manipulation of α7 nicotinic acetylcholine receptor signaling in primary ovine fetal microglia cultures
Neuroinflammation in utero may result in life-long neurological disabilities. Microglia play a pivotal role, but the mechanisms are poorly understood. No early postnatal treatment strategies exist to enhance neuroprotective potential of microglia. An important step toward that goal is establishing adequate pathophysiological models where microglia function in a large, well gyrated mammalian brain can be studied. Here we present the protocol to manipulate ovine fetal microglia in primary cultures in vitro without or with previous exposure to endotoxin (lipopolyssacharide, LPS) in vivo. We present the approach of blocking or stimulating the α7 nicotinic acetylcholine receptor (α7nAChR) in fetal microglia which allows to enhance or reduce their pro-inflammatory phenotype when incubated with LPS. The measurement of cytokine IL-1beta in supernatant and the RNAseq pipeline are presented.
Figure 1
Posted 01 Feb, 2018
Manipulation of α7 nicotinic acetylcholine receptor signaling in primary ovine fetal microglia cultures
Posted 01 Feb, 2018
Neuroinflammation in utero may result in life-long neurological disabilities. Microglia play a pivotal role, but the mechanisms are poorly understood. No early postnatal treatment strategies exist to enhance neuroprotective potential of microglia. An important step toward that goal is establishing adequate pathophysiological models where microglia function in a large, well gyrated mammalian brain can be studied. Here we present the protocol to manipulate ovine fetal microglia in primary cultures in vitro without or with previous exposure to endotoxin (lipopolyssacharide, LPS) in vivo. We present the approach of blocking or stimulating the α7 nicotinic acetylcholine receptor (α7nAChR) in fetal microglia which allows to enhance or reduce their pro-inflammatory phenotype when incubated with LPS. The measurement of cytokine IL-1beta in supernatant and the RNAseq pipeline are presented.
Figure 1
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