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Optogenetic Analysis of Area‐Specific Glial‐Neuronal Signalling
Author(s) -
Tang Feige,
Figueiredo Melina F.,
Gourine Alex V,
Teschemacher Anja G,
Kasparov Sergey
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.1064.19
Subject(s) - cnqx , depolarization , optogenetics , neuroscience , antagonist , chemistry , locus coeruleus , medicine , ampa receptor , receptor , endocrinology , nmda receptor , biology , microbiology and biotechnology , nucleus
We studied glia‐neuronal signalling in two brainstem areas, the retrotrapezoid nucleus (RTN) and the locus coeruleus (LC). RTN and LC neurons were visualized using adenoviral expression of DsRed2 under control of PRSx8 promoter. For optogenetic selective stimulation of astrocytes we have generated an adenoviral vector AVV‐sGFAP‐ChR2(H134R)‐Katushka1.3. We first patched AVV‐sGFAP‐ChR2(H134R)‐Katushka1.3‐transduced astrocytes and found that flashing 470 nm light induced precisely timed depolarisations (+18.8 ± 2.0 mV, n=7, p<0.001). We then patched RTN and LC neurons and stimulated adjacent astrocytes using light. This evoked long lasting depolarization of both RTN (+3.57 ± 0.5 mV, n=9, p<0.001) and LC neurons (+ 3.08 ± 0.33 mV, n=14, p<0.001). Responses of the RTN neurons were markedly reduced by the ATP receptor antagonist MRS2179 (10 μm), depolarisation +0.7mV, n=7, p<0.001). In contrast, neither MRS 2179 nor TNP‐ATP (10 μm) had any effect on of the responses in LC neurons (n=6, p=0.71 for MRS2179 and n=9, p=0.86 for TNP‐ATP). At the same time depolarisations in LC neurons were prevented by an NMDA receptor antagonist dAPV (50 μm, n=8, p<0.01) and AMPA receptor antagonist CNQX (10 μm, n=5, p<0.001). Therefore, glia‐neuronal information transfer occurs in area‐ and transmitter‐dependent manner.