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ASCB | EMBO San Diego 2018
Annual Meeting of the American Society for Cell Biology and the European Molecular Biology Organization

Dec 08-12, 2018, San Diego, CA, USA
Booth Nr. 1137 (of 89Nort / Chroma Technology)

 

BPS Baltimore 2019
63rd Annual Meeting of the Biophysical Society

Mar 2-6, 2019, Baltimore, Maryland, USA
Booth Nr. 503

 

NWG Göttingen 2019
13th Meeting of the German Neuroscience Society

Mar 20-23, 2019, Göttingen, Germany
Booth Nr. 8

Response of Isolated Olfactory Receptor Neurons from Ano2+/+ and Ano2-/- Mice to Photoreleased Ca2+ or 8-Br-cAMP

(a) Typical current responses to a sudden [Ca2+] increase in isolated olfactory sensory neurons (OSNs) from Ano2+/+ and Ano2-/- mice in whole-cell patch-clamp experiments under symmetrical Cl-. Cells were clamped  to -50 mV, 0 mV and +50 mV. Arrows indicate the flash that releases Ca2+ from DMNP-EDTA. Inset, larger magnification to reveal the currents remaining in Ano2-/- OSNs.

(b) Typical currents elicited by flash release of 8-Br-cAMP (arrows) in isolated OSNs from Ano2+/+ and Ano2-/- mice held at -50 mV. Mean amplitudes were -445 ± 111 pA and -43.2 ± 9.9 pA for Ano2+/+ and Ano2-/- OSNs, respectively (±s.e.m., P = 0.0016).

(c) Peak currents elicited by uncaging Ca2+ as function of holding voltage, averaged from eight Ano2+/+ and seven Ano2-/- OSNs with three or more mice per genotype. Error bars, s.e.m. Significance levels (two sample t-test): *, P < 0.05; **, P < 0.01.

 

Experimental information:

 

Caged compounds: [6,7-Bis(carboxymethoxy)coumarin-4-yl] methyl-8-bromoadenosine-3',5'-cyclic monophosphate (BCMCM)-caged 8-Br-cAMP and DMNP-EDTA caged Ca2+.

 

Equipment:

 

Uncaging of 8-Br-cAMP:  UV flash lamp JML-C2 (Rapp OptoElectronic GmbH) (settings: 200 V at 1 mF ) coupled to a Zeiss Axiovert 200, 100X Fluar objective.

Ca2+ uncaging: SP-20 UV flash lamp (Rapp OptoElektronic GmbH) (settings: 300 V at 3 mF) coupled to an Olympus BX51WI microscope, 60X LUMPlanFl objective.

 

From: Billig G. M. et al., (2011) Ca2+-Activated Cl- Currents Are Dispensable for Olfaction, Nat. Neurosci. 14, 763-769.

 

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