Translucence Biosystems is honored to present at the 7th Annual BRAIN Initiative Investigators Meeting. Our presentation is entitled, "Measurement of Npas 4 and cFos for Brain-Wide Snapshots of Neuronal Activity". Access our presentation poster here.
Recent advances in optical clearing and light sheet imaging have opened an exciting new avenue for brain-wide, cellular resolution immunostaining at the forefront of a dimensional shift from 2D to 3D histology. Translucence Biosystems has developed the Mesoscale Imaging SystemTM which adapts the ZEISS Lightsheet Z.1 microscope, designed for imaging small live organisms, to image large intact tissues with unprecedented speed and resolution. Traditional immediate-early gene (IEG) products such as cFos are turned on by neuronal activity, but also by cAMP and various paracrine factors. Npas4 is unique among IEGs in that it is neuron-specific and exquisitely tuned to Ca2+ signaling.
We developed a recombinant rabbit monoclonal anti-NPAS4 antibody (Activity Signaling: www.npas4.com) that we are using to generate cellular-resolution brain-wide signatures of neuronal activity. Using a modified version of iDISCO, we are optically clearing and immunostaining intact brains to mark neurons expressing cFos and Npas4. Our machine-learning enabled software, 3TK, surveys 3D- rendered whole brains to identify individual stained cells, map them to the Allen Brain Reference Atlas and quantify their expression across 100’s regions.
Our data show that cFos is expressed in a large number of neurons in control brains, whereas Npas4 expression at baseline is much lower. Behavioral manipulations that increase neuronal activity drive the expression of both cFos and Npas4, but the lower baseline expression of Npas4 provides for a higher fidelity readout of neuronal activity.
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