FoxJ1-GCaMP8.1 transgenic mice express GCaMP8.1 under control of the FoxJ1 locus promoter/enhancer regions within the BAC transgene. FoxJ1-GCaMP8.1 mice from founder line B35-3 exhibit moderate to high expression levels of GCaMP8.1 expression levels in ciliated epithelial cells in lung airway, trachea, testis and oviduct - consistent with endogenous FoxJ1. Additional details described further below. FOXJ1 is a transcription factor involved in ciliogenesis and is required for motile ciliated cell differentiation. Calcium is a key molecular signal for cell functions including heart, smooth muscle, vessel, airway contraction, lung secretion, autonomic neurotransmission and immunocyte function. GCaMP8.1 is a sensitive GCaMP variant with high dynamic range and fast response kinetics, as well as improved calcium-binding affinity. In the absence of calcium binding, low/baseline EGFP fluorescence is observed. Following calcium binding, bright EGFP fluorescence is expected at several-fold greater levels than baseline (see additional GCaMP8 information below). For these FoxJ1-GCaMP8.1 transgenic mice, the donating investigator reports that GCaMP8.1 expression patterns are consistent with that of endogenous FoxJ1 expression (ciliated epithelial cells in lung airway, trachea, testis and oviduct). Of all the founder lines, line 35-3 also had the highest level of message in the testis (based on qPCR analysis). Specifically, GCaMP8.1 expression in bronchioles of the lung, trachea, testis and oviduct was demonstrated via direct fluorescence and anti-GFP antibody staining. They have not tested expression in other tissues to date (July 2019). While expected to do so, the donating investigator has not yet confirmed if calcium concentration changes are detected by the GCaMP8.1 molecule in these FoxJ1-GCaMP8.1 mice (July 2019). Mice hemizygous for the FoxJ1-GCaMP8.1 transgene are viable and fertile with normal breeding, and no reported gross phenotypic abnormalities. To date (July 2019), it has not been attempted to make this strain homozygous. The genetically encoded calcium indicator GCaMP8.1 is a codon-optimized variant of GCaMP8 having one additional mutation (M144I) in its calmodulin region for improved calcium-binding affinity (lowers the Kd from 235 nM to 188 nM). GCaMP8 is a sensitive GCaMP variant with high dynamic range and fast response kinetics (<a href="http://www.ncbi.nlm.nih.gov/pubmed/23240011">Ohkura et al. 2012 PLoS One 7:e51286</a>). Specifically, the high signal amplitude and low baseline fluorescence of GCaMP8 results in ability to detect single Ca2+ spikes / single neuronal action potentials. Compared to GCaMP3, the GCaMP8 has ~3-fold greater dynamic range, similar response kinetics, ~2-fold more rapid decay kinetics and lower baseline fluorescence. In the absence of calcium binding, dim EGFP fluorescence is expected as there is a pore from the outside of its barrel into the chromophore. Upon calcium binding, this pore becomes occupied and bright EGFP fluorescence is observed at several-fold greater levels than baseline (e.g., ~16-fold greater in neurons ex vivo). In the presence of Ca2+, GCaMP8 has absorbance and emission peaks of ~488 nm and ~510 nm, respectively. This mouse model is available by way of a collaborative effort between Cornell/National Heart Lung Blood Resource for Optogenetic Mouse Signaling (<a href="http://chromus.vet.cornell.edu">CHROMus</a>) and The Jackson Laboratory.

FoxJ1-GCaMP8.1 transgenic mice express GCaMP8.1 in ciliated epithelial cells in lung airway, trachea, testis and oviduct. GCaMP8.1 is a sensitive GCaMP variant with high dynamic range and fast response kinetics, as well as improved calcium-binding affinity. Upon GCaMP8.1 binding to calcium, increased EGFP fluorescence is expected. These FoxJ1-GCaMP8.1 mice may be useful to examine calcium signaling in ciliated epithelial cells both in vivo and in vitro.

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