Figure 5. Rostral rhombic lip neurons project to paramotor nuclei and the preBötC.

(A) Intersectional strategy to label only those neurons with a history of Atoh1 (FlpO) and En1 (Cre) expression. Representative whole-brain image of P21 Atoh1^FlpO;En1^Cre;Ai65/+ mouse on the right. (B) Cell bodies in the PBC are tdTomato⁺. (C) tdTomato⁺ puncta overlap with the synapse-marker synapsin. (D) Schematic of the respiratory circuitry. Solid red PBC is the only respiratory nucleus expressing tdTomato. Dotted nuclei are the nuclei in which tdTomato⁺ projections were observed. TdTomato⁺ puncta near cell bodies were observed in the (E) ITR, (F) RTN, and (G) preBötC. No tdTomato⁺puncta were observed in the (H) NTS, or in respiratory motor nuclei (I) mV, (J) mVII, (K), NA, (L), XII. Abbreviations: PBC, parabrachial complex; ITR, intertrigeminal region; RTN, retrotrapezoid nucleus; preBötC, preBötzinger Complex; NTS, nucleus tractus solitarius; mV, trigeminal motor nucleus; mVII, facial motor nucleus; NA, nucleus ambiguus; XII, hypoglossal motor nucleus.

Figure 5—figure supplement 1. Generation of Atoh1^FlpO mice.

(A) A FlpO-lox2722-PGK-Neo-lox2722 cassette was cloned in between the 5’ and 3’ homologues arm of the Atoh1 locus in a pBlueScript II KS+ vector. Through homologous recombination this sequence was targeted to the endogenous Atoh1 locus. Sox2^Cre was used to excise the PGK-Neo cassette from the target sequence. (B) Representative PCR genotyping reaction from wild-type and Atoh1^FlpO/+ mice.