, 2008) We speculate that the conditioned maturation of commissu

, 2008). We speculate that the conditioned maturation of commissural Selleck Doxorubicin axon output synapses might limit the expected detrimental effects of uncrossed commissural axons on the cognitive and sensory abilities of HGPPS patients. Therefore, the conditioned functional maturation of a commissural output synapse uncovered here might contribute to compensatory mechanisms that help HGPPS patients to develop near-normal degrees of sensory and motor functions, despite the fact that major commissural systems are un-crossed. We used the Krox20::Cre mouse line ( Voiculescu et al., 2000; gift of Patrick Charnay,

Paris, France), which drives Cre expression in the lower auditory brainstem ( Han et al., 2011; Maricich et al., 2009), to recombine the floxed Robo3 allele and to suppress axon midline crossing in the calyx of Held projection ( Renier et al., 2010). Heterozygous Krox20Cre/+ mice were crossed with Robo3lox/lox mice ( Renier et al., 2010); the early expression onset of Cre-recombinase under the Krox20 promoter (∼E10; Voiculescu et al., 2000) ensured that the floxed Robo3 allele was recombined early enough to Anti-diabetic Compound Library prevent axon midline crossing. Additional breeding methods, and methods regarding the CAGGS::CreERTM approach are given in Supplemental Experimental Procedures.

Transverse 200 μm thick slices of the brainstem containing the MNTB and LSO nuclei were made according to standard procedures with a LEICA VT1000S vibratome, using Robo3 cKO and control mice of three different age groups: P9–P12, P20–P25, and P90–P110. Protocols of mouse handling and animal experimentation were approved by the Veterinary office of the Canton of Vaud, Switzerland (authorization # 2063). The extracellular recording solutions were bicarbonate-based Ringer solutions containing 2 mM CaCl2 and 1 mM MgCl2 (see Supplemental Experimental Procedures for detailed composition). For the recordings only of fiber stimulation-evoked

EPSCs (Figures 2 and 6), bicuculline (10 μM) and strychnine (2 μM) were added to the extracellular solution. For recordings of IPSCs in LSO principal neurons (Figure 8), NBQX (10 μM) was added. For paired recordings (Figure 5), tetraethylammonium chloride (TEA, 10 mM), tetrodotoxin (TTX, 1 μM), D-AP5 (50 μM), γ-D-glutamylglycine (γ-DGG, 2 mM), cyclothiazide (CTZ, 100 μM), bicuculline (10 μM), and strychnine (2 μM) were added to the extracellular solution. We used four different pipette solutions: (1) a Cs-gluconate based solution with 5 mM EGTA, for recording of MNTB neurons in the voltage clamp mode (Figures 2, 3, 5, 6, and 7); (2) a Cs-gluconate based solution with 0.

Leave a Reply

Your email address will not be published. Required fields are marked *

*

You may use these HTML tags and attributes: <a href="" title=""> <abbr title=""> <acronym title=""> <b> <blockquote cite=""> <cite> <code> <del datetime=""> <em> <i> <q cite=""> <strike> <strong>