Journal: Nature Communications

Article Title: GABAergic interneurons form transient layer-specific circuits in early postnatal neocortex

doi: 10.1038/ncomms10584

Figure Lengend Snippet: ( a ) R26::P2X2 allele construct. Grey boxes, Rosa26 homology sequences; yellow, CAG hybrid promoter; blue triangles, loxP sites; red, open-reading frame; white, selection marker. ( b ) Conditional expression of the rat P2X2 receptor (rP2X2R) in Nkx2-1 interneurons. Inset top: an EGFP+ cell (arrow) showing expression of rP2X2R (middle); inset bottom: combined image; scale bar, 15 μm. ( c ) Distribution of EGFP+ cells across the depth of the cortex (green histogram bars) with corresponding percentage of EGFP+/P2X2R+ (white circles) and P2X2R+/EGFP+ (black circles). ( n =631 cells; n =5 brains, P7–8). ( d ) Laser intensity was adjusted to give focal ATP uncaging an effective resolution of 50 μm. Top panel: recording across our standard LSPS grid ( n =153 laser target spots) with only one suprathreshold response (black trace) observed when firing the laser directly at the recorded interneuron soma; blue line, ultraviolet laser. Bottom panel: the dendritic morphology of the same cell superimposed with observed depolarisation colour coded according to the scale bar shown below. ( e ) The average laser power necessary to evoke a consistent, focal presynaptic response over the period of early development studied; Student's t -test P13–15, FS versus NFS P =0.264, t(10)=1.1828). ( f ) Direct suprathreshold responses recorded in cell-attached (top trace) and whole-cell patch-clamp (bottom trace) configuration ( n =5 cells). ( g ) The average time course of ATP-evoked presynaptic APs in response to calibrated ultraviolet laser pulses at early (top graph; n =7 cells) and late (bottom; n =6) ages. ( h ) Top trace: synaptic currents recorded across the extent of the LSPS grid in a P5 PYR neuron in the presence of glutamatergic antagonists CNQX and AP-5 (both 30 μM); bottom trace: GABA A receptor antagonist picrotoxin (50 μM) abolished laser-evoked PSCs. ( i ) Postsynaptic currents in a P10 PYR cell were abolished following incubation with 1 μm TTX (bottom trace).

Article Snippet: The following primary antibodies were used: rabbit anti-SST (1:600, AB5494, Millipore), rat anti-SST (1:400, MAB354, Millipore), mouse anti-PV (1:400, MAB1572, Millipore); rabbit anti-P2X2 receptor (extracellular; 1:400 detects rat and mouse; APR-025, Alomone labs, Israel), rabbit anti-P2X2 receptor (1:400, detects mouse; ab48864, Abcam plc, UK), rabbit anti-Nkx6.2 (1:600, ab58708, Abcam plc), chicken anti-GFP (1:400, ab13970, Abcam plc), rabbit anti-calretinin (1:600, AB5054, Millipore) and mouse anti-calretinin (1:400, MAB1568, Millipore).

Techniques: Construct, Selection, Marker, Expressing, Patch Clamp, Incubation

Journal: Cell and Tissue Research

Article Title: Time-of-day-dependent expression of purinergic receptors in mouse suprachiasmatic nucleus

doi: 10.1007/s00441-017-2634-8

Figure Lengend Snippet: Antibodies used

Article Snippet: Rabbit anti-P2X2 , GeneTex, GTX10266 , 1:500.

Techniques: Plasmid Preparation