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. 2019 Jul 3;103(1):102–117.e5. doi: 10.1016/j.neuron.2019.04.022

Figure 1.

Figure 1

Physiological and Neurochemical Characteristics of Dorsal Horn GRP and GRPR Neurons

(A) Transverse section of the lumbar dorsal horn of a Grp::eGFP mouse immunostained against eGFP, Pax2, and Lmx1b. Arrowheads indicate Lmx1b-positive Grp-eGFP neurons. Scale bars, 100 μm (overview) and 20 μm (high-magnification images). Bar chart: percentage of Grp-eGFP neurons positive for Lmx1b or Pax2 (n = 9 sections from 3 mice).

(B) In situ hybridization for vGluT2 and vGAT mRNA on dorsal horn sections of Grp::eGFP mice (merged image, DAPI in blue). Filled arrowheads, co-expression of vGluT2 with Grp-eGFP and Grp mRNA; open arrowheads, lack of vGAT expression in Grp-eGFP neurons. Scale bar, 20 μm. Bar chart: percentage of Grp-eGFP neurons positive for vGluT2 (43 out of 43 cells from 2 mice) and vGAT mRNA (0 out of 49 cells from 2 mice).

(C) Left: experimental setup used for targeted recordings from dorsal horn neurons identified by eGFP fluorescence, and superimposition of a bright field and an epifluorescence image showing two Grp-eGFP neurons and a recording pipette. Scale bar, 10 μm. Right: voltage traces recorded from a Grp-eGFP neuron during somatic current injection. Bar chart: incidence of different firing patterns (n = 31 neurons from 12 animals). Ib, initial burst; T, tonic; D, delayed; G, gap; P, phasic firing.

(D) Same as (A) but GRPR neurons (n = 15 sections from 5 mice). Open and filled arrowheads indicate Lmx1b-positive and Pax2-positive Grpr-eGFP neurons, respectively.

(E) Same as (B) but Grpr-eGFP neurons. Bar chart: incidence of Grpr-eGFP neurons positive for vGluT2 (29 out of 36 cells from 4 mice) and vGAT mRNA (5 out of 27 cells from 4 mice). Filled and open arrowheads indicate Grpr-eGFP neurons positive for vGluT2 or vGAT, respectively.

(F) Same as (C) but GRPR neurons (n = 91 cells from 61 mice).