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. 2020 Jul 23;14:45. doi: 10.3389/fncir.2020.00045

Figure 1.

Figure 1

Cre-dependent identification of CS-Parv neurons in the mouse auditory cortex. (A) Schematic depicting injection site using the Parv-Cre-tdTomato transgenic mouse line to identify CS-Parv neurons in the auditory cortex. Bottom, striatum: green AAV1-Flex-GFP injection site; yellow Parv somata were coexpressing GFP and tdTomato; red Parv-tdTomato-positive interneurons. Top, auditory cortex: yellow CS-Parv somata coexpressing GFP and tdTomato; red Parv-tdTomato-positive “interneurons.” (B) Epifluorescence images of Parv tdTomato-positive neurons. Left, top: bright-field image of a slice containing the striatum injection site of AAV1-Flex-GFP in the Parv-Cre.tdTomato transgenic mouse line. Left, middle: tdTomato-expressing Parv neurons in the Parv-Cre-tdTomato transgenic mouse line. Left, bottom: GFP-positive Parv neurons in the dorsal striatum containing the viral injection of AAV1-Flex-GFP in the Parv-Cre-tdTomato transgenic mouse line. The dashed line indicates the striatum boundaries containing the striatum injection site of AAV1-Flex-GFP. Right, top: higher magnification of the bright-field image of a slice containing the striatum injection site of AAV1-Flex-GFP in the Parv-Cre.tdTomato transgenic mouse line. Right, middle: higher magnification image of tdTomato-expressing Parv neurons in the Parv-Cre-tdTomato transgenic mouse line. Right, bottom: higher magnification image of GFP-positive Parv neurons in dorsal striatum containing the viral injection of AAV1.GFP.Flex in the Parv-Cre-tdTomato transgenic mouse line. (C) Left: overlay image of GFP-positive Parv neurons in the auditory cortex identified by viral injection of AAV1.GFP.Flex and Parv neurons in the Parv-Cre-tdTomato transgenic mouse line. The dashed box and the arrows indicate the location of the somata of CS-Parv neurons. Top, right: tdTomato-expressing Parv neurons in the Parv-Cre-tdTomato transgenic mouse line. Middle, right: GFP-positive CS-Parv neurons in the auditory cortex retrogradely identified by viral injection of AAV1.GFP.Flex in the dorsal striatum of the Parv-Cre-tdTomato transgenic mouse line. Bottom right: overlay of GFP and tdTomato images. The arrow indicates the location of the CS-Parv neurons. (D) Schematic depicting the injection site of red RetroBeads using the Parv-Cre/eYFP transgenic mouse line to identify CS-Parv neurons in the auditory cortex. Bottom, striatum: red RetroBeads injection site; Top: auditory cortex: red beads retro-labeled CS-Parv neurons identified by YFP expression (green). (E) Left: overlay image of red-positive neurons in the auditory cortex identified by injection of retrograde beads in the dorsal striatum and Parv eYFP neurons in the Parv-Cre/eYFP transgenic mouse line. Middle: high magnification epifluorescence images of Parv red-beads-positive neurons. Middle, top: eYFP-positive Parv neurons in the auditory cortex in the Parv-Cre-tdTomato transgenic mouse line. Middle, center: CS-Parv neurons identified by anatomical retrograde labeling in the Parv-Cre-eYFP transgenic mouse line. Middle, bottom: overlay of eYFP and retrograde beads labeled CS-Parv neurons. The arrows indicate the location of the red beads in the CS-Parv neurons. Top, right: CS-Parv neurons immunostained with anti-Parv. Middle, right: CS-Parv neurons identified by anatomical retrograde labeling in the Parv-Cre-eYFP transgenic mouse line. Bottom, right: overlay of CS-Parv neurons immunostained with anti-parv and retrograde beads labeled CS-Parv neurons. The arrows indicate the location of the red beads in the CS-Parv neuron.