Table 2.

Recommended parameters for H129-G4 tracer

Animal	Brain region	Coordinates (mm)a			H129-G4 tracer		Perfusion time (dpi)c
		AP	ML	DV	Dose (pfu)	Vol. (μL)b
C57BL/6	Retina [16]	NA	NA	NA	5.0 × 106	1.00	4
C57BL/6	OB [16]	+ 4.28	+ 0.50	− 2.50	2.0 × 106	0.40	3
C57BL/6	M1 [16]	+ 1.54	− 1.70	− 1.75	1.0 × 106	0.20	3
C57BL/6	AD	− 0.82	− 0.75	− 2.75	4.0 × 105	0.08	3
C57BL/6	CeA [23]	− 1.00	− 2.64	− 4.30	3.0 × 105	0.06	1.5
C57BL/6	αBLA [22]	− 1.34	+ 3.40	− 4.80	5.0 × 105	0.10	1–2
C57BL/6	VPM [16]	− 1.82	+ 1.40	− 3.62	1.0 × 106	0.20	3
C57BL/6	CA1 [34]	− 2.00	+ 1.50	− 1.60	2.9 × 105	0.12	1.5
C57BL/6	LGN [16]	− 2.30	+ 2.13	− 2.75	5.0 × 105	0.10	3
C57BL/6	pBLA [22]	− 2.30	+ 3.40	− 4.85	5.0 × 105	0.10	1–2
C57BL/6	AuD	− 2.80	− 4.13	− 1.68	2.0 × 105	0.04	3
C57BL/6	V2L	− 2.80	− 3.15	− 1.25	2.0 × 105	0.04	3
C57BL/6	V1 [16]	− 3.52	+ 2.30	− 1.15	5.0 × 105	0.10	3
C57BL/6	IC	− 5.20	+ 1.40	− 2.00	2.0 × 105	0.04	3
SD rat	M1 [24]	+ 3.00	− 2.60	− 2.20	5.0 × 105 / 2.5 × 106	0.10 / 0.50	3
Tree shrew	M1 [16]	+ 1.85	− 2.60	− 2.10	1.5 × 106	0.30	6

^aCoordinates according to the mouse brain in stereotaxic coordinates [31].

^bInjection volume should not exceed 0.5 μL for brain regions.

^cExtending the waiting time before perfusion normally results in higher labeling intensity, wider tracer spread, and more orders of anterograde transmission. However, the incidence of animal deaths is also increased due to H129-G4 toxicity [16].