Table 1. A summary of the previous literature focusing on adenoviral-mediated gene delivery to motor neurons via intramuscular injections.

Authors	Volume and Concentration of Adenovirus	Species/Strain	Age at Injection	Post-operational Time	Promoter	Gene of Interest	Injection Site	Transduction Site
Ghadge et al.21	5–70 μl of 0.5–1.0 × 1010 pfu/ml,	SJL/J Mice	4–6 weeks old	1–10 days	CAG	LacZ	Tibialis Ant., Tongue	Lumbar ventral horn, Hypoglossal Nucleus
Petrof et al.62	200 μl of 3×1012 pfu/ml	MDX and C57Bl6 mice	5–8 weeks old	1 week	RSV, CMV	Luc, LacZ	2 or 3 separate injections in the hemidiaphragm	Only muscle was analysed
Giménez y Ribotta et al.69	4 μl of 1.2–4 × 107 pfu/ml	SD Rats	Post natal day 1	7 days	RSV	LacZ, BDNF, GDNF	Nasolabial or lower lip muscles	Facial Nuclei
Haase et al.23	100 μl of 1 × 109 pfu/ml	PMN Mice	3–5 days old	Up to 88 days	RSV	Luc, NT-3	Gastrocnemius, Triceps Brachii, Dorsal Trunk muscles	Muscles and L2-L4 Spinal Cord Segments
Baumgartner and Shine22	10 μl of 1.9 × 108 pfu/ml	SD Rats	Newborn	7 days	RSV	β-Gal, BDNF, CNTF, GDNF	Facial muscles of the cheek, lower lip and whisker pad	Facial nucleus
Baumgartner and Shine26	10 μl of 1.9 × 108 pfu/ml 10 μl of 5 × 108 pfu/ml	SD Rats	1 day old	7 days, 21 days or 42 days	RSV	NT-3, CNTF, BDNF, GDNF, β-Gal,	Three different sites in the Gastrocnemius, Flexor Longus Digitorum and Tibialis Anterior muscles	Lumbar ventral horn
Baumgartner and Shine27	15 μl of 3 × 108 pfu/ml	SD Rats	1 day old	At 2 days facial nerve was severed. 4 or 20 weeks	RSV	GDNF, β-Gal,	Nasiolabialis, Frontalis and Auricularis Anterior muscles	Facial nucleus
Haase et al.74	1 × 109 pfu/ml	PMN Mice	3–5 days old	40–88 days	RSV	NT-3	Gastrocnemius, Triceps Brachii, Dorsal Trunk muscles	Lumbar ventral horn
Glatzel et al.24	3 μl of 2.5 × 107 pfu/ml	C57BL/6 Mice	Unknown	7–21 Days	CMV, RSV	LacZ	Tibialis Cranialis	Not observed
Soudais et al.70	3–5 × 1012 pfu	Swiss OF1 Mice	4 days old	24 Days	CMV	GFP	Gastrocnemius	Sacral Dorsolumbar Rachis Ventral Horn
Yamashita et al.45	5–10 μl of 0.5–1 × 1011 pfu/ml	SOD1-G93A and B6SJL Mice	1–35 weeks old	2–5 days, 1, 2 and 4 weeks	CAG	LacZ, Bcl-2, Cre	Middle or right side of the tongue	Hypoglossal Nucleus
Acsadi et al.46	5 μl of 5 × 109 pfu/ml	SOD1-G93A Mice	5–7 days old	2, 3 and 4 months	CMV	GDNF, eGFP, LacZ	Tibialis anterior, Gastrocnemius, Quadriceps and Paraspinal muscles	Lumbar ventral horn
Martinov et al.28	2–8 μl of 1.0 × 1012 pfu/ml	NMRI Mice, Wistar Rats	Adult	6–41 days	CMV	eGFP, Luc	Soleus, extensor digitorum longus	Lumbar ventral horn
Millecamps et al.71	108 pfu/μl	C57BL/6, SOD1-G93A mice	45 or 130 days old	8 days	PGK, RSV	Luc, β-Gal,	4 sites in the tongue, 2 sites in Triceps Brachii and 2 sites in Gastrocnemius	Hypoglossal Nucleus
Yamashita et al.72	5–10 μl of 0.3×1011 pfu/ml	SOD1-G93A and B6SJL mice	10 weeks	25 weeks	CAG	Bcl-2, Cre	Middle or right side of the tongue	Hypoglossal Nucleus
Tsai et al.73	10 μl of 6.0 × 108 pfu/ml,	FVB/NJ mice	12 day old	4–9 days	PGK	eGFP	Gastrocnemius	Lumbar ventral horn
Nakajima et al.31	100 μl of 5.0 × 108 pfu/ml	Wistar Rats	12–14 weeks	3 days-4 weeks	CAG	LacZ	Middle section of the superficial layer of the Sternomastoid muscle	Medulla-C8
Nakajima et al.32	100 μl of 5.0 × 108 pfu/ml	SD Rats	8–10 weeks	3 days-4 weeks	CAG	BDNF, LacZ	Middle section of the superficial layer of the Sternomastoid muscle	Medulla-C8
Uchida et al.33	2.5 μl of 5 × 1010 pfu/ml	Twy mice	16 weeks old	1–4 weeks	CAG	NT-3, LacZ	Middle belly of the superficial layer of the Sternomastoid muscle	Medulla- C7
Nakajima et al.29	100 μl of 5.0 × 109 pfu/ml	SD Rats	8–10 weeks	1–6 weeks	CAG	BDNF, LacZ	Middle belly of the superficial layer of the Sternomastoid muscle	Medulla-C7
Uchida et al.25	25 μl of 1.0 × 108 pfu/ml	Twy mice	18 week old	4 weeks	CAG	BDNF, LacZ	Middle belly of the superficial layer of the Sternomastoid muscle	Medulla- C3

This table highlights the diversity in adenoviral-mediated gene delivery methods utilised in the last 20 years. β-Gal: beta-galactosidase; BDNF: brain-derived neurotrophic factor; CAG: CMV-enhancer chicken β–actin hybrid promoter; CMV: cytomegalovirus immediately early promoter; CNTF: cilliary-derived neurotrophic factor; GDNF: glial-derived neurotrophic factor; Luc: luciferase; LacZ: lactose operon Z; NGF: nerve growth factor; NT-3: neurotrophin-3; RSV: Rous-sarcoma virus long terminal repeat promoter.