Figure 2.

Construction of an HSV-1 viral vector to overexpress GH. (A) The full-length gene for presomatotropin was cloned into an HSV-1 amplicon under the control of the HSV α-4 promoter. eGFP was co-expressed via the HSV α-22 promoter. (B) GH protein expression was confirmed in vitro. Vero cells were infected with GH virus at increasing MOIs. As the MOI increased, progressively higher levels of both GH and eGFP were detected. (C) The viral vector was infused into the dorsal hippocampus of rats. A representative infection, showing high levels of expression in pyramidal cells of CA1, and sparse infection in the granule cell layer of the dentate gyrus, is shown. Scale bar = 100 microns. (D) GH protein expression was quantified in infected dorsal hippocampal slices four days following virus delivery. GH levels were expressed as a percentage, relative to the NS control average. Viral overexpression of GH led to an approximate doubling of GH protein. ^*Indicates p < 0.05.