Levatte M A, Weaver L C, York I A, Johnson D, Dekaban G A
John P. Robarts Research Institute, University of Western Ontario, London, Canada.
Neuroscience. 1995 Jun;66(3):737-50. doi: 10.1016/0306-4522(94)00617-e.
Two recombinant herpes simplex type 1 viruses expressing beta-galactosidase (encoded by the Escherichia coli lacZ gene) inserted into the unique long 41 (encoding virus host shutoff) or unique short 5 (encoding glycoprotein J) open reading frames were generated. Purified recombinants or wild-type herpes simplex type 1 were injected into the left adrenal gland of hamsters. Three days later, virus-infected neurons were detected in spinal cord sections from all infected hamsters. Neurons were visualized with beta-galactosidase histochemistry in spinal cord sections from hamsters infected with either of the recombinants but not with the wild-type virus. Wild-type virus could only be detected with immunocytochemistry. Insertional mutagenesis into the unique long 41 or unique short 5 regions of the herpes simplex genome by lacZ did not disrupt the neurotropic properties of the virus. Both recombinant viruses labelled the central nervous system sympathoadrenal preganglionic neurons as well as brainstem neurons. Because the virus host shutoff recombinant more readily crossed synapses to reach the brainstem compared to the glycoprotein J recombinant, the presence of glycoprotein J may facilitate cell to cell transmission in vivo. Both recombinants may be useful for the study of synaptic organization of neural circuits. Our recombinant viruses were less lytic yet neurovirulent after mutation of either glycoprotein J or virus host shutoff of herpes simplex virus type 1 wild-type. These recombinant viruses express the bacterial beta-galactosidase which is readily detectable using simple histochemistry. Inoculation of the adrenal gland or kidney with these viruses led to clear labelling of spinal cord cells. These viruses may be useful markers of specific neural circuits.
构建了两种重组单纯疱疹病毒1型，它们分别将表达β-半乳糖苷酶（由大肠杆菌lacZ基因编码）的基因插入到独特长41（编码病毒宿主关闭蛋白）或独特短5（编码糖蛋白J）开放阅读框中。将纯化的重组体或野生型单纯疱疹病毒1型注射到仓鼠的左肾上腺中。三天后，在所有受感染仓鼠的脊髓切片中检测到病毒感染的神经元。在用任一重组体感染的仓鼠的脊髓切片中，通过β-半乳糖苷酶组织化学可使神经元可视化，但野生型病毒感染的仓鼠则不行。野生型病毒只能通过免疫细胞化学检测到。通过lacZ插入单纯疱疹病毒基因组的独特长41或独特短5区域进行插入诱变，并未破坏病毒的嗜神经特性。两种重组病毒均标记了中枢神经系统交感肾上腺节前神经元以及脑干神经元。由于与糖蛋白J重组体相比，病毒宿主关闭蛋白重组体更容易穿过突触到达脑干，因此糖蛋白J的存在可能促进体内细胞间的传播。两种重组体都可能有助于研究神经回路的突触组织。在单纯疱疹病毒1型野生型的糖蛋白J或病毒宿主关闭蛋白发生突变后，我们的重组病毒的裂解性降低但仍具有神经毒性。这些重组病毒表达细菌β-半乳糖苷酶，使用简单的组织化学即可轻松检测到。用这些病毒接种肾上腺或肾脏会导致脊髓细胞的清晰标记。这些病毒可能是特定神经回路的有用标记物。
