Subject(s)
Behavior/physiology , Neurosciences/history , History, 20th Century , History, 21st Century , Humans , United StatesABSTRACT
We have inoculated a herpes simplex virus type 1 (HSV-1) vector into a variety of sites in the mouse brain and assayed the regions of latency and expression of a beta-glucuronidase (GUSB) cDNA from the latency-associated transcript promoter. Injection sites used were somatosensory cortex, visual cortex, striatum, dorsal hippocampus, and CSF spaces. Latent vector was detected in regions at a distance from the respective injection sites, consistent with axonal transport of vector. Regions of GUSB activity varied by injection site and included cerebral cortex, striatum, thalamus, hypothalamus, substantia nigra, hippocampus, midbrain, pons, medulla, cerebellum, and spinal cord. After a single injection, GUSB enzymatic activity reached wild-type levels in several brain regions. GUSB was found in some areas without any detectable vector, indicative of axonal transport of GUSB enzyme. GUSB-deficient mice, which have the lysosomal storage disease mucopolysaccharidosis (MPS) VII, have lysosomal storage lesions in cells throughout the brain. Adult MPS VII mice treated by injection of vector into a single site on each side of the brain had correction of storage lesions in a large volume of brain. The potential for long-term, widespread correction of lysosomal storage diseases with HSV-1 vectors is discussed.
Subject(s)
Genetic Therapy , Genetic Vectors/therapeutic use , Glucuronidase/metabolism , Herpesvirus 1, Human/genetics , Lysosomes/enzymology , Mucopolysaccharidosis VII/therapy , Animals , Brain/metabolism , Brain/pathology , Brain/virology , Chlorocebus aethiops , Female , Genetic Vectors/administration & dosage , Glucuronidase/analysis , Mice , Mice, Inbred C3H , Mice, Mutant Strains , Models, Anatomic , Mucopolysaccharidosis VII/genetics , Mucopolysaccharidosis VII/metabolism , Tissue Distribution , Vero CellsABSTRACT
To study the fascicular anatomy of peripheral nerves, three different groups of retrograde axonal tracers were evaluated: fluorophores, horseradish peroxidase conjugated to subunit B of cholera toxin (CT-HRP), and adeno-associated virus (AAV). The hindlimb nerves in rats served as a model to identify the most efficient tracer in regard to labeling axons within peripheral nerves. The rat's tibial and common peroneal nerves were injected with the different tracers and the sciatic nerve was subsequently examined for evidence of labeled axons. The CT-HRP clearly provided the best results in this rat model. Subsequently, CT-HRP was injected into the recurrent laryngeal nerve (RLN) of two horses in order to identify the location and distribution pattern of the RLN axons within the course of the cervical vagus nerve trunk. No labeling could be observed in either of the two horses.
Subject(s)
Axons/drug effects , Cholera Toxin/pharmacology , Fluorescent Dyes/pharmacology , Genetic Vectors/pharmacology , Horseradish Peroxidase/pharmacology , Horses/anatomy & histology , Peripheral Nerves/anatomy & histology , Animals , Dependovirus , Male , Models, Animal , Peripheral Nerves/drug effects , Rats , Rats, Sprague-Dawley , Recurrent Laryngeal Nerve/anatomy & histology , Recurrent Laryngeal Nerve/drug effectsABSTRACT
Previous studies indicated that within the ventromedial hypothalamus (VMH), the estrogen receptor alpha (ERalpha)-containing neurons express substance P (SP), but do not comprise the majority of projection neurons. The present study tested the hypothesis that projection neurons within the VMH express SP receptors (NK1), allowing responsiveness to signals from ERalpha-containing neurons. Pseudorabies virus was transneuronally transported from the lordosis-relevant lumbar epaxial muscles to the VMH, labeling 28% of the NK1-containing neurons in the VMH and surrounding area. Thus, SP may influence sexual behavior through its release from the ERalpha-containing neurons, perhaps synaptically affecting NK1 receptor-labeled lordosis-relevant projection neurons within the VMH.
