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Project TitleNeurotrophic Factor Function in Neurodegenerative Disease
Principal InvestigatorB. J. Wilcox
LaboratoryLaboratory of Immunology; Division of Therapeutic Proteins; Office of Therapeutics Research and Review
Project SummaryOur laboratory studies the function of neurotrophic factors (NTF) in the brain and their potential for use as therapeutic agents in treatment of human neurodegenerative disease. Animal models of neurodegeneration are being used to study the potential therapeutic value of neurotrophic factors in treatment of neurodegenerative disease. Careful characterization of each animal model including analysis of the mechanisms and time course of neuron death is an important step in evaluating novel therapeutic interventions. Our work is focused on characterization of the neonatal facial nerve axotomy model of motor neuron degeneration. This model is often used in preclinical studies to support clinical trials for testing of therapeutic agents in treatment of adult motor neuron disease. Transection of the rat facial nerve on the day of birth results in apoptosis of neurons in the facial nucleus (FN) within seven days. Our preliminary investigations of the mechanism of cell death in this model revealed numerous profiles in the FN exhibiting morphological characteristics of apoptosis within 5-7 days of axotomy. To confirm biochemically that apoptosis occurs in neonatal FN neurons after axotomy, DNA was extracted from the axotomized and normal FN and analyzed by gel electrophoresis. Results revealed the "ladder" effect characteristic of apoptosis in samples from both lesioned and unlesioned nuclei 6 days after axotomy or sham surgery, respectively. This result suggests that ongoing apoptosis is present in the normal neonatal facial nucleus independent of axotomy. Therefore, we hypothesized that developmental cell death in the FN persists during the first postnatal week, much later than previously reported. To test this hypothesis, we performed a developmental study in rats (ages E18-P15) using unbiased sterology to quantify the total number of motor neurons at each age. Results show a 40% loss in motor neurons in the FN between E20 and P15. These results show that developmental cell death continues through the first 2 postnatal weeks and suggests that the neonatal facial nucleus is not fully developed. Therefore, the neonatal axotomy model may not be appropriate for predicting efficacy of novel therapies for adult motor neuron disease. To understand why adult motor neurons are less vulnerable to injury than neonatal motor neurons, we used in situ hybridization to measure the levels of acidic fibroblast growthfactor (aFGF) mRNA in the FN at selected developmental ages ( E20-adult). Results show that aFGF mRNA is strongly expressed in motor neurons in the adult FN but very little is found before P3. At P15, aFGF mRNA levels approach levels seen in the adult. Therefore, aFGF may function as an endogenous source of trophic support for motor neuron after injury. Due to reduced resources, time spent on this project has been significantly reduced. The work is expected to be completed within FY2002.
Last Updated: 4/1/2002
Date created: September 25, 2003 |
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