I continue to evaluate factors affecting the internalization, degradation and transport of lectin-HRP conjugates in the central and peripheral nervous system of rats. Such factors probably also regulate the interneuronal transfer of these probes. The progress in the past year has resulted in the development of methods which permit the immunochemical detection of species of lectin-HRP conjugates, such as wheat germ agglutinin conjugated to HRP (WGHRP), following internalization and transport in the nervous system in rats. These methods have shown that WGHRP is rapidly degraded by neurons as the lectin-HRP conjugates accumulate in neuronal perikarya following retrograde transport. The relative resistance of WGHRP to degradation following internalization at the level of the neuronal perikaryon versus the axon terminals of a neuron may account for the ability of WGHRP to undergo interneuronal transfer following orthograde, but not retrograde, transport to the visual artex. I will determine the forms of WGHRP which are internalized by retinal ganglion cells and compare them with the species of WGHRP which arrive at the axon terminals of reinal ganglion cells following orthograde transport in the two neuronatomical systems used to study this important phenomenon. In the coming year these studies will be extended using immunochemical methods to identify species of WGHRP which reach the axon terminals of neurons. After reaching the axon terminals of retinal ganglion cells, I have shown that WGHRP (and/or its degradative products) is released to undergo another cycle of internalization and orthograde transport by these same neurons. Comparisons will be made with transported native HRP since native HRP does not undergo interneuronal transfer. These studies should enable me to specifically identify the form or forms in which WGHRP exists as it is released from retinal ganglion cell axon terminals for uptake by other neurons in the superior colliculus and lateral geniculate body. These studies will provide insights into the mechanisms which determine or regulate the interneuronal transfer of exogenous proteins in the nervous system.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Academic/Teacher Award (ATA) (K07)
Project #
5K07NS000762-04
Application #
3078115
Study Section
Neurological Disorders Program Project Review B Committee (NSPB)
Project Start
1983-01-01
Project End
1987-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
4
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Christen, B; Trojanowski, J Q; Pietra, G G (1987) Immunohistochemical demonstration of phosphorylated and nonphosphorylated forms of human neurofilament subunits in human pulmonary carcinoids. Hum Pathol 18:997-1001
Kuwayama, Y; Terenghi, G; Polak, J M et al. (1987) A quantitative correlation of substance P-, calcitonin gene-related peptide- and cholecystokinin-like immunoreactivity with retrogradely labeled trigeminal ganglion cells innervating the eye. Brain Res 405:220-6
Trojanowski, J Q (1987) Neurofilament proteins and human nervous system tumors. J Histochem Cytochem 35:999-1003
Trojanowski, J Q; Walkenstein, N; Lee, V M (1986) Expression of neurofilament subunits in neurons of the central and peripheral nervous system: an immunohistochemical study with monoclonal antibodies. J Neurosci 6:650-60
Schmidt, M L; Trojanowski, J Q (1986) Enzymatic detection of native and derivatized horseradish peroxidase in sodium dodecyl sulfate polyacrylamide gels. Anal Biochem 155:371-5
Trojanowski, J Q; Atkinson, B; Lee, V M (1986) An immunocytochemical study of normal and abnormal human cerebrospinal fluid with monoclonal antibodies to glial fibrillary acidic protein. Acta Cytol 30:235-9
Lee, V M; Carden, M J; Trojanowski, J Q (1986) Novel monoclonal antibodies provide evidence for the in situ existence of a nonphosphorylated form of the largest neurofilament subunit. J Neurosci 6:850-8
Trojanowski, J Q; Stone, R A; Lee, V M (1985) Presence of an alpha-melanocyte-stimulating hormone-like epitope in the 150,000 dalton neurofilament subunit from diverse regions of the central nervous system: an immunohistochemical and immunoblot study in guinea pig. J Histochem Cytochem 33:900-4
Leff, E L; Brooks, J S; Trojanowski, J Q (1985) Expression of neurofilament and neuron-specific enolase in small cell tumors of skin using immunohistochemistry. Cancer 56:625-31
Trojanowski, J Q; Lee, V M (1985) Expression of neurofilament antigens by normal and neoplastic human adrenal chromaffin cells. N Engl J Med 313:101-4

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