Recovery of cholinergic phenotype in the injured rat neostriatum: roles for endogenous and exogenous nerve growth factor.
Polyclonal antibodies against recombinant human nerve growth factor (rhNGF) potently inhibited PC12 neurite outgrowth, blocked high-affinity 125I-rhNGF binding but not its receptor, and cross-reacted with rat, mouse, and human nerve growth factor (NGF) but not with brain-derived neurotrophic factor, neurotrophin-3, ciliary neurotrophic factor, insulin-like growth factor, epidermal growth factor, or activin A. Immunocytochemistry revealed many NGF-positive neurons in the rat neostriatum. The NGF-positive neurons disappeared by 3 days after mechanical injury to the neostriatum and were replaced by intensely NGF- and glial fibrillary acidic protein-positive astrocytes. Enzyme-linked immunosorbent assay measurements revealed that the NGF content of the injured striatum was elevated by eightfold 3 days postinjury and by twofold 2 weeks later. The high-affinity choline uptake (HACU) into cholinergic nerve terminals was decreased by 23% at 2 and 4 weeks postinjury, yet choline acetyltransferase (ChAT) activity in these neurons was unchanged at 2 weeks and decreased by 14% at 4 weeks. Daily infusion of 1 microgram of rhNGF into the injury area did not alter the loss of HACU. However, this treatment elevated ChAT activity by 23-29% above intact neostriatal levels and by 53-65% relative to HACU at both survival times. Thus, lesion-induced increases in NGF levels within astrocytes are associated with maintenance of striatal ChAT activity at normal levels following cholinergic injury, even with decreases in HACU. Pharmacologic doses of rhNGF can further augment ChAT activity in damaged cholinergic neurons, showing the usefulness of exogenous NGF even when endogenous NGF is elevated in response to injury.
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Developmental Biology, Genentech, Inc., South San Francisco, California.
Armanini
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Dugich-Djordjevic
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Bennett
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Williams
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R
Feinglass
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Anicetti
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Sinicropi
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Bakhit
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C
D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000276:immunology; D000918:Antibody Specificity; D001253:Astrocytes / Q000737:chemistry / Q000201:enzymology; D002794:Choline / Q000378:metabolism; D002795:Choline O-Acetyltransferase / Q000378:metabolism* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D007457:Iodine Radioisotopes; D008297:Male; D017072:Neostriatum / Q000737:chemistry / Q000201:enzymology* / Q000293:injuries*; D009414:Nerve Growth Factors / Q000276:immunology / Q000494:pharmacology* / Q000502:physiology*; D009474:Neurons / Q000737:chemistry / Q000201:enzymology; D016716:PC12 Cells; D010641:Phenotype; D051381:Rats; D011916:Rats, Inbred F344; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology; D013314:Stress, Mechanical
D000906:Antibodies; D005904:Glial Fibrillary Acidic Protein; D007457:Iodine Radioisotopes; D009414:Nerve Growth Factors; D011994:Recombinant Proteins; D002795:Choline O-Acetyltransferase; D002794:Choline