Functional blockade of tyrosine kinase A in the rat basal forebrain by a novel antagonistic anti-receptor monoclonal antibody.
We have exploited a new monoclonal antibody against the tyrosine kinase A (TrkA) nerve growth factor (NGF) receptor to block the NGF-TrkA interaction in the rat basal forebrain. The monoclonal antibody MNAC13 is a potent antagonist that prevents the binding of NGF to TrkA in a variety of systems. This antibody was used to study the maintenance of the cholinergic phenotype in the rat basal forebrain in vivo, by the implant of antibody-secreting cells. Basal forebrain cholinergic neurons (BFCNs) are greatly affected by the antibody treatment, both in terms of cell number and of cell soma size. When antibody-secreting cells are implanted at postnatal day 2 (P2), the effects observed at P8 are as severe as those obtained with anti-NGF antibodies and, interestingly, are observed also if anti-TrkA cells are implanted at P8, when anti-NGF antibodies, delivered by the same route, are no longer effective (). The effects induced by anti-TrkA, as those induced by anti-NGF, are reversible, but the time required for recovery and the critical period in the sensitivity of BFCNs to the functional inactivation of TrkA is twice as long than that observed when NGF is intercepted. These results demonstrate that BFCNs are more sensitive to the block of TrkA activation than they are to the block of NGF. The cloning of MNAC13 variable regions and their assembly into a functional polypeptide of reduced size (single chain Fv fragment) will allow its use in gene transfer applications.
Cattaneo
A
A
Neuroscience Programme, International School for Advanced Studies, 34014 Trieste, Italy. cattaneo@sissa.it
Capsoni
S
S
Margotti
E
E
Righi
M
M
Kontsekova
E
E
Pavlik
P
P
Filipcik
P
P
Novak
M
M
D016475:3T3 Cells; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002795:Choline O-Acetyltransferase / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007074:Immunoglobulin G / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D009414:Nerve Growth Factors / Q000494:pharmacology / Q000502:physiology; D016501:Neurites / Q000502:physiology / Q000648:ultrastructure; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D016548:Prosencephalon / Q000502:physiology*; D051381:Rats; D020917:Receptor, trkA / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D020813:Receptor, trkB / Q000187:drug effects / Q000502:physiology; D013377:Substantia Innominata / Q000502:physiology*; D014162:Transfection
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D009414:Nerve Growth Factors; D002795:Choline O-Acetyltransferase; D020917:Receptor, trkA; D020813:Receptor, trkB