p75 and TrkA receptors are both required for uptake of NGF in adult sympathetic neurons: use of a novel fluorescent NGF conjugate

被引:21
作者
Gatzinsky, KP
Haugland, RP
Thrasivoulou, C
Orike, N
Budi-Santoso, AW
Cowen, T
机构
[1] Univ Gothenburg, Dept Anat & Cell Biol, S-40530 Gothenburg, Sweden
[2] Univ Gothenburg, Dept Clin Neurosci, S-40530 Gothenburg, Sweden
[3] Mol Probes Inc, Eugene, OR 97402 USA
[4] UCL Royal Free & Univ Coll, Med Sch, Dept Anat & Dev Biol, London NW3 2PF, England
基金
英国惠康基金;
关键词
rat; NGF transport; superior cervical ganglion; antibody receptor blockade; confocal microscopy;
D O I
10.1016/S0006-8993(01)03099-2
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
we have developed and tested the biological activity, and specificity of a novel fluorescent dextran-Texas Red-nerve growth factor (DTR-NGF) conjugate. DTR-NGF was found to promote survival and neurite outgrowth in cultured dissociated sympathetic neurons similarly to native NGF. The conjugate was taken up and transported retrogradely by terminal sympathetic nerves innervating the iris to neurons in the ipsilateral superior cervical ganglion (SCG) of young adult rats. Uptake and transport was assessed by Counting numbers of labelled neurons and by measuring intensity of neuronal labelling using confocal microscopy and image analysis. DTR-NGF labelling in SCG neurons was shown to be dose-dependent with an EC(50) of 75 ng. Similar concentrations of unconjugated DTR resulted in no neuronal labelling. DTR-NGF uptake was competed off using a 50-fold excess of native NGF, resulting in a 73% reduction in numbers of labelled neurons. Pretreatment of nerve terminals with function-blocking antibodies against the low (p75) and high (TrkA) affinity NGF receptors resulted in a large (85-93%) reduction in numbers of DTR-NGF labelled neurons. Anti-p75 and anti-TrkA antibodies had comparable effects which were concentration-dependent. These findings indicate that both receptors are required for uptake of NGF in adult rat sympathetic neurons. In particular, the results provide strong evidence that the p75 receptor plays a more active role in transducing the NGF signal than has been proposed. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:226 / 238
页数:13
相关论文
共 78 条
[1]  
Andrews TJ, 1996, J COMP NEUROL, V368, P33
[2]   DISRUPTION OF NGF BINDING TO THE LOW-AFFINITY NEUROTROPHIN RECEPTOR P75(LNTR) REDUCES NGF BINDING TO TRKA ON PC12 CELLS [J].
BARKER, PA ;
SHOOTER, EM .
NEURON, 1994, 13 (01) :203-215
[3]  
BATTLEMAN DS, 1993, J NEUROSCI, V13, P941
[4]   DIFFERENTIAL EXPRESSION OF NERVE GROWTH-FACTOR RECEPTORS LEADS TO ALTERED BINDING-AFFINITY AND NEUROTROPHIN RESPONSIVENESS [J].
BENEDETTI, M ;
LEVI, A ;
CHAO, MV .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (16) :7859-7863
[5]  
Bhattacharyya A, 1997, J NEUROSCI, V17, P7007
[6]  
BOTHWELL M, 1995, ANNU REV NEUROSCI, V18, P223, DOI 10.1146/annurev.ne.18.030195.001255
[7]   p75(NTR) and apoptosis: Trk-dependent and Trk-independent effects [J].
Bredesen, DE ;
Rabizadeh, S .
TRENDS IN NEUROSCIENCES, 1997, 20 (07) :287-290
[8]   p75(NGFR)and TrkA receptors collaborate to rapidly activate a p75(NGFR)-associated protein kinase [J].
Canossa, M ;
Twiss, JL ;
Verity, AN ;
Shooter, EM .
EMBO JOURNAL, 1996, 15 (13) :3369-3376
[9]   Selective activation of NF-kappa B by nerve growth factor through the neurotrophin receptor p75 [J].
Carter, BD ;
Kaltschmidt, C ;
Kaltschmidt, B ;
Offenhauser, N ;
BohmMatthaei, R ;
Baeuerle, PA ;
Barde, YA .
SCIENCE, 1996, 272 (5261) :542-545
[10]  
Casaccia-Bonnefil P, 1999, MICROSC RES TECHNIQ, V45, P217, DOI 10.1002/(SICI)1097-0029(19990515/01)45:4/5<217::AID-JEMT5>3.0.CO