Yamamoto H, Gurney M E
Department of Cell, Molecular, and Structural Biology, Northwestern University Medical School, Chicago, Illinois 60611.
J Neurosci. 1990 Nov;10(11):3469-78. doi: 10.1523/JNEUROSCI.10-11-03469.1990.
Neurotrophic support to peripheral sensory neurons is provided by 2 factors of related sequence, NGF and brain-derived neurotrophic factor (BDNF). NGF is present in peripheral target tissues, while BDNF has only been reported in the CNS. We now report the biological characterization and molecular cloning of a cDNA for BDNF from human platelets. BDNF in human platelets has biological activities very similar to those of BDNF obtained from adult porcine brain in neuron-enriched cultures prepared from peripheral ganglia of chick embryos at 8-12 d of incubation. BDNF from human platelets promoted the survival and neurite outgrowth of placodal and neural crest-derived sensory neurons, but not to parasympathetic or sympathetic neurons. Activity of the factor was additive to that of NGF in dorsal root ganglia (DRG) neuron cultures and is equivalent to porcine brain BDNF in nodose ganglion neuron cultures. On SDS-PAGE, BDNF from human platelets is recovered at an apparent molecular weight equivalent to porcine brain BDNF (13,000 D). A BDNF cDNA fragment was amplified from human platelet RNA by using a coupled reverse transcriptase-polymerase chain reaction. Molecular cloning and DNA sequence analysis of the amplified cDNA fragment revealed complete identity for the deduced amino acid sequences of human and porcine BDNF [amino acid (aa) 10-108 of the mature factor]. Thus, human platelets might provide an important source of BDNF for regenerating peripheral sensory neurons at the site of nerve injury.
神经生长因子和脑源性神经营养因子(BDNF)这两种相关序列的因子为外周感觉神经元提供神经营养支持。神经生长因子存在于外周靶组织中,而脑源性神经营养因子仅在中枢神经系统中有报道。我们现在报告从人血小板中克隆出的脑源性神经营养因子cDNA的生物学特性和分子克隆。在从孵化8 - 12天的鸡胚外周神经节制备的富含神经元的培养物中,人血小板中的脑源性神经营养因子具有与从成年猪脑中获得的脑源性神经营养因子非常相似的生物学活性。人血小板中的脑源性神经营养因子促进了基板和神经嵴衍生的感觉神经元的存活和神经突生长,但对副交感或交感神经元没有作用。在背根神经节(DRG)神经元培养物中,该因子的活性与神经生长因子的活性相加,在结状神经节神经元培养物中与猪脑脑源性神经营养因子相当。在SDS - PAGE上,人血小板中的脑源性神经营养因子以与猪脑脑源性神经营养因子(13,000 D)相当的表观分子量回收。通过使用偶联的逆转录酶 - 聚合酶链反应从人血小板RNA中扩增出脑源性神经营养因子cDNA片段。对扩增的cDNA片段进行分子克隆和DNA序列分析,结果显示人和猪脑源性神经营养因子推导的氨基酸序列完全相同[成熟因子的氨基酸(aa)10 - 108]。因此,人血小板可能为神经损伤部位外周感觉神经元的再生提供脑源性神经营养因子的重要来源。