Calvet S, Doherty P, Prochiantz A
Centre National de la Recherche Scientifique, Unité de Recherche Associée 1414, Ecole Normale Supérieure, 75230 Paris Cedex 05, France.
J Neurosci. 1998 Dec 1;18(23):9751-65. doi: 10.1523/JNEUROSCI.18-23-09751.1998.
In two earlier reports we demonstrated that natural heparan sulfate, but not dermatan or chondroitin sulfate glycosaminoglycans, stimulate axonal elongation and inhibit dendrite growth in vitro (Lafont et al., 1992). The latter specific effect on dendrite elongation was reproduced by chemically synthesized heparan sulfates and by SR 80037A, a purified sulfated and hexanoylated heparin fragment (Lafont et al., 1994). Adding radioactive SR 80037A to purified neurons demonstrated the existence, at the neuronal surface, of heparan sulfate-specific and saturable binding sites, suggesting that SR 80037A activates specific signal transduction pathways. In the present study, using rat or mouse neurons from the embryonic cortex, we show that SR 80037A signaling involves one or several G-coupled receptor or receptors, small GTPases rhoA and/or rhoC, and one or several PKCs. We also demonstrate that the rapid soma rounding elicited by SR 80037A does not require protein synthesis but that the long-term effect on dendrite initiation requires protein synthesis in a short period after the addition of the heparan sulfate. Finally, by preparing membranes from the somatodendritic or axonal compartments we demonstrate that the identified signaling pathway is activated by SR 80037A primarily in the somatodendritic compartment and is not sensitive to the addition of a dermatan sulfate glycosaminoglycan that does not induce the axonal phenotype by impairing dendrite initiation and elongation.
在之前的两份报告中,我们证明了天然硫酸乙酰肝素而非硫酸皮肤素或硫酸软骨素糖胺聚糖能在体外刺激轴突伸长并抑制树突生长(拉方特等人,1992年)。化学合成的硫酸乙酰肝素以及一种纯化的硫酸化和己酰化肝素片段SR 80037A再现了对树突伸长的后一种特定作用(拉方特等人,1994年)。向纯化的神经元中添加放射性SR 80037A证明,在神经元表面存在硫酸乙酰肝素特异性和可饱和的结合位点,这表明SR 80037A激活了特定的信号转导途径。在本研究中,我们使用来自胚胎皮质的大鼠或小鼠神经元,表明SR 80037A信号传导涉及一个或多个G偶联受体、小GTP酶rhoA和/或rhoC以及一个或多个蛋白激酶C。我们还证明,SR 80037A引起的快速胞体变圆不需要蛋白质合成,但对树突起始的长期影响需要在添加硫酸乙酰肝素后的短时间内进行蛋白质合成。最后,通过制备来自胞体树突或轴突区室的膜,我们证明所确定的信号通路主要在胞体树突区室被SR 80037A激活,并且对添加硫酸皮肤素糖胺聚糖不敏感,后者不会通过损害树突起始和伸长来诱导轴突表型。