Feron F, Bianco J, Ferguson I, Mackay-Sim A
Eskitis Institute for Cell and Molecular Therapies, Griffith University, Nathan, QLD 4111, Australia.
Brain Res. 2008 Feb 27;1196:13-21. doi: 10.1016/j.brainres.2007.12.003. Epub 2007 Dec 8.
Published reports of neurotrophin expression in the olfactory system are incomplete because of missing data and conflicting results. Previous studies used a variety of fixation procedures and antibodies on different species and different ages. The aim of the present study was to examine expression of neurotrophins and their receptors using optimized methodologies: five methods of fixation, multiple antibodies, a variety of immunochemical protocols, and RT-PCR. We show here that (i) transcripts for all neurotrophins and their receptors are found in the adult olfactory epithelium; (ii) all neurotrophins are expressed in the supporting cells and the neuronal layers of the undisturbed adult olfactory epithelium while NT4 is found additionally in the horizontal basal cells; (iii) neurotrophin immunoreactivity required a fixative that included parabenzoquinone (not used in previous studies of olfactory tissue); (iv) TrkB and TrkC are restricted to the globose basal cell and neuron layers while TrkA is found in the horizontal basal cells and in the supporting cells where it co-localizes with the low affinity receptor for NGF (p75NTR). These findings confirm that neurotrophins are produced within the olfactory epithelium, suggesting autocrine and paracrine regulation of olfactory neurogenesis.
由于数据缺失和结果相互矛盾,已发表的关于嗅觉系统中神经营养因子表达的报告并不完整。先前的研究在不同物种和不同年龄的样本上使用了多种固定程序和抗体。本研究的目的是使用优化的方法来检测神经营养因子及其受体的表达:五种固定方法、多种抗体、多种免疫化学方案以及逆转录聚合酶链反应(RT-PCR)。我们在此表明:(i)在成年嗅觉上皮中发现了所有神经营养因子及其受体的转录本;(ii)所有神经营养因子均在未受干扰的成年嗅觉上皮的支持细胞和神经元层中表达,而神经营养因子4(NT4)还存在于水平基底细胞中;(iii)神经营养因子免疫反应性需要一种含有对苯醌的固定剂(嗅觉组织先前的研究未使用);(iv)酪氨酸激酶受体B(TrkB)和酪氨酸激酶受体C(TrkC)局限于球状基底细胞层和神经元层,而酪氨酸激酶受体A(TrkA)存在于水平基底细胞和支持细胞中,在支持细胞中它与神经生长因子(NGF)的低亲和力受体(p75NTR)共定位。这些发现证实神经营养因子是在嗅觉上皮内产生的,提示嗅觉神经发生存在自分泌和旁分泌调节。