von Bartheld C S, Patterson S L, Heuer J G, Wheeler E F, Bothwell M, Rubel E W
Hearing Development Laboratories RL-30, University of Washington School of Medicine, Seattle 98195.
Development. 1991 Oct;113(2):455-70. doi: 10.1242/dev.113.2.455.
The expression of nerve growth factor receptors (NGFRs) was studied in the developing inner ear with in situ hybridization in chick embryos and with immunocytochemistry in rat embryos to determine sites of possible functions of NGF or NGF-like molecules in inner ear development. NGFR expression in the chick otocyst and acoustic ganglion is compared with epithelial differentiation and the onset of afferent innervation as determined with fluorescent carbocyanine tracers. In the inner ear of the chick embryo, NGFR mRNA expression shows an alternating pattern in mesenchymal and epithelial tissues. NGFR mRNA is heavily expressed in the mesenchyme surrounding the otocyst (E2-3), ceases at E3-5, and reappears in a thin layer of mesenchymal cells surrounding the membraneous epithelia (E5-13). In the otocyst epithelium, NGFR mRNA expression develops in one anterior and one posterior focus at E3-4.5. NGFR mRNA is expressed in the primordia of the ampullary cristae (E5-7) and possibly the anlage of the utricle; label transiently concentrates in the planum semilunatum of the cristae ampullares and in superior portions of the semicircular canals at E9, but is not seen in differentiating hair cells. In the acoustic ganglion, NGFR mRNA expression begins at E4; at the same time, the first peripheral acoustic nerve processes penetrate the otic epithelium (E4-4.5). The acoustic ganglia remain weakly NGFR mRNA-labeled in the posthatch animal. In the rat embryo, NGFR immunoreactivity is present in the auditory placode at E9, in the periotic mesenchyme at E9-10, and in the medial half of the otocyst at E10-11. At E12, epithelial NGFR expression becomes restricted anteriorly and posteriorly in a pattern similar to that of the chick otocyst and ceases at E13. NGFR immunoreactivity appears transiently in pillar cells of the cochlea in the third week of gestation. NGFR and NGFR mRNA is expressed after E11 in the acoustic ganglia. While NGFR transcripts are expressed in the cochlear ganglion cell bodies, NGFR protein becomes restricted to neuronal processes by the third week of gestation. The vestibular, but not the cochlear (spiral) ganglia remain NGFR-labeled in the adult rat. Onset of NGFR mRNA expression in the acoustic ganglion during the period of afferent fiber ingrowth into the otocyst epithelium is consistent with the hypothesis that NGF-like molecules may have a neurotrophic function for acoustic ganglion cells. Transient expression of NGFRs in secretory cells of the vestibular endorgan and pillar cells in the organ of Corti implicate a role for neurotrophins in the differentiation of these epithelial cell types.
利用鸡胚原位杂交和大鼠胚免疫细胞化学技术研究了发育中内耳神经生长因子受体(NGFRs)的表达,以确定NGF或NGF样分子在内耳发育中可能发挥作用的部位。将鸡胚耳囊和听神经节中的NGFR表达与用荧光碳菁染料示踪剂确定的上皮分化及传入神经支配的起始进行比较。在鸡胚内耳中,NGFR mRNA表达在间充质和上皮组织中呈交替模式。NGFR mRNA在耳囊周围的间充质中大量表达(E2 - 3),在E3 - 5时停止表达,并在膜性上皮周围的一层薄间充质细胞中重新出现(E5 - 13)。在耳囊上皮中,NGFR mRNA表达在E3 - 4.5时在一个前部和一个后部焦点处开始发育。NGFR mRNA在壶腹嵴原基(E5 - 7)以及可能的椭圆囊原基中表达;在E9时,标记物短暂集中在壶腹嵴的半月平面和半规管的上部,但在分化的毛细胞中未见表达。在听神经节中,NGFR mRNA表达在E4时开始;与此同时,第一批外周听神经纤维穿透耳上皮(E4 - 4.5)。在孵化后的动物中,听神经节仍有较弱的NGFR mRNA标记。在大鼠胚胎中,E9时听觉基板中有NGFR免疫反应性,E9 - 10时在耳周间充质中有,E10 - 11时在耳囊的内侧半部有。在E12时,上皮NGFR表达在前后部受到限制,其模式与鸡胚耳囊相似,并在E13时停止。在妊娠第三周,NGFR免疫反应性短暂出现在耳蜗的支柱细胞中。E11后,听神经节中表达NGFR和NGFR mRNA。虽然NGFR转录本在耳蜗神经节细胞体中表达,但到妊娠第三周时,NGFR蛋白局限于神经突起。成年大鼠中,前庭神经节而非耳蜗(螺旋)神经节仍有NGFR标记。在传入纤维长入耳囊上皮期间,听神经节中NGFR mRNA表达的起始与NGF样分子可能对听神经节细胞具有神经营养功能的假说一致。NGFR在前庭终器分泌细胞和柯蒂器支柱细胞中的短暂表达表明神经营养因子在这些上皮细胞类型的分化中起作用。