Department of Neurobiology, David Geffen School of Medicine at Los Angeles, University of California at Los Angeles, Los Angeles, California, 90095-1763.
J Comp Neurol. 2014 Apr 15;522(6):1411-43. doi: 10.1002/cne.23521.
There are few neurochemical markers that reliably identify retinal ganglion cells (RGCs), which are a heterogeneous population of cells that integrate and transmit the visual signal from the retina to the central visual nuclei. We have developed and characterized a new set of affinity-purified guinea pig and rabbit antibodies against RNA-binding protein with multiple splicing (RBPMS). On western blots these antibodies recognize a single band at 〜24 kDa, corresponding to RBPMS, and they strongly label RGC and displaced RGC (dRGC) somata in mouse, rat, guinea pig, rabbit, and monkey retina. RBPMS-immunoreactive cells and RGCs identified by other techniques have a similar range of somal diameters and areas. The density of RBPMS cells in mouse and rat retina is comparable to earlier semiquantitative estimates of RGCs. RBPMS is mainly expressed in medium and large DAPI-, DRAQ5-, NeuroTrace- and NeuN-stained cells in the ganglion cell layer (GCL), and RBPMS is not expressed in syntaxin (HPC-1)-immunoreactive cells in the inner nuclear layer (INL) and GCL, consistent with their identity as RGCs, and not displaced amacrine cells. In mouse and rat retina, most RBPMS cells are lost following optic nerve crush or transection at 3 weeks, and all Brn3a-, SMI-32-, and melanopsin-immunoreactive RGCs also express RBPMS immunoreactivity. RBPMS immunoreactivity is localized to cyan fluorescent protein (CFP)-fluorescent RGCs in the B6.Cg-Tg(Thy1-CFP)23Jrs/J mouse line. These findings show that antibodies against RBPMS are robust reagents that exclusively identify RGCs and dRGCs in multiple mammalian species, and they will be especially useful for quantification of RGCs.
几乎没有神经化学标记物能够可靠地识别视网膜神经节细胞（RGC），RGC 是一个异质细胞群体，它们整合并将视觉信号从视网膜传递到中央视觉核。我们开发并表征了一组新的针对 RNA 结合蛋白多剪接（RBPMS）的亲和纯化豚鼠和兔抗体。在 Western blot 上，这些抗体识别出一个约 24 kDa 的单一条带，对应于 RBPMS，并且它们强烈标记小鼠、大鼠、豚鼠、兔和猴视网膜中的 RGC 和移位的 RGC（dRGC）体。通过其他技术鉴定的 RBPMS 免疫反应性细胞和 RGC 具有相似的体直径和面积范围。在小鼠和大鼠视网膜中，RBPMS 细胞的密度与以前对 RGC 的半定量估计相当。RBPMS 主要在神经追踪剂（NeuroTrace）、DAPI、DRAQ5 和 NeuN 染色的中等和大直径 DAPI、DRAQ5、NeuroTrace 和 NeuN 染色的细胞层（GCL）中表达，并且 RBPMS 不在内核层（INL）和 GCL 中的 syntaxin（HPC-1）免疫反应性细胞中表达，与它们作为 RGC 的身份一致，而不是移位的无长突细胞。在小鼠和大鼠视网膜中，视神经挤压或 3 周时横断后，大多数 RBPMS 细胞丢失，所有 Brn3a、SMI-32 和黑视蛋白免疫反应性 RGC 也表达 RBPMS 免疫反应性。RBPMS 免疫反应性定位于 B6.Cg-Tg（Thy1-CFP）23Jrs/J 小鼠系中 CFP 荧光 RGC 的青色荧光蛋白（CFP）荧光。这些发现表明，针对 RBPMS 的抗体是一种强大的试剂，可特异性识别多种哺乳动物物种中的 RGC 和 dRGC，它们对于 RGC 的定量特别有用。
