Schrödl Falk, Kaser-Eichberger Alexandra, Trost Andrea, Strohmaier Clemens, Bogner Barbara, Runge Christian, Bruckner Daniela, Krefft Karolina, Kofler Barbara, Brandtner Herwig, Reitsamer Herbert A
Department of Ophthalmology and Optometry, Paracelsus Medical University, Muellner Hauptstrasse 48, 5020, Salzburg, Austria; Department of Anatomy, Paracelsus Medical University, Strubergasase 21, 5020, Salzburg, Austria.
Department of Ophthalmology and Optometry, Paracelsus Medical University, Muellner Hauptstrasse 48, 5020, Salzburg, Austria.
Exp Eye Res. 2015 Feb;131:63-8. doi: 10.1016/j.exer.2014.12.007. Epub 2014 Dec 9.
Extrinsic and intrinsic sources of the autonomic nervous system contribute to choroidal innervation, thus being responsible for the control of choroidal blood flow, aqueous humor production or intraocular pressure. Neuropeptides are involved in this autonomic control, and amongst those, alarin has been recently introduced. While alarin is present in intrinsic choroidal neurons, it is not clear if these are the only source of neuronal alarin in the choroid. Therefore, we here screened for the presence of alarin in human cranial autonomic ganglia, and also in rat, a species lacking intrinsic choroidal innervation. Cranial autonomic ganglia (i.e., ciliary, CIL; pterygopalatine, PPG; superior cervical, SCG; trigeminal ganglion, TRI) of human and rat were prepared for immunohistochemistry against murine and human alarin, respectively. Additionally, double staining experiments for alarin and choline acetyltransferase (ChAT), tyrosine hydroxilase (TH), substance P (SP) were performed in human and rat ganglia for unequivocal identification of ganglia. For documentation, confocal laser scanning microscopy was used, while quantitative RT-PCR was applied to confirm immunohistochemical data and to detect alarin mRNA expression. In humans, alarin-like immunoreactivity (alarin-LI) was detected in intrinsic neurons and nerve fibers of the choroidal stroma, but was lacking in CIL, PPG, SCG and TRI. In rat, alarin-LI was detected in only a minority of cranial autonomic ganglia (CIL: 3.5%; PPG: 0.4%; SCG: 1.9%; TRI: 1%). qRT-PCR confirmed the low expression level of alarin mRNA in rat ganglia. Since alarin-LI was absent in human cranial autonomic ganglia, and only present in few neurons of rat cranial autonomic ganglia, we consider it of low impact in extrinsic ocular innervation in those species. Nevertheless, it seems important for intrinsic choroidal innervation in humans, where it could serve as intrinsic choroidal marker.
自主神经系统的外在和内在来源都对脉络膜神经支配有贡献,从而负责控制脉络膜血流、房水生成或眼压。神经肽参与这种自主控制,其中,阿拉瑞林(alarin)最近被引入。虽然阿拉瑞林存在于脉络膜内在神经元中,但尚不清楚这些是否是脉络膜中神经元阿拉瑞林的唯一来源。因此,我们在此筛查了人类颅部自主神经节以及大鼠(一种缺乏脉络膜内在神经支配的物种)中阿拉瑞林的存在情况。分别制备了人类和大鼠的颅部自主神经节(即睫状神经节,CIL;翼腭神经节,PPG;颈上神经节,SCG;三叉神经节,TRI)用于针对小鼠和人类阿拉瑞林的免疫组织化学检测。此外,在人类和大鼠神经节中进行了阿拉瑞林与胆碱乙酰转移酶(ChAT)、酪氨酸羟化酶(TH)、P物质(SP)的双重染色实验,以明确神经节的鉴定。为了记录,使用了共聚焦激光扫描显微镜,同时应用定量逆转录聚合酶链反应(qRT-PCR)来确认免疫组织化学数据并检测阿拉瑞林mRNA表达。在人类中,在脉络膜基质的内在神经元和神经纤维中检测到阿拉瑞林样免疫反应性(alarin-LI),但在CIL、PPG、SCG和TRI中未检测到。在大鼠中,仅在少数颅部自主神经节中检测到alarin-LI(CIL:3.5%;PPG:0.4%;SCG:1.9%;TRI:1%)。qRT-PCR证实了大鼠神经节中阿拉瑞林mRNA的低表达水平。由于人类颅部自主神经节中不存在alarin-LI,且仅在大鼠颅部自主神经节的少数神经元中存在alarin-LI,我们认为它在这些物种的眼外神经支配中的影响较小。然而,它似乎对人类脉络膜内在神经支配很重要,在人类中它可作为脉络膜内在标记物。