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Sensory innervation of the anterior eye segment in rats: a retrograde tracing study

Haixia LIU MD , Zhiwang LI MD , Min YANG MD , Xiang TIAN MS , Chaoying LI PhD , Lei PEI PhD ,

《医学前沿（英文）》 2009年第3卷第3期页码 352-356 doi: 10.1007/s11684-009-0065-y

摘要： This study aimed to investigate the sensory innervation of the anterior eye segment in rats by retrograde tracing with 1,1-dioleyl-3,3,3,3-tetrameth-ylindocarbocyanine, 4-chlorobenesulfonate (FAST Dil) injected into the anterior chamber. In our study, the sensory innervation of distinct elements of the anterior segment of the rat’s eye, i.e. the cornea, ciliary body, iris, and trabecular meshwork, were studied by retrograde tracing using FAST Dil as a tracer. FAST Dil was injected into the anterior chambers of the rat’s eyes. The animals were sacrificed at different time points, i.e., 2, 3, 4, and 5 days after the injection. FAST Dil localization in trigeminal ganglions was studied with a fluorescent microscope. Two days after FAST Dil injection into the anterior chambers, the cornea, the ciliary body, the iris, and the trabecular meshwork were heavily labeled. Neurons in the ipsilateral trigeminal ganglion were also consistently labeled. The number of labeled cells increased over time until 4 days after FAST Dil injection. FAST Dil-labeled neurons could be divided into two parts. Most of the Dil-labeled neurons were concentrated in a sharp, longitudinal, spindle-like stripe, located in the dorso-medial side of the trigeminal ganglion, approximately two thirds of the dorsal portion. The other part of Dil-labeled neurons scattered laterally to the stripe, but just in one third of the dorsal region. Thus, with our preliminary results, we conclude that the primary afferent sensory neurons innervating the rat’s anterior eye segments aggregate in the dorso-medial part of the ipsilateral trigeminal ganglion. It is feasible to identify them using retrograde tracing with FAST Dil anterior chamber injection.

关键词： trigeminal ganglion retrograde tracing

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A modified chronic ocular hypertension rat model for retinal ganglion cell neuroprotection

null

《医学前沿（英文）》 2013年第7卷第3期页码 367-377 doi: 10.1007/s11684-013-0266-2

摘要：

This study aimed to modify a chronic ocular hypertension (OHT) rat model to screen for potential compounds to protect retinal ganglion cells (RGCs) from responding to increased intraocular pressure (IOP). A total of 266 rats were prepared and randomly grouped according to different time-points, namely, weeks 3, 8, 16, and 24. Rats were sedated and eye examination was performed to score as the corneal damage on a scale of 1 to 4. The OHT rat model was created via the injection of a hypertonic saline solution into the episcleral veins once weekly for two weeks. OHT was identified when the IOP at week 0 was≥6 mmHg than that at week -2 for the same eye. Viable RGCs were labeled by injecting 4% FluoroGold. Rats were sacrificed, and the eyes were enucleated and fixed. The fixed retinas were dissected to prepare flat whole-mounts. The viable RGCs were visualized and imaged. The IOP (meanβ±βSD) was calculated, and data were analyzed by the paired t-test and one-way ANOVA. The OHT model was created in 234 of 266 rats (87.97%), whereas 32 rats (12.03%) were removed from the study because of the absence of IOP elevation (11.28%) and/or corneal damage scores over 4 (0.75%). IOP was elevated by as much as 81.35% for 24 weeks. The average IOP was (16.68β±β0.98)βmmHg in non-OHT eyes (n = 234), but was (27.95±0.97)βmmHg in OHT eyes (n = 234). Viable RGCs in the OHT eyes were significantly decreased in a time-dependent manner by 29.41%, 38.24%, 55.32%, and 59.30% at weeks 3, 8, 16, and 24, respectively, as compared to viable RGCs in the non-OHT eyes (P<β0.05). The OHT model was successfully created in 88% of the rats. The IOP in the OHT eyes was elevated by approximately 81% for 24 weeks. The number of viable RGCs was decreased by 59% of the rats in a time-dependent manner. The modified OHT model may provide an effective and reliable method for screening drugs to protect RGCs from glaucoma.

关键词： chronic ocular hypertension intraocular pressure retinal ganglion cells neuroprotection glaucoma