Normal Human Choroid Research Articles

One target to quench many

Inhibiting the GTPase ARF6 chokes the diverse output of mutant G αq proteins in uveal melanoma.

ECRG-4 expression in normal and neoplastic choroid plexus

The choroid plexus is a major site of gene expression of esophageal cancer-related gene (ECRG)-4 during development, suggesting that its gene product may be involved in cerebrospinal fluid (CSF) homeostasis. Yet, ECRG-4 is also a novel candidate tumor suppressor gene whose expression is downregulated and is inversely associated with a worse prognosis in several different cancers. Reduced expression of ECRG-4 has been demonstrated in most tumors, including colorectal carcinoma and malignant glioma, to be mediated by hypermethylation of its promoter. Materials and methods In this study, samples of normal human choroid plexus (both fetal and adult) and choroid plexus neoplasms (WHO grade I papilloma, grade II atypical papilloma, and grade III carcinoma) were stained with antibodies that we generated to augurin, the gene product of ECRG4. DNA was then extracted from the tissue, treated with bisulfite, and subjected to PCR using a 217-base pair region encompassing the ECRG-4 promotor to detect methylation. Results Both fetal and adult human choroid plexus cells demonstrated a robust positive immunostaining at the apical surface that is consistent with our prior results in human, rat, and mouse brains. In contrast, there was a near-complete absence of immunostaining in all of the choroid plexus neoplasms examined. The choroid plexus carcinoma demonstrated significant methylation of the ECRG-4 promotor region. Conclusions Taken together, these data suggest that ECRG-4 is downregulated in neoplasms of the choroid plexus just as has been observed in other central nervous system (CNS) and non-CNS cancers. This is likely due to hypermethylation of the ECRG-4 promotor, as shown in the choroid plexus carcinoma. Further analysis is underway to determine the (1) physiologic and (2) pathophysiologic consequences of ECRG-4 over- and under- expression in the choroid plexus on CSF formation, function, and composition.

The Normal Human Choroid Is Endowed with a Significant Number of Lymphatic Vessel Endothelial Hyaluronate Receptor 1 (LYVE-1)–Positive Macrophages

Lymphatic vessel endothelial hyaluronic acid receptor (LYVE-1) is a newly discovered lymphatic endothelium-specific marker that is also expressed by a subpopulation of macrophages. To date, there is no report on its expression in the posterior human uvea. The purpose of this study was to investigate the expression of LYVE-1 in normal human choroids. Eyes of body/cornea donors (55-89 years of age; 4-9 hours postmortem) were obtained. Choroids were dissected and prepared for cryosections followed by immunohistochemistry with anti-human LYVE-1 antiserum and immunogold labeling. In addition, anti-human antibodies against macrophage markers (CD68, MHC class II) and lymphatic (podoplanin) and blood vascular endothelium (CD31, vWF) were used. For documentation, light-, fluorescence-, confocal laser scanning-, and electron-microscopy were used. The normal human choroidal stroma contained 274 +/- 86 LYVE-1 positive cells/mm(2). The cells displayed irregular shapes with a relatively uniform diameter of 32 mum. Costaining with CD68 and negativity for CD31, podoplanin, and melan-A/HMB45, as well as electron microscopic features, suggest these LYVE-1(+) cells to be macrophages. Besides that, no classic LYVE-1(+)/podoplanin(+) lymphatic vessels were detected within the normal adult human choroid. The normal adult human choroid does not contain typical lymph vessels, but is endowed with a significant number of LYVE-1 positive macrophages. These cells may be involved in choroidal hyaluronic acid metabolism or contribute to temporary formation of lymphatic channels under inflammatory conditions.

Choroid plexus hyperplasia: surgical treatment and immunohistochemical results

Diffuse villous hyperplasia of the choroid plexus is a rare but potential source of nonobstructive hydrocephalus. In addition to discussing the authors' staged surgical approach and medical management decisions in a patient with this rare and challenging condition, immunohistochemical studies of the choroid plexus epithelium are presented to examine the pathophysiological factors involved in abnormal cerebrospinal fluid (CSF) production in this disease. The patient, a 15-month-old girl born at 36 weeks' gestation, underwent a bilateral craniotomy with resection of the choroid plexus to treat her villous hyperplasia. Immunohistochemical studies of the resected choroid plexus were conducted for the purpose of examining the carbonic anhydrase II (CAII) enzyme and the aquaporin 1 (AQP1) membrane protein. Results were compared with immunohistochemical studies conducted in a small series of autopsy specimens of normal human choroid plexuses. There was no change in the immunoreactivity of CAII in the patient with villous hyperplasia compared with normal controls, whereas AQP1 immunoreactivity was significantly weaker in the patient compared with normal controls. Postoperatively, the patient's CSF overproduction resolved and her neurological symptoms improved over time. Shunting techniques and presently available pharmaceutical treatments alone do not provide adequate treatment of high-output CSF conditions. Surgical removal of the affected choroid plexus is a feasible and effective treatment. Results of the immunohistochemical studies reported here support the suggestion that the CAII enzyme is retained in villous hyperplasia of the choroid plexus. However, there appears to be decreased expression and perhaps downregulation of AQP1 in villous hyperplasia compared with normal choroid plexus. Future studies may elucidate the significance of these observations.

Micro Serial Analysis of Gene Expression in Normal Human Choroid and Retinal Pigment Epithelial Transcriptomes

To investigate the gene expression profile of the normal human choroid/retinal pigment epithelium(RPE) tissues. Micro serial analysis of gene expression (Micro SAGE) was performed. A SAGE library was constructed from 110 microg of total RNA of normal human choroid/RPE tissue, and cloned tag concatemers transformed to E.coli were sequenced. The sequence data were analyzed by SAGE software and matched to GenBank and UniGene public databases. The sequence data were also compared with the choroid/RPE cDNA library of NEIBank. A total of 12 070 tags were sequenced; 3627 tags were unique. Of these 3627 tags, 2508 tags were encoded genes and 1119 tags were unknown tags in the UniGene database. The most frequently expressed tag was TCCCTATTAA, but the gene corresponding to this tag has not been identified yet. Other frequently expressed tags encoded a tissue inhibitor of matrix metalloproteinase 3, insulin-like growth factor binding protein-related protein 1, and transthyretin. These genes are notably different, with high expression frequencies when compared to the cDNA library of NEIBank. This gene expression profile of the normal human choroid/RPE tissue should provide further understanding of the biological function of the choroid and the pathogenesis of diseases in which the choroid and RPE play a role, such as choroidal neovascularization.

HLA-DR-positive dendritic cells of the normal human choroid plexus: A potential reservoir of HIV in the central nervous system

In a previous study of choroid plexus (CPx) from patients with the acquired immunodeficiency syndrome (AIDS), we found a population of stromal cells infected with the human immunodeficiency virus (HIV). To determine whether these represented antigen-presenting dendritic cells, we examined the phenotype of normal human choroid plexus by light and electron microscopy (EM) and established the HIV-infected cell type by immunohistochemistry in AIDS cases with HIV-infected CPx. Monoclonal antibodies were used to detect class II major histocompatibility antigens (MHC), S-100 and S-100β protein, lymphocytes, monocytes/macrophages, and HIV glycoprotein. A variable number of stromal cells had slightly elongated nuclei and long branching processes that were strongly immunoreactive for class II MHCs, rarely reactive for S-100 and S-100β and immunonegative for monocyte/macrophage markers. Phagocytic activity was absent by EM and immunomarkers. They were numerous in the subepithelial region, and their processes occasionally extended toward the stromal capillaries or between the CPx epithelial cells. The HIV-infected cells were intensely inununoreactive for class II MHC markers and often displayed a dendritic morphology. These results document the presence of dendritic cells in the normal human CPx whose morphology and immunophenotype closely resemble those of DCs elsewhere in the body. They also show that these immunoreactive MHC class II cells are the cell type infected by HIV. We suggest that the functional activity of the CPx DCs is similar to that of antigen-presenting dendritic cells elsewhere in the body. This includes the potential to harbor HIV during the prolonged period of clinical latency, acting as a central nervous system reservoir of infection before the onset of AIDS.

Nerves of the normal human choroid.

The innervation of the iris and ciliary body by antagonistic autonomic elements of the long and short ciliary nerves is well known to all ophthalmologists (Adler¹). It is also known that branches of the ciliary nerves supply the choroid (Duke-Elder²). However, no detailed informations about the structure, arrangement, and distribution of nerves in the human choroid could be found in the literature. It is the purpose of this paper to fill this gap. Normal human choroid was fixed in brom formalin (ammonium bromide formalin, known as Cajal's solution). The choroid was cut in flat sections on the freezing microtome and stained with the nerve fiber stain of del Rio Hortega (double impregnation without reduction).³ These stains showed that there are two general types of nerve fibers in the human choroid: (1) fibers which branch off the ciliary nerves in small bundles and run in the intervascular spaces