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F L Chan

Publications and source records attributed to F L Chan.

At least 55 records · Page 3Linked to original sources
PubMed1999

Contribution of nitric oxide and K+ channel activation to vasorelaxation of isolated rat aorta induced by procaine.

The endothelium-dependent and -independent relaxant effect of procaine was examined in isolated rat aortic rings. Procaine induced relaxation of arteries precontracted with phenylephrine or with 60 mM K+ in a concentration-dependent manner (0.01-3 mM). Procaine (1 mM) inhibited the transient contraction induced by caffeine (10 mM) in Ca2+-free Krebs solution. Removal of the endothelium caused a rightward shift of the concentration-response curve for procaine. N(G)-Nitro-L-arginine (L-NNA, 10-100 microM), N(G)-nitro-L-arginine methyl ester (L-NAME, 100 microM) and methylene blue (1-10 microM) significantly attenuated the procaine-induced relaxation without affecting the maximal response. L-Arginine (1 mM) partially but significantly antagonized the effect of L-NAME (100 microM). Pretreatment of endothelium-intact aortic rings with procaine (1 mM) or with acetylcholine (10 microM) significantly elevated the tissue contents of cyclic GMP and this increase was inhibited in the presence of 100 microM L-NNA. Tetrapentylammonium ions (1-3 microM) reduced the procaine-induced relaxation in both endothelium-intact and -denuded arteries. Tetrapentylammonium ions (3 microM) did not affect the procaine-induced relaxation of 60 mM K+-contracted arteries. Tetraethylammonium ions (3 mM) inhibited the procaine-induced relaxation. In contrast, iberiotoxin (100 nM), glibenclamide (3 microM), 4-aminopyridine (3 mM) and indomethacin (10 microM) had no effect. These results indicate that the procaine-induced relaxation may be mediated through multiple mechanisms. A substantial portion of the procaine-induced relaxation in rat aorta was caused by nitric oxide but not by other endothelium-derived factors. The activation of tetrapentylammonium- and tetraethylammonium-sensitive K+ channels contributes in part to the procaine-induced vasorelaxation. Besides, procaine may directly inhibit both external Ca2+ entry and internal Ca2+ release in aortic smooth muscle cells.

Anesthetics, Local↗
PubMed1999

Comparative study of glycoconjugates of the rat prostatic lobes by lectin histochemistry.

BACKGROUND: The rat prostate gland is composed of several distinct lobes: the ventral, lateral, dorsal, and anterior (also called the coagulating gland). Each prostatic lobe is different in its morphology, secretions, and response to hormones. METHODS: The present study characterized and compared the expression of glycoconjugates in three different lobes (ventral, lateral, and dorsal) of the Noble rat prostate gland by lectin histochemistry, using a battery of 30 different lectins. RESULTS: The results indicate that different prostatic lobes elaborate and secrete different glycoconjugates. The dorsal prostatic epithelium secretes mainly in an apocrine mode, as characterized by the presence of apical cytoplasmic blebs which are absent in the other two lobes, though merocrine secretion is also suggested. These cytoplasmic blebs were rich in Man, GlcNAc, Gal/GalNac, Fuc, NeuAc(alpha2,6)Gal, and oligosaccharides. Two types of cytoplasmic belbs were also identified according to their lectin bindings: blebs stained by S-Con A, LCA, PWA, UEA-II, RCA-I, DBA, ECA, WFA, Jacalin, MPA, SJA, LTA, UEA-I, PHA-E, and PHA-L, and some blebs stained by S-WGA, SBA, HAA, HPA, VAA, BPA, MAA, and SNA. The lectins, which reacted with the second type of blebs, also revealed some cytoplasmic granules in the dorsal prostatic epithelial cells. The epithelium of the lateral prostate showed weak or negative reactions to lectins, suggesting that it may express fewer glycoconjugates as compared to the other two lobes. The epithelial Golgi region of the ventral prostate is the most developed among the three lobes, and contained abundant GlcNAc, alpha/beta-GalNAc, and NeuAc(alpha2,6)Gal residues, as shown by its intense reactions to S-WGA, DBA, SBA, HAA, HPA, WFA, and SNA. GS-I-B4 reacted specifically with the basal epithelial cells in all three lobes, indicating that glycoconjugates with terminal alpha-Gal/GalNAc were commonly expressed in these basal cells. The basal cells in ventral prostate might express more complex glycoconjugates, as they were also stained by PWA, RCA-I, ECA, and PHA-L, in addition to GS-I-B4, suggesting that these basal cells contained more Gal(beta1,4)GlcNAc and Gal(beta1,4)GlcNAc(alpha1,2)Man residues. GS-I-B4 and PHA-L also demonstrated a differential staining of the stromal smooth muscles and their associated extracellular matrix in the three prostatic lobes, suggesting that these structural components in different prostatic lobes were different in their complex oligosaccharide contents, which probably has an influence on the epithelial-stromal interaction in the prostate. CONCLUSIONS: In summary, the present study has established for the first time the expression patterns of glycoconjugates in the three lobes of the Noble rat prostate gland and suggests that certain lectins are useful as histochemical markers for the three individual prostatic lobes.

Animals↗
PubMed1999

Ultrastructural and biochemical observations on the early changes in apoptotic epithelial cells of the rat prostate induced by castration.

The present study describes the sequential ultrastructural changes in the apoptotic cells of the rat ventral and dorsal prostates during the early period of 1-3 days postcastration. The major morphological changes include: (1) condensation of heterochromatin along the nuclear envelope and fragmentation into crescent-shaped micronuclei; (2) formation of membrane-bound cytoplasmic spherical bodies, which contain various organelles and micronuclei, within the apoptotic cells; (3) formation of non-membrane-bound autolytic vacuoles by autolysis of cytoplasm; (4) focal rupture of outer mitochondrial membrane; and (5) phagocytosis of the fragmented cytoplasmic spherical bodies and apoptotic cells by macrophages. The occurrence of both cytoplasmic apoptotic bodies and autolytic vacuoles in apoptotic cells suggests that the cytoplasm of the apoptotic cells could be destroyed by different means. The responsiveness of different prostatic lobes to androgen withdrawal and the time course of the transitory apoptotic activity in different lobes were analyzed by counting the indices of the TUNEL-labeled apoptotic cells against the postcastration periods. The results showed that the ventral lobe responded more rapidly to castration than the lateral and dorsal lobes. The dorsal lobe was the slowest in response to castration among the three lobes. Analysis of protease activities by zymography has identified two Ca(2+)-independent proteases of apparent MW 20 and 24 kDa (expressed in both ventral and dorsolateral lobes), and one Ca(2+)-dependent protease of MW 66.5 kDa (expressed only in the dorsolateral lobe) which became activated at day 3 postcastration. Their expression patterns were different from that of CPP-3 in the castrated prostates, suggesting that the activated proteases were enzymes other than CPP-3. The association of their highest activities with the maximum apoptotic activity at day 3 postcastration and also their loss of activity at day 15 suggest that these protease activities might be related to apoptosis or glandular involution.

Animals↗
PubMed1999

CT appearance of midgut volvulus with malrotation in a young infant.

Computed tomography (CT) appearances of small bowel malrotation and midgut volvulus (MGV) have rarely been described in paediatric patients. We present spiral CT images of a surgically proven case in a young infant. The literature on imaging techniques to diagnose these conditions is reviewed. Radiation doses of upper gastrointestinal series (UGI) and spiral CT are estimated and compared.

Humans↗
PubMed1999

Glycoconjugates of the rat ciliary body epithelium: a lectin histochemical and protein blotting study.

The present study sought to identify and partially characterize the glycoconjugates specific to the double-layered ciliary body epithelium of the rat eye by lectin histochemistry and lectin blottings. Hydrated paraffin sections of Carnoy-fixed Sprague-Dawley rat eyes were stained with a panel of 21 different biotinylated lectins, followed by streptavidin-peroxidase and the glucose oxidase-diaminobenzidine-nickel staining procedure. The results of lectin histochemistry revealed that the inner epithelial layer was rich in GlcNAc(beta1,4)GlcNAc, alpha-Gal, Gal(beta1,3)GalNAc, GalNAc(alpha1,3)GalNAc/Gal, GalNAc(alpha1,6)Gal, Fuc(alpha1,2)Gal(beta1,4)GlcNAc and Gal(beta1,4)GlcNAc(beta1,2)Man(alpha1,6) sugar residues as shown by its positive reactivities with S-WGA, PWA, DSA, GS-I-B4, PNA, DBA, SBA, WFA, UEA-I, LTA and PHA-E. The reactivities of GS-I-B4, PNA, DBA and SBA were restricted to the inner layer at the tips of the ciliary processes. On the other hand, the outer epithelial layer was stained evenly by DSA and Jacalin, and partly by MAA, showing that this epithelial layer was rich in GlcNAc(beta1,4)GlcNAc, Gal(beta1,3)GalNAc and NeuAc(alpha2,3)Gal disaccharides. These lectin binding patterns of the ciliary body epithelium suggest a topographical and functional difference in this double cell-layered epithelium. Their possible roles in the secretion of aqueous humour and production of ciliary zonule are discussed. Some identified lectin markers specific to these two cell layers may be useful for further experimental studies. Glycoproteins extracted from the dissected ciliary body were separated by SDS-PAGE electrophoresis and analyzed by protein blottings with 8 different lectins. The results showed that at least 10 major membrane-bound glycoproteins, with molecular weights ranging from 30 to 150 kD, rich in beta-GlcNAc, beta-Gal, alpha/beta-GalNAc and NeuAc(alpha2,6)Gal residues, were present in the microsomal fraction.

Acetylgalactosamine↗
PubMed1999

cDNA, genomic cloning, and gene expression analysis of mouse PSP94 (prostate secretory protein of 94 amino acids).

The potential use of prostate secretory protein of 94 amino acids (PSP94) as a diagnostic biomarker or a therapeutic agent for prostate cancer has been reported. In order to establish an animal model to further elucidate on its biological role, we cloned the mouse PSP94 cDNA (approximately 500 bp) by reverse transcriptase-polymerase chain reaction (RT-PCR) and disclosed its genomic structure. The whole mouse PSP94 gene (approximately 23 kb) was amplified by long and accurate-PCR and also cloned by screening of a mouse embryo stem-cell genomic library. Computational and statistical analyses have demonstrated several highly conserved characteristics of PSP94 among different species. Comparison of PSP94 from human, two primates, pig, and rodents revealed that the most significant feature is that PSP94 is rich in cysteines (10% of the total sequence) and their positions are highly conserved. The three intron-four exon structure of the human PSP94 gene and the consensus sequence (....GT-intron-AG...) for mRNA splicing are also strongly conserved. A high divergence in cDNA sequence in the protein-coding region and also in the genomic sequence of PSP94 was also observed among these species. Comparing with alpha-globin, a typical evolutionally conserved gene, with the PSP94 gene, the rate of nonsynonymous changes per site per year (kN) is 2 to 6 times higher, indicating that PSP94 gene has been under far fewer evolutionary constraints than other genes and has a potential role as a species barrier in reproductive biology. In order to test this hypothesis, we investigated the gene expression of PSP94 and its tissue distribution in various rodent tissues by RT-PCR and in situ hybridization (ISH). Gene expression was found only in the prostate, suggesting that PSP94 is probably more tissue specific in the prostate of rodents than in mammals. The ISH analysis also revealed a prostate lobe-specific expression of the PSP94 gene in both mice and rats. It was strongly expressed in the lateral prostate, but the findings were negative in the dorsal and ventral lobe. Therefore, it is hypothesized that one of the primary functions of rodent PSP94, as a major prostate secretory protein, is related to reproductive biology.

Amino Acid Sequence↗
PubMed1999

Pulmonary sequestration diagnosed by contrast enhanced three-dimensional MR angiography.

Pulmonary sequestration is a congenital bronchopulmonary foregut malformation in which a segment of lung parenchyma is not connected to the tracheobronchial tree. This abnormal segment receives a blood supply from the systemic circulation. Multiple imaging modalities have been used to demonstrate the vascular anatomy of the sequestration. Different magnetic resonance angiography (MRA) techniques have been employed in the identification of these anomalous vessels. We report a case of pulmonary sequestration diagnosed by MRI with the use of contrast enhanced three-dimensional MRA.

Adult↗
PubMed1999

Imaging features of focal intrahepatic extramedullary haematopoiesis.

The imaging findings of focal intrahepatic extramedullary haematopoiesis (EMH) in a 51-year-old woman with beta-thalassaemia intermedia are described with particular reference to MRI and CT. Bone marrow colloid scintigraphy was unhelpful in confirming the diagnosis, which was made from fine needle aspiration. This is the first description of stellate scars occurring in an EMH lesion. A review of the radiological appearances of this rare condition is presented.

Female↗
PubMed1999

MR cholangiography and CT cholangiography of pediatric patients with choledochal cysts.

OBJECTIVE: We report our experience using MR cholangiography and CT cholangiography in pediatric patients with choledochal cysts. SUBJECTS AND METHODS: Fourteen patients (two boys, 12 girls; mean age, 7.8 years) with either a preoperative diagnosis of choledochal cyst or a surgical finding of choledochal cyst underwent non-breath-hold MR cholangiography using T2-weighted fat-suppressed fast spin-echo sequences with a 1.5-T magnet, and CT cholangiography with IV infusion of meglumine iodoxamic acid. Radiologic findings were correlated with findings from surgery, operative cholangiography, or percutaneous transhepatic cholangiography. RESULTS: The biliary tree was visualized in all 14 patients with MR cholangiography and in 13 patients (92.9%) with CT cholangiography. In the 11 preoperative cases of choledochal cyst, MR cholangiography correctly showed all cysts and CT cholangiography showed 10 cysts (90.9%). The quality of images on CT cholangiography and MR cholangiography was comparable. The sensitivity of CT cholangiography and MR cholangiography in revealing intrahepatic stones was 83.3% and 66.7%, respectively; the specificity for both techniques was 100%. The rate of detecting the pancreatic duct and the common channel by CT cholangiography and MR cholangiography was 63.6% and 45.5% respectively. After surgery, CT cholangiography was superior to MR cholangiography in revealing the location of biliary-enteric anastomosis and the extent of anastomotic narrowing. CONCLUSION: Because non-breath-hold MR cholangiography is not invasive and does not use ionizing radiation and potentially toxic contrast agents, it is recommended as the imaging technique of choice in children with choledochal cysts. CT cholangiography can be considered as an adjunct after surgery and in patients in whom MR cholangiography is unsatisfactory.

Adolescent↗
PubMed1998

Clinical outcome of micrometastasis in the lung in stage IA persistent gestational trophoblastic disease.

BACKGROUND: Computed tomography (CT) of the thorax can be used in the staging of persistent gestational trophoblastic disease (PGTD). However, the prognostic significance of micrometastasis in the lung detected by CT of the thorax has not been well documented. The aim of the study is to define the effect of micrometastasis on the clinical course of the disease. METHODS: Thirty-five patients who had nonmetastatic GTD underwent CT thorax examination before treatment in the Department of Obstetrics and Gynaecology, University of Hong Kong. All patients had workups which showed no evidence of metastasis and were diagnosed as FIGO stage IA. They all received methotrexate (MTX) infusion therapy. RESULTS: Three groups of patients were identified based on the thorax CT findings. Sixteen patients (45.7%) showed no evidence of micrometastasis on CT thorax. Two of them (12.5%) had poor response to MTX with unsatisfactory fall in serum hCG levels requiring change of chemotherapy to actinomycin D. Nine patients had suspicious micrometastasis and one (11.1%) of them needed change of MTX. Ten patients had micrometastasis and one (10%) of them needed change of MTX. There was only one recurrence and it was in the suspicious micrometastasis group (11.1%). There was no statistically significant difference in the rate of poor drug response or recurrence among the three groups of patients. CONCLUSIONS: Micrometastases in the lung do not affect the clinical outcome of patients with FIGO stage IA disease. CT thorax is not essential in the staging of GTD.

Adult↗
PubMed1998

Gadolinium-infusion magnetic resonance angiogram: a new, noninvasive, and accurate method of preoperative localization of impalpable undescended testes.

PURPOSE: This study investigates the usefulness of Gadolinium (Gd)-infusion magnetic resonance angiogram (MRA) in the detection of impalpable undescended testes. METHODS: Magnetic resonance (MRI) examination was performed in 14 patients with 17 impalpable undescended testes (unilateral, n = 11; bilateral, n = 3). MRAwasthen performed as follows: Gadolinium-DTPA-BMA (Omniscan) at 0.3 mmol/kg body weight was injected intravenously, and dynamic coronal examination using fast spoiled gradient recalled sequences was obtained with images taken at early arterial and delay venous phases. The testis was located by detection of the enhanced pampiniform venous plexus. RESULTS: Of the 17 undescended testes, 11 canalicular hypoplastic testes and 3 intraabdominal testes were detected in both routine MRI and MRA. Three atrophic testes were found in the scrotum by MRA but not detected by routine MRI. The detection rates of impalpable testes by MRI and MRA were 82.4% and 100%, respectively. All imaging findings were confirmed by surgical exploration. There was no morbidity associated with MRA. CONCLUSION: Gd-infusion MRA is an accurate and sensitive method of preoperative localization of impalpable testes and is superior to conventional MRI in the detection of atrophic testes.

Adolescent↗
PubMed1998

Prejunctionally mediated inhibition of neurotransmission by isoprenaline in rat vas deferens.

Effects of isoprenaline on monophasic contractions evoked by electric field stimulation were studied in rat isolated prostatic vas deferens. Isoprenaline reduced electrically evoked contractions (EC50: 0.27 +/- 0.05 microM), and propranolol concentration-dependently antagonized the effect of isoprenaline. In contrast, isoprenaline (0.3-3 microM) did not affect the contractile response induced by exogenous noradrenaline or ATP, while forskolin (100 nM) attenuated agonist-induced contraction. In some tissues, adrenergic and purinergic components of the electrically evoked contraction were isolated by exposure to alpha,beta-methylene ATP (3 microM) and prazosin (3 microM), respectively. Isoprenaline induced a greater inhibition of purinergic than adrenergic component of the electrically evoked contraction. Iberiotoxin (50 nM), glibenclamide (3 microM), 4-aminopyridine (0.3 mM) and tetraethylammonium ions (1 mM) attenuated the effect of isoprenaline. These results indicate that isoprenaline-induced inhibition of the electrically evoked (both purinergic and adrenergic) contraction was mediated primarily through activation of prejunctional beta-adrenoceptors, which probably inhibited release of contractile transmitters from sympathetic nerves supplying vas deferens. Lack of effect of isoprenaline on agonist-induced contraction does not favour a functional role of beta-adrenoceptors in vas smooth muscle.

Adrenergic beta-Agonists↗
PubMed1998

Characterization of glycoconjugates of guinea pig seminal vesicle by lectin histochemistry.

In the present study, the expression of glycoconjugates in the guinea pig seminal vesicle was localized and partially characterized by lectin histochemistry using a battery of 30 different lectins specific for different carbohydrate residues. The results indicate that the glandular epithelium of the guinea pig seminal vesicle exhibits complex glycoconjugates rich in Man, beta-GlcNAc, beta-Gal, alpha/beta-GalNAc, Fuc and complex NeuAc(alpha2,6)Gal/GalNAc residues, as shown by its positive reactions to most lectins used. The Golgi region of the luminal secretory epithelial cells expresses a complex glycoconjugate pattern, as shown by its strong reactions to Man-(PSA, GNA), beta-GlcNAc-(S-WGA, PWA, DSA, UDA), beta-Gal-(RCA-I and -II), alpha/beta-GalNAc-(SBA, Jac, VVA, BPA) and complex NeuAc-(SNA) specific lectins, indicating that the secretory epithelial cells are active in glycosylation and secretion process. It was also shown in the present study that the basal and luminal epithelial cells are different in their glycoconjugates. The basal epithelial cells are rich in NeuAc(alpha2,3)Gal residues as they are stained specifically by MAA. The fibroblasts in the epithelial-smooth muscle interface and the smooth muscle cells close to the glandular epithelium are shown to express more glycoconjugates as they are stained intensely by GS-I-B4, GS-II and SBA. However, their role in the epithelial-stromal interaction in the seminal vesicle remains to be elucidated. In summary, the present study reports for the first time on the lectin binding patterns of the guinea pig seminal vesicle, and the results show that the seminal vesicle epithelium elaborates and secretes glycoconjugates in a complex pattern. Some of the lectins might be useful as histochemical markers for the secretory activity and specific structural components in the guinea pig seminal vesicle.

Acetylgalactosamine↗
PubMed1998

Adrenal extramedullary haemopoiesis: diagnosis by a non-invasive method.

Bilateral adrenal masses were discovered incidentally in a patient with beta-thalassaemia intermedia. Endocrine investigations showed that the adrenal lesion was hormonally inactive. Extramedullary haemopoiesis involving the adrenal glands was suggested by the presence of reticuloendothelial tissue as demonstrated by bone marrow scintigraphy using Technetium Tc-99m nanocolloid. This report illustrates the use of non-invasive functional imaging techniques in the management of adrenal 'incidentalomas', which in this case turned out to be a rare presentation of extramedullary haemopoiesis.

Adrenal Gland Neoplasms↗
PubMed1998

Lumbar spine magnetic resonance imaging: comparison between fast spin echo proton density and spin echo T1 axial scans.

Spin echo (SE) T1 axial scans are routinely obtained in magnetic resonance imaging of the lumbar spine in many centres. This study directly compared matched SE T1 and fast SE (FSE) proton density (PD) axial scans. Both SE T1 and FSE PD axial scans of the lumbar spine were obtained in 116 consecutive patients. The imaging parameters (field-of-view, slice thickness, interslice gap, number of excitations and matrix size) and scan levels were identical for each pair of sequences. At two selected levels, L4/5 and L5/S1, various structures were independently graded by two observers. In 232 lumbar levels analysed, the bone marrow, epidural fat, disc, extradural nerve root and facet joint were equally well seen on both sequences by both observers (combined mean grades of 2.93-2.99). The thecal sac was marginally better depicted on FSE PD than on SE T1 images, with mean grades of 2.96 and 2.88, respectively. The psoas muscle was adequately visualized for diagnostic purposes on both sequences (mean grades of 2.30-2.32). The cauda equina were better seen on FSE PD (mean grade 1.92) than on SE T1 (mean grade 1.00) images. In conclusion, FSE PD scans are comparable to and may potentially replace SE T1 axial MR scans of the lumbar spine.

Humans↗
PubMed1997

Evaluation of cervical nodal necrosis in nasopharyngeal carcinoma by computed tomography: incidence and prognostic significance.

PURPOSE: The purpose was to study the prognostic value of contrast-enhanced computed tomography (CT) nodal necrosis in nasopharyngeal carcinoma. PATIENTS AND METHODS: One hundred sixty-one patients with newly diagnosed nasopharyngeal carcinoma and nodal metastases were reviewed. Forty patients also received cisplatin-based neoadjuvant chemotherapy in addition to radiotherapy. Nodal necrosis was defined as presence of hypodense areas in more than 33% of the node. Nodal response rate to chemotherapy, overall nodal control rate, local control rate, distant failure rate, overall relapse-free survival rate, and overall and cause-specific survival rates were compared between patients with and without nodal necrosis. Multivariate analysis was also performed. RESULTS: The incidence of nodal necrosis was 22.9%. Overall nodal response rates to chemotherapy were 88.9% (8/9) in patients with nodal necrosis and 74.2% (23/31) in those without. No significant differences in nodal control rate, local control rate, distant failure rate, and overall and cause-specific survival rates were found. Five-year overall relapse-free survival rate was lower in patients with cervical nodal necrosis (36%) as compared with those without (53%, p = .04). Multivariate analysis, however, did not confirm cervical nodal necrosis to be an independent prognostic factor. CONCLUSIONS: Presence of nodal necrosis in nasopharyngeal carcinoma does not affect nodal response to chemotherapy and nodal control by radiotherapy with or without chemotherapy. Cervical nodal necrosis does not appear to be an independent factor in predicting treatment outcome. Further studies to correlate nodal density with oxygenation status as well as tumor cell kinetics are warranted.

Adult↗
PubMed1997

Hyaluronan and chondroitin sulfate proteoglycans are colocalized to the ciliary zonule of the rat eye: a histochemical and immunocytochemical study.

In previous studies, chondroitin sulfate proteoglycans have been localized to the periphery of the zonular fibers and the individual zonular fibrils (or microfibrils) after Cuprolinic blue staining in conjunction with chondroitinase digestions and immunogold labelling with 2-B-6 antibody. In the present study, we wished to determine if these proteoglycans are linked to hyaluronan to form a large multimolecular aggregate. To accomplish this, we localized the hyaluronan using a biotinylated hyaluronan-binding protein fragment of chondroitin sulfate proteoglycan, containing also the link protein, purified from bovine nasal cartilage. The results showed that the ciliary zonule of the rat eye was reactive with the biotinylated hyaluronan-binding probe as demonstrated by streptavidin-peroxidase-diaminobenzidine staining and streptavidin-gold labelling. Hyaluronan-gold labelling showed that the gold particles were mostly localized on the periphery of the zonular fibers, which was similar to the localization pattern of the zonule associated-proteoglycans. This hyaluronan-binding probe also strongly labelled the sites of zonule insertion over the basement membrane of the inner ciliary epithelium at the pars plana and the lens capsule at the equatorial region, which suggests its probable role in the attachment of ciliary zonule to the basement membranes. To demonstrate whether these two molecules are linked to one another, ultrastructural colocalization of both hyaluronan and chondroitin sulfate proteoglycans was performed on the same sections by double-gold labelling, and combined Cuprolinic blue staining and hyaluronan-gold labelling. Gold particles of 15 and 10 nm in sizes labelling both hyaluronan and chondroitin 4-sulfate, were colocalized to the surface of the zonular fibers. The combined Cuprolinic blue staining and hyaluronan-gold labelling showed that the gold particles were localized towards the ends of the Cuprolinic blue-stained rodlets, which strongly suggests that these chondroitin sulfate proteoglycans are linked to the hyaluronan chain to form a large aggregate surrounding the periphery of the zonular fibers. These ciliary zonule-associated proteoglycan-hyaluronan aggregates may play a role in organizing the individual zonular fibrils (microfibrils) into bundles of zonular fibers.

Animals↗