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1.
Quant Imaging Med Surg ; 12(10): 4734-4746, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36185050

RESUMO

Background: The purpose of the present proof-of-concept study was to use large-area in vivo confocal laser scanning microscopy (CLSM) mosaics to determine the migration rates of nerve branching points in the human corneal subbasal nerve plexus (SNP). Methods: Three healthy individuals were examined roughly weekly over a total period of six weeks by large-area in vivo confocal microscopy of the central cornea. An in-house developed prototype system for guided eye movement with an acquisition time of 40 s was used to image and generate large-area mosaics of the SNP. Kobayashi-structures and nerve entry points (EPs) were used as fixed structures to enable precise mosaic registration over time. The migration rate of 10 prominent nerve fiber branching points per participant was tracked and quantified over the longitudinal period. Results: Total investigation times of 10 minutes maximum per participant were used to generate mosaic images with an average size of 3.61 mm2 (range: 3.18-4.42 mm2). Overall mean branching point migration rates of (46.4±14.3), (48.8±15.5), and (50.9±13.9) µm/week were found for the three participants with no statistically significant difference. Longitudinal analyses of nerve branching point migration over time revealed significant time-dependent changes in migration rate only in participant 3 between the last two measurements [(63.7±12.3) and (43.0±12.5) µm/week, P<0.01]. Considering individual branching point dynamics, significant differences in nerve migration rate from the mean were only found in a few exceptions. Conclusions: The results of this proof-of-concept study have demonstrated the feasibility of using in vivo confocal microscopy to study the migration rates of corneal subbasal nerves within large areas of the central human cornea (>1 mm2). The ability to monitor dynamic changes in the SNP opens a window to future studies of corneal nerve health and regenerative capacity in a number of systemic and ocular diseases. Since corneal nerves are considered part of the peripheral nervous system, this technique could also offer an objective diagnostic tool and biomarker for disease- or treatment-induced neuropathic changes.

2.
Diagnostics (Basel) ; 11(2)2021 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-33494468

RESUMO

In vivo large-area confocal laser scanning microscopy (CLSM) of the human eye using EyeGuidance technology allows a large-scale morphometric assessment of the corneal subbasal nerve plexus (SNP). Here, the SNP of a patient suffering from diabetes and associated late complications was analyzed. The SNP contained multiple clusters of large hyperintense, stellate-shaped, cellular-like structures. Comparable structures were not observed in control corneas from healthy volunteers. Two hypotheses regarding the origin of these atypical structures are proposed. First, these structures might be keratocyte-derived myofibroblasts that entered the epithelium from the underlying stroma through breaks in Bowman's layer. Second, these structures could be proliferating Schwann cells that entered the epithelium in association with subbasal nerves. The nature and pathophysiological significance of these atypical cellular structures, and whether they are a direct consequence of the patient's diabetic neuropathy/or a non-specific secondary effect of associated inflammatory processes, are unknown.

3.
Invest Ophthalmol Vis Sci ; 60(15): 4972-4984, 2019 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-31790560

RESUMO

Purpose: The subbasal nerve plexus (SNP) is the densest and most recognizable component of the mammalian corneal innervation; however, the anatomical configuration of the SNP in most animal models remains incompletely described. The purpose of the current study is to describe in detail the SNP architecture in eight different mammals, including several popular animal models used in cornea research. Methods: Corneal nerves in mouse, rat, guinea pig, rabbit, dog, macaque, domestic pig, and cow eyes were stained immunohistochemically with antiserum directed against neurotubulin. SNP architecture was documented by digital photomicrography and large-scale reconstructions, that is, corneal nerve maps, using a drawing tube attached to a light microscope. Results: Subbasal nerve fibers (SNFs) in mice, rats, guinea pigs, dogs, and macaques radiated centrally from the corneoscleral limbus toward the corneal apex in a whorl-like or spiraling pattern. SNFs in rabbit and bovine corneas swept horizontally across the ocular surface in a temporal-to-nasal direction and converged on the inferonasal limbus without forming a spiral. SNFs in the pig cornea radiated centrifugally in all directions, like a starburst, from a focal point located equidistant between the corneal apex and the superior pole. Conclusions: The results of the present study have demonstrated for the first time substantial interspecies differences in the architectural organization of the mammalian SNP. The physiological significance of these different patterns and the mechanisms that regulate SNP pattern formation in the mammalian cornea remain incompletely understood and warrant additional investigation.


Assuntos
Anatomia Comparada , Córnea/inervação , Nervo Oftálmico/anatomia & histologia , Animais , Bovinos , Cães , Cobaias , Macaca , Camundongos , Microscopia Confocal , Modelos Animais , Fibras Nervosas , Coelhos , Ratos , Suínos , Gânglio Trigeminal/anatomia & histologia
4.
Vet Ophthalmol ; 19(3): 230-6, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26076886

RESUMO

OBJECTIVE: The lacrimal gland (LG) and the third eyelid gland (TELG) are two intraorbital glands that, in dogs, secrete the aqueous component of the tear film. Despite the central importance of these structures for maintaining ocular surface health, the gross anatomy of the glands remains understudied. We investigated the macroscopic morphometric characteristics of the LG and TELG in three different dog breeds. PROCEDURES: Twenty-six dog heads were dissected to expose the LG and TELG; the length, width, thickness, and weight of each were measured. During the dissections, the relationships between the glands and adjacent ocular structures and the blood and nerve supplies to the LG were photo-documented. RESULTS: The LG had a flat and irregular shape with morphological variations among dogs. The LG was located on the dorsolateral aspect of the globe underneath the orbital ligament. The average length, width, and thickness (SEM) of the LG (mm) were 16.5 ± 0.7, 12.5 ± 0.4, and 2.7 ± 0.1 and of the TELG 10.5 ± 0.6, 11.0 ± 0.3, and 3.3 ± 0.1, respectively. The mean weights (SD) of the LG and TELG (mg) were 315.7 ± 21.1 and 263.3 ± 13.2, respectively. Beagles were observed to have significantly smaller LGs compared to pit bull terriers and pointer mixed-breed dogs. CONCLUSIONS: The present study provides detailed normative anatomical and morphometric data for the LG and TELG. These data will aid researchers investigating alterations induced by disease states and should inform strategies for the local delivery of pharmacologic and cellular therapeutics.


Assuntos
Cães/anatomia & histologia , Aparelho Lacrimal/anatomia & histologia , Membrana Nictitante/anatomia & histologia , Animais , Feminino , Masculino
5.
Invest Ophthalmol Vis Sci ; 56(13): 8125-40, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26720465

RESUMO

PURPOSE: In an effort to elucidate possible neural mechanisms underlying diminished tearing in dry eye disease, this study sought to determine if hyperosmolar tears, a ubiquitous sign of dry eye disease, produce functional changes in corneal nerve responses to drying of the cornea and if these changes correlate with alterations in corneal nerve morphology. METHODS: In vivo extracellular electrophysiological recordings were performed in rat trigeminal ganglion neurons that innervated the cornea before, and up to 3 hours after, the ocular application of continuous hyperosmolar tears or artificial tears. In corollary experiments, immunohistochemical staining was performed to compare corneal nerve morphology in control and in eyes treated with hyperosmolar solutions. RESULTS: Our previous studies identified a population of corneal afferents, dry-sensitive neurons that are strongly excited by corneal dessication ("dry response"), a response thought to trigger the lacrimation reflex. In the present study, we found that the dry responses of corneal dry-sensitive neurons were depressed or even completely abolished by hyperosmolar tears in a time- (30 minutes to 3 hours) and dose (450- to 1000-mOsm solutions)-dependent manner. Furthermore, eyes treated with hyperosmolar tears for 3 hours contained large numbers of morphologically abnormal (granular, fragmented, or prominently beaded) subbasal nerves that appeared to be undergoing degeneration. CONCLUSIONS: These results demonstrate that tear hyperosmolarity, considered to be a "core" mechanism of dry eye disease, significantly decreases physiological sensitivity and morphologic integrity of the corneal nerves important in tear production. These alterations might contribute to the diminished tearing seen clinically in dry eye patients.


Assuntos
Córnea/inervação , Síndromes do Olho Seco/metabolismo , Neurônios Aferentes/fisiologia , Lágrimas/química , Gânglio Trigeminal/fisiologia , Animais , Modelos Animais de Doenças , Síndromes do Olho Seco/patologia , Síndromes do Olho Seco/fisiopatologia , Fenômenos Eletrofisiológicos , Imuno-Histoquímica , Masculino , Concentração Osmolar , Ratos , Ratos Sprague-Dawley
6.
Am J Pathol ; 184(6): 1652-9, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24828391

RESUMO

Peripheral neuropathy is the most frequent neurological complication of HIV infection, affecting more than one-third of infected patients, including patients treated with antiretroviral therapy. Although emerging noninvasive techniques for corneal nerve assessments are increasingly being used to diagnose and monitor peripheral neuropathies, corneal nerve alterations have not been characterized in HIV. Here, to determine whether SIV infection leads to corneal nerve fiber loss, we immunostained corneas for the nerve fiber marker ßIII tubulin. We developed and applied both manual and automated methods to measure nerves in the corneal subbasal plexus. These counting methods independently indicated significantly lower subbasal corneal nerve fiber density among SIV-infected animals that rapidly progressed to AIDS compared with slow progressors. Concomitant with decreased corneal nerve fiber density, rapid progressors had increased levels of SIV RNA and CD68-positive macrophages and expression of glial fibrillary acidic protein by glial satellite cells in the trigeminal ganglia, the location of the neuronal cell bodies of corneal sensory nerve fibers. In addition, corneal nerve fiber density was directly correlated with epidermal nerve fiber length. These findings indicate that corneal nerve assessment has great potential to diagnose and monitor HIV-induced peripheral neuropathy and to set the stage for introducing noninvasive techniques to measure corneal nerve fiber density in HIV clinical settings.


Assuntos
Infecções por HIV , HIV-1 , Doenças do Sistema Nervoso Periférico , Síndrome de Imunodeficiência Adquirida dos Símios , Vírus da Imunodeficiência Símia , Animais , Córnea/inervação , Córnea/metabolismo , Córnea/patologia , Epiderme/inervação , Epiderme/metabolismo , Epiderme/patologia , Proteína Glial Fibrilar Ácida/metabolismo , Infecções por HIV/metabolismo , Infecções por HIV/prevenção & controle , Macaca nemestrina , Fibras Nervosas/metabolismo , Fibras Nervosas/patologia , Doenças do Sistema Nervoso Periférico/metabolismo , Doenças do Sistema Nervoso Periférico/patologia , Síndrome de Imunodeficiência Adquirida dos Símios/metabolismo , Síndrome de Imunodeficiência Adquirida dos Símios/patologia
7.
Invest Ophthalmol Vis Sci ; 54(11): TFOS71-97, 2013 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-24058137

RESUMO

This report characterizes the neurobiology of the ocular surface and highlights relevant mechanisms that may underpin contact lens-related discomfort. While there is limited evidence for the mechanisms involved in contact lens-related discomfort, neurobiological mechanisms in dry eye disease, the inflammatory pathway, the effect of hyperosmolarity on ocular surface nociceptors, and subsequent sensory processing of ocular pain and discomfort have been at least partly elucidated and are presented herein to provide insight in this new arena. The stimulus to the ocular surface from a contact lens is likely to be complex and multifactorial, including components of osmolarity, solution effects, desiccation, thermal effects, inflammation, friction, and mechanical stimulation. Sensory input will arise from stimulation of the lid margin, palpebral and bulbar conjunctiva, and the cornea.


Assuntos
Lentes de Contato/efeitos adversos , Oftalmopatias/etiologia , Grupos Focais , Humanos , Neurobiologia
8.
Vet Ophthalmol ; 16(4): 282-8, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22958361

RESUMO

OBJECTIVE: To describe clinical, in vivo confocal microscopic, histopathologic, and immunohistochemical features of a dog with metaherpetic corneal disease that developed subsequent to a protracted episode of canine herpesvirus-1 (CHV-1) dendritic ulcerative keratitis. CASE DESCRIPTION: A 7-year-old, spayed-female, Miniature Schnauzer was treated for bilateral CHV-1 dendritic ulcerative keratitis. Following resolution of ulcerative keratitis, sectoral peripheral superficial corneal gray opacification, vascularization, and pigmentation slowly migrated centripetally to the axial cornea of both eyes. Corneal sensitivity measured with a Cochet-Bonnet esthesiometer was dramatically and persistently reduced. In vivo corneal confocal microscopic examination revealed regions of epithelium with a conjunctival phenotype. In these areas, the surface epithelium was thin, disorganized, and composed of hyper-reflective epithelial cells. Goblet cells and Langerhans cells were frequent, and the subbasal nerve plexus was completely absent or markedly diminished. Histopathologic abnormalities in the globes were restricted to the superficial cornea and included sectoral corneal conjunctivalization, increased anterior stromal spindle cells, and vascularization. Immunohistochemical evaluation of the corneas with anti-neurotublin antibody demonstrated attenuation of the epithelial and subbasal nerve plexuses with marked stromal hyperinnervation and increased numbers of morphologically abnormal neurites. CONCLUSIONS: Similar to herpes simplex virus keratitis in humans, CHV-1 ulcerative keratitis may be associated with the development of chronic degenerative corneal disease in dogs. In the described dog, this chronic corneal disease included progressive corneal opacification because of partial limbal stem cell deficiency and neurotrophic keratitis. Long-term monitoring of dogs following resolution of active CHV-1 keratitis may be indicated, particularly when ulcerations persist for an extended period.


Assuntos
Doenças do Cão/etiologia , Infecções por Herpesviridae/veterinária , Herpesvirus Canídeo 1/isolamento & purificação , Ceratite/veterinária , Células-Tronco/patologia , Animais , Doenças do Cão/patologia , Doenças do Cão/virologia , Cães , Feminino , Infecções por Herpesviridae/complicações , Infecções por Herpesviridae/patologia
9.
Exp Eye Res ; 90(4): 478-92, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20036654

RESUMO

The anatomy of the human corneal innervation has been the subject of much investigation; however, a comprehensive description remains elusive. The purpose of the present study was to provide a detailed description of the human corneal innervation using a novel approach involving immunohistochemically stained anterior-cornea whole mounts. Sixteen donor corneas aged 19-78 years were cut with a 6.0 mm trephine into a central plug and two peripheral rims. Each specimen was sectioned serially on a cryostat to produce several 100 microm-thick stromal sections and a 100-140 microm-thick anterior-cornea whole mount that contained the entire corneal epithelium and much of the anterior stroma. The corneal innervation was stained with a primary antibody against beta neurotubulin and subjected to rigorous quantitative and qualitative analyses. The results showed that a mean of 71.3 +/- 14.3, uniformly spaced, main stromal nerve bundles entered the cornea at the corneoscleral limbus. The bundles averaged 20.3 +/- 7.0 microm in diameter, were separated by a mean spacing of 0.49 +/- 0.40 mm, and entered the cornea at a mean distance of 293 +/- 106 microm from the ocular surface. Each stromal bundle gave rise through repetitive branching to a moderately dense midstromal plexus and a dense subepithelial plexus (SEP). The SEP was comprised of modest numbers of straight and curvilinear nerves, most of which penetrated Bowman's membrane to supply the corneal epithelium, and a more abundant and anatomically complex population of tortuous, highly anastomotic nerves that remained largely confined in their distribution to the SEP. SEP density and anatomical complexity varied considerably among corneas and was less dense and patchier in the central cornea. A mean of 204 +/- 58.5 stromal nerves penetrated Bowman's membrane to supply the central 10 mm of corneal epithelium (2.60 nerves/mm(2)). The density of Bowman's membrane penetrations was greater peripherally than centrally. After entering the epithelium, stromal nerves branched into groups of up to twenty subbasal nerve fibers known as epithelial leashes. Leashes in the central and intermediate cornea anastomosed extensively to form a dense, continuous subbasal nerve plexus, while leashes in the peripheral cornea demonstrated fewer anastomoses and were less complex anatomically. Viewed in its entirety, the subbasal nerve plexus formed a gentle, whorl-like assemblage of long curvilinear subbasal fibers, 1.0-8.0 mm in length, that converged on an imaginary seam or gentle spiral (vortex) approximately 2.51 +/- 0.23 mm inferonasal to the corneal apex. Mean subbasal nerve fiber density near the corneal apex was 45.94 +/- 5.20 mm/mm(2) and mean subbasal and interconnecting nerve fiber diameters in the same region were 1.51 +/- 0.74 microm and 0.69 +/- 0.26 microm, respectively. Intraepithelial terminals originated exclusively as branches of subbasal nerves and terminated in all epithelial layers. Nerve terminals in the wing and squamous cell layers were morphologically diverse and ranged in total length from 9 to 780 microm. The suprabasal layers of the central corneal epithelium contained approximately 605.8 terminals/mm(2). The results of this study provide a detailed, comprehensive description of human corneal nerve architecture and density that extends and refines existing accounts. An accurate, detailed model of the normal human corneal innervation may predict or help to understand the consequences of corneal nerve damage during refractive, cataract and other ocular surgeries.


Assuntos
Córnea/inervação , Nervo Oftálmico/anatomia & histologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Substância Própria/inervação , Epitélio Corneano/inervação , Humanos , Técnicas Imunoenzimáticas , Pessoa de Meia-Idade , Fibras Nervosas , Nervo Oftálmico/metabolismo , Tubulina (Proteína)/metabolismo , Adulto Jovem
10.
Invest Ophthalmol Vis Sci ; 49(3): 910-6, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18326711

RESUMO

PURPOSE: To determine the effect of aging on corneal epithelial nerve density in an animal model. METHODS: Corneal whole mounts from rats aged 6, 12, 18, and 24 months were stained immunohistochemically with antisera against the pan-neuronal marker neurotubulin. Epithelial nerve terminals and subbasal nerves in standardized 1-mm(2) central and peripheral zones from each cornea were drawn using a drawing tube attached to a light microscope. Images were scanned, and nerve densities were calculated as the percentage of each 1-mm(2) area occupied by nerves. The diameters of subbasal nerves in 6- and 24-month old animals were measured. Subbasal nerve vortices were analyzed qualitatively with reference to location, morphologic appearance, and directionality. RESULTS: Epithelial nerve terminal density decreased by approximately 50% between 6 and 24 months. The rate of decline was roughly linear and similar in both central and peripheral cornea. In contrast, subbasal nerve density increased by more than 50% between 6 and 24 months in both central and peripheral cornea. The mean diameter of corneal subbasal nerves decreased approximately 30% (0.384 microm vs. 0.271 microm) between 6 and 24 months. The morphologic appearance and directionality of the subbasal nerve vortex demonstrated considerable interanimal variability and did not correlate with age. CONCLUSIONS: Rat corneal nerve terminal density decreases, but corneal subbasal nerve density increases, as a function of age. The age-related loss of nerve terminal density seen in the rat cornea is in keeping with the decreased corneal sensitivity reported in elderly humans and may contribute to the pathogenesis of dry eye disease in aged persons.


Assuntos
Envelhecimento/fisiologia , Epitélio Corneano/inervação , Nervo Oftálmico/citologia , Animais , Técnicas Imunoenzimáticas , Terminações Nervosas/citologia , Fibras Nervosas , Nervo Oftálmico/metabolismo , Ratos , Ratos Endogâmicos F344 , Tubulina (Proteína)/metabolismo
11.
Exp Eye Res ; 80(5): 633-42, 2005 May.
Artigo em Inglês | MEDLINE | ID: mdl-15862170

RESUMO

Nerve growth factor in the tear film and corneal epithelium is hypothesized to play an important role in ocular surface maintenance and corneal wound healing. The purpose of this study was to determine the expression of nerve growth factor and its high affinity (trkA) receptor in tears, cornea, and lacrimal glands of normal dogs, the modulation of nerve growth factor and its trkA receptor during corneal wound healing, and the effect of topical nerve growth factor application on canine corneal epithelial wound healing. In the first of three experiments, the nerve growth factor content of tears, corneal epithelium, lacrimal gland, and 3rd eyelid gland was determined in normal dogs by enzyme-linked immunosorbent assay and the expression of nerve growth factor and its trkA receptor were evaluated in the cornea and lacrimal glands by immunohistochemistry. In a second experiment, unilateral corneal epithelial defects were created, and tissues were evaluated for changes in nerve growth factor or trkA expression for 1 week. In a third experiment, bilateral corneal epithelial defects were created and the right eyes in each animal were treated 4 times daily with either recombinant human nerve growth factor, murine nerve growth factor, or nerve growth factor-blocking antibody. The results of this study showed that nerve growth factor levels in normal dog tears, corneal epithelium, third eyelid gland and lacrimal gland were 15.4+/-4.6 ng ml(-1), 33.5+/-12.3, 52.4+/-17.4 and 48.8+/-9.4 ng g(-1), respectively. NGF and trkA receptors were identified by immunohistochemistry in all tissues examined. After unilateral corneal wounding, nerve growth factor concentration increased in the tears bilaterally for 3 days, especially in the wounded eye, and then returned to pre-wounding values. Nerve growth factor content, and immunohistochemical staining for nerve growth factor and trkA, increased significantly in the ipsilateral cornea epithelium following unilateral wounding. Nerve growth factor concentrations in lacrimal and third eyelid glands also increased bilaterally (p<0.01) after unilateral wounding. Time to wound closure and rate of epithelial migration did not differ significantly between nerve growth factor-treated, nerve growth factor antibody-treated, and control eyes. In conclusion, nerve growth factor is present under resting physiologic conditions in normal canine tears, and nerve growth factor and its trkA receptor are present under resting conditions in normal canine corneal epithelium, lacrimal gland and third eyelid gland. Nerve growth factor is elevated in the tears, cornea, and lacrimal glands after corneal epithelial wounding; however, topical application of nerve growth factor, or its blocking antibody does not modulate corneal wound healing in the normal dog eye.


Assuntos
Epitélio Corneano/química , Epitélio Corneano/lesões , Fator de Crescimento Neural/análise , Cicatrização/fisiologia , Animais , Anticorpos Bloqueadores/administração & dosagem , Cães , Epitélio Corneano/patologia , Pálpebras/química , Feminino , Humanos , Imunoensaio/métodos , Imuno-Histoquímica/métodos , Aparelho Lacrimal/química , Fator de Crescimento Neural/imunologia , Receptor trkA/análise , Proteínas Recombinantes/uso terapêutico , Lágrimas/química
12.
Mol Vis ; 11: 169-78, 2005 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-15765049

RESUMO

PURPOSE: Plasma membrane Ca2+-ATPases (PMCAs) are integral membrane proteins essential to the control of intracellular Ca2+ ([Ca2+]i) concentration. Four genes encode PMCA proteins termed PMCA1-PMCA4. Little is known about the expression of these isoforms in corneal epithelium (CE). The purpose of this investigation is to characterize the expression and distribution of PMCAs in human CE (hCE). METHODS: PMCA mRNA expression was examined by RT-PCR analysis of total RNA from native hCE using PMCA gene specific primers. PMCA isoform expression at the protein level in native hCE was examined by immunoblotting using isoform specific antibodies (Abs) and a panPMCA Ab that recognizes all PMCAs. Distribution of PMCAs in postmortem and surgical sections of hCE was determined by immunohistochemistry with the same Abs. RESULTS: Immunoblot analysis with the panPMCA Ab yielded an intense band of approximately 135 kDa and several faintly staining bands above and below this major band. The isoform specific Abs labeled one or more bands that corresponded to bands detected with the panPMCA Ab. RT-PCR analysis of total RNA from hCE yielded PCR DNAs that were identified by sequencing as products of PMCA1, PMCA2, PMCA3, and PMCA4, thus confirming the immunoblot data. Immunohistochemistry demonstrated localization of PMCAs in all layers of hCE. PMCA4 was the predominant isoform, and was expressed along the plasma membrane of cells in all layers of CE, except with a notable absence along the basal cell membranes adjacent to the stroma. PMCA1 and PMCA2 were found mainly on basal and wing cells. In contrast to PMCA4, PMCA1 immunoreactivity (IR) was located on portions of basal cell plasma membranes adjacent to the stroma. PMCA2 IR was detected cytoplasmically within basal and wing cells in both central cornea and limbus. PMCA3 IR was located in basal cell nuclei in central cornea, but in a perinuclear location in the limbal, basal, and wing cells. CONCLUSIONS: Human CE expresses multiple PMCA isoforms that are differentially expressed and localized among the layers and cells that comprise the CE. We propose that the differential expression of multiple PMCA isoforms affords CE the requisite flexibility to respond to the demands for Ca2+ regulation required during renewal and regeneration of its multiple cell types.


Assuntos
ATPases Transportadoras de Cálcio/genética , Proteínas de Transporte de Cátions/genética , Epitélio Corneano/enzimologia , Regulação Enzimológica da Expressão Gênica , Idoso , Anticorpos Monoclonais , Cálcio/metabolismo , ATPases Transportadoras de Cálcio/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Membrana Celular/enzimologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Immunoblotting , Isoenzimas , Masculino , Pessoa de Meia-Idade , ATPases Transportadoras de Cálcio da Membrana Plasmática , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
13.
Exp Eye Res ; 76(5): 521-42, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12697417

RESUMO

This review provides a comprehensive analysis of the structure, neurochemical content, and functions of corneal nerves, with special emphasis on human corneal nerves. A revised interpretation of human corneal nerve architecture is presented based on recent observations obtained by in vivo confocal microscopy (IVCM), immunohistochemistry, and ultrastructural analyses of serial-sectioned human corneas. Current data on the neurotransmitter and neuropeptide contents of corneal nerves are discussed, as are the mechanisms by which corneal neurochemicals and associated neurotrophins modulate corneal physiology, homeostasis and wound healing. The results of recent clinical studies of topically applied neuropeptides and neurotrophins to treat neurotrophic keratitis are reviewed. Recommendations for using IVCM to evaluate corneal nerves in health and disease are presented.


Assuntos
Córnea/inervação , Animais , Córnea/ultraestrutura , Humanos , Ceratite/etiologia , Microscopia Confocal , Fibras Nervosas/ultraestrutura , Neuropeptídeos/fisiologia , Neurotransmissores/fisiologia , Cicatrização
14.
Am J Vet Res ; 63(6): 906-15, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12061541

RESUMO

OBJECTIVE: To determine effects of cyclophotocoagulation via administration of 100 J with a neodymium:yttrium aluminum garnet (Nd:YAG) laser on corneal touch threshold (CTT), intraocular pressure (IOP), aqueous tear production, and corneal nerve morphology in eyes of dogs. ANIMALS: 15 dogs. PROCEDURE: Noncontact Nd:YAG laser was transsclerally applied (10 applications; 25 W for 0.1 seconds for each application to each of 4 quadrants) to the ciliary body of the left eye of 15 dogs; the right eye was the control eye. Corneal integrity, CTT, tear production as measured by the Schirmer tear test (STT), and IOP were evaluated for 14 days following laser treatment. On day 14, dogs were euthanatized, eyes harvested, and corneas stained with gold chloride. Major nerve bundles were analyzed by use of a drawing tube attached to a light microscope, and maximum diameters were measured by use of image analysis software. RESULTS: All laser-treated eyes had significantly higher CTT values, compared with control eyes. Six of 15 laser-treated eyes developed ulcerative keratitis. On most days, IOP was significantly lower in laser-treated eyes in both morning and evening. Laser-treated eyes had a significant decrease of approximately 1 nerve bundle/corneal quadrant. Values for STT or nerve bundle diameters did not differ significantly. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of 100 J with a Nd:YAG laser effectively reduced IOP while increasing CTT and caused a significant decrease in number, but not diameter, of major corneal nerve bundles. Nerve damage and corneal hypoesthesia are etiologic factors in ulcerative keratitis following Nd:YAG cyclophotocoagulation.


Assuntos
Córnea/fisiologia , Cães/fisiologia , Pressão Intraocular/fisiologia , Fotocoagulação a Laser/veterinária , Animais , Córnea/inervação , Córnea/cirurgia , Cães/cirurgia , Feminino , Compostos de Ouro/química , Fotocoagulação a Laser/efeitos adversos , Masculino , Nervo Oftálmico/patologia , Lágrimas/metabolismo
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