Molecular Vision最新文献

{"title":"Evaluation of <i>MYRF</i> as a candidate gene for primary angle closure glaucoma.","authors":"Xiaowei Yu,&nbsp;Nannan Sun,&nbsp;Congcong Guo,&nbsp;Zhenni Zhao,&nbsp;Meifang Ye,&nbsp;Jiamin Zhang,&nbsp;Jian Ge,&nbsp;Zhigang Fan","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Primary angle-closure glaucoma (PACG) is a leading cause of blindness. Despite tremendous human effort and financial input, no definitive causative gene has been identified either through genome-wide association or Mendelian family studies. In the current study, novel candidate genes for PACG were investigated by studying the variants of nanophthalmos-associated genes.</p><p><strong>Methods: </strong>A case-control study was conducted that included 45 PACG patients and 12 normal controls with short axial length (AL, less than 23.5 mm but more than 20.5 mm). Whole-exome sequencing (WES) was performed to screen the variants in previously identified nanophthalmos-associated genes, as well as other risk genes.</p><p><strong>Results: </strong>The age range of the 45 PACG patients was 24 to 80 years, with an average AL of 21.87±0.65 mm (range: 20.54-23.45 mm) in the right eye and 21.89±0.64 mm (range 20.60-23.23 mm) in the left eye. Four novel myelin regulatory factor (<i>MYRF</i>) gene missense variants (p.G117S, p.H1057R, p.H230R, and p.R316C) were identified in four out of the 45 enrolled PACG patients, respectively. No <i>MYRF</i> or other nanophthalmos-associated gene variants were detected in the 12 normal controls.</p><p><strong>Conclusions: </strong>An appropriate approach was adopted to investigate the genetics of PACG through nanophthalmos-associated genes. A genetic variant, <i>MYRF</i>, was identified in four out of 45 PACG patients, which might be a novel candidate gene for PACG.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"734-740"},"PeriodicalIF":2.2,"publicationDate":"2021-12-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/50/48/mv-v27-734.PMC8763663.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39777518","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recombinant adenovirus causes prolonged mobilization of macrophages in the anterior chambers of mice.","authors":"Kacie J Meyer, Danielle Pellack, Adam Hedberg-Buenz, Nicholas Pomernackas, Dana Soukup, Kai Wang, John H Fingert, Michael G Anderson","doi":"","DOIUrl":"","url":null,"abstract":"&lt;p&gt;&lt;strong&gt;Purpose: &lt;/strong&gt;Ocular tissues of mice have been studied in many ways using replication-deficient species C type 5 adenovirus (Ad5) as a tool for manipulating gene expression. Whereas refinements to injection protocols and tropism have led to several advances in targeting cells of interest, there remains a relative lack of information concerning how Ad5 may influence other ocular cell types capable of confounding experimental interpretation. Here, a slit lamp is used to thoroughly photodocument the sequelae of intraocular Ad5 injections over time in mice, with attention to potentially confounding indices of inflammation.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Methods: &lt;/strong&gt;A cohort of C57BL/6J mice was randomly split into three groups (Virus, receiving unilateral intracameral injection with 5×10&lt;sup&gt;7&lt;/sup&gt; plaque-forming units (pfu) of a cargo-less Ad5 construct; Saline, receiving unilateral balanced salt solution injection; and Naïve, receiving no injections). From this initial experiment, a total of 52 eyes from 26 mice were photodocumented via slit lamp at four time points (baseline and 1, 3, and 10 weeks following initiation of the experiment) by an observer masked to treatments and other parameters of the experimental design. Following the last in vivo exam, tissues were collected. Based on the slit-lamp data, tissues were studied via immunostaining with the macrophage marker F4/80. Subsequently, three iterations of the original experiment were performed with otherwise identical experimental parameters testing the effect of age, intravitreal injection, and A195 buffer, adding slit-lamp photodocumentation of an additional 32 eyes from 16 mice.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Results: &lt;/strong&gt;The masked investigator could use the sequential images from each mouse in the initial experiment to assign each mouse to its correct treatment group with near perfect fidelity. Virus-injected eyes were characterized by corneal damage indicative of intraocular injection and a prolonged mobilization of clump cells on the surface of the iris. Saline-injected eyes had only transient corneal opacities indicative of intraocular injections, and Naïve eyes remained normal. Immunostaining with F4/80 was consistent with ascribing the clump cells visualized via slit-lamp imaging as a type of macrophage. Experimental iterations using Ad5 indicate that all virus-injected eyes had the distinguishing feature of a prolonged presence of clump cells on the surface of the iris regardless of injection site. Mice receiving an intraocular injection of Ad5 at an advanced age displayed a protracted course of corneal cloudiness that prevented detailed visualization of the iris at the last time point.&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Conclusions: &lt;/strong&gt;Because the eye is often considered an \"immune privileged site,\" we suspect that several studies have neglected to consider that the presence of Ad5 in the eye might evoke strong reactions from the innate immune system. Ad5 injection caused a sustained mobilization of cl","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"741-756"},"PeriodicalIF":2.2,"publicationDate":"2021-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/fb/c6/mv-v27-741.PMC8763664.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41139128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ligustrazine protects against chronic hypertensive glaucoma in rats by inhibiting autophagy via the PI3K-Akt/mTOR pathway.","authors":"Hong-Yan Du,&nbsp;Rong Wang,&nbsp;Jian-Liang Li,&nbsp;Huang Luo,&nbsp;Xiao-Yan Xie,&nbsp;Ran Yan,&nbsp;Yue-Ling Jian,&nbsp;Jin-Ying Cai","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Glaucoma is a leading cause of global irreversible blindness, and characterized by the progressive loss of retinal ganglion cells (RGCs). Ligustrazine (TMP) is a natural product that has shown beneficial effects on various diseases. This study aimed to determine whether ligustrazine produces a therapeutic effect on glaucoma and to investigate its underlying mechanisms.</p><p><strong>Methods: </strong>A rat chronic hypertensive glaucoma model was induced by episcleral vein cauterization (EVC). Adult Sprague-Dawley (SD) rats were intraperitoneally administered TMP at a dose of 80 mg/kg once a day, from two days before EVC to one month after EVC. To elucidate the role of the mammalian target of rapamycin (mTOR) and phosphoinositide 3-kinase (PI3K), TMP-treated experimental rats were co-treated with the mTOR inhibitor rapamycin (5 mg/kg) or the PI3K inhibitor Ly294002 (10 mg/kg). The intraocular pressure (IOP) of the experimental and control rats was measured every six days. Retinal cells were examined by hematoxylin-eosin and terminal deoxynucleotidyltransferase-mediated biotinylated UTP nick end labeling (TUNEL) staining, as well as transmission electron microscopy. Immunohistochemistry and western blot analysis were performed to measure proteins involved in apoptosis and autophagy.</p><p><strong>Results: </strong>Ligustrazine protected retinal cells from death in experimental glaucoma rats, which was not due to the lowering of IOP, but could be attributable to direct suppression of retinal cell apoptosis. In glaucoma rats, autophagy was markedly activated in retina cells, as evidenced by increased numbers of autophagosomes and the expression of autophagy-related proteins (ATG5 and LC3-II/I). Notably, such alterations in glaucoma rats were almost completely reversed by ligustrazine. The suppressive effects of ligustrazine on apoptosis and autophagy of retina cells were markedly attenuated by the mTOR inhibitor rapamycin or the PI3K inhibitor Ly294002. Additionally, ligustrazine significantly increased the protein levels of phosphorylated PI3K (p-PI3K), protein kinase B (p-Akt), and mTOR (p-mTOR) in glaucoma rats, whereas such increases were attenuated by rapamycin or Ly294002.</p><p><strong>Conclusions: </strong>These results demonstrate that ligustrazine is protective in experimental glaucoma by inhibiting autophagy via the activation of the PI3K-Akt/mTOR pathway, providing compelling evidence that ligustrazine is potentially therapeutic for patients with glaucoma.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"725-733"},"PeriodicalIF":2.2,"publicationDate":"2021-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/13/96/mv-v27-725.PMC8711580.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39701907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prevalence of primary mutations in Leber hereditary optic neuropathy: A five-year report from a tertiary eye care center in India.","authors":"Srilekha Sundaramurthy,&nbsp;Ambika Selvakumar,&nbsp;Vidhya Dharani,&nbsp;Nagasamy Soumittra,&nbsp;Jayaprakash Mani,&nbsp;Karthiyayini Thirumalai,&nbsp;Porkodi Periyasamy,&nbsp;Sinnakaruppan Mathavan,&nbsp;Sarangapani Sripriya","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Genetic testing for primary mutations m.3460G>A, m.11778G>A, and m.14484T>C in <i>ND1, ND4,</i> and <i>ND6</i> genes of mitochondrial DNA is the recommended assay for Leber hereditary optic neuropathy (LHON; OMIM 535000). This report discusses the outcome of molecular genetic screening for these three primary mutations in suspected LHON cases in India.</p><p><strong>Methods: </strong>Two hundred and seventy-eight unrelated presumed LHON patients who were seen at the neuro-ophthalmology clinic of a tertiary eye care center from 2014-2018 were analyzed. They were genotyped for the three common variants by polymerase chain reaction-based direct sequencing, and their plasmy status was also determined by restriction enzyme digestion.</p><p><strong>Results: </strong>Eighty two of 278 patients were positive for one of the 3 common mutations with m.11778G>A in <i>ND4</i> gene more frequently distributed (N=72) in homoplasmic state (N=59/82). The mean onset age of visual loss was 21.1years (SD, 9.8 years; range, 5-58 years) in patients harboring the primary mutation. The most common clinical presentation was bilateral sequential painless vision loss with central and cecocentral scotomas in the visual field due to optic disc atrophy.</p><p><strong>Conclusions: </strong>The study subjects are a sample of a much larger number of suspected LHON cases tested for primary mutations in India. (N= 278) and 29.4% (82/278) of patients harbour one of the 3 common mutations. Screening the entire mitochondrial genome and the other nuclear genes encoding mitochondrial protein, would probably aid in identifying the other less common mtDNA mutations causing LHON in Indian population.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"718-724"},"PeriodicalIF":2.2,"publicationDate":"2021-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/8a/8d/mv-v27-718.PMC8711579.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39687133","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Collagen matrix perturbations in corneal stroma of Ossabaw mini pigs with type 2 diabetes.","authors":"Nishant R Sinha, Praveen K Balne, Filiz Bunyak, Alexandria C Hofmann, Rayne R Lim, Rajiv R Mohan, Shyam S Chaurasia","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Diabetes mellitus (DM) is a metabolic disorder that affects over 450 million people worldwide. DM is characterized by hyperglycemia, causing severe systemic damage to the heart, kidneys, skin, vasculature, nerves, and eye. Type 2 diabetes (T2DM) constitutes 90% of clinical cases and is the most common cause of blindness in working adults. Also, about 70% of T2DM patients show corneal complications including delayed wound healing, often described as diabetic keratopathy (DK). Despite the increasing severity of DM, the research on DK is bleak. This study investigated cellular morphology and collagen matrix alterations of the diabetic and non-diabetic corneas collected from Ossabaw mini pigs, a T2DM animal model with a \"thrifty genotype.\"</p><p><strong>Methods: </strong>Pig corneas were collected from six-month-old Ossabaw miniature pigs fed on a western diet (WD) for ten weeks. The tissues were processed for immunohistochemistry and analyzed using hematoxylin and eosin staining, Mason Trichrome staining, Picrosirus Red staining, Collage I staining, and TUNEL assay. mRNA was prepared to quantify fibrotic gene expression using quantitative reverse-transcriptase PCR (qRT-PCR). Transmission electron microscopy (TEM) was performed to evaluate stromal fibril arrangements to compare collagen dynamics in WD <i>vs.</i> standard diet (SD) fed Ossabaw pig corneas.</p><p><strong>Results: </strong>Ossabaw mini pigs fed on a WD for 10 weeks exhibit classic symptoms of metabolic syndrome and hyperglycemia seen in T2DM patients. We observed significant disarray in cornea stromal collagen matrix in Ossabaw mini pigs fed on WD compared to the age-matched mini pigs fed on a standard chow diet using Masson Trichome and Picrosirius Red staining. Furthermore, ultrastructure evaluation using TEM showed alterations in stromal collagen fibril size and organization in diabetic corneas compared to healthy age-matched corneas. These changes were accompanied by significantly decreased levels of Collagen IV and increased expression of matrix metallopeptidase 9 in WD-fed pigs.</p><p><strong>Conclusions: </strong>This pilot study indicates that Ossabaw mini pigs fed on WD showed collagen disarray and altered gene expression involved in wound healing, suggesting that corneal stromal collagens are vulnerable to diabetic conditions.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"666-678"},"PeriodicalIF":2.2,"publicationDate":"2021-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/8a/46/mv-v27-666.PMC8684810.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39888915","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mutational screening of <i>AGRN</i>, <i>SLC39A5</i>, <i>SCO2</i>, <i>P4HA2, BSG</i>, <i>ZNF644</i>, and <i>CPSF1</i> in a Chinese cohort of 103 patients with nonsyndromic high myopia.","authors":"Yi-Han Zheng,&nbsp;Xue-Bi Cai,&nbsp;Lu-Qi Xia,&nbsp;Fang-Yue Zhou,&nbsp;Xin-Ran Wen,&nbsp;De-Fu Chen,&nbsp;Fang Han,&nbsp;Kai-Jing Zhou,&nbsp;Zi-Bing Jin,&nbsp;Wen-Juan Zhuang,&nbsp;Bing Lin","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>High myopia (HM) is one of the leading causes of irreversible vision loss in the world. Many myopia loci have been uncovered with linkage analysis, genome-wide association studies, and sequencing analysis. Numerous pathogenic genes within these loci have been detected in a portion of HM cases. In the present study, we aimed to investigate the genetic basis of 103 patients with nonsyndromic HM, focusing on the reported causal genes.</p><p><strong>Methods: </strong>A total of 103 affected individuals with nonsyndromic HM were recruited, including 101 patients with unrelated sporadic HM and a mother and son pair. All participants underwent comprehensive ophthalmic examinations, and genomic DNA samples were extracted from the peripheral blood. Whole exome sequencing was performed on the mother and son pair as well as on the unaffected father. Sanger sequencing was used to identify mutations in the remaining 101 patients. Bioinformatics analysis was subsequently applied to verify the mutations.</p><p><strong>Results: </strong>An extremely rare mutation in <i>AGRN</i> (c.2627A>T, p.K876M) was identified in the mother and son pair but not in the unaffected father. Another two mutations in <i>AGRN</i> (c.4787C>T, p.P1596L/c.5056G>A, p.G1686S) were identified in two unrelated patients. A total of eight heterozygous variants potentially affecting the protein function were detected in eight of the remaining 99 patients, including c.1350delC, p.V451Cfs*76 and c.1023_1024insA, p.P342Tfs*41 in <i>SLC39A5</i>; c.244_246delAAG, p.K82del in <i>SCO2</i>; c.545A>G, p.Y182C in <i>P4HA2</i>; c.415C>T, p.P139S in <i>BSG</i>; c.3266A>G, p.Y1089C in <i>ZNF644</i>; and c.2252C>T, p.S751L and c.1708C>T, p.R570C in <i>CPSF1</i>. Multiple bioinformatics analyses were conducted, and a comparison to a group with geographically matched controls was performed, which supported the potential pathogenicity of these variants.</p><p><strong>Conclusions: </strong>We provide further evidence for the potential role of <i>AGRN</i> in HM inheritance and enlarged the current genetic spectrum of nonsyndromic HM by comprehensively screening the reported causal genes.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"706-717"},"PeriodicalIF":2.2,"publicationDate":"2021-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/7f/d0/mv-v27-706.PMC8684808.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39888918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Increased lacrimal inflammatory mediators in patients with keratoconus.","authors":"Gustavo Souza Moura,&nbsp;Albert Santos,&nbsp;Marcos Antonio Cenedeze,&nbsp;Meire Ioshie Hiyane,&nbsp;Niels Olsen Saraiva Camara,&nbsp;Luciene Barbosa de Sousa,&nbsp;Lauro Augusto de Oliveira","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>This study aimed to characterize the tear film immunologic profile in keratoconus (KC) patients compared with healthy individuals (control group) and to investigate the correlation between the tear film immunologic profile and atopy, disease severity, and disease status over time.</p><p><strong>Methods: </strong>The study involved 30 KC patients and 18 healthy individuals. Tear collection was obtained using microcapillary tubes. Tear film levels of fractalkine, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon (IFN)-γ, interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17A, IL-21, IL-23, interferon-inducible T-cell alpha chemoattractant (ITAC), macrophage inflammatory protein-1 alpha (MIP-1α), MIP-1β, MIP-3α, and tumor necrosis factor (TNF)-α were detected. Keratometric measurements and topographic patterns were used to diagnose and define disease progression. Tear immunologic profiles were compared, emphasizing the presence or absence of ocular allergy. Correlations between the cytokine profile, disease severity, and disease status were also analyzed longitudinally in the KC patients.</p><p><strong>Results: </strong>Lacrimal cytokine concentrations were higher in the KC patients than they were in the controls in 14 of 21 cytokines analyzed. IL-6 was the most relevant cytokine found in KC patients, especially when associated with ocular allergy. There was no correlation between KC progression and the level of inflammatory cytokines when analyzed longitudinally. KC severity correlated with IL-6 concentration, where the more severe KC presented a higher IL-6 concentration in tears.</p><p><strong>Conclusions: </strong>Inflammatory activity seems to be involved in the pathogenesis of KC. Out of 21 cytokines, 14 were more concentrated in the tears of KC patients than healthy subjects. IL-6 was significantly higher in KC patients' tears and was related to disease severity. Disease progression did not correlate with cytokine levels when analyzed longitudinally.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"656-665"},"PeriodicalIF":2.2,"publicationDate":"2021-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/38/ff/mv-v27-656.PMC8684811.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39888914","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lacrimal gland homeostasis is maintained by the <i>AQP5</i> pathway by attenuating endoplasmic reticulum stress inflammation in the lacrimal gland of <i>AQP5</i> knockout mice.","authors":"Shaohua Hu,&nbsp;Guohu Di,&nbsp;Xin Cao,&nbsp;Yaning Liu,&nbsp;Yihui Wang,&nbsp;Hui Zhao,&nbsp;Dianqiang Wang,&nbsp;Peng Chen","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>AQP5^-/- mice spontaneously exhibit dry eye symptoms. The purpose of this study was to assess the endoplasmic reticulum (ER) stress-mediated inflammation generated by a deficiency of aquaporin 5 (AQP5) in the lacrimal gland.</p><p><strong>Methods: </strong>Hematoxylin and eosin (H&E) staining, Oil Red O staining, and transmission electron microscopy (TEM) analysis were performed to identify structural changes in lacrimal gland epithelial cells because of AQP5 deficiency. Corneal epithelial defects were assessed with sodium fluorescein staining. The expression profiles of mRNA and proteins were determined by quantitative real-time reverse transcription PCR (qRT-PCR) and western blot. Mice in the quercetin group were injected intraperitoneally with 40 mg/kg of quercetin, and the control group was injected with an equal volume of dimethyl sulfoxide (DMSO) for 4 weeks.</p><p><strong>Results: </strong>Aqueous tear secretion fell at about 50% in 1- and 6-month-old AQP5^-/- mice compared with that of AQP5^+/+ mice. TEM showed that the ER structure was damaged. ER stress was significantly increased in the lacrimal gland of AQP5^-/- mice. Lipid droplets accumulated in the matrix and acinar cells, and changes occurred in the lipid metabolism and gene expression levels for <i>PPARα</i>, <i>CPT1α</i>, and <i>CPT2</i> in the AQP5^-/- mice. Immune cell infiltration and increases in the gene expression levels of the chemokines <i>CXCL1</i>, <i>CXCL2</i>, and <i>CCL5</i> were found in the lacrimal gland of AQP5^-/- mice. Quercetin partially reversed ER stress levels, inflammation, and lipid accumulation, and it inhibited tear secretion.</p><p><strong>Conclusions: </strong>The study data indicated that a deficiency of AQP5 induced pathophysiological changes and functional decompensation of the lacrimal gland. Quercetin may improve the inflammation in the lacrimal glands of AQP5^-/- mice by regulating the ER stress levels.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"679-690"},"PeriodicalIF":2.2,"publicationDate":"2021-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/9d/8b/mv-v27-679.PMC8684812.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39888916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tissue plasminogen activator rescues steroid-induced outflow facility reduction via non-enzymatic action.","authors":"Sofya Gindina,&nbsp;Arturo O Barron,&nbsp;Yan Hu,&nbsp;Antonios Dimopoulos,&nbsp;John Danias","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Tissue plasminogen activator (tPA) prevents steroid-induced reduction in aqueous humor outflow facility; however, its mechanism of action at the trabecular meshwork (TM) remains unclear. Enzymatic and non-enzymatic domains allow tPA to function as both an enzyme and a cytokine. This study sought to determine whether cytokine activity is sufficient to rescue steroid-induced outflow facility reduction.</p><p><strong>Methods: </strong>Outflow facility was measured in C57BL/6J mice following triamcinolone acetonide exposure and either transfection of the TM using adenoviral vectors, encoding for enzymatically active and inactive tPA, or administration of the respective proteins. Protein injections were also administered to tPA deficient (<i>Plat</i>KO) and <i>Mmp-9</i> deficient (<i>Mmp-9</i>KO) mice to determine the potential to rescue reductions in outflow facility and determine downstream mechanisms. Gene expression of matrix metalloproteinases (<i>Mmp-2, -9</i>, and <i>-13</i>) was measured in angle ring tissues containing the TM.</p><p><strong>Results: </strong>Enzymatically active and inactive tPA (either produced after TM transfection or after direct administration) were equally effective in attenuating steroid-induced outflow facility reduction in C57BL/6J mice. They were also equally effective in rescuing outflow reduction in <i>Plat</i>KO mice and causing enhanced expression of matrix metalloproteinases. However, both enzymatically active and enzymatically inactive tPA did not improve outflow reduction in <i>Mmp-9</i>KO mice or increase the baseline outflow facility in naïve C57BL/6J mice.</p><p><strong>Conclusions: </strong>tPA enzymatic activity is not necessary in the regulation of aqueous humor outflow. tPA can increase the expression of matrix metalloproteinases in a cytokine-mediated fashion. This cascade of events may eventually lead to extracellular matrix remodeling at the TM, which reverses outflow facility reduction caused by steroids.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"691-705"},"PeriodicalIF":2.2,"publicationDate":"2021-12-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/c4/3e/mv-v27-691.PMC8684809.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39888917","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Five novel copy number variations detected in patients with familial exudative vitreoretinopathy.","authors":"Jia Luo,&nbsp;Jing Li,&nbsp;Xiang Zhang,&nbsp;Jia-Kai Li,&nbsp;Hao-Jie Chen,&nbsp;Pei-Quan Zhao,&nbsp;Ping Fei","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Purpose: </strong>Familial exudative vitreoretinopathy (FEVR) is an inherited retinal vascular disease genetically heterogeneous with multiple causative genes. The aim of this study is to report five novel copy number variation (CNV) regions in FEVR patients and to investigate the possible contributions of novel CNVs to FEVR.</p><p><strong>Methods: </strong>In this study, 824 FEVR families were collected. All cases were performed using the targeted next generation sequencing (NGS) assay, and families with no definite pathogenic mutations in FEVR genes were screened for CNVs according to the NGS results. Droplet digital polymerase chain reaction (ddPCR) testing was introduced to validate the screened CNV regions. We also reviewed the clinical presentations of the probands and affected family members associated with the novel CNVs and conducted segregation analysis.</p><p><strong>Results: </strong>Five CNVs in five patients were detected in this study: heterozygous deletions of kinesin family member 11 (<i>KIF11</i>) exons 2-4, KIF11 exon 11, KIF11 exons 1-10, tetraspanin-12 (<i>TSPAN12</i>) exons 1-3, and low-density lipoprotein receptor-related protein 5 (<i>LRP5</i>) exons 19-21. Among the five affected families, <i>TSPAN12</i> exons 1-3 heterozygous deletion and <i>LRP5</i> exons 19-21 heterozygous deletion originate from the mother and the father of the proband, respectively. No other family members manifested as FEVR except for the probands. The correlation between disease severity and CNV loci seems uncertain.</p><p><strong>Conclusions: </strong>Five novel CNV loci in FEVR patients were uncovered in this study, including one maternally-inherited and one paternally-inherited CNV region. Though there is no evidence of co-segregation between these CNVs and FEVR, our findings suggest novel genetic risk factors for FEVR.</p>","PeriodicalId":18866,"journal":{"name":"Molecular Vision","volume":"27 ","pages":"632-642"},"PeriodicalIF":2.2,"publicationDate":"2021-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/56/75/mv-v27-632.PMC8645187.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39850982","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}