Data_for_Descriptions

Chloroplast number in leaf stomatal GCs of Col, Ler, and Ws. Measurements of chloroplast number in arbitrarily selected pairs of GCs ('cell A' and 'cell B') are shown

Data_for_Fig6

Chloroplast number in leaf stomatal GCs of WT, arc5, arc6, and atminE1. Measurements of chloroplast number in arbitrarily selected pairs of GCs ('cell A' and 'cell B') are shown

Gene Expression Analysis of the Irrigation Solution Samples Collected during Vitrectomy for Idiopathic Epiretinal Membrane

<div><p>Purpose</p><p>The analysis of gene expression in idiopathic epiretinal membranes (iERMs) may help elucidate ERM formation and its pathology. Here, we conducted a case-control study, in order to determine the expression levels of cytokines and other genes in eyes with macular hole (MH) or iERM.</p><p>Methods</p><p>Twenty eyes, obtained from seven male and 13 female patients, were included in the study. The average age of the study subjects was 69.1 ± 7.67 years, and 15 eyes had iERM, while five eyes had MH. Irrigation solution samples were collected during vitrectomy, centrifuged, and the levels of cytokine and other mRNAs in the sediment were assessed using real-time PCR. The expression level of 11 cytokine genes, four transcription factor genes, two cytoskeletal genes, and genes encoding two extracellular matrix proteins in eyes with MH or iERM were determined and compared.</p><p>Results</p><p>The expression levels of interleukin 6 (<i>IL6</i>), tumor growth factor B2 (<i>TGFB2)</i>, vascular endothelial growth factor A (<i>VEGFA</i>), chemokine C-X-C motif ligand 1 (<i>CXCL1</i>), v-rel avian reticuloendotheliosis viral oncogene homolog A (<i>RELA</i>), glial fibrillary acidic protein (<i>GFAP</i>), and tenascin C (<i>TNC</i>) were significantly higher in eyes with iERM than in eyes with MH. The expression of these genes was not associated with the preoperative visual acuity of the investigated patients.</p><p>Conclusions</p><p>The obtained results indicate that real-time PCR analysis of irrigation solution samples collected during vitrectomy can help assess the expression levels of several genes, and that iERM is associated with the expression of pro-inflammatory genes and the genes expressed during angiogenesis and wound healing process (<i>IL6</i>, <i>TGFB2</i>, <i>VEGFA</i>, <i>CXCL1</i>, <i>RELA</i>, <i>GFAP</i>, and <i>TNC</i>).</p></div

Plastid phenotypes in leaf PCs of <i>Arabidopsis</i> WT, <i>arc5</i>, <i>arc6</i>, and <i>atminE1</i>.

<p>(A–J) Images of plastid-targeted YFP in the third-fourth leaf petioles of 4-week-old WT (A), <i>arc5</i> (B, E, H), <i>arc6</i> (C, F, I), and <i>atminE1</i> (D, G, J) seedlings. CLSM images of maximal intensity projection are shown. Giant plastids (arrows), putative dividing plastids (arrowheads), stomata (double arrowheads), and mini-plastids (asterisks) are indicated. Bar = 10 μm.</p

Expression Levels of Cytokines and Other Investigated Genes in the Solid Component of the Irrigation Solution.

<p>Expression Levels of Cytokines and Other Investigated Genes in the Solid Component of the Irrigation Solution.</p

Plastid phenotypes in leaf stomatal GCs in <i>Arabidopsis</i> WT, <i>arc5</i>, <i>arc6</i>, and <i>atminE1</i> plants.

<p>(A–I) Images of plastid-targeted YFP in the third-fourth leaf petioles of 4-week-old seedlings from the WT (A), <i>arc5</i> (B–D), <i>arc6</i> (E–G), and <i>atminE1</i> (H, I). CLSM images of maximal intensity projection (top panels) and merged with DIC (bottom panels) are shown. (J, K) Images of YFP-labeled plastids in GCs reconstructed from a series of optical sections generated by CLSM, taken at 0.3 or 0.6 μm intervals. The GC pairs in panels (J) and (K) are identical to those in panels (D) and (G), respectively. (L) Epifluorescence microscopy images of GCs in the third-fourth leaf petioles of 4-week-old seedlings from <i>arc6</i>. Images of YFP (top panels) and DIC (bottom panels) are shown. Bar = 10 μm.</p

The <i>Arabidopsis arc5</i> and <i>arc6</i> mutations differentially affect plastid morphology in pavement and guard cells in the leaf epidermis

<div><p>Chloroplasts, or photosynthetic plastids, multiply by binary fission, forming a homogeneous population in plant cells. In <i>Arabidopsis thaliana</i>, the division apparatus (or division ring) of mesophyll chloroplasts includes an inner envelope transmembrane protein ARC6, a cytoplasmic dynamin-related protein ARC5 (DRP5B), and members of the FtsZ1 and FtsZ2 families of proteins, which co-assemble in the stromal mid-plastid division ring (FtsZ ring). FtsZ ring placement is controlled by several proteins, including a stromal factor MinE (AtMinE1). During leaf mesophyll development, <i>ARC6</i> and <i>AtMinE1</i> are necessary for FtsZ ring formation and thus plastid division initiation, while <i>ARC5</i> is essential for a later stage of plastid division. Here, we examined plastid morphology in leaf epidermal pavement cells (PCs) and stomatal guard cells (GCs) in the <i>arc5</i> and <i>arc6</i> mutants using stroma-targeted fluorescent proteins. The <i>arc5</i> PC plastids were generally a bit larger than those of the wild type, but most had normal shapes and were division-competent, unlike mutant mesophyll chloroplasts. The <i>arc6</i> PC plastids were heterogeneous in size and shape, including the formation of giant and mini-plastids, plastids with highly developed stromules, and grape-like plastid clusters, which varied on a cell-by-cell basis. Moreover, unique plastid phenotypes for stomatal GCs were observed in both mutants. The <i>arc5</i> GCs rarely lacked chlorophyll-bearing plastids (chloroplasts), while they accumulated minute chlorophyll-less plastids, whereas most GCs developed wild type-like chloroplasts. The <i>arc6</i> GCs produced large chloroplasts and/or chlorophyll-less plastids, as previously observed, but unexpectedly, their chloroplasts/plastids exhibited marked morphological variations. We quantitatively analyzed plastid morphology and partitioning in paired GCs from wild-type, <i>arc5</i>, <i>arc6</i>, and <i>atminE1</i> plants. Collectively, our results support the notion that ARC5 is dispensable in the process of equal division of epidermal plastids, and indicate that dysfunctions in ARC5 and ARC6 differentially affect plastid replication among mesophyll cells, PCs, and GCs within a single leaf.</p></div

Chloroplast length in leaf stomatal GCs of the WT, <i>arc5</i>, <i>arc6</i>, and <i>atminE1</i>¹.

<p>Chloroplast length in leaf stomatal GCs of the WT, <i>arc5</i>, <i>arc6</i>, and <i>atminE1</i><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0192380#t001fn001" target="_blank">¹</a>.</p

Distribution of chloroplasts in leaf stomatal GCs of <i>Arabidopsis</i> WT, <i>arc5</i>, <i>arc6</i>, and <i>atminE1</i> plants.

<p>(A, B) Measurements of chloroplast number in arbitrarily selected pairs of GCs (‘cell A’ and ‘cell B’) are shown in graph (A) and table (B) form.</p

Demographics of Patients Included in Macular Hole and Idiopathic Epiretinal Membrane Groups.

<p>Demographics of Patients Included in Macular Hole and Idiopathic Epiretinal Membrane Groups.</p