sample-drift-correction-following-4d-confocal-time-lapse-imaging

Sample Drift Correction Following 4D Confocal Time-lapse Imaging

The generation of four-dimensional (4D) confocal datasets; consisting of 3D image sequences over time; provides an excellent methodology to capture ...

Monash University

&#x751F;&#x6210;&#x7684;&#x8132;&#x8BF1;&#x53D8;&#x7684;&#x6591;&#x9A6C;&#x9C7C;&#x7A81;&#x53D8;&#x4F53;&#x4E3A;&#x5939;&#x5C42;&#x7684;&#x53D1;&#x5C55;&#x8FDB;&#x7A0B;&#xFF0C;&#x5305;&#x62EC;&#x5668;&#x5B98;&#x5206;&#x5316;&#x7684;&#x57FA;&#x56E0;&#x8C03;&#x63A7;&#x63D0;&#x4F9B;&#x4E86;&#x4E00;&#x4E2A;&#x5F3A;&#x5927;&#x7684;&#x5DE5;&#x5177;&#x3002;&#x4E00;&#x4E2A;&#x6591;&#x9A6C;&#x9C7C;&#x7A81;&#x53D8;&#xFF0C;&quot;flotte &#x4E50;&#x5929;&quot;&#xFF08;flo&#xFF09;&#xFF0C;&#x80A0;&#x3001; &#x809D;&#x3001; &#x80F0;&#x817A;&#x4E0E;&#x773C;&#x775B;&#x5F62;&#x6210; 78 &#x5C0F;&#x65F6; postfertilization (hpf) &#x5728;&#x663E;&#x793A;&#x660E;&#x663E;&#x7684;&#x7F3A;&#x9677;&#x3002;&#x5728;&#x6B64;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x8BD5;&#x56FE;&#x8BC6;&#x522B; flo &#x7684;&#x57FA;&#x7840;&#x7A81;&#x53D8;&#x7684;&#x57FA;&#x56E0;&#x5E76;&#x94FE;&#x63A5;&#x5230;&#x5176;&#x8868;&#x578B;&#x7684;&#x9057;&#x4F20;&#x75C5;&#x53D8;&#x90E8;&#x4F4D;&#x3002;

&#x5F02;&#x5E38;&#x6838;&#x5B54;&#x9699;&#x5F62;&#x6210;&#x89E6;&#x53D1; elys &#x7F3A;&#x6591;&#x9A6C;&#x9C7C;&#x80A0;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#x51CB;&#x4EA1;&#x3002;

Zebrafish mutants generated by ethylnitrosourea-mutagenesis provide a powerful tool for dissecting the genetic regulation of developmental processes, including organogenesis. One zebrafish mutant, "flotte lotte" (flo), displays striking defects in intestinal, liver, pancreas, and eye formation at 78 hours postfertilization (hpf). In this study, we sought to identify the underlying mutated gene in flo and link the genetic lesion to its phenotype.

Abnormal nuclear pore formation triggers apoptosis in the intestinal epithelium of elys-deficient zebrafish.

Mutants de poisson-z&#xE8;bre g&#xE9;n&#xE9;r&#xE9;s par mutagen&#xE8;se &#xE9;thylnitrosour&#xE9;e-fournir un outil puissant pour diss&#xE9;quer la r&#xE9;gulation g&#xE9;n&#xE9;tique des processus de d&#xE9;veloppement, y compris l&#x26;#39;organogen&#xE8;se. Un poisson z&#xE8;bre mutant, &#x26;quot;Flotte lotte&#x26;quot; (flo), affiche des d&#xE9;fauts frappants dans l&#x26;#39;intestin, le foie, le pancr&#xE9;as, et la formation de l&#x26;#39;&#x153;il &#xE0; 78 heures post-f&#xE9;condation (HPF). Dans cette &#xE9;tude, nous avons cherch&#xE9; &#xE0; identifier le g&#xE8;ne mut&#xE9; dans sous-jacente flo et lier la l&#xE9;sion g&#xE9;n&#xE9;tique &#xE0; son ph&#xE9;notype.

La formation anormale pore nucl&#xE9;aire d&#xE9;clenche l&#x26;#39;apoptose dans l&#x26;#39;&#xE9;pith&#xE9;lium intestinal des Elys-d&#xE9;ficient poisson z&#xE8;bre

Zebrafisch-Mutanten von Ethylnitrosourea-Mutagenese erzeugt bieten ein leistungsf&auml;higes Werkzeug f&uuml;r sezieren der genetischen Steuerung der Entwicklungsprozesse, einschlie&szlig;lich der Organogenese. Ein Zebrafisch mutant, &quot;Flotte Lotte&quot; (Flo), zeigt auffallende M&auml;ngel im Darm, Leber, Bauchspeicheldr&uuml;se und Auge-Formation bei 78 Stunden Postfertilization (hpf). In dieser Studie wollten wir identifizieren das zugrunde liegende mutierte Gen in Flo und genetische L&auml;sion mit seinen Ph&auml;notyp verkn&uuml;pfen.

Abnorme nukleare Porenbildung l&ouml;st Apoptosis in der intestinalen Epithel Elys-defizienten Zebrab&auml;rbling.

Zebrafish mutanti generati da etilnitrosourea-mutagenesi forniscono un potente strumento per la regolazione genetica dei processi di sviluppo, tra cui organogenesi di dissezione. Uno zebrafish mutanti, &quot;flotte lotte&quot; (flo), consente di visualizzare sorprendenti difetti nell&#39;intestino, fegato, pancreas e la formazione di occhio a postfertilization 78 ore (hpf). In questo studio, abbiamo cercato di identificare il gene mutato sottostante in flo e linkare la lesione genetica suo fenotipo.

Formazione anomala poro nucleare innesca apoptosi nell&#39;epitelio intestinale di zebrafish elys-carenti.

&#x30A8;&#x30C1;&#x30EB;&#x30CB;&#x30C8;&#x30ED;&#x30BD;&#x30A6;&#x30EC;&#x30A2;&#x3001;&#x7A81;&#x7136;&#x5909;&#x7570;&#x306B;&#x3088;&#x308A;&#x751F;&#x6210;&#x3055;&#x308C;&#x305F;&#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x5909;&#x7570;&#x4F53;&#x306F;&#x3001;&#x5668;&#x5B98;&#x5F62;&#x6210;&#x3092;&#x542B;&#x3080;&#x767A;&#x751F;&#x904E;&#x7A0B;&#x306E;&#x907A;&#x4F1D;&#x7684;&#x8ABF;&#x7BC0;&#x3092;&#x89E3;&#x5256;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x5F37;&#x529B;&#x306A;&#x30C4;&#x30FC;&#x30EB;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002; One&#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x5909;&#x7570;&#x4F53;&#x306F;&#x3001; &#x26;quot;Flotte&#x306E;&#x30ED;&#x30C3;&#x30C6;&#x26;quot;&#xFF08;FLO&#xFF09;&#x306F;&#x3001;&#x8178;&#x3001;&#x809D;&#x81D3;&#x3001;&#x81B5;&#x81D3;&#x3001;78&#x6642;&#x9593;postfertilization&#xFF08;HPF&#xFF09;&#x306E;&#x76EE;&#x306E;&#x5F62;&#x6210;&#x306B;&#x8457;&#x3057;&#x3044;&#x6B20;&#x9665;&#x304C;&#x8868;&#x793A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x305D;&#x3053;&#x3067;&#x3001;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;&#x3001;FLO&#x306E;&#x57FA;&#x790E;&#x3068;&#x306A;&#x308B;&#x5909;&#x7570;&#x907A;&#x4F1D;&#x5B50;&#x3092;&#x540C;&#x5B9A;&#x3057;&#x3001;&#x305D;&#x306E;&#x8868;&#x73FE;&#x578B;&#x306B;&#x907A;&#x4F1D;&#x7684;&#x75C5;&#x5909;&#x3092;&#x30EA;&#x30F3;&#x30AF;&#x3057;&#x3088;&#x3046;&#x3068;&#x3057;&#x305F;&#x3002;

&#x7570;&#x5E38;&#x306A;&#x6838;&#x819C;&#x5B54;&#x5F62;&#x6210;&#x306F;Elys&#x6B20;&#x640D;&#x30BC;&#x30D6;&#x30E9;&#x30D5;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x306E;&#x8178;&#x4E0A;&#x76AE;&#x306B;&#x304A;&#x3051;&#x308B;&#x30A2;&#x30DD;&#x30C8;&#x30FC;&#x30B7;&#x30B9;&#x3092;&#x8A98;&#x767A;

Zebrafish &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; ethylnitrosourea mutagenesis&#xC5D0; &#xC758;&#xD574; &#xC0DD;&#xC131; &#xB41C; organogenesis&#xB97C; &#xD3EC;&#xD568; &#xD558; &#xC5EC; &#xAC1C;&#xBC1C; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC758; &#xC720;&#xC804;&#xC790; &#xC870;&#xC808; &#xBD80;&#xB97C; &#xC704;&#xD55C; &#xAC15;&#xB825;&#xD55C; &#xB3C4;&#xAD6C;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xD55C; zebrafish &#xB3CC;&#xC5F0;&#xBCC0;&#xC774;, &quot;&#xB85C;&#xD2B8; &#xB86F;&#xB370;&quot; (flo) &#xCC3D; &#xC790;, &#xAC04;, &#xCDCC; &#xC7A5; &#xBC0F; 78 &#xC2DC;&#xAC04; postfertilization (hpf)&#xC5D0;&#xC11C; &#xB208; &#xD615;&#xC131;&#xC5D0; &#xB208;&#xC5D0; &#xB744;&#xB294; &#xACB0;&#xD568;&#xC744; &#xD45C;&#xC2DC; &#xD569;&#xB2C8;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC;&#xAC00; &#xD50C;&#xB85C;&#xC5D0; &#xAE30;&#xBCF8; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xC720;&#xC804;&#xC790;&#xB97C; &#xC2DD;&#xBCC4; &#xD558; &#xACE0; &#xADF8; &#xD45C;&#xD604; &#xD615; &#xC720;&#xC804; &#xBCD1; &#xBCC0; &#xC5F0;&#xACB0; &#xD558;&#xACE0;&#xC790; &#xD588;&#xB2E4;.

&#xBE44;&#xC815;&#xC0C1;&#xC801;&#xC778; &#xD575; &#xAE30; &#xACF5; &#xD615;&#xC131; elys &#xBD88;&#xCDA9;&#xBD84; &#xD55C; zebrafish&#xC758; &#xCC3D; &#xC790; &#xC0C1;&#xD53C;&#xC5D0; apoptosis&#xB97C; &#xD2B8;&#xB9AC;&#xAC70;&#xD569;&#xB2C8;&#xB2E4;.

Pez cebra mutantes generadas por mutag&#xE9;nesis etilnitrosourea-proporcionan una herramienta poderosa para la disecci&#xF3;n de la regulaci&#xF3;n gen&#xE9;tica de los procesos de desarrollo, incluyendo la organog&#xE9;nesis. Un pez cebra mutante, &#x26;quot;Flotte Lotte&#x26;quot; (FLO), muestra notables defectos en el h&#xED;gado intestinal, el p&#xE1;ncreas y la formaci&#xF3;n de ojos en 78 horas postfertilizaci&#xF3;n (HPF). En este estudio, hemos tratado de identificar el gen mutado en el subyacente flo y vincular la lesi&#xF3;n gen&#xE9;tica de su fenotipo.

Formaci&#xF3;n anormal de los poros nucleares desencadena la apoptosis en el epitelio intestinal de Elys deficiente en pez cebra

Optically transparent zebrafish embryos provide an excellent vertebrate model system in which to reveal specific mRNA and protein expression patterns during development. Whole-mount preparations can be used to generate three-dimensional color or fluorescent readouts of the expression pattern of a given gene (or genes), matched with a bright-field image of all the tissues in the developing embryo. Whole-mount mRNA in situhybridization (WISH) has long been the method of choice for revealing gene expression patterns in zebrafish because this method depends only on being able to identify a relatively short region of nucleotide sequence unique for the gene of interest. In contrast, the scarcity of antibodies that are specific to or cross-react with zebrafish proteins has limited the widespread use of immunocytochemical applications, though this situation will improve in the future. The elucidation of the specific expression patterns of Wnt pathway genes in zebrafish has made a major contribution to our current understanding of their roles in vertebrate development.

Determination of mRNA and protein expression patterns in zebrafish.

The zebrafish provides an ideal model for the study of vertebrate organogenesis, including the formation of the digestive tract and its associated organs. Despite optical transparency of embryos, the internal position of the developing digestive system and its close juxtaposition with the yolk initially made morphological analysis relatively challenging, particularly during the first 3 d of development. However, methodologies have been successfully developed to address these problems and comprehensive morphologic analysis of the developing digestive system has now been achieved using a combination of light and fluorescence microscope approaches-including confocal analysis-to visualize wholemount and histological preparations of zebrafish embryos. Furthermore, the expanding number of antibodies that cross-react with zebrafish proteins and the generation of tissue-specific transgenic green fluorescent protein reporter lines that mark specific cell and tissue compartments have greatly enhanced our ability to successfully image the developing zebrafish digestive system.

Morphologic analysis of the zebrafish digestive system.

Ribosome biogenesis underpins cell growth and division. Disruptions in ribosome biogenesis and translation initiation are deleterious to development and underlie a spectrum of diseases known collectively as ribosomopathies. Here, we describe a novel zebrafish mutant, titania (tti(s450)), which harbours a recessive lethal mutation in pwp2h, a gene encoding a protein component of the small subunit processome. The biochemical impacts of this lesion are decreased production of mature 18S rRNA molecules, activation of Tp53, and impaired ribosome biogenesis. In tti(s450), the growth of the endodermal organs, eyes, brain, and craniofacial structures is severely arrested and autophagy is up-regulated, allowing intestinal epithelial cells to evade cell death. Inhibiting autophagy in tti(s450) larvae markedly reduces their lifespan. Somewhat surprisingly, autophagy induction in tti(s450) larvae is independent of the state of the Tor pathway and proceeds unabated in Tp53-mutant larvae. These data demonstrate that autophagy is a survival mechanism invoked in response to ribosomal stress. This response may be of relevance to therapeutic strategies aimed at killing cancer cells by targeting ribosome biogenesis. In certain contexts, these treatments may promote autophagy and contribute to cancer cells evading cell death.

Article	Year
Abnormal nuclear pore formation triggers apoptosis in the intestinal epithelium of elys-deficient zebrafish. Gastroenterology\| PubMed ID: 19073184	2009
Determination of mRNA and protein expression patterns in zebrafish. Methods in molecular biology (Clifton, N.J.)\| PubMed ID: 19109715	2008
Morphologic analysis of the zebrafish digestive system. Methods in molecular biology (Clifton, N.J.)\| PubMed ID: 19378111	2009
Autophagy induction is a Tor- and Tp53-independent cell survival response in a zebrafish model of disrupted ribosome biogenesis. PLoS genetics\| PubMed ID: 23408911	2013

Adam Parslow

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