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Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei
Park, Kyunghyuk,Frost, Jennifer M.,Adair, Adam James,Kim, Dong Min,Yun, Hyein,Brooks, Janie S.,Fischer, Robert L.,Choi, Yeonhee Korean Society for Molecular and Cellular Biology 2016 Molecules and cells Vol.39 No.10
The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.
Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei
Yeonhee Choi,Kyunghyuk Park,Jennifer M. Frost,Adam James Adair,Dong Min Kim,Hyein Yun,Janie S. Brooks,Robert L. Fischer 한국분자세포생물학회 2016 Molecules and cells Vol.39 No.10
The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dis-section followed by the derivation of central cell proto-plasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.
Duenas-Osorio, Leonardo,Park, Joonam,Towashiraporn, Peeranan,Goodno, Barry J.,Frost, David,Craig, James I.,Bostrom, Ann Techno-Press 2004 Structural Engineering and Mechanics, An Int'l Jou Vol.17 No.3
Consequence-Based Engineering (CBE) is a new paradigm proposed by the Mid-America Earthquake Center (MAE) to guide evaluation and rehabilitation of building structures and networks in areas of low probability - high consequence earthquakes such as the central region of the U.S. The principal objective of CBE is to minimize consequences by prescribing appropriate intervention procedures for a broad range of structures and systems, in consultation with key decision makers. One possible intervention option for rehabilitating unreinforced masonry (URM) buildings, widely used for essential facilities in Mid-America, is passive energy dissipation (PED). After the CBE process is described, its application in the rehabilitation of vulnerable URM building construction in Mid-America is illustrated through the use of PED devices attached to flexible timber floor diaphragms. It is shown that PED's can be applied to URM buildings in situations where floor diaphragm flexibility can be controlled to reduce both out-of-plane and in-plane wall responses and damage. Reductions as high as 48% in roof displacement and acceleration can be achieved as demonstrated in studies reported below.
The Effect of Glycolated Alkyl-group Including Long Spacer in Organic Electrochemical Transistor
김진관,Ellasia Tan,Katherine Stewart,Charalampos Pitsalidis,권순철,Nicholas Siemons,김제한,Yifei Jiang,Jarvist M. Frost,Drew Pearce,James E. Tyrrell,Jenny Nelson,Roisin M. Owens,김지선,김윤희 한국고분자학회 2022 한국고분자학회 학술대회 연구논문 초록집 Vol.47 No.2
Brain mu-opioid receptor binding: relationship to relapse to cocaine use after monitored abstinence.
Gorelick, David A,Kim, Yu Kyeong,Bencherif, Badreddine,Boyd, Susan J,Nelson, Richard,Copersino, Marc L,Dannals, Robert F,Frost, J James Springer-Verlag 2008 Psychophamacology Vol.200 No.4
<P>Cocaine users have increased regional brain mu-opioid receptor (mOR) binding which correlates with cocaine craving. The relationship of mOR binding to relapse is unknown.</P>