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3-D imaging draws new picture of Golgi during cell division

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Resolving a fundamental question in cell biology and showing off the powers of new high-resolution 3-D imaging, National Institutes of Health (NIH) scientists have discovered where the Golgi apparatus, which sorts newly synthesized proteins for transport inside and outside the cell, goes when it disassembles during cell division, according to research presented at the American Society for Cell Biology annual meeting in New Orleans.

Conventional microscopy techniques only allow biologists to watch as the Golgi dissolve into tiny “puncta” and an unresolvable haze. But powerful new imaging techniques allowed the NIH researchers – Dr. Dylan Burnette, Dr. Prabuddha Sengupta and Dr. Jennifer Lippincott-Schwartz of the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD) – to follow the Golgi through its “choreographed disassembly process,” which now appears tightly linked to the endoplasmic reticulum (ER) during cell division.

Cell division by mitosis is the complicated yet critical process by which a mother cell divides into two daughter cells. But first, the mother cell has to pack up her cellular household contents, disassembling and dividing up everything for her soon-to-be-formed daughters.


Despite exhaustive study, mysteries about cell division remain.


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How cells manage division has been exhaustively studied for over a century, and yet basic mysteries remained. Scientists knew that some organelles such as the ER are pulled apart before division but keep their tubular membrane structure intact. Other organelles such as the Golgi go to pieces after the prophase of mitosis through choreographed disassembly.

But where does the Golgi go once it is in pieces? To answer the question, the NIH researchers started with two plausible theories: In the ER-linked hypothesis, the Golgi puncta and enzyme haze are closely held by the ER; in the non-ER-linked model, the puncta and haze float about on their own, waiting for cytokinesis, when the two daughter cells separate and the Golgi body reappears as stacks of membrane-bound cisternae, ready to sort proteins from the reassembled ER.

Powered by their new imaging technologies, which gave them far greater resolution than previously possible, the researchers saw clear support of the ER-linked model: The enzyme haze stuck close to ER markers, and the puncta clustered near ER exits.

For a second line of proof, the NICHD researchers followed up with a pharmacologically based trapping assay that showed Golgi enzymes being held tightly by the ER during mitosis. The results indicate that Golgi enzymes redistribute into the ER during mitosis, and that they must follow an ER export pathway to reform the Golgi at the end of mitosis.

This study not only resolves a basic cellular question but also shows what new solutions await as these new technologies give us keener vision and sharper tools.

Published: March 2014
Glossary
cell
1. A single unit in a device for changing radiant energy to electrical energy or for controlling current flow in a circuit. 2. A single unit in a device whose resistance varies with radiant energy. 3. A single unit of a battery, primary or secondary, for converting chemical energy into electrical energy. 4. A simple unit of storage in a computer. 5. A limited region of space. 6. Part of a lens barrel holding one or more lenses.
haze
An aggravated form of fog in a polished surface caused by the scattering of light. The defects causing haze are larger than those causing fog, but singly are not large enough to be seen by the unaided eye.
3-DAmerican Society for Cell BiologyAmericasassayBethesdabiologyBiophotonicsBioScanCellcisternaedivisionendoplasmic reticulumenzymeerhazehigh-resolutionImagingMarylandmembraneMicroscopymitosisNational Institute of Child Health and Human DevelopmentNational Institutes of HealthNew OrleansNIHorganellesprocessproteinsscientistsstructuretransporttrappingtubularGolgiapparatussynthesizedpunctaDylan BurnettePrabuddha SenguptaJennifer Lippincott-SchwartzEunice Kennedy ShriverNICHDchoreographeddisassemblycytokinesispharmacologically

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