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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 PLoS+One
2016 ; 11
(10
): e0165162
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gab.com Text
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English Wikipedia
Loukoumasomes Are Distinct Subcellular Structures from Rods and Rings and Are
Structurally Associated with MAP2 and the Nuclear Envelope in Retinal Cells
#MMPMID27798680
Noble JW
; Hunter DV
; Roskelley CD
; Chan EK
; Mills J
PLoS One
2016[]; 11
(10
): e0165162
PMID27798680
show ga
"Rods and rings" (RR) and loukoumasomes are similarly shaped, subcellular
macromolecular structures with as yet unknown function. RR, so named because of
their shape, are formed in response to inhibition in the GTP or CTP synthetic
pathways and are highly enriched in the two key enzymes of the nucleotide
synthetic pathway. Loukoumasomes also occur as linear and toroidal bodies and
were initially inferred to be the same as RR, largely due to their shared shape
and size and the fact that it was unclear if they shared the same subcomponents.
In human retinoblastoma tissue and cells we have observed toroidal, perinuclear,
macromolecular structures of similar size and antigenicity to those previously
reported in neurons (neuronal-loukoumasomes). To further characterize the
subcomponents of the retinal-loukoumasomes, confocal analysis following
immunocytochemical staining for alpha-tubulin, beta-III tubulin and detyrosinated
tubulin was performed. These studies indicate that retinal-loukoumasomes are
enriched for beta-III tubulin and other tubulins associated with microtubules.
Immunofluorescence together with the in situ proximity ligation assay (PLA),
confirmed that beta-III tubulin colocalized with detyrosinated tubulin within
loukoumasomes. Our results indicate that these tissues contain only loukoumasomes
because these macromolecular structures are immunoreactive with an anti-tubulin
antibody but are not recognized by the prototype anti-RR/inosine monophosphate
dehydrogenase (IMPDH) antibody (It2006). To further compare the RR and
retinal-loukoumasomes, retinoblastoma cells were exposed to the IMPDH-inhibitor
ribavirin, a drug known to induce the formation of RR. In contrast to RR, the
production of retinal-loukoumasomes was unaffected. Coimmunostaining of Y79 cells
for beta-III tubulin and IMPDH indicate that these cells, when treated with
ribavirin, can contain both retinal-loukoumasomes and RR and that these
structures are antigenically distinct. Subcellular fractionation studies indicate
that ribavirin increased the RR subcomponent, IMPDH, in the nuclear fraction of
Y79 cells from 21.3 ± 5.8% (0 mM ribavirin) to 122.8 ± 7.9% (1 mM ribavirin)
while the subcellular localization of the retinal-loukoumasome subcomponent
tubulin went unaltered. Further characterization of retinal-loukoumasomes in
retinoblastoma cells reveals that they are intimately associated with lamin folds
within the nuclear envelope. Using immunofluorescence and the in situ PLA in this
cell type, we have observed colocalization of beta-III tubulin with MAP2. As MAP2
is a microtubule-associated protein implicated in microtubule crosslinking, this
supports a role for microtubule crosslinkers in the formation of
retinal-loukoumasomes. Together, these results suggest that loukoumasomes and RR
are distinct subcellular macromolecular structures, formed by different cellular
processes and that there are other loukoumasome-like structures within retinal
tissues and cells.