A distinct class of intracellular storage vesicles, identified by expression of the glucose transporter GLUT4.

Herman, Gary; Bonzelius, Frank; Cieutat, Anne-Marie; Kelly, Regis B.

doi:10.1073/pnas.91.26.12750

Cited by 83 publications

(101 citation statements)

References 43 publications

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“…Conversely, anti rabaptin 5-coated and control LAMP II-coated beads retrieved SLMV antigens to a similar extent ( Figure 5E). These findings are consistent with previous reports (Clift-O'Grady et al, 1990;Linstedt and Kelly, 1991b;Bonzelius et al, 1994;Herman et al, 1994;Lichtenstein et al, 1998;de Wit et al, 1999de Wit et al, , 2001) and demonstrate that our isolated PC12 SLMV are reasonably devoid of other contaminant vesicles.To determine whether different populations of vesicles existed within the SLMV pool, we used magnetic beads coated with antibodies against the tails of either ZnT3, synaptophysin, or VAMP II to isolate vesicles from our PC12 SLMV (see SLVM pool in Figure 4B). The protein composition of the isolated vesicles was assessed by immunoblot and compared with the contents of the pooled purified SLMV.…”

supporting

confidence: 83%

The Zinc Transporter ZnT3 Interacts with AP-3 and It Is Preferentially Targeted to a Distinct Synaptic Vesicle Subpopulation

Salazar

Love

Werner

et al. 2004

MBoC

111

178

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Synaptic vesicles (SV) are generated by two different mechanisms, one AP-2 dependent and one AP-3 dependent. It has been uncertain, however, whether these mechanisms generate SV that differ in molecular composition. We explored this hypothesis by analyzing the targeting of ZnT3 and synaptophysin both to PC12 synaptic-like microvesicles (SLMV) as well as SV isolated from wild-type and AP-3-deficient mocha brains. ZnT3 cytosolic tail interacted selectively with AP-3 in cell-free assays. Accordingly, pharmacological disruption of either AP-2-or AP-3-dependent SLMV biogenesis preferentially reduced synaptophysin or ZnT3 targeting, respectively; suggesting that these antigens were concentrated in different vesicles. As predicted, immuno-isolated SLMV revealed that ZnT3 and synaptophysin were enriched in different vesicle populations. Likewise, morphological and biochemical analyses in hippocampal neurons indicated that these two antigens were also present in distinct but overlapping domains. ZnT3 SV content was reduced in AP-3-deficient neurons, but synaptophysin was not altered in the AP-3 null background. Our evidence indicates that neuroendocrine cells assemble molecularly heterogeneous SV and suggests that this diversity could contribute to the functional variety of synapses. INTRODUCTIONThe molecular diversity in total brain synaptic vesicle (SV) composition is generally presumed to result from differential expression of synaptic vesicle membrane proteins in different brain regions. However, the possibility that synaptic vesicles differ in composition because of different biogenesis mechanisms has not been explored. Different vesiculation pathways could result in molecularly diverse synaptic vesicles. Vesiculation mechanisms are known to produce distinct cargo carriers from a population of donor membranes (Bonifacino and Dell'Angelica, 1999;Springer et al., 1999;Boehm and Bonifacino, 2001). This process is achieved by adaptor complexes that recognize and concentrate specific membrane proteins in the donor membranes (Bonifacino and Dell 'Angelica, 1999;Kirchhausen, 1999). PC12 cells have been shown to possess two such adaptor-dependent pathways for the assembly of synaptic vesicles, also known as synaptic-like microvesicles (SLMV) (Shi et al., 1998;de Wit et al., 1999;Jarousse and Kelly, 2001). In one pathway, SLMV are generated from the plasma membrane by a vesiculation mechanism that requires the adaptor AP-2, clathrin, and the GTPase dynamin (Shi et al., 1998). In the second pathway, SLMV are generated from endosomes by the adaptor complex AP-3 (Faundez et al., 1998;Blumstein et al., 2001) and the GTPase ARF1 (Faundez et al., 1997). In contrast to the plasma membrane pathway, the endosomederived mechanism is highly sensitive to brefeldin A (BFA) (Shi et al., 1998).Phenotypes observed in the AP-3-deficient mocha mouse are consistent with a role for the AP-3-dependent, endosome-derived pathway in neurons (Kantheti et al., 1998). The mocha mossy fibers are devoid of both the synaptic vesiclespecific zinc transpor...

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supporting

confidence: 83%

The Zinc Transporter ZnT3 Interacts with AP-3 and It Is Preferentially Targeted to a Distinct Synaptic Vesicle Subpopulation

Salazar

Love

Werner

et al. 2004

MBoC

111

178

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“…Despite considerable efforts to characterize the intracellular GLUT4 compartment in muscle and fat cells it is still unclear whether GLUT4 is targeted to a unique secretory compartment or to a subcompartment within the pre-existing endosomal system [3,4,9,37,38]. In the present study we have utilized the TfR as a prototypic marker of the endosomal\recycling system in an effort to distinguish between each of these possibilities.…”

Section: Discussionmentioning

confidence: 99%

“…The data presented in Figure 7 again confirm that a large pool of GLUT4 has not been affected by the ablation reaction. It is noteworthy that a transferrin-negative intracellular pool of GLUT4 has also been identified following expression of this transporter in PC12 cells by stable transfection [38], suggesting that this compartment may not be unique to insulin-sensitive cells.…”

Section: Okadaic Acid On Deglc Transport and Glut4 Ablation In 3t3-l1mentioning

confidence: 99%

Compartment ablation analysis of the insulin-responsive glucose transporter (GLUT4) in 3T3-L1 adipocytes

Livingstone

James

Rice

et al. 1996

Biochemical Journal

138

159

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The translocation of a unique facilitative glucose transporter isoform (GLUT4) from an intracellular site to the plasma membrane accounts for the large insulin-dependent increase in glucose transport observed in muscle and adipose tissue. The intracellular location of GLUT4 in the basal state and the pathway by which it reaches the cell surface upon insulin stimulation are unclear. Here, we have examined the colocalization of GLUT4 with the transferrin receptor, a protein which is known to recycle through the endosomal system. Using an anti-GLUT4 monoclonal antibody we immunoisolated a vesicular fraction from an intracellular membrane fraction of 3T3-L1 adipocytes that contained 90 % of the immunoreactive GLUT4 found in this fraction, but only 40 % of the transferrin receptor (TfR). These results suggest only a limited degree of colocalization of these proteins. Using a technique to cross-link and render insoluble (' ablate ') intracellular compartments containing the TfR by means of a transferrin-horseradish peroxidase conjugate (Tf-HRP), we further examined the relationship between the endosomal recycling pathway and the intracellular compartment containing GLUT4 in these cells. Incubation of non-stimulated cells with Tf-HRP for 3 h at 37 mC resulted in quantitative ablation of the intracellular TfR, GLUT1 and mannose-6-phosphate receptor and a shift in the density of Rab5-positive membranes. In contrast, only 40 % of intracellular GLUT4 was ablated under the same conditions. Ablation was specific for the endosomal system as there was no significant

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“…Another possibility is that it is associated instead with a novel Ca# + -regulated exocytotic vesicle. Evidence for such an organelle comes from the observation of a third uncharacterized vesicle type in PC12 cells that expresses GLUT4 after transfection [26], and the fact that fibroblasts can demonstrate Ca# + -regulated exocytosis via some unknown vesicle population [13,14]. The data presented here suggest, in contrast, that Doc2 is associated…”

Section: Discussionmentioning

confidence: 65%

Doc2 is not associated with known regulated exocytotic or endosomal compartments in adrenal chromaffin cells

Charvin

Williams

Burgoyne

1999

Biochemical Journal

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Doc2 is a C2-domain-containing protein that is highly expressed in the nervous system and has a constitutively expressed isoform. It has been implicated as a potential Ca2+ sensor in regulated exocytosis, and has been suggested to be associated with synaptic vesicles. To examine whether Doc2 is associated with synaptic-like microvesicles (SLMVs) or dense-core granules in neuroendocrine cells, we examined the distribution of Doc2 in subcellular fractionation of chromaffin cells of the adrenal medulla and in PC12 cells. Doc2 did not co-distribute with SLMVs from either cell type, but did appear to co-distribute with dense-core granules from PC12 cells. In contrast, it was not associated with the dense-core granules during subcellular fractionation of the adrenal medulla, and nor did it appear to be associated with endosomes, cis-Golgi or the trans-Golgi network. In contrast, Doc2 co-distributed under all conditions with a mitochondrial marker. We conclude that Doc2 is not a general component of regulated secretory vesicles, but may instead be associated with mitochondria.

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A distinct class of intracellular storage vesicles, identified by expression of the glucose transporter GLUT4.

Cited by 83 publications

References 43 publications

The Zinc Transporter ZnT3 Interacts with AP-3 and It Is Preferentially Targeted to a Distinct Synaptic Vesicle Subpopulation

The Zinc Transporter ZnT3 Interacts with AP-3 and It Is Preferentially Targeted to a Distinct Synaptic Vesicle Subpopulation

Compartment ablation analysis of the insulin-responsive glucose transporter (GLUT4) in 3T3-L1 adipocytes

Doc2 is not associated with known regulated exocytotic or endosomal compartments in adrenal chromaffin cells

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