Gap junction assembly in the preimplantation mouse conceptus is independent of microtubules, microfilaments, cell flattening, and cytokinesis.

Kidder, Gerald M.; Rains, J.; McKeon, Joanie

doi:10.1073/pnas.84.11.3718

Cited by 32 publications

(10 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The plaque regions are in continuous motion, moving around at considerable speed, changing shape, fusing frequently with each other, and separating into smaller patches. This is in agreement with the view that neither microfilaments nor microtubules are needed for gap junction assembly (Kidder et al 1987;Feldman et al 1997;George et al 1999), although certain aspects of clustering and stability may be regulated by these filament systems (e.g., Rassat et al 1982;Wang and Rose 1995), and connexins may associate with cortical linker and/or signaling molecules (e.g., Giepmans and Moolenaar 1998;Toyofuku et al 1998). However, in contrast to Jordan et al (1999) who were only able to record fluorescence for up to 37.3 min, we should like to stress that the gap junctions formed in our cells were highly motile.…”

Section: Discussionsupporting

confidence: 92%

Visualization of gap junction mobility in living cells

Windoffer¹,

Beile²,

Leibold³

et al. 2000

Cell and Tissue Research

View full text Add to dashboard Cite

In order to study the dynamics of gap junctions in living cells, a cDNA was expressed in hepatocellular carcinoma-derived PLC cells coding for chimerical polypeptide Cx.EGFP-1, which consists of rat connexin32 and enhanced green fluorescent protein (EGFP). Cx.EGFP-1 was integrated into gap junctions, and the emitted epifluorescence reliably reported the distribution of the chimera. Therefore, stably transfected PLC clone PCx-9 was used to examine the dynamic behavior of gap junctions by time-lapse fluorescence microscopy. The pleomorphic fluorescent junctional plaques were highly motile within the plasma membrane. They often fused with each other or segregated into smaller patches, and fluctuation of fluorescence was detected within individual gap junctions. Furthermore, the uptake of junctional fragments into the cytoplasm of live cells was documented as originating from dynamic invaginations that form long tubulovesicular structures that pinch off. Endocytosis and subsequent lysosomal degradation, however, appeared to contribute only a little to the rapid gap junction turnover (determined half-life of 3.3 h for Cx.EGFP-1), since most cytoplasmic Cx.EGFP-1 fluorescence did not colocalize with the endocytosed fluid phase marker horseradish peroxidase or the receptor-specific endocytotic ligand transferrin and since it was distinct from lysosomes. Disassembly of gap junctions was monitored in the presence of the translation-inhibitor cycloheximide and showed increased endocytosis and continuous reduction of junctional plaques. Highly motile cytoplasmic microvesicles, which were detectable as multiple, weakly fluorescent puncta in all movies, are proposed to contribute significantly to gap junction morphogenesis by the transport of small subunits between biosynthetic, degradative, and recycling compartments.

show abstract

Section: Discussionsupporting

confidence: 92%

Visualization of gap junction mobility in living cells

Windoffer¹,

Beile²,

Leibold³

et al. 2000

Cell and Tissue Research

View full text Add to dashboard Cite

show abstract

“…Accumulation of both Cxn43 mRNA and protein begins at the 4-cell stage (Nishi et al, 1991;Valdimarsson et al, 1991), but insertion of gap junctions into the plasma membranes does not occur until the 8-cell stage, when embryo compaction begins. Inhibition of transcription and protein synthesis at the 4-cell stage does not prevent the formation of gap junctions at the correct time (Kidder and McClachlin, 1985;Kidder et al, 1987), implying that cytoplasmic stores of Cxn43 detectable at the 4-cell stage (De Sousa et al, 1993) are sufficient for preliminary gap junction assembly. Some oogenic Cxn32 protein has been reported to persist during early mouse development (Barron et al, 1989).…”

Section: Connexins In the Preimplantation Mouse Embryo: A Brief Overviewmentioning

confidence: 99%

Role of gap junctions in the development of the preimplantation mouse embryo

Becker

Davies

1995

Microscopy Res & Technique

View full text Add to dashboard Cite

We have taken several approaches to study the role of gap junctional communication during preimplantation mouse development. Firstly, the normal expression pattern of gap junctions has been characterized using immunostaining in conjunction with laser scanning confocal microscopy. Changes in junctional distribution have been correlated with developmental events. We have gone on to study development and junctional organization in mice which naturally exhibit reduced cell to cell communication (DDK syndrome), and in normal mice in which gap junction permeability has been artificially manipulated. Furthermore, anti-peptide antibodies have been tested for their ability to block gap junction communication and for the effects of such a block on subsequent development. Collectively, the results demonstrate that gap junctional communication plays an important role in the maintenance of compaction and the differentiation of an organized epithelium within an embryo, features which are vital for preimplantation development to progress successfully.

show abstract

“…How the junctions are anchored is not yet clear. It has been claimed a number of times that gap junctions interact with cytoskeletal elements (3,9), although this is not universally accepted (5) . The well anchored plaques may be located in regions where the lipid has not been completely removed, or be plaques that have strong connections to cytoskeletal or other cellular structures, either naturally or as a result of fixation.…”

Section: And Discussionmentioning

confidence: 99%