QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online 19 October 2004
doi: 10.1242/jcs.01424

This Article Summary Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Thomas, F. C. Articles by Fleming, T. P. Search for Related Content PubMed PubMed Citation Articles by Thomas, F. C. Articles by Fleming, T. P. Social Bookmarking                         What's this?

Contribution of JAM-1 to epithelial differentiation and tight-junction biogenesis in the mouse preimplantation embryo

¹ School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton, SO16 7PX, UK
² Developmental Origins of Health and Disease Division, School of Medicine, University of Southampton, Coxford Road, Southampton, SO16 5YA, UK
³ Laboratory of Vascular Biology, Istituto di Ricerche Farmacologiche, Mario Negri, 20157 Milano, Italy

View larger version (58K): [in a new window]   Fig. 1. (A) RT-PCR analysis of the expression of JAM-1-encoding mRNA in egg and embryo stages. The 600 bp cDNA product is indicated by arrows. (Left) Produced using poly(A)+ RNA with five eggs or embryos per sample. (Right) Total RNA using one egg or embryo equivalent per sample. `Co' indicates the control without the reverse transcription step. (B) Immunoblot analysis of JAM-1 protein expression in eggs and embryos using BV20 antibody, showing approximately 500 eggs or embryos per lane. Mouse lung tissue (18 µg protein) was used as positive tissue control (TC). The stages are unfertilized eggs (UF); fertilized eggs (FE) and embryos at two-cell (2C), four-cell (4C), precompact eight-cell (8C), compact eight-cell (C8), early morula (EM), late morula (LM), early blastocyst (EB) and late blastocyst (LB) stages. AbC, secondary-antibody-only control.

View larger version (108K): [in a new window]   Fig. 2. Immunofluorescence confocal microscopy of E-cadherin (A-H) and JAM-1 (I-P) staining patterns during preimplantation development. The stages are unfertilized eggs (UF) and embryos at two-cell (2C), four-cell (4C), precompact eight-cell (8C), compact eight-cell (C8C), morula (Mor), early blastocyst (EB) and late blastocyst (LB) stages. Bar, 20 µm.

View larger version (72K): [in a new window]   Fig. 3. (A) Co-localization of JAM-1 and actin (phalloidin staining) at the apical pole (arrows) in compact eight-cell embryos. (B) JAM-1 localization in representative eight-cell embryos at 0 hours, 3 hours, 6 hours and 9 hours after division in z-axis series (z) and single mid-plane (mp) sections. Bar, 20 µm. (C) Timing of cell-cell contact and apical polar staining of JAM-1 in eight-cell embryos at different times during the fourth cell cycle (n=30-40 embryos per time point; experiment repeated four times).

View larger version (35K): [in a new window]   Fig. 4. (A) JAM-1 localization in representative 2/8 couplets at 0 hours, 3 hours, 6 hours and 9 hours after division from 1/4 blastomeres. E-cadherin and negative control (secondary antibody only) are shown at the bottom. Bar, 20 µm. (B) Timing of cell-cell contact and apical polar staining of JAM-1 in 2/8 couplets at different times after division from 1/4 cells (data from single experiment comprising 39 couplets).

View larger version (84K): [in a new window]   Fig. 5. (A) Localization of PKC isoforms , / and within representative compact eight-cell embryos (C8). Arrows indicate immunostaining at the apical membrane domain of polarized blastomere for PKC and . (B) Immunolocalization of nectin-2 (arrows) within representative compact eight-cell embryo (C8), 16-cell morula (16-c) and blastocyst (Blast; left, mid-plane showing staining at trophectoderm junctions and at ICM cell contacts; right, z-axis series showing trophectoderm in tangential plane; Bl, blastocoel). n=10-20 embryos per stage or treatment in three replicates. Bar, 20 µm.

View larger version (12K): [in a new window]   Fig. 6. (A) Effect of embryo culture during the eight-cell stage in T6 plus BSA plus BV11 or BV12 anti-JAM-1 antibodies compared with control IgG2b antibody (all at 5 µg ml–1) on timing of compaction (mean of 89 embryos per treatment). (B) Effect of embryo culture from compaction in media as above on timing of cavitation (mean of 50 embryos per treatment). Rates of cavitation significantly lower than in control medium are indicated with asterisks: *P<0.005; **P<0.0001.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

Twitter