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

First published online 16 May 2007
doi: 10.1242/dev.002642

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 Billon, N. Articles by Dupin, E. Search for Related Content PubMed PubMed Citation Articles by Billon, N. Articles by Dupin, E. Social Bookmarking                         What's this?

The generation of adipocytes by the neural crest

Nathalie Billon^1,*, Palma Iannarelli², Miguel Caetano Monteiro¹, Corinne Glavieux-Pardanaud^3,, William D. Richardson², Nicoletta Kessaris², Christian Dani¹ and Elisabeth Dupin^3,

¹ Institut de Recherche, Signalisation, Biologie du Développement et Cancer, CNRS UMR 6543, Centre de Biochimie, Faculté des Sciences, Université Nice Sophia-Antipolis, Nice, France.
² Wolfson Institute for Biomedical Research and Department of Biology, University College London, Gower Street, London WC1E 6BT, UK.
³ Laboratoire d'Embryologie Cellulaire et Moléculaire, CNRS UMR 7128, Nogent sur Marne, France.

View larger version (48K): [in this window] [in a new window]   Fig. 1. Development of adipocytes from ES cell-derived neuroepithelial precursors. Sox2-ßgeo/oct4-tk genetically engineered ES cells were treated with RA and then selected between day 6 (D6) and day 10 (D10) to enrich for neuroepithelial cells and to eliminate residual undifferentiated ES cells. They were then induced to differentiate towards the adipocyte lineage. (A) At various times, the cells were processed for quantitative PCR analysis using Oct4, Sox1, Sox2, Sox9, Sox10, FoxD3 or GAPDH probes. Data were normalised relative to GAPDH amplification and the highest expression was defined as 100%. Similar results were obtained in two independent experiments. +sel, with selection; -sel, no selection. (B) After selection (D10), neuroepithelial precursors were stained with anti-FoxD3, anti-Sox9 or anti-Sox10 antibody (red) to identify NC-like cells, and with bisbenzimide to identify cell nuclei (blue). 14 days after induction of adipocyte differentiation (+), adipocytes were identified either using their characteristic morphology (C) or by RT-PCR to detect FABP4 mRNA (D). The results in D are shown for two independent experiments. Scale bar: 100 µm.

View larger version (45K): [in this window] [in a new window]   Fig. 2. Development of adipocytes in primary cultures of quail cephalic NC cells (CNCC). (A,B) Primary cultures of CNCC were obtained from mes-rhombencephalon of HH stage 9 quail embryo in explant culture and expanded for 5 days in cloning medium. Adipocyte differentiation was then induced using different media (cloning, L1 and hmads) and adipogenic treatments (DIF1 or DIF2). Adipocytes were identified after 15 days. (A) Typical adipocytes show lipid droplet-filled cytoplasm (left) and are stained with Oil Red O, which reveals neutral lipids (right). Scale bar: 100 µm. (B) Quantification of CNCC primary cultures containing adipocytes after treatment with the mentioned media and adipogenic treatments. A total of 100 cultures were analysed. (C,D) Secondary cultures of quail CNCC were isolated from 48-hour primary cultures. Adipocyte differentiation was then induced at day 6 using the mentioned media and adipogenic treatments. (C) Quantification of secondary CNCC cultures containing adipocytes after 15 days of treatment. A total of 84 cultures were analysed. (D) Expression of CEBP, PPAR, FABP4 and 18s RNAs after 3 days in cloning medium+DIF1. The results are shown for two independent CNCC cultures out of 10 distinct experiments.

View larger version (47K): [in this window] [in a new window]   Fig. 3. Development of adipocytes in secondary cultures of quail trunk NCC (TNCC). TNCC that had migrated from cultured thoracic neural tubes were replated into secondary cultures and treated as in Fig. 2. (A) Typical adipocytes showing Oil Red O-stained lipid droplets (left) and SMP-positive glial cells (right) were identified after 20 days in adipogenic conditions. Scale bar: 100 µm. (B) Quantification of TNCC cultures containing adipocytes after treatment with the mentioned media and adipogenic treatments. A total of 42 cultures were analysed.

View larger version (106K): [in this window] [in a new window]   Fig. 4. Development of adipocytes from the NC during mouse development. Permanent genetic lineage labelling of pre- and post-migratory NC was achieved by crossing transgenic mice carrying a Sox10-Cre construct into a R26-YFP reporter. Double immunolabelling of P28 Sox10-Cre/R26-YFP offspring with anti-GFP (green) and anti-perilipin (red) antibodies was used to identify NC derivatives and adipocytes, respectively. Bisbenzimide was used to identify cell nuclei (blue). Sections show salivary gland and ear (A-D), peri-ovarial (E-H) and trunk subcutaneous (I-L) regions. There is almost complete colocalisation of YFP and perilipin in the salivary gland area, whereas no overlapping can be detected in the ovary and trunk subcutaneous adipose depots. Scale bar: 50 µm.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

Twitter