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First published online March 31, 2005
Journal of Experimental Biology 208, 1551-1561 (2005)
Published by The Company of Biologists 2005
doi: 10.1242/jeb.01567

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Influence of environmental conditions on early development of the hydrothermal vent polychaete Alvinella pompejana

Florence Pradillon^1,*, Nadine Le Bris², Bruce Shillito¹, Craig M. Young³ and Françoise Gaill¹

¹ UMR CNRS 7138, Systématique, Adaptation et Evolution, Université Pierre et Marie Curie, 7 quai Saint-Bernard, 75252 Paris Cedex 05, France,
² IFREMER, Direction des Recherches Océaniques, Département Environnement Profond, BP 70, Plouzané F-29280, France
³ Oregon Institute of Marine Biology, University of Oregon, PO Box 5389, Charleston, OR 97420, USA

View larger version (99K): [in a new window]   Fig. 1. ELSA (13°N/EPR), the instrumented chimney where incubators were deployed. (A) Lateral view of the ELSA chimney with its faunal coverage, described according to the dominant species observed on video recordings. Positions of incubators I1, I2 and I3 deployed on the chimney are indicated. (B) Incubator I1 (arrowhead) deployed on oxidised sulphide blocks at the base of the chimney. (C) Close-up view of incubator I2 (arrowhead) inside the Riftia pachyptila clump. (D) Close-up view of the upper part of the chimney with incubator I3 (arrowhead) and the autonomous probe deployment (black arrow).

View larger version (84K): [in a new window]   Fig. 2. Early development of Alvinella pompejana after in vitro fertilisation and incubation at 26 MPa. (A) Oocyte just after fertilisation. (B) Fertilisation envelope. (C) First polar lobe. (D) 2-cell stage. (E) 4-cell stage. (F) 8-cell stage. (G) 12-cell stage. (H) >12-cell stage. A and B are from cultures incubated at atmospheric pressure, 5 and 30 min after fertilisation, respectively. Embryos in C, D, E and G were obtained after in situ incubation at the bottom of the ocean at 5°C for 5 days. Embryos in F and H were obtained after incubation in pressure vessels at 10°C for 72 h and 96 h, respectively. Scale bars, 50 µm.

View larger version (129K): [in a new window]   Fig. 3. Abnormal morphologies observed during in vitro development in Alvinella pompejana. Observations were made after 96 h of incubation at 10°C (A,B) and after 24 h ofincubation at 20°C (C,D), at 1 atmosphere. (A) Embryo with abnormal general shape and bubbles (arrowhead). (B) Embryo with abnormal cleavage giving one large blastomere and many very small cells. (C) Bubble formation (arrowhead) at the surface of the oocyte, possibly responsible for subsequent breaking of the cell. (D) Oocyte with broken plasma membrane and cytoplasm spreading into the perivitelline space. B was obtained using SEM; other pictures of live embryos before fixation were recorded on board ship. Scale bars, 50 µm.

View larger version (35K): [in a new window]   Fig. 4. Alvinella pompejana embryos cultured at 2–27°C and 1 atm. Histograms show the frequency distribution of developmental stages achieved by at least 20 embryos sampled in the main culture. Embryos were observed 12 h, 24 h, 48 h and 72 h after fertilisation except in two cases: * observation 16 h after fertilisation, and ** observation 63 h after fertilisation. U, uncleaved embryos; 2c, 2-cell stage; 4c, 4-cell stage; 8c, 8-cell stage; 16c, 16-cell stage and more; D, degrading embryos.

View larger version (9K): [in a new window]   Fig. 5. Effect of temperature on cleavage rates in Alvinella pompejana embryos incubated at 1 atm. Cleavage times were estimated from the frequency of developmental stages presented in Fig. 4.

View larger version (14K): [in a new window]   Fig. 6. Alvinella pompejana embryos cultured at 2–20°C and 250 atm. Histograms show the frequency distribution of developmental stages achieved by at least 20 embryos sampled in the main culture. Embryos were observed after 24 h, 48 h, 72 h and 8 days of incubations. U, uncleaved embryos; 2c, 2-cell stage; 4c, 4-cell stage; 8c, 8-cell stage; 16c, 16-cell stage and more; D, degrading embryos.

View larger version (10K): [in a new window]   Fig. 7. Alvinella pompejana embryos cultured at the ELSA hydrothermal vent site, positions I1, I2 and I3 (see Fig. 1). Incubators were deployed for 5 days before recovery by the submersible. U, uncleaved embryos; 2c, 2-cell stage; 4c, 4-cell stage; 8c, 8-cell stage; >8c, above 8-cell stage; D, degrading embryos.

View larger version (33K): [in a new window]   Fig. 8. Temperature recorded over the duration of the experiment by autonomous probes. The higher temperature recordings were taken at the surface of the alvinellid colony about 20 cm from incubator I3, and the lower ones at the base of the chimney a few dm from incubator I1.