Clothilde HEUDE - Université de Caen Basse Normandie

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Clothilde HEUDE
http://w3.unicaen.fr/ufr/ibfa/biomea/spip.php?article54
Clothilde HEUDE
- Fiches - Fiches personnels - Enseignants chercheurs -
FRE3484 BioMEA
Date de parution : 1er janvier 2012
Date de mise en ligne : lundi 16 novembre 2009
Mise à jour le : dimanche 1er janvier 2012
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Sommaire
• Responsabilités administratives
• Responsabilités pédagogiques
• Responsabilités scientifiques
• Participation à des programmes
• Thèmes de recherche
• Liste complète des publications
Clothilde HEUDE
Enseignant chercheur, Maître de Conférences Université de Caen Basse-Normandie Section de Recherche : CNU
68 Equipe de rattachement : Physiologie de la reproduction des Mollusques Marins (PhyMR).
E mail : clothilde.heude@unicaen.fr Tel : 02 31 56 51 14 Adresse postale CNRS INEE - FRE3484 BioMEA Biologie
des Mollusques Marins et des Ecosystèmes Associés Esplanade de la Paix IBFA - UNIVERSITE DE CAEN
BASSE-NORMANDIE 14032 CAEN CEDEX
Responsabilités administratives :
" Membre du vivier interne de constitution du Comité de Sélection de 68ème section (UCBN) " Membre extérieur du
comité de sélection (68ème section), Université du Havre.
Responsabilités pédagogiques
" Responsable pédagogique de la première année d'étude de l'ESIX AgroA " Présidente du jury d'examen de première
année ESIX AgroA " Membre du Conseil de Perfectionnement de l'ESIX AgroA " Membre des jurys d'examen de
deuxième année ESIX AgroA et licence professionnelle « Innovation et Assemblages Culinaires »
Responsabilités scientifiques :
" Programme de Recherche Vème Plan Régional : Appui scientifique aux filières professionnelles de la pêche et des
cultures marines. Coordinatrice de l'Action 3 : Bases d'étude de la reproduction des mollusques marins en vue d'une
exploitation durable par les professionnels de la côte ouest du Cotentin.
Participation à des programmes
" Projet européen REPROSEED (2009-2012). Research to improve Production of seed of established and emerging
bivalve species in European hatcheries " Programme de Recherche Vème Plan Regional (2008-2013). Appui
scientifique aux filières professionnelles de la pêche et des cultures marines. Action 4 : Comparaison d'huîtres
diploïdes et triploïdes : développement de marqueurs liés à la reproduction et à la mise en réserve énergétique pour
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l'exploitation de Crassostrea gigas.
Thèmes de recherche
Clothilde HEUDE
" Mots clés : Mollusques - Gamétogenèse - Spermatogenèse - Proliférations cellulaires - Cellules somatiques
intragonadiques
La thématique développée tend à explorer la physiologie de la reproduction des mollusques bivalves avec comme
espèce modèle l'huître creuse Crassostrea gigas. La régulation des proliférations cellulaires au cours de la
gamétogenèse est un axe important de recherche abordé par des approches cellulaires, immunologiques et
moléculaires. Les premiers tris de cellules germinales d'huître (spermatides et spermatogonies/spermatocytes) sont
en cours de développement.
Liste complète des publications
2013
Jouaux, A., Blin, J. L., Adeline, B., Heude-Berthelin, C., Sourdaine, P., Mathieu, M., et al. (2013).
Impact of energy storage strategies on gametogenesis and reproductive effort in diploid and triploid
Pacific oysters Crassostrea gigas . Involvement of Insulin signaling. Aquaculture, .
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2012
Jouaux A, Franco A., Heude-Berthelin C, Sourdaine P, Blin J.L, Mathieu M, et al. (2012).
Identification of Ras, Pten and P70S6K homologs in the Pacific oyster Crassostrea gigas and diet
control of insulin pathway. General and Comparative Endocrinology., 176(1), 28-38.
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2011
Clothilde HEUDE
Franco, A., Kellner K., Goux D., Mathieu M., & Heude Berthelin C. (2011). Intragonadal Somatic
Cells (ISCs) in the male oyster Crassostrea gigas: Morphology and contribution in germinal
epithelium structure. Micron, 42(7), 718-725.
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Franco, A., Kellner, K., Mathieu, M., Lelong, C., Goux, D., & Heude-Berthelin, C. (2011). Male
germ cells of the Pacific oyster Crassostrea gigas: flow cytometric analysis, cell sorting and
molecular expression. Aquatic Living Ressources, 24, 237-245.
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Heude-Berthelin, C., Hégron-Macé, L., Legrand, V., Jouaux, A., Adeline, B., Mathieu, M., et al.
(2011). Growth and reproduction of the common whelk Buccinum undatum in west Cotentin
(Channel), France. Aquat. Living Resour, 24, 317-327.
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2010
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Clothilde HEUDE
Franco, A., Jouaux, A., Mathieu, M., Sourdaine, P., Lelong, C., Kellner, K., et al. (2010).
Proliferating cell nuclear antigen in gonad and associated storage tissue of the Pacific oyster
Crassostrea gigas: seasonal immunodetection and expression in laser microdissected tissues. Cell
and Tissue Research, 340(1), 201-210.
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Jouaux, A., Heude-Berthelin, C., Sourdaine, P., Mathieu, M., & Kellner, K. (2010). Gametogenic
stages in triploid oysters Crassostrea gigas: irregular locking of gonial proliferation and
subsequent reproductive effort. JEMBE, 395, 162-170.
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2008
Franco, A., Heude Berthelin, C., Goux, D., Sourdaine, P., & Mathieu, M. (2008). Fine structure of
the early stages of spermatogenesis in the Pacific oyster, Crassostrea gigas (Mollusca, Bivalvia).
Tissue & cell, 40(4), 251-260.
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Résumé: The aim of this study is to describe the early stages of spermatogenesis of the Pacific
oyster Crassostrea gigas using both light and electron microscopy. The gonad is formed by
gonadal tubules invaginated in a connective tissue constituting a storage tissue. Myoepithelial cells
surround each gonadal tubule and are associated with an acellular matrix delimiting the outer part
of the tubule, the inner part is composed by intragonadal somatic cells associated with germinal
lineage. Two types of spermatogonia are identified, where type I spermatogonia (Spg I) are large,
scarce and pale cells leaned against the base of the tubule (nuclear diameter: 5.5+/-0.5 microm).
Type II spermatogonia (Spg II) are clustered and dark cells which appear smaller than type I
(nuclear diameter: 4.3+/-0.3 microm). The aspect of nuage-like material in cytoplasm is described
from pale spermatogonia to primary spermatocytes (nuclear diameter: pachytene 3.6+/-0.3
microm, diplotene 3.4+/-0.3 microm), while no structure related to a chromatoid body was observed
in oyster spermatocytes and spermatids.
Mots-Clés: huitre; stages of spermatogenesis; structure
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2006
Clothilde HEUDE
Hanquet-Dufour, A. - C., Kellner, K., Heude, C., Naimi, A., Mathieu, M., & Poncet, J. - M. (2006).
Cryopreservation of Crassostrea gigas vesicular cells: Viability and metabolic activity. Cryobiology,
53(1), 28-36.
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Résumé: Cryopreservation is widely used for long-term conservation of various tissues, embryos
or gametes. However, few studies have described cryopreservation of invertebrate primary cell
cultures and more particularly of marine invertebrate somatic cells. This technique would however
be of great interest to facilitate the study of various metabolic processes which vary seasonally.
The aim of the present study was to develop a protocol for cryopreservation of Crassostrea gigas
vesicular cells. Different parameters were adjusted to improve recovery of cells after freezing. The
most efficient cryoprotectant agent was a mix of Me2SO, glycerol, and ethylene glycol (4% each).
The optimal cooling rate was -1 °C min-1 down to -70 °C before transfer into liquid nitrogen. In
these conditions the percentage of viable cells reached 70% of the control. The glucose
metabolism of thawed cells was evaluated using radioactive glucose as a tracer. Immediately after
thawing, glucose uptake involving membrane transporters was greatly reduced (24% of control)
whereas glucose incorporation into glycogen was less affected (68% of control).
Mots-Clés: Crassostrea gigas; Cryopreservation; Glycogen metabolism; Mollusc; Oyster;
Vesicular cells
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2003
Heude Berthelin, C., Fievet, B., Leclerc, G., Germain, P., Kellner, K., & Mathieu, M. (2003). In vivo
and in vitro approaches to the analysis of glycogen metabolism in the pacific oyster Crassostrea
gigas. Journal of Shellfish Research, 22(3), 715-720.
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Clothilde HEUDE
Kellner, K., Heude-Berthelin, C., & Mathieu, M. (2003). Transport du glucose dans les cellules
vésiculeuses d'huître creuse Crassostrea gigas. Haliotis, 32, 31-40.
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Résumé: Chez Crassostrea gigas, huître creuse du Pacifique, le cycle saisonnier de gestion
des réserves sous forme de glycogène est très strictement corrélé au cycle de
reproduction. En effet, ce glycogène, stocké dans les cellules vésiculeuses du tissu de
réserve, sert de support énergétique à la gamétogenèse. Les travaux de Heude-Berthelin
(2000) ont montré que le métabolisme du glycogène devait être soumis à une régulation
stricte. La mise au point d'un test biologique in vitro utilisant un analogue non métabolisable du
D-glucose, le 3-O-Méthyl-D-Glucose (3-OMeG) a permis de quantifier l'entrée brute de glucose
dans les cellules vésiculeuses isolées. L'effet de la concentration extracellulaire de 3-OMeG
sur l'entrée de 3-OMeG montre l'existence de plusieurs composantes dans l'entrée de glucose
dans la cellule: une composante saturable avec la concentration en substrat et une composante de
diffusion passive non saturable. Afin de mieux définir la composante saturable, l'effet d'un
inhibiteur non spécifique des systèmes de transport facilités, la phlorétine, a montré que la
composante de diffusion simple est négligeable aux concentrations physiologiques de glucose
extracellulaire. Une recherche de transporteurs de type GluT (Glucose Transporteur) a été
entreprise par des techniques immunologiques. En immunofluorescence indirecte avec un
anticorps anti-GluT4 de vertébrés, un marquage est visualisé d'une part sur les cellules
vésiculeuses dissociées, d'autre part sur des coupes de tissu de réserve.
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2002
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Clothilde HEUDE
Kellner, K., Heude-Berthelin, C., & Mathieu, M. (2002). Immunocytochemical demonstration of
glucagon-like peptides in Mytilus edulis cerebral ganglia and an in vitro effect of vertebrate
glucagon on glycogen metabolism. Tissue and Cell, 34(2), 109-116.
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Résumé: Immunological detection of glucagon-like peptides was performed in the cerebral
ganglia of the mussel Mytilus edulis using an anti-vertebrate glucagon antibody. Two clusters of
positive neurosecretory cells were observed, as well as stained nervous fibers. The effect of
vertebrate glucagon on glucose incorporation into glycogen of reserve cells was tested using an in
vitro microplate bioassay. Optimal incubation conditions were previously defined and an inhibitory
effect of porcine glucagon was obtained for concentrations ranging from 10-6 to 10-9 M. It is
postulated that the glucagon-like peptide may be implicated in the regulation of glucose
metabolism in bivalves.
Mots-Clés: cerebral ganglia; glucagon; glycogen metabolism; immunocytochemistry; mollusca;
Mytilus edulis
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2001
Heude-Berthelin, C., Laisney, J., Espinosa, J., Martin, O., Hernandez, G., Mathieu, M., et al.
(2001). Storage and reproductive strategy in Crassostrea gigas of two different growing area
(Normandy and the Atlantic coast, France). Invertebrate Reproduction and Development, 40(1),
79-86.
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Résumé: The reproductive cycle of the Pacific oyster (Crassostrea gigas) is compared in two
different areas (Saint-Vaast la Hougue, Normandy, and Marennes-Oléron, Charentes). Major
differences were observed with respect to the success of spawning: although the general course of
gametogenesis appeared to be similar, gonial mitoses were delayed on the Normandy shore and
the timing of the subsequent expansion of gonadal tubules was different. However, spawning time
was the same in both areas. This spawning was massive in Atlantic oysters and only partial in
Normandy oysters. Storage tissue development in the gonadal area was also investigated and
related to the process of gametogenesis.
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2000
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Clothilde HEUDE
Berthelin-Heude, C., Kellner, K., & Mathieu, M. (2000). Histological Characterization and Glucose
Incorporation into Glycogen of the Pacific Oyster Crassostrea gigas Storage Cells. Marine
Biotechnology, 2(2), 136-145.
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Résumé: Abstract: In order to investigate glycogen metabolism in the oyster Crassostrea gigas,
the distribution of storage cells in the whole animal was studied before histological and biochemical
characterization. These cells were found mainly in the labial palps, the mantle, and gonadal area
and also in gills and the digestive area. Storage cells from palps, mantle, and gonad presented the
same morphological features and the same seasonal glycogen variations. Storage cells were
isolated from the labial palps and the mantle plus gonadal area of the oyster by enzymatic
dispersion and centrifugation through discontinuous Percoll gradient. These cells have a modal
density of 1.043 g/ml. An ultrastructural study confirmed that glycogen is present in the cytoplasm
either as fine particles or sequestered within vesicles. Glucose incorporation into glycogen was
evaluated in vitro using [U-14C]glucose: the incorporation in isolated cells increased linearly for at
least 8 hours, was proportional to the cell concentration, and showed saturation kinetics with
respect to the exogenous glucose concentration.
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Clothilde HEUDE
Berthelin-Heude, C., Kellner, K., & Mathieu, M. (2000). Storage metabolism in the Pacific oyster (
Crassostrea gigas) in relation to summer mortalities and reproductive cycle (West Coast of
France). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology,
125(3), 359-369.
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Résumé: We describe seasonal changes in the biochemical composition of digestive gland,
adductor muscle and gonad and surrounding mantle area in Crassostrea gigas from the Western
Atlantic coast of France. Seasonality in histology of storage tissues and glycogen storage capacity
in isolated vesicular cells were also studied. Proteins, the main muscle components did not
contribute to the gametogenetic effort. Glycogen and lipids were stored in the digestive gland,
gonad and surrounding mantle area during the wintering period and the gonad and surrounding
mantle area represented the main storage compartment supplying the reproductive effort.
Gametogenesis in spring and summer was associated with an increase in lipid and protein
contents and took place at the expense of glycogen reserves. Histological study of storage tissue
in the gonad led us to define four seasonal stages of storage tissue development. In vitro, glycogen
storage capacity in isolated vesicular cells was high from November to March and markedly
reduced during gametogenesis, decreasing below detectable levels after spawning. This
physiological state should be taken into account with relation to summer mortalities occurring in
commercial growing areas.
Mots-Clés: Bivalve; Crassostrea gigas; Gametogenesis; Glycogen; Pacific oyster; Storage tissue;
Summer mortalities; Vesicular cells
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