M.J. DARIAS*, D. MAZURAIS, G. KOUMOUNDOUROS, E. University of Patras, Greece. * Present address
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1 M.J. DARIAS*, D. MAZURAIS, G. KOUMOUNDOUROS, E. GISBERT, C. CAHU, J.L. ZAMBONINO-INFANTE IFREMER Centre de Brest, Frnce University of Ptrs, Greece * Present ddress
2 lrvi 2009 MALFORMATIONS NUTRITION Lrve need more VM Mix VITAMINS A An overdose produce thn juveniles A smll mount of VM growth dely nd induce cephlic mlformtions nd induce mlformtions D C one verter less (Mzuris et l., 2008) (Gisert et l., 2005;?? Villeneuve et l., 2005, 2006; Mzuris et l., 2009)
3 lrvi 2009 Clcium nd phosphte homeostsis Protection of skeletl integrity Co-sustrte for hydroxylse nd oxygense enzymes involved in the iosynthesis of pro-collgen Antioxidnt Pro-oxidnt Hilton & Ferguson, 1982 Grff et l., 2002 Hg et l., 2004 Hlver, 1957,1989 Andrews & Muri, 1975 Lim & Lovell, 1978 Chávez de Mrtínez et l., 1990 Solimn et l., 1986 No studies t lrvl stge Studies t lrvl stge Drowski, 1990 Gpsin et l., 1998
4 lrvi 2009 Poor minerliztion Pughedness Deformities of the cudl fin Verterl deformities (Kyphosis, scoliosis) Brnchiostegl rys deformities Poor minerliztion Crtilge dmge Pughedness One vertere lost Crtilginous vertere Heml rch not formed Epurls, uroneurl, specilized neurl rch Minerliztion dely Poor minerliztion Verterl deformities (kyphosis, scoliosis) Epurls, uroneurl, specilized neurl rch Brnchiostegl rys deformities Deformities of dentry Deformities of the dorsl nd nl fin Supernumerry vertere 8 times the requirements of juvenil fish (NRC, 1993) evel / lrve) Mlfo ormtions (%) sifiction le pixel numer / Frec cuency (%) Oss (Totl p vertere 25 vertere CONTROL VC-15 VD-0 VC-30 VD-19.2 VC-50 VD-38.4 VC-400 VD-140 VD-0 VD-19.2 VD-38.4 VD CONTROL VC-15 VC-30 VC-50 VC-400
5 lrvi 2009 Mlformtio ons (%) evel / lrve) sifiction le pixel numer / Oss (Totl p VD-0 VD-19.2 VD-38.4 VD VD-0 VD-19.2 VD-38.4 VD VC-15 VC-30 VC-50 VC-400 VC-15 VC-30 VC-50 VC-400
6 MOLECULAR PATHWAYS INVOLVED IN MINERALIZATION lrvi 2009 VD 3 Vitmin D Receptor (VDR)? VC INTESTIN Clcium sorption TRPV-6 SVCT-1 VDR BONE Regultion of osteolsts differentition, prolifertion nd minerliztion indirectly directly BMP-4/PPAR γ IGF-1, RARγ Osteoclcin Regultion of one minerliztion Mlformtions?
7 lrvi 2009 CONTROL OF INTESTINAL ABSORPTION TRPV-6 C 2+ sorption Expre ession of TR RPV-6 1,6 1,2 0,8 04 0,4 VD-0 VD-19.2 VD-38.4 VD-140 c c Dy 11 Dy 22 Dy 45 Ex xpression of TRPV-6 1,6 1,2 0,8 0,4 VC-0 VC-5 VC-15 VC-30 VC-50 VC-400 Dy 11 Dy 25 Dy 45 INDIRECT EFFECT EFFECT ON GROWTH ON BONE AND MINERALIZATION DEVELOPMENT
8 lrvi 2009 CONTROL OF INTESTINAL ABSORPTION SVCT-1 VC sorption Totl pix xel numer / lrve Strong lue Ligth Blue Dy 45 VC-15 VC-30 VC-50 VC-400 ression of SVCT-1 Exp 1,4 1,2 1,0 0,8 0,6 0,4 0,2 VC-0 VC-5 VC-15 VC-30 VC-50 VC-400 Dy 11 Dy 15 Dy 25 Dy 45
9 lrvi 2009 CONTROL OF BONY TISSUE DEVELOPMENT IGF-1 Osteolsts prolifertion nd stimultion of its function Exp pression of IGF-1 3,0 2,5 2,0 1,5 1,0 0,5 c VD-0 VD-19.2 VD-38.4 VD-140 Ex xpression of IGF ,5 1,0 0,5 VC-0 VC-15 VC-50 VC-5 VC-30 VC-400 c c Dy 11 Dy 22 Dy 45 Dy 11 Dy 25 Dy 45
10 lrvi 2009 CONTROL OF BONY TISSUE DEVELOPMENT Osteoclcin Osteolsts minerliztion Expres ssion of oste eoclcin 1,00 0, VD-0 VD-19.2 VD VD-140 Expr ression of os steoclcin 100 1,00 VC-0 VC-5 VC-15 VC-30 VC-50 VC-400 0,10 c 1 c Dy 11 Dy 22 Dy 45 0 Dy 11 Dy 25 Dy 45
11 lrvi 2009 CONTROL OF BONY TISSUE DEVELOPMENT VDR Stimultion of ftrpv6 TRPV-6 nd osteoclcin expression Expr ression of VDRβ 1,4 1,2 1,0 0,8 0,6 0,4 0,2 * * * * VD-0 VD-19.2 VD-38.4 VD-140 ** ** ** ** ** *** *** *** Expression of VDRβ 1,8 1,6 1,4 1,2 1,0 0,8 0,6 0,4 0,2 * * * * * * * VC-0 VC-5 VC-15 VC-30 VC-50 VC-400 * * ** ** c ** c c * ** ** ** Dy 11 Dy 22 Dy 45 Dy 11 Dy 25 Dy 45
12 lrvi 2009 CONTROL OF BONY TISSUE DEVELOPMENT BMP-4 Osteolsts differentition MP-4 Expr ression of B 1,5 1,0 0,5 1,5 VC-0 VC-5 VC-15 VC-30 VC-50 VC-400 VD-0 VD ,0 VD-38.4 VD-140 Dy 11 Dy 22 Dy 45 Exp pression of BMP-4 0,5 Dy 11 Dy 15 Dy 25 Dy 45
13 lrvi 2009 CONTROL OF CARTILAGE ADIPOCITIC TISSUE DEVELOPMENT RARγ PPARγ Adipocyte differentition Crtilge formtion Expr pression of of PPARγ RARγ 2,0 1,6 1,2 12 1,2 0,8 0,8 0,4 04 0,4 VC-0 VC-5 VD-0 VC-15 VD-19.2 VC-30 VC-50 VD-38.4 VC-400 VD-140 xpression of RARγ E 2,0 VC-0 VC-5 VC-15 VC-30 1,6 VC-50 VC ,2 0,8 04 0,4 c Dy 11 Dy Dy 1522 Dy Dy 25 45Dy 45 Dy 11 Dy 25 Dy 45
14 Before dy 22 After dy 22 lrvi 2009 Skeletl And elements Skeletl lso elements those tht developed tht tht developed during lter this lter during on period on were this were period more were sensitive sensitive less to sensitive to low low to VDhigh 3 levels to 3 levels high VD 3 levels VD 3 levels pughedness Verterl rnchiostegl & column cudl fin & rnchiostegl deformities rys (8 mmtl, (5 mm rys 25 dph) TL, deformities 11 dph) Multipotent cells BMP-4 + OSTEOBLASTS + + IGF-1 OSSIFICATION DELAY + RARγ ACCELERATION OSSIFICATION 59% MALFORMATIONS 49% VITAMIN D VDR OST OPTIMAL OSSIFICATION 20% ENTEROCYTES TRPV-6 C +2 ~14 IU g -1 ~16 IU g IU g IU g IU g -1
15 lrvi 2009 Skeletl elements Skeletl Skeletl tht And elements developed lso elements those tht lter developed tht on developed were erly more erly lter were sensitive on were less to Beforedy45 Atdy22 sensitive high VC to levels Epurls & specilized high to low VC VC levels neurl rch Epurls, specilized neurl At dy 45 Pughedness rch, dentry, & heml dorsl rches & nl fins, supernumerry vertere OSSIFICATION 50% ADIPOCYTES PPARγγ DELAY Multipotent cells + BMP-4 OSTEOBLASTS IGF-1 + RARγ MALFORMATIONS VITAMIN C VDR OST OPTIMAL 30% OSSIFICATION ENTEROCYTES > 30 mg Kg -1 TRPV-6 C mg kg -1 SVCT-1 VC 400 mg kg -1
16 CONCLUSIONS lrvi 2009 Mturtion of the intestinl functions delyed => effect on lrvl development Disruption of intestinl C 2+ sorption (TRPV-6) Disruption of the expression of genes involved in skeletogenesis (BMP-4, IGF-1, RARγ) nd one minerliztion (VDR, osteoclcin) No evident effect on lrvl development Disruption of intestinl C 2+ & VC sorption (TRPV-6, SVCT-1) Disruption of the expression of genes involved in skeletogenesis (IGF-1, RARγ) nd one minerliztion (VDR, osteoclcin) in fvor of dipocytic tissue formtion (PPARγ) Skeletl elements tht developed in erly nd lter stges were eqully sensitive to low VD 3 levels Skeletl elements tht developed in erly nd lter stges were eqully sensitive to low VC levels Skeletl l elements tht t developed d in erly Skeletl l elements tht t developed d in erly stges were more resistnt to high VD 3 thn stges were more resistnt to high VC thn those tht developed lter on those tht developed lter on Optiml VD 3 is very restricted Optiml VC is restricted 27.6 IU VD 3 /g diet MALFORMARTIONS ~30% 50 mg VC/kg diet (11.5 x dose of juveniles; NRC,1993) (0.5 x dose of juveniles; NRC, 1993)
17 PERSPECTIVES lrvi 2009 Pughedness osteoclcin Deformities of the cudl fin RARg Verterl deformities osteoclcin Brnchiostegl rys deformities osteoclcin Crtilge dmge Crtilginous vertere Pughedness Heml rch not formed One vertere lost osteoclcin PPARg Epurls, specilized neurl rch Verterl deformities osteoclcin Brnchiostegl rys deformities osteoclcin DISRUPTIONS Epurls, specilized neurl rch osteoclcin Deformities of dentry PPARg Deformities of the dorsl nd nl fin DIFFERENT FUNCTION DIFFERENT MODE OF ACTION DIFFERENT ACTION ON THE RATE OF BONE MINERALIZATION CHONDRAL DIFFERENT TYPE OF BONE MINERALIZATION INTRAMEMBRANOUS STUDY OF THE SPECIFIC MOLECULAR MARKERS OF EACH TYPE OF OSSIFICATION ADAPTATION OF THE AMOUNT OF VITAMINS TO THE DEVELOPMENTAL STAGE
18 ACKNOWLEDGEMENTS L of Adpttion ti Reproduction & Nutrition of mrine fish lrvi 2009 Centre de Brest Frnce Unit of Fish Biology & Qulity in Aquculture Biology Deprtment University of Ptrs Greece Thnk You for Your Attention!
19 Before dy 22 After dy 22 lrvi 2009 Skeletl And Skeletl elements lso elements those tht tht tht developed during lter lter during on this on were period this were period were more were very sensitive sensitive less sensitive to low to to high VD low to 3 levels VD high 3 levels VD 3 levels pughedness Verterl rnchiostegl & column cudl & fin rnchiostegl rys deformities (8 mmtl, (5 rys 25 mm dph) deformities TL, 11 dph) Multipotent cells BMP-4 + OSTEOBLASTS + + IGF-1 OSSIFICATION DELAY + RARγ ACCELERATION OSSIFICATION 59% MALFORMATIONS 49% VITAMIN D VDR SCL OST? OSTEOCYTES OPTIMAL OSSIFICATION 20% ENTEROCYTES TRPV-6 C +2 ~14 IU g -1 ~16 IU g IU g IU g IU g -1
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