title: Leptin has concentration and stage-dependent effects on embryonic development in vitro.
abstract: There is accumulating evidence that leptin may be directly involved in pre-implantation embryonic development, however, it is unclear whether there is a concentration and stage-dependent regulatory pattern. In this study, the addition of 10 ng/ml human recombinant leptin to the culture medium significantly increased the percentage of two-cell mouse embryos that developed into blastocysts and hatched blastocysts, whereas in the presence of 100 ng/ml leptin, the development rate was significantly inhibited. The total cell numbers in the hatched blastocysts were significantly higher in the presence of 10 ng/ml leptin compared with controls and higher concentrations. The differential sensitivity to leptin was found to vary among embryos at different stages of development. Supplementation of leptin (10 ng/ml) to culture medium at two- to eight-cell stages resulted in a consistent stimulatory effect on embryo development. Most interestingly, the inhibitory effect of high leptin concentration (100 ng/ml) on embryo development was diminished when it was added to the culture medium at the eight-cell stage of development. The concentration-dependent regulation pattern was confirmed using sheep embryos, under similar conditions although sheep embryos appeared to be more sensitive in responding to leptin. Having established the effect of exogenous leptin on embryo development, the expression pattern of leptin and its receptors were also investigated. Leptin mRNA was not detected in mouse two-, four-, eight-cell and blastocyst stage embryos, whereas three isoforms of leptin receptor (Ob-Ra, Ob-Rb and Ob-Re) were identified in these cells, indicating that leptin is likely to modulate embryo development via a paracrine signalling system.
authors.affiliation: Animal Physiology, University of New England, Armidale NSW 2350, Australia. muren.herrid@csiro.au
authors.forename: Muren
authors.initials: M
authors.lastname: Herrid
authors.forename: Van Ly
authors.initials: VL
authors.lastname: Nguyen
authors.forename: Geoff
authors.initials: G
authors.lastname: Hinch
authors.forename: James R
authors.initials: JR
authors.lastname: McFarlane
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000737:chemistry / Q000187:drug effects / Q000502:physiology*; D002970:Cleavage Stage, Ovum / Q000737:chemistry / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004586:Electrophoresis; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects*; D005260:Female; D020738:Leptin / Q000494:pharmacology*; D051379:Mice; D008815:Mice, Inbred Strains; D011247:Pregnancy; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D011956:Receptors, Cell Surface / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D054411:Receptors, Leptin; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012756:Sheep
chemical_list: C515448:LEPR protein, human; D020738:Leptin; D020033:Protein Isoforms; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D054411:Receptors, Leptin; D011994:Recombinant Proteins; C515447:leptin receptor, mouse