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MODO ( MOD el O rganisms) fellowship

Stazione Zoologica Anton Dohrn. MODO ( MOD el O rganisms) fellowship. Identification of thyroid hormon precursor(s) in Ciona intestinalis endostyles. Maria Salzano. Tutor: Doct. Antonietta Spagnuolo Cellular and Developmental Biology Group. Satoh, 2003.

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MODO ( MOD el O rganisms) fellowship

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  1. Stazione Zoologica Anton Dohrn MODO (MODel Organisms) fellowship Identification of thyroid hormon precursor(s)in Ciona intestinalis endostyles Maria Salzano Tutor: Doct. Antonietta Spagnuolo Cellular and Developmental Biology Group Satoh, 2003 MODO meeting: 27th of June 2013

  2. The link between Endostyle and Thyroid Anatomical and molecular studies confirmed that a THYROID-LIKE FUNCTION lies within the ENDOSTYLES also of Urochordates and Cephalochordates. (Ogasawara et al., 1999) Specific endostyle cells have been shown to concentrate radioiodine from sea water (Barrington, 1957), and to produce monoiodiotyrosine (MIT), diiodiotyrosine (DIT) and thyroxine (Roche et al., 1959); (Kennedy, 1966); (Dunn, 1974). Thyroidhormones play a criticalrole in amphibians[Furlowet al.,2006; Tata, 2005], urochordate[Patricoloet al., 2001], amphioxus[Paris and Laudet, 2008] and lamprey[Manzonet al., 2001] metamorphosis. In vertebrates TG is the protein precursor of T3 and T4 PROJECT AIM Identification of the protein(s) used as scaffold for THs biosynthesis in Ciona genome, no homologus of TG gene has been found

  3. V D As negative control.. Preliminary results Free T4 Endostyles total proteins extracts LC analysis on column Proteins are dried and digested (pronase) T3/T4 bound to protein/s analysis Proteins are removed (ice EtOH; -20°C) free T3/T4 analysis * in collaboration with A. Amoresano’s group Selected Reaction Monitoring mass spectroscopy analysis

  4. Preliminary results (2) PEPTIDE IDENTIFICATION Software for identification, characterisation and quantization of proteins using mass spectrometry data. Proteins are dried and digested (pronase) T3/T4 bound to Protein analysis * in collaboration with A. Amoresano’s group

  5. A parallel strategy… IP anti T4-Ab Sup NB Endostyles TE Endostyles total proteins extracts immunoprecipitated with anti T4-Ab IP Anti T4-Ab: Sup NB : Supernatant of proteins Not Ab - Bound Protein Immuno – Precipitated with anti T4-Ab Endostylus : TE Endostyles proteins Total Extract 75 Kda 50 Kda 37 Kda 25 Kda * * in collaboration with Prof. Pucci’s group

  6. Next analysis Selected Reaction Monitoring mass spectroscopy analysis 1) 2) blood + KClO4 Is there T4? Is T4 free or bound to proteins? What are the effects on T4 production in endostyles??

  7. THANKS FOR YOUR ATTENTION

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