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Holly’s Work Progress in 08-177

Holly’s Work Progress in 08-177. Dec 2008 - July 2009. Outline. ELISA assays of 6 inbred strains of mice 08-177 B6C3H F2 work progress. ~35 inbred strains in plan, 17% completion. ELISA assays of 6 inbred strains of mice.

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Holly’s Work Progress in 08-177

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  1. Holly’s Work Progress in 08-177 Dec 2008 - July 2009

  2. Outline • ELISA assays of 6 inbred strains of mice • 08-177 B6C3H F2 work progress

  3. ~35 inbred strains in plan, 17% completion ELISA assays of 6 inbred strains of mice

  4. At baseline sRANKL level varies among different strains: C3H and B6 are much higher than NOD/Ltj, A/J, 129/SvimJ, and CAST/Ei is the lowest Following 3wks 100ppm F treatment, only A/J see significant dose response decrease by 64% (p<0.01) Plasma sRANKL ELISA Assays of Inbred Strains of mice

  5. At baseline, A/J and 129/SvimJ PTH level is higher than that of C3H and B6 Following 3wks 100ppm F treatment, A/J sees dose response decrease by 31% (p<0.1), but no significant changes in B6, C3H or 129/SvimJ. Plasma Intact PTH ELISA assays of inbred strains of mice

  6. Serum PTH of F treated (50ppm, 60days) A/J is higher than that of untreated Serum Intact PTH ELISA Assays

  7. Old F2 Genotyping Samples Preparation • 187 genomic DNA samples prepared from previous F2 mice livers/spleens • 140 of 187 samples sent to Jackson Lab in Jun2009

  8. 08-177 B6C3HF2 work progress

  9. B6C3H F2 Treatment Plan

  10. 08-177 B6C3HF2 – Genotyping • 212 genomic DNA samples prepared from 08-177 B6C3HF2 mice livers/Spleens • 156 of 212 samples sent to HPCPG in July 09; the rest of 56 ready to be sent in Aug 09

  11. 08-177 B6C3HF2 – sRANKL ELISA Assays See Hongyou’s presentation

  12. Results & Discussion • At baseline, sRANKL level or intact PTH level varies among different strains; • Following 3 wks of 100ppm F treatment, different strains response differently. Among 6 inbred strains only A/J see significant dose response decrease of sRANKL by 64% (p<0.01) and decrease of intact PTH by 31% (p<0.1), which did not show the similar trend of previous results (Dong Y 2007) – different anesthesia? Serum vs Plasma? F in diet? different length of treatment?...

  13. Future work • Continue work on more inbred strains and B6C3HF2s • Mini study to check the impact of anaethesia (Ketamine/Xylazine or Avertin) on serum or plasma PTH level using ALPCO PTH ELISA kit and verify the effect of fluoride treatment on PTH level. • 40 male C3H mice, 8 groups of 5.

  14. Questions …? Thanks!

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