ÐоÑÑико-ÑÑÑиаÑалÑнÑе клеÑоÑнÑе кÑлÑÑÑÑÑ Ð±Ñли опÑимизиÑÐ¾Ð²Ð°Ð½Ñ Ñ ÑелÑÑ ÑмоделиÑоваÑÑ Ð±Ð¾Ð»ÐµÐ·Ð½Ñ Ð¥Ð°Ð½ÑингÑона in vitro. ÐеÑодом иммÑноÑиÑÐ¾Ñ Ð¸Ð¼Ð¸ÑеÑкого окÑаÑÐ¸Ð²Ð°Ð½Ð¸Ñ Ð¿Ð¾ÐºÐ°Ð·Ð°Ð½Ð° ÑлиминаÑÐ¸Ñ Ñипиков нейÑонов ÑÑÑиаÑÑма на 21 Ð´ÐµÐ½Ñ ÐºÑлÑÑивиÑÐ¾Ð²Ð°Ð½Ð¸Ñ Ð² кÑлÑÑÑÑÐ°Ñ Ð¼ÑÑанÑного Ñипа YAC128, не наблÑÐ´Ð°ÐµÐ¼Ð°Ñ Ð² кÑлÑÑÑÑÐ°Ñ Ð´Ð¸ÐºÐ¾Ð³Ð¾ Ñипа (WT). РазÑабоÑан колиÑеÑÑвеннÑй анализ ÑеноÑипиÑеÑкого пÑоÑÐ²Ð»ÐµÐ½Ð¸Ñ Ð±Ð¾Ð»ÐµÐ·Ð½Ð¸ пÑÑÑм подÑÑÑÑа и оÑенки ÑоÑÐ¼Ñ Ñипиков на поÑÑÑинапÑиÑеÑкой ÑÑоÑоне Ñ Ð¿Ð¾Ð¼Ð¾ÑÑÑ ÐºÐ¾Ð¼Ð¿ÑÑÑеÑного пÑогÑаммного обеÑпеÑÐµÐ½Ð¸Ñ Neuron Studio. ÐÑодемонÑÑÑиÑовано ÑвелиÑение плоÑноÑÑи Ñипиков в кÑлÑÑÑÑÐ°Ñ Ð¼ÑÑанÑного Ñипа пÑи добавлении в кÑлÑÑÑÑалÑнÑÑ ÑÑÐµÐ´Ñ Ð°Ð³Ð¾Ð½Ð¸ÑÑа Ñигма-1 ÑеÑепÑоÑа 3-Ð Ð Ð . ÐокдаÑн Ñигма-1 ÑеÑепÑоÑа вÑзвал ÑлиминаÑÐ¸Ñ Ñипиков в кÑлÑÑÑÑÐ°Ñ Ð´Ð¸ÐºÐ¾Ð³Ð¾ Ñипа и далÑнейÑее ÑменÑÑение плоÑноÑÑи Ñипиков в YAC128 кÑлÑÑÑÑÐ°Ñ . ÐÑи ÑÑом оÑмеÑено изменение ÑооÑноÑÐµÐ½Ð¸Ñ ÑазлиÑнÑÑ Ð²Ð¸Ð´Ð¾Ð² Ñипиков. ÐÑÑÑÑÑÑвие ÑÑÑекÑа воÑÑÑÐ°Ð½Ð¾Ð²Ð»ÐµÐ½Ð¸Ñ Ð¿Ð»Ð¾ÑноÑÑи Ñипиков поÑле Ð´Ð¾Ð±Ð°Ð²Ð»ÐµÐ½Ð¸Ñ 3-РРРв кÑлÑÑÑÑалÑнÑÑ ÑÑÐµÐ´Ñ Ð½ÐµÐ¹Ñонов Ñ Ð½Ð¾ÐºÐ´Ð°Ñном Ñигма-1 ÑеÑепÑоÑа доказало ÑпеÑиÑиÑноÑÑÑ ÐµÐ³Ð¾ воздейÑÑÐ²Ð¸Ñ Ð¸Ð¼ÐµÐ½Ð½Ð¾ на иÑÑледÑемÑй ÑеÑепÑоÑ. С помоÑÑÑ Ð¾ÑмÑвки акÑивиÑÑÑÑего агенÑа Ñ Ð¿Ð¾ÑледÑÑÑей ÑикÑаÑией клеÑок и подÑÑÑÑом плоÑноÑÑи Ñипиков опÑеделена длиÑелÑноÑÑÑ ÑÑÑекÑа акÑиваÑии Ñигма-1 ÑеÑепÑоÑа, оказавÑаÑÑÑ Ñавной пÑимеÑно 8 ÑаÑам в ÑÐ°Ð¼ÐºÐ°Ñ Ð¸ÑÑледÑемой модели. ÐолÑÑеннÑе ÑезÑлÑÑаÑÑ ÑÑаÑиÑÑиÑеÑки обÑабоÑÐ°Ð½Ñ Ð¸ пÑедÑÑÐ°Ð²Ð»ÐµÐ½Ñ Ð² виде гÑаÑиков и ÑÑолбиковÑÑ Ð´Ð¸Ð°Ð³Ñамм. Cortico-striatal cell culture was optimized to model Huntington's disease in vitro. On the 21st day of in vitro cultivation spine elimination of striatal neurons was shown by the method of immunocytochemistry in mutant cultures (YAC128) but not in wild type (WT) ones. Quantitative analysis was worked out to evaluate phenotypic expression of the disease. Spine counting and its shape analysis were performed automatically by Neuron Studio program. Spine rescue in YAC128 cultures was shown after sigma-1 receptor's agonist 3-PPP exposure. Knockdown of sigma-1 receptor caused spine elimination both in YAC128 and WT cultures. Herein change in the ratio of different spine types was marked. No spine rescue effect after 3-PPP exposure to knockdown cell cultures evidenced for its specific action on sigma-1 receptor. Washing 3-PPP out and cell fixing with further spine counting made it possible to estimate time of spine rescue effect which occurred to be 8 hours approximately within the framework of the working model. The results were statistically processed and presented as graphs and bar charts.
