| Autor: |
Odongoo B; Department of Functional Pathology, Faculty of Medicine, Shimane University , Izumo, Japan., Ohara H; Department of Functional Pathology, Faculty of Medicine, Shimane University , Izumo, Japan., Ngarashi D; Department of Functional Pathology, Faculty of Medicine, Shimane University , Izumo, Japan.; Department of Physiology, School of Medicine, Muhimbili University of Health and Allied Sciences , Dar Es Salaam, Tanzania., Kaneko T; Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University , Kyoto, Japan.; Laboratory of Animal Reproduction and Development, Faculty of Science and Engineering, Iwate University , Morioka, Japan., Kunihiro Y; Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University , Kyoto, Japan.; Institute of Experimental Animal Sciences, Graduate School of Medicine, Osaka University , Osaka, Japan., Mashimo T; Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University , Kyoto, Japan.; Institute of Experimental Animal Sciences, Graduate School of Medicine, Osaka University , Osaka, Japan.; Laboratory Animal Research Center, the Institute of Medical Science, The University of Tokyo , Tokyo, Japan., Nabika T; Department of Functional Pathology, Faculty of Medicine, Shimane University , Izumo, Japan. |
| Abstrakt: |
Genetic approach using rat congenic lines between SHRSP/Izm and WKY/Izm identified stromal interaction molecule 1 ( Stim1 ), an essential component of store-operated Ca 2+ entry (SOCE), as a promising candidate gene responsible for the exaggerated sympathetic response to stress in SHRSP. Since SHRSP has a nonsense mutation in Stim1 resulting in the expression of a truncated form of STIM1 that caused reduction of SOCE activity in primary cultured cerebral astrocytes, we created SHRSP/Izm knocked-in with the wild-type Stim1 (KI SHRSP) by the CRISPR/Cas9 method to investigate whether the functional recovery of STIM1 would mitigate sympatho-excitation to stress in vivo in SHRSP. No potential off-target nucleotide substitutions/deletions/insertions were found in KI SHRSP. Western blotting and fluorescent Ca 2+ imaging of astrocytes confirmed wild-type STIM1 expression and restored SOCE activity in astrocytes from KI SHRSP, respectively. Blood pressure (BP) measured by the tail-cuff method at 12, 16, and 20 weeks of age did not significantly differ between SHRSP and KI SHRSP, while the heart rate of KI SHRSP at 16 and 20 weeks of age was significantly lower than that of age-matched SHRSP. Unexpectedly, the sympathetic response to stress (evaluated with urinary excretion of norepinephrine under cold stress and BP elevation under cold/restraint stress) did not significantly differ between SHRSP and KI SHRSP. The present results indicated that the functional deficit of STIM1 was not a genetic determinant of the exaggerated sympathetic response to stress in SHRSP and that it would be necessary to explore other candidates within the congenic fragment on chromosome 1. |
