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Rotenone-Induced Dysregulation of Synaptosomal Ca²⁺ And Hypokinetic Behavior in A Rat Model of Parkinsonism

Zayniyeva MakhbubaNаtiоnаl Univеrsitу оf Uzbеkistаn named after M. Ulugbek, Tashkent, Almazar 100174, UzbekistanIzzatillaeva SabinaNаtiоnаl Univеrsitу оf Uzbеkistаn named after M. Ulugbek, Tashkent, Almazar 100174, UzbekistanTajiyeva OyimjanUrgench Abu Ali ibn Sina Public Health Technical School, Khorezm, UzbekistanKоzоkоv IslomInstitute of Biophysics and Biochemistry, National University of Uzbekistan, 100174, Tashkent, UzbekistanDеdаbоеv JobirInstitute of Biophysics and Biochemistry, National University of Uzbekistan, 100174, Tashkent, UzbekistanMukhtоrоv AlisherNаtiоnаl Univеrsitу оf Uzbеkistаn named after M. Ulugbek, Tashkent, Almazar 100174, UzbekistanKhоshimоv NozimInstitute of Biophysics and Biochemistry, National University of Uzbekistan, 100174, Tashkent, Uzbekistan
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Background: Rotenone, a lipophilic mitochondrial complex I inhibitor, is widely used to model Parkinsonian neurodegeneration and synaptic failure. Objective: To quantify rotenone-associated behavioral changes and determine whether they co-occur with altered glutamate-evoked synaptosomal Ca²⁺ dynamics. Methods: Adult male Wistar rats were randomized into control and rotenone groups (2.5 mg/kg/day, i.p., 11 consecutive days; n=6/group). Exploratory behavior was assessed in an open-field/hole-board ar ena (42 × 42 cm; 42-floor grid; 3 min; ~100 lux). Crude synaptosomes (P2 fraction) were isolated from whole brain tissue, loaded with Fluo‑4AM (final 5 µM, 30 min, 37°C), and stimulated with L‑glutamate (50 µM). Ca²⁺ responses were summarized as resting signal, peak amplitude, area under the curve (AUC), and clearance time constant (τ). Results: Rotenone reduced horizontal locomotion (72±6 vs 14±4 crossings), vertical activity (38±5 vs 8±3 rearings), and hole‑poking (16±2 vs 6±1; all p<0.05). Synaptosomes from rotenone-treated rats displayed a higher resting Ca²⁺-related fluorescence (+21%) and enhanced glutamate-evoked Ca²⁺ responses (peak +18%, AUC +25%), alongside faster decay (τ −17%) relative to controls (p<0.05). Conclusions: Subchronic rotenone exposure produces a reproducible hypokinetic phenotype that parallels presynaptic Ca²⁺ dysregulation, supporting a mechanistic link between mitochondrial stress and abnormal glutamate-triggered Ca²⁺ signaling.

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