Ene 2017 MiPschool Obergurgl
Ene HM, Robicsek O, Karry R, Ben-Yehuda R, Yitzhaki O, Wiener I, Ben-Shachar D (2017)
Event: MiPschool Obergurgl 2017
Malfunction of mitochondria, key players in various essential cell processes, has been repeatedly reported in schizophrenia (SZ) . Recent studies have reported functional improvement and cellular viability following mitochondrial transplantation in several diseases . Here we aimed to study (A) the effect of transfer of isolated active normal mitochondria (IAN-MIT) into SZ-derived iPSCs on their impaired differentiation into glutamatergic neurons ; and (B) the effect of IAN-MIT transfer into the medial prefrontal cortex (mPFC) in adolescence on SZ-related behavior and mitochondrial function in adulthood, using the poly I:C rat model of SZ. In this model, maternal immune activation by poly I:C leads to SZ-relevant behavioral and morphological deficits in adult offspring . Notably, a significant decrease in mitochondrial ATP production was observed in poly I:C-treated mice, mainly attributed to a lower complex I activity.
In the in-vitro study, IAN-MIT were transferred into differentiating SZ-derived iPSCs into glutamatergic neurons. At the end of the differentiation period, the efficiency of neuronal differentiation was assessed by the expression of neuronal and glutamatergic markers (β3-Tubulin, Tbr1, PSD-95 and Synapsin1) using immunofluorescence staining. In addition, glutamate (Glu) and glutamine (Gln) metabolism were evaluated by high-pressure liquid chromatography (HPLC). In the in-vivo study, pregnant dams were injected with poly I:C or saline. On postnatal days 34-47, their offspring were administered with a single bilateral stereotaxic injection of freshly prepared IAN-MIT or vehicle into the mPFC. In adulthood, attentional function was assessed by the latent inhibition (LI) test, and mitochondrial function was evaluated in fronto-cortical freshly isolated JC-1 stained neurons by measuring mitochondrial membrane potential (Δym) and mitochondrial network dynamics, using confocal microscopy.
In the in-vitro study, IAN-MIT transfer improved differentiation of SZ-derived iPSCs into neurons, by increased expression of neuronal and glutamatergic markers β3-Tubulin, synapsin1 and Tbr1 and by an activation of the Glu-Gln cycle. In the in-vivo study, medial prefrontal IAN-MIT transfer in adolescence had a prolonged effect on animal behavior and mitochondrial function. In adult poly I:C-exposed offspring, IAN-MIT prevented disrupted LI and increased the reduced mitochondrial Δym. In control offspring however, IAN-MIT injection disrupted LI and increased mitochondrial Δym. IAN-MIT transfer did not affect mitochondrial cell distribution and network connectivity.
These findings demonstrate a beneficial effect of IAN-MIT transfer in-vitro and in-vivo, suggesting its therapeutic potential in diseases with bioenergetic and neurodevelopmental abnormalities such as SZ. The results suggest that in-vivo IAN-MIT transfer is beneficial when mitochondrial function is impaired, while under normal conditions it interferes with the homeostasis of mitochondrial diverse functions. Moreover, this study provides evidence for a possible relationship between mitochondria and attention, as an opposite pattern was obtained in controls and poly-I:C offspring for both mitochondrial and attentional functions.
• Bioblast editor: Kandolf G
Organism: Rat Tissue;cell: Nervous system
Event: B2, Oral
- Ene HM(1), Robicsek O(1), Karry R(1), Ben-Yehuda R(2), Yitzhaki O(1), Wiener I(2) Ben-Shachar D(1)
- Dept Neurosci, Psychobiol Res Lab, Rambam Medical Center, Bruce Rappaport Fac Med, Technion IIT, Israel
- Dept Socsci, TAU Univ, Isreal. - [email protected]
Figure 1. Glutamatergic differentiation following early and late IAN-MIT transfer. A. Bright-field images of neurons. B. Immunofluorescence staining and D. quantification of β3-Tubulin and Tbr1, both decreased in SZ neurons. Early and late IAN-MIT transfer increased β3–Tubulin, while Tbr1 increased only following late transfer. GLM analysis showed a time-dependent increase in both (β3–Tubulin– F=20.16, P<0.001. Tbr1-F=116.8, P<0.001). C. Immunofluorescence staining and E. quantification of synapsin1 and PSD-95 and their co-localization, which were decreased in SZ-neurons. Early and Late IAN-MIT transfer had a minor effect on PSD-95 and co-localization, yet profoundly affected synapsin1, which following late transfer reached control levels. GLM showed a time-dependent increase in synapsin1 (Synapsin1-F=90.15, P<0.002). Values are means±SEM of 2 controls and 2 patients (one iPSCs clone for each) from 2 experiments in duplicates with 4-6 cells in each. Scale bars A,B-100Im, C-25Im. F. Late, but not early, IAN-MIT transfer almost normalized Gln release and ameliorated abnormal Glu consumption. Values are means±SEM of 2 controls and 2 patients (one iPSCs clone for each) from 2 experiments in triplicates. *P<0.01 SZ vs. Cont; +P<0.008 SZ-Mit vs. Cont; #P<0.03 SZ-Mit vs. SZ.
Figure 2. mPFC IAN-MIT transfer prevents disrupted LI and dissipated Δψm in poly-I:C exposed offspring. A. LI is manifested as shorter log times to complete licking criteria after tone onset of PE compared with NPE groups. Values are means±SEM. *P<0.0001 PE vs. NPE in saline-vehicle exposed offspring and in poly-I:C+IAN-MIT exposed offspring. B. Representative images of JC-1 stained cortical neurons. JC-1 in cytosol (green), in active mitochondria (red). Scale bars-5Im. C. Quantification of Δψm, which is reduced in poly-I:C rats and increased by IAN-MIT injection in both poly-I:C.
- Ben-Shachar D (2016) Mitochondrial multifaceted dysfunction in schizophrenia; complex I as a possible pathological target. Schizophr Res. In press.
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- Piontkewitz Y, Arad M, Weiner I (2011) Risperidone administered during asymptomatic period of adolescence prevents the emergence of brain structural pathology and behavioral abnormalities in an animal model of schizophrenia. Schizophr Bul. 37:1257-69.
- Recepient of a MitoEAGLE scholarship.
- Selected mentor: Prof. Sandi Carmen