Mitochondrial dysfunction as a mediator of hippocampal apoptosis in a model of hepatic encephalopathy.

BUSTAMANTE, J.; LORES-ARNAIZ, S.; TALLIS, S.; ROSELLÓ, D.; LAGO, N.; LEMBERG, A.; BOVERIS, A. ; PERAZZO, JC.

MOLECULAR AND CELLULAR BIOCHEMISTRY

SPRINGER

Año: 2011

0300-8177

Abstract. In this study here we describe the presence of apoptosis, associated with a mitochondrial dysfunction in the hippocampus of animals in a model of minimal hepatic encephalopathy (MHE). This experimental model was studied after 10 days of induced portal vein calibrated stricture, leading to portal hypertension and to a moderate hyperammonemia, without the presence of other evident central nervous system changes. The molecular mechanisms here proposed, indicate the presence of apoptosis intrinsic pathways, that point to hippocampal mitochondria as an important mediator of the hypertension and slight hyperammonemia observed. In this model of MHE, the presence of DNA fragmentation in positive GFAP cells is documented by 2.3-times increased number of TUNEL-positive cells. These findings together with 5 times higher expression of the proapoptotic member of the Bcl-2 family, Bax, associated to the outer mitochondrial membrane, indicate the presence of apoptosis in this experimental model. A detailed analysis of the hippocampal mitochondrial physiology was performed after mitochondrial isolation. The determination of the respiratory rate in the presence of malate plus glutamate and ADP showed 45% decrease in respiratory control in MHE animals as compared with the Sham group. A marked decrease of cytochrome oxidase (complex IV of the electron transport chain) was also observed, showing 46% less activity in hippocampal mitochondria from MHE animals. In addition, mitochondria from these animals showed less ability to maintain membrane potential (īĎm) which was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. mitochondrial dysfunction in the hippocampus of animals in a model of minimal hepatic encephalopathy (MHE). This experimental model was studied after 10 days of induced portal vein calibrated stricture, leading to portal hypertension and to a moderate hyperammonemia, without the presence of other evident central nervous system changes. The molecular mechanisms here proposed, indicate the presence of apoptosis intrinsic pathways, that point to hippocampal mitochondria as an important mediator of the hypertension and slight hyperammonemia observed. In this model of MHE, the presence of DNA fragmentation in positive GFAP cells is documented by 2.3-times increased number of TUNEL-positive cells. These findings together with 5 times higher expression of the proapoptotic member of the Bcl-2 family, Bax, associated to the outer mitochondrial membrane, indicate the presence of apoptosis in this experimental model. A detailed analysis of the hippocampal mitochondrial physiology was performed after mitochondrial isolation. The determination of the respiratory rate in the presence of malate plus glutamate and ADP showed 45% decrease in respiratory control in MHE animals as compared with the Sham group. A marked decrease of cytochrome oxidase (complex IV of the electron transport chain) was also observed, showing 46% less activity in hippocampal mitochondria from MHE animals. In addition, mitochondria from these animals showed less ability to maintain membrane potential (īĎm) which was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. mitochondrial dysfunction in the hippocampus of animals in a model of minimal hepatic encephalopathy (MHE). This experimental model was studied after 10 days of induced portal vein calibrated stricture, leading to portal hypertension and to a moderate hyperammonemia, without the presence of other evident central nervous system changes. The molecular mechanisms here proposed, indicate the presence of apoptosis intrinsic pathways, that point to hippocampal mitochondria as an important mediator of the hypertension and slight hyperammonemia observed. In this model of MHE, the presence of DNA fragmentation in positive GFAP cells is documented by 2.3-times increased number of TUNEL-positive cells. These findings together with 5 times higher expression of the proapoptotic member of the Bcl-2 family, Bax, associated to the outer mitochondrial membrane, indicate the presence of apoptosis in this experimental model. A detailed analysis of the hippocampal mitochondrial physiology was performed after mitochondrial isolation. The determination of the respiratory rate in the presence of malate plus glutamate and ADP showed 45% decrease in respiratory control in MHE animals as compared with the Sham group. A marked decrease of cytochrome oxidase (complex IV of the electron transport chain) was also observed, showing 46% less activity in hippocampal mitochondria from MHE animals. In addition, mitochondria from these animals showed less ability to maintain membrane potential (īĎm) which was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. mitochondrial dysfunction in the hippocampus of animals in a model of minimal hepatic encephalopathy (MHE). This experimental model was studied after 10 days of induced portal vein calibrated stricture, leading to portal hypertension and to a moderate hyperammonemia, without the presence of other evident central nervous system changes. The molecular mechanisms here proposed, indicate the presence of apoptosis intrinsic pathways, that point to hippocampal mitochondria as an important mediator of the hypertension and slight hyperammonemia observed. In this model of MHE, the presence of DNA fragmentation in positive GFAP cells is documented by 2.3-times increased number of TUNEL-positive cells. These findings together with 5 times higher expression of the proapoptotic member of the Bcl-2 family, Bax, associated to the outer mitochondrial membrane, indicate the presence of apoptosis in this experimental model. A detailed analysis of the hippocampal mitochondrial physiology was performed after mitochondrial isolation. The determination of the respiratory rate in the presence of malate plus glutamate and ADP showed 45% decrease in respiratory control in MHE animals as compared with the Sham group. A marked decrease of cytochrome oxidase (complex IV of the electron transport chain) was also observed, showing 46% less activity in hippocampal mitochondria from MHE animals. In addition, mitochondria from these animals showed less ability to maintain membrane potential (īĎm) which was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. mitochondrial dysfunction in the hippocampus of animals in a model of minimal hepatic encephalopathy (MHE). This experimental model was studied after 10 days of induced portal vein calibrated stricture, leading to portal hypertension and to a moderate hyperammonemia, without the presence of other evident central nervous system changes. The molecular mechanisms here proposed, indicate the presence of apoptosis intrinsic pathways, that point to hippocampal mitochondria as an important mediator of the hypertension and slight hyperammonemia observed. In this model of MHE, the presence of DNA fragmentation in positive GFAP cells is documented by 2.3-times increased number of TUNEL-positive cells. These findings together with 5 times higher expression of the proapoptotic member of the Bcl-2 family, Bax, associated to the outer mitochondrial membrane, indicate the presence of apoptosis in this experimental model. A detailed analysis of the hippocampal mitochondrial physiology was performed after mitochondrial isolation. The determination of the respiratory rate in the presence of malate plus glutamate and ADP showed 45% decrease in respiratory control in MHE animals as compared with the Sham group. A marked decrease of cytochrome oxidase (complex IV of the electron transport chain) was also observed, showing 46% less activity in hippocampal mitochondria from MHE animals. In addition, mitochondria from these animals showed less ability to maintain membrane potential (īĎm) which was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. mitochondrial dysfunction in the hippocampus of animals in a model of minimal hepatic encephalopathy (MHE). This experimental model was studied after 10 days of induced portal vein calibrated stricture, leading to portal hypertension and to a moderate hyperammonemia, without the presence of other evident central nervous system changes. The molecular mechanisms here proposed, indicate the presence of apoptosis intrinsic pathways, that point to hippocampal mitochondria as an important mediator of the hypertension and slight hyperammonemia observed. In this model of MHE, the presence of DNA fragmentation in positive GFAP cells is documented by 2.3-times increased number of TUNEL-positive cells. These findings together with 5 times higher expression of the proapoptotic member of the Bcl-2 family, Bax, associated to the outer mitochondrial membrane, indicate the presence of apoptosis in this experimental model. A detailed analysis of the hippocampal mitochondrial physiology was performed after mitochondrial isolation. The determination of the respiratory rate in the presence of malate plus glutamate and ADP showed 45% decrease in respiratory control in MHE animals as compared with the Sham group. A marked decrease of cytochrome oxidase (complex IV of the electron transport chain) was also observed, showing 46% less activity in hippocampal mitochondria from MHE animals. In addition, mitochondria from these animals showed less ability to maintain membrane potential (īĎm) which was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. In this study here we describe the presence of apoptosis, associated with a mitochondrial dysfunction in the hippocampus of animals in a model of minimal hepatic encephalopathy (MHE). This experimental model was studied after 10 days of induced portal vein calibrated stricture, leading to portal hypertension and to a moderate hyperammonemia, without the presence of other evident central nervous system changes. The molecular mechanisms here proposed, indicate the presence of apoptosis intrinsic pathways, that point to hippocampal mitochondria as an important mediator of the hypertension and slight hyperammonemia observed. In this model of MHE, the presence of DNA fragmentation in positive GFAP cells is documented by 2.3-times increased number of TUNEL-positive cells. These findings together with 5 times higher expression of the proapoptotic member of the Bcl-2 family, Bax, associated to the outer mitochondrial membrane, indicate the presence of apoptosis in this experimental model. A detailed analysis of the hippocampal mitochondrial physiology was performed after mitochondrial isolation. The determination of the respiratory rate in the presence of malate plus glutamate and ADP showed 45% decrease in respiratory control in MHE animals as compared with the Sham group. A marked decrease of cytochrome oxidase (complex IV of the electron transport chain) was also observed, showing 46% less activity in hippocampal mitochondria from MHE animals. In addition, mitochondria from these animals showed less ability to maintain membrane potential (īĎm) which was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. īĎm) which was 13% lower than the Sham group. Light scattering experiments showed that mitochondria from MHE animals were more sensitive to swell in the presence of increased calcium concentrations as compared with the Sham group. In addition, in vitro studies performed in mitochondria from Sham animals showed that mitochondrial permeability transition (MPT) could be a mitochondrial mediator of the apopototic signaling in the presence of NH4 + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. + and calcium. 4 + and calcium.and calcium.