Kynurenic Acid Metabolism in Different Types of Brain ...

Kynurenic Acid Metabolism in Different Types of Brain ...

2nd International Conference on HIV/AIDS, STDs & STIs October 27-29, 2014 Las Vegas, USA Alteration of choline acetyltransferase activity in piglet brain after EncephaloMyoCarditis Virus (EMCV) infection. Kepplinger B., Baran H., Draxler M., Schmoll F., Nowotny N., Schuh M. and Hofecker G. Karl Landsteiner Research Institute for Neurochemistry, Neuropharmacology &, Neurorehabilitation, Neuropsychiatric Hospital Mauer Institute of Pharmacology und Toxicology, Institute of Physiology, IInd. Med. Clinic, Institute of Pathology, Clinical Virology, Veterinary Medical University Vienna, Austria The work is supported by Austrian Fonds der Wissenschaftlichen Forschung. FWF Project No. 15371 to Prof. H. Baran, PhD Introduction o EMCV (Encephalo-Myo-Carditis-Virus) infection in piglets mainly affects heart tissue and leads to CNS alterations (Noack and Liebermann, 1992),

however no reliable data are available. o Clinically, EMCV causes a sudden death syndrome in piglets due to acute myocarditis, however the mechanism(s) has not been elaborated yet. o In hippocampal cell culture EMCV leads to neurodegeneration (Weiping Su, 1999). Alteration of choline acetytranferase activity in piglet brain after EMCV infection Acetylcholine excitatory neurotransmitter a well known endogenous agonist of the nicotinic and muscarinic cholinergic receptors, is synthesised in cholinergic neurons is involved in neurodegenerative disorders (as Alzheimerss) Alteration of choline acetyltransferase activity in piglet brain after EMCV infection

Synthesis and degradation of Acetylcholine Cholinergic neurons in the rat brain si substancia innominata tpp pedunculopotin nuclei ms medial septal nucleus bas nucleus basalis poma magnucellular preoptic field dltn laterodorsal tegmental nuclei ICj islands of Calleja Aim of the study To estimate whether EMC-Virus infection induces molecular changes of the marker of cholinergic neurons, Choline acetyltransferase (ChAT) was selected and activity of enzyme was investigated in the CNS in piglets at 4 days after EMC-Virus infection.

Material and Methods Piglets at an age of 8 weeks were used. Infected with 8 ml suspension 107/ml of EMC-virus - strain B279/95; nasal and oral, each 4 ml (5 controls & 7 EMCV). 4 days after EMCV infection animals were sacrificed and brain regions were prepared immediately. ChAT activity was determined by radioenzymatic method. Dissected regions: 1 Frontal Cortex 2 - Parietal Cortex 3 - Temporal Cortex 4 - Hippocampus 5 - Caudate Nucleus 6 - Putamen 7 - Globus Pallidus

Substantia Nigra 8 - Thalamus 9 - Hypothalamus 10 - Substantia Nigra Putamen 11 - Cerebellum 12 - Bulbus olfactorius Globus Pallidus 13 - Midbrain 14 - Pons 15 - Medulla Caudatus Nucleus Choline acetyltransferase (ChAT) in control pig brain

Regions: Control value of ChAT activity in various brain p=0.046 regions p=0.013 p=0.024 ChAT (nmol/mg wet tissue wight/h) P=0.045 p=0.014 16 14 12 10 8

6 4 2 0 1 2 3 4 5 6 7 8 Region

9 10 11 12 13 14 15 P=0.045 1 Frontal cortex 2 - Parietal cortex 3 - Temporal cortex 4 - Hippocampus 5 - Caudate nucleus 6 - Putamen 7 - Globus pallidus 8 - Thalamus 9 - Hypothalamus 10 - Substantia nigra 11 - Cerebellum 12 - Bulbus olfactorius 13 - Midbrain 14 - Pons

15 - Medulla Comparison between ChAT activity in the rat and pig brains: ChAT (nmol/mg wet tissue weight/h) Rat Frontal Cortex: Temporal Cortex: Hippocampus: Caudate Nucleus : Pig 16,32 15,86 17,10 26,46 1,72 1,82

2,13 10,45 Kinetic analysis of ChAT in the hippocampus revealed a value of Km = 0,023 mM for Acetyl-CoA and a Vmax =2,22 (nmol/mg wet tissue weight/h). Choline acetyltransferase (ChAT) activity in the pig brain 4 days after infection control Regions: EMCV ChAT (nmol/mg wet tissue weight) 16,0 14,0 12,0 10,0 8,0

* 6,0 * 4,0 2,0 0,0 1 2 3 4 5

6 7 8 9 10 11 12 13 14 15 1 Frontal cortex 2 - Parietal cortex 3 - Temporal cortex 4 - Hippocampus 5 - Caudate nucleus 6 - Putamen 7 - Globus pallidus 8 - Thalamus 9 - Hypothalamus

10 - Substantia nigra 11 - Cerebellum 12 - Bulbus olfactorius 13 - Midbrain 14 - Pons 15 - Medulla Observation ChAT in control pig brain Within investigated brain regions the highest activity of ChAT was seen in the putamen and the caudate nucleus and the lowest ChAT activity was found in the cerebellum. Enzymatic analyses of ChAT show Km value of 0.02 mM for Acetyl-CoA, which is comparable to other species including human. Observation ChAT at 4 days after EMCV infection reduced activity was measured in the midbrain (54 % of control) and in the medulla (56 % of

control). in putamen and caudate nucleus ChAT activity was moderately lowered. In most other regions a tendency of enzyme activity reduction was found. Conclusion EMCV infection in piglets causes a significant reduction of ChAT activity in the midbrain and in the medulla 4 days after inoculation. Reduction of ChAT activity would suggest an affection of the cholinergic system. ChAT reduction may progress very likely with time and intensity of the disease. Reduction of ChAT is an indicator for an neurodegenerative process. Remark EMCV infection in piglets leads not only to ChAT activity reduction in the midbrain and in the medulla ..but also to:

significant alterations of kynurenic acid metabolism ! visit . Poster: Halina Baran et al., KYNURENIC ACID METABOLISM in PIGLETS after ENCEPHALO-MYOCARDITIS-VIRUS (EMCV) INFECTION Thank You for Your attention!

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