Department of Neurochemistry

• Chronic pain
• Depression
• Addiction
• Neurodegeneration
• Neuroprotection
• The mechanisms of action of antidepressant and neuroprotective drugs

The long-term goal of our research is to understand the molecular mechanisms regulating neuroplastic processes in the endogenous cannabinoid system. The endocanabinoid system is involved in the control of motivational processes, reward seeking and pain perception. An approach that combines pain phenomena and the activity of the mesolimbic reward system, as well as an involvement of learning mechanisms in the maintenance of the effects of severe pain are another interesting lines of research in our lab. Understanding the role of endocannabinoids in pain transmission at the reward circuitry level may provide new insight in the search for effective analgesics. In order to investigate the central nervous system (CNS) dysfunction in chronic pain in the context of the reward system and learning mechanisms and the influence of the endocannabinoid system we employ a variety of research techniques which have been used in the Department of Neurochemistry for many years.

A primary goal of experimental work in the field of neuropharmacology and also in our Department is an identification of new drugs with enhanced therapeutic efficacy and the least side effects. A search for more effective therapy of psychiatric (depression, addiction) and neurodegenerative disorders, most of which have no extant biomarker or identified biological background, requires prior knowledge of the relevant mechanisms leading to the disease. Research aimed at better understanding the mechanisms that cause the disease and the potential drug action is carried out predominantly in animal models, which enable the researcher to identify CNS dysfunctions that may underlie the condition. Despite intensive research, the etiology of serious psychiatric disorders such as depression, drug addiction, as well as neurodegenerative diseases (Parkinson's disease, Alzheimer's disease) has not yet been identified. The diagnosis of these diseases is possible only in the late, irreversible stages, and the available symptomatic therapies are not effective and cause undesirable side effects. Using various animal models we investigate the processes that lead to depressive behaviors, addictions and neurotoxicity. The course of neurodegenerative processes and the assessment of antidepressant or anti-addictive potential of substances is evaluated on three independent levels: behavioral, biochemical and molecular. Our research also aims to identify biomarkers which enable effective detection of different stages of Parkinson's disease, the molecular mechanisms leading to neurotoxicity and depression, and most of all, new drugs that can prevent these adverse processes in the brain.

Furthermore, we are interested in developing new methods for the treatment of osteoarthritis (OA) and pain relief. The studies are focused on the role of the synovial membrane activity in the development of OA associated pain and the effects of mesenchymal stem cells on the tissue regeneration in patients with osteoarthritis.

Research methods

• Behavioral: automatic measurement of locomotor activity using actometers; observation of rat and mouse behavior: open field test, catalepsy test, forced swim test (FST), tail suspension test – as behavioral models to test pro and anti-depressant activity; experimental animal models of chronic (neuropathic) pain and pain associated with osteoarthritis; assessment of chronic pain symptoms in animal models: assay to test touch perception / mechanical sensitivity, knee pressure test, dynamic weight bearing test in freely moving animals.
• Brain stereotactic operations in rats.
• Intracerebral administration of drugs.
• Biochemical methods: analysis of biogenic amines and their metabolites, the rate of monoamines metabolism analysis in different rat brain structures by HPLC methodology with ED, assessment of excitatory amino acids concentration by coulometric HPLC with UV detection.
• In vivo biochemical methods: microdialysis study and analysis of exytracellular concentrations of neurotransmitters and their metabolites.
• Radioisotope methods: displacement analysis of radiolabeled ligands, measurement of the density and affinity of receptors (Bmax, Kd), monoamines uptake in brain sections, enzyme activity.
• Analysis of gene expression on mRNA level:
  - RNA isolation and purification from animal material and cell cultures
  - qPCR method (real-time PCR)
  - microarray analysis of mRNA.
• Analysis of gene expression on the protein level:
  - quantitative Western blot analysis of protein levels in the biological material
  - preparation of animal material for immunofluorescence staining (perfusion, fixation and tissue sectioning)
  - qualitative determination of proteins in the biological material by immunofluorescence.
• In vitro assays:
  Primary cell cultures of chondrocytes and osteoblasts
  Biochemical tests:
  - Cell activation assay (Griess test)
  - Cell viability assay (MTT)
  - Cell proliferation assay (BrdU)
  - Cytotoxicity assay (LDH).

The most important discoveries in the last 3 years

Demonstration and promotion of neuroprotective, anti-addictive and antidepressant properties of an endogenous amine, 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ). Based on our recently published research results 1MeTIQ was introduced to the international catalog of Sigma-Aldrich (cat. No. 4965- 09-7) in 2015.

Demonstration, that OMDM-198, the compound targeting two molecular targets (FAAH activity inhibitor and TRPV1 receptor blocker) has a higher analgesic efficacy than substances acting on one receptor system. The discovery may help in developing new, more effective and accessible chronic pain therapy (Malek et al., PAIN, 2015).

Cooperation with national research centers

1. Prof. dr hab. Zbigniew Madeja and dr hab. Justyna Drukała, Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology of the Jagiellonian University.
2. Dr Piotr Duda, Laboratorium Mikrotomografii, Zakład Komputerowych Systemów Biomedycznych, Instytut Informatyki, Wydział Informatyki i Nauki o Materiałach, University of Silesia.
3. Dr hab. Ewa Zuba-Surma, Laboratory of Stem Cell Research, Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology of the Jagiellonian University..

Cooperation with foreign research centers

1. Professor Vincenzo Di Marzo, Institute of Biomolecular Chemistry of the National Research Council (ICB-CNR), Pozzuoli, Neapol, Italy / Canada Research Excellence, Laval University, Quebeck, Canada.
2. Professor Stephen McMahon, Wolfson Centre For Age-Related Diseases, King’s College London, UK.
3. Professor Roger G. Pertwee School of Medical Sciences Institute of Medical Sciences University of Aberdeen, UK.
4. Professor Ganesh A. Thakur, Department of Pharmaceutical Sciences, School of Pharmacy, Bouvé College of Health Sciences, Northeastern University, Boston, USA.