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SEMINARIO BIOMOL.pptx
- 1. Amalia Medina Martínez 3rd semester Universidad Pontificia Bolivariana
- 2. INTRODUCTION NLRP1 INFLAMMASOME • Innate immune receptor that assembles into an inflammasome • Multiprotein complex • Potent activator of inflammation
- 3. INTRODUCTION • METHAMPHETAMINE - Potent central nervous system stimulant - Mainly used as a recreational drug • RELATION WITH COGNITIVE - Several clinical studies have demonstrated that METH use disorder is associated with a wide range of cognitive deficits, including attention, executive function, language/verbal fluency, learning and memory
- 4. OBJECTIVE • The objective of this study is to evaluate the effect of the NLRP1 inflammasome on methamphetamine induced cognitive impairment on rats
- 5. METHODS • 2.4: New object recognition (NOR) test - Three stages including an adaptation, training and testing session - Day 1: 10 min alone - Day 2: 10 min with two identical objects - Day 3: one of the objects was changed • 2.7: Quantitative real-time PCR (q-PCR) - RNA diluted with DEPC water - Purified RNA was reversely transcribed - cDNA was quantified by quantitative reverse transcription-PCR
- 6. METHODS • 2.8 Western blotting (WB) - Analytical technique used to detect a specific protein - Using antibodies (proteins produced by immune system) - They detect foreign proteins (antigens) and specifically bind to them. • 2.9 Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labelling (TUNEL) - Dark buffy nuclei staining was shown in TUNEL-positive cells, while blue nuclei was observed in TUNEL-negative cells. The total number of TUNEL-positive cells per field was counted and then divided by the field area. - Apoptosis was detected
- 8. RESULTS
- 9. DISCUSSION Authors Comment The article Liu et al It has been shown that METH-induced toxicity could also be GSDME-dependent via the endoplasmic reticulum stress pathway in HT-22 cells Dos Santos et al DEX decreased cognitive and memory capacity by diminishing neuronal activity and potentiating some aspects of neuroinflammation in experimental allergic encephalomyelitis Zhang et al Interestingly, the glucocorticoid receptor antagonist mifepristone (RU486) had protective effects on chronic DEX-induced hippocampal neuron injury and NLRP1 inflammasome activation DISCUSSION
- 10. CONCLUSIONS • Molecular biology studies can be important for us to know the impact of certain drugs such as METH in our bodies and how these affect us • Molecular biology can help us with researches to search for treatments
- 11. MIND MAP
Editor's Notes
- NLRP1: associated to neuroinflammatory processes
- Methamphetamine: It reverses transport through the dopamine transporter, thereby increasing the extracellular level of dopamine in the brain, which is associated with the rewarding effect
- NOR: is used to evaluate cognition, particularly recognition memory, in rodent models of CNS disorders Quantitative real-time PCR (q-PCR): this technique is used to detect, characterize and quantify nucleic acids for numerous applications.
- Western blot: to detect specific proteins in a sample of tissue homogenate or extract Terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labelling (TUNEL): is a method used to detect fragmented DNA Western blot: is an analytical technique used to detect a specific protein in a crude extract, using antibodies. Antibodies are proteins produced by an animal's immune system (immunoglobulins) that are able to detect foreign proteins (antigens) and specifically bind to them.
- Beta actina es el control, la prueba se evaluó tanto con salina y el otro con la metanfetamina. En los de arriba se evaluaron los NLRP1, pro-caspase 1, etc… y en el de abajo el N-GSDMD. Se evidencia que con la presencia de la metanfetamina se aumenta la expresión de los componentes del lado izq y que con salina no se ve tan aumentada la expresión
- FIGURA A: The effect of NLRP1 blockade by ATL on the NLRP/Caspase-1 signaling pathway in the hippocampus of METH-treated rats FIGURA C: Quantitative results of TUNEL experiment in the hippocampus of rats
- Dos Santos: moreover, 3-day treatment of 1.5 μМ DEX significantly increased the expression of NF-κB in hippocampal neurons. Zhang: there´s a mediation in a prot