Basics Of Brain And Cognitive Sciences Research PDF

“Guide to Research Techniques in Neuroscience” by Carter and Shieh  Goal  Describe techniques for visualizing activity and function in fixed tissue  Describe nonelectrophysiological methods of measuring neural activity  Describe techniques for visualizing protein function 2 Basics of...

“Guide to Research Techniques in Neuroscience” by Carter and Shieh  Goal  Describe techniques for visualizing activity and function in fixed tissue  Describe nonelectrophysiological methods of measuring neural activity  Describe techniques for visualizing protein function 2 Basics of Brain and Cognitive Sciences Research  Techniques Covered: √ √  Static measures of activity and function: measuring neural activity 1 using immediate early genes, measuring cell proliferation with thymidine analogs, measuring protein trafficking with pulse-chase 1 labeling  Visualizing dynamic neural activity: voltage sensors (voltage-sensitive dyes, genetically encoded voltage indicators), calcium sensors (calcium-sensitive dyes, genetically encoded calcium indicators), synaptic transmission sensors (FM dyes, synaptopHluorin)  Visualizing protein function: reporter genes, fluorescence resonance energy transfer (FRET), bimolecular fluorescence complementation (BiFC), fluorescence recovery after photobleaching (FRAP), photoactivation/ photoconversion 3 Basics of Brain and Cognitive Sciences Research  not alvays possible i… Static Markers of Activity  There are two fundamental approaches to examining neural activity after it has already occurred:  (1) measuring activity indirectly by measuring byproducts that ) scientzts accumulate during specific processes in active neurons examine of snapshots  (2) incorporating a marker into cells that can indicate the dynamic procen presence of activity during subsequent histological examination. 5 Basics of Brain and Cognitive Sciences Research  ω Assaying Neural Activity in Fixed Tissue  Neural activity leads to the transient and rapid (within minutes) transcription of a group of genes known as immediate early 」 ~ genes (IEGs). d 3  These genes encode a diverse range of proteins, including transcription factors (Fos), cytoskeletal-interacting proteins 이를 볼어스된 」 (Arc), and phosphorylated ribosomal subunits (pS6). obready fixed  IEG patterns can also be used to screen neurons for the. presence of activities that correlate with specific behaviors. sleep regios 1 ex ) bron mnea 1 thatuac aninald usgleep - irstquies provide 6 Basics of Brain and Cognitive Sciences Research  IEG and neural activity FoS MRNA.. ㅇ  In the striatum and hippocampus, Fos expression inflex. is mediated by ERK/MAPK- ^ dependent phosphorylation σ of SRF and phosphorylation, via ribosomal S6 kinase (RSK), of CREB on the Fos promoter.  VSCC: voltage-sensitive ) 이런 calcium channels yere priteinen V - 알 수있 iof ~ 7 Basics of Brain and Cognitive Sciences Research  「 freqsddictireding. Nicotine-induced activation of IPN (interpeduncular nucleus) cont Uh mice에서 거의 작동X " Nicotre? 이 경도 inilo, Habenular α5 nicotinic receptor 원래 receptor for subunit knockout acetylcholive. mice I xrecaprespond compored ovicdtyr. Habenula-IPN pathway is important in the regulation of nicotine addiction. This effect of the high nicotine dose was almost completely abolished in knockout mice. 8 Basics of Brain and Cognitive Sciences Research  O Assaying Cellular Function in Fixed Tissue  To measure some functional processes in fixed tissue, a scientist can introduce a marker while the process occurs in live tissue, then detect this marker in subsequent histological experiments.  Two biological functions commonly assayed using these methods include cell proliferation and protein trafficking. Torporate m arkers 」 9 Basics of Brain and Cognitive Sciences Research  Assaying Cell Proliferation with Thymidine Analogs  BrdU (bromo-deoxyuridine), a synthetic analog of the DNA base thymidine, or radioactive tritiated thymidine (3H- thymidine), can be injected into an animal or introduced into cells or tissue in culture media. cells with DNA - " ∞  BrdU can be detected using IHC, and 3H-thymidine can be preseace f 밖이 detected using autoradiography.. - → onect jeeton. cell proliferation → assay " □ between ijection , gap } ine 10 Basics of Brain and Cognitive Sciences Research ㄴ  Assaying Cell Proliferation with Thymidine Analogs jection 1 BrdVf.