Fluorescently labeled polyamine uptake (confocal microscopy)
By onMethod visualizes polyamine uptake in cells using fluorescently labeled polyamines and confocal microscopy.
Collection of mouse cerebrospinal fluid from the cisterna magna
By onThis protocol describes the collection of mouse cerebrospinal fluid from the cisterna magna.
Preparation of total cell lysates
By onThis protocol outlines a step-by-step method for preparing cell lysates.
RNA isolation and cDNA synthesis
By onThis protocol details the isolation of RNA, purification, and subsequent cDNA synthesis for downstream applications such as qPCR.
RT-qPCR (primary cells)
By onProtocol details qPCR method for Atp13a4 expression in astrocytes and neurons using Fast SYBR Green Master Mix in a 96-well format. Normalization to 18S gene ensures accurate quantification for reliable gene expression analysis.
Developmental milestones assessment in newborn mice (P1–P21)
By onBehavioral tests for newborn mice from P1 to P21 assess weight gain, reflexes, and movements with a focus on developmental milestones.
Lentiviral vector production and primary astrocyte transduction
By onProtocol for lentivirus production & transduction into astrocytes is outlined, allowing efficient transduction & stable selection. Facilitates research on primary gene function in astrocytes.
Astrocyte morphology assessment in astrocyte-neuron co-cultures
By onThis protocol outlines the process for analyzing astrocyte morphology in astrocyte-neuron co-cultures using immunostaining and Sholl analysis.
Genotyping Atp13a4(KO) mice
By onProtocol for DNA extraction from mouse biopsies & genotyping Atp13a4(KO) mice involves alkaline lysis method, PCR with specific primers, & agarose gel analysis to differentiate genotypes.
Multiplexed immunofluorescence and RNA-FISH
By onProtocol detects Atp13a4 mRNA in astrocytes of visual cortex using immunofluorescence and RNA-FISH. Sections from Aldh1L1-EGFP mice are stained, hybridized with probes, and imaged for spatial analysis of Atp13a4 expression in postnatal development.
Metabolomic analysis of polyamines
By onProtocol outlines metabolite extraction, derivatization, and mass spectrometry for various sample types. It covers sample preparation, LC-MS parameters, and data analysis for polyamine profiling.
Glia-Free Cortical Neuronal Feeding Schedule – Neuronal Morphology
By onThis protocol outlines the treatment of cortical neurons with astrocyte-conditioned medium (ACM) or spermidine and subsequent analysis of dendrite length.
24-Hour Home Cage Activity Monitoring in Mice
By onProtocol assesses 24-hour home cage activity in adult mice using infrared beam-break technology. Beam crossings indicate horizontal locomotion continuously throughout light-dark cycle.
Astrocyte Isolation and Transfection for Astrocytes/Neuron co-culture assay
By onProtocol for isolating rat cortical astrocytes and producing astrocyte conditioned media for synaptogenesis assays.
Morris Water Maze: Spatial Learning and Memory Assessment in Mice
By onThe Morris Water Maze task assesses spatial learning and memory in adult mice by training them to find a hidden platform using spatial cues. Probe trials are conducted weekly to evaluate memory consolidation.
Open Field Exploration: Locomotor Activity and Anxiety Assessment in Mice
By onProtocol assesses locomotor activity and anxiety in mice using an open-field arena. Path length & corner crossings indicate activity level, center entries reflect anxiety levels or conflict resolution.
Accelerating Rotarod: Motor Coordination and Balance Assessment in Mice
By onProtocol assesses motor coordination in adult mice using an accelerating rotarod. Mice trained at constant speed before four test trials on accelerating rod to measure latency to fall.
DigiGait: Quantitative Gait Analysis in Mice
By onProtocol details use of DigiGait system for high-speed ventral-plane imaging & gait analysis in adult mice, quantifying spatiotemporal parameters during treadmill locomotion.
PTZ Seizure Challenge: Assessment of Seizure Sensitivity in Mice
By onProtocol assesses seizure sensitivity in adult mice using PTZ injection at 1% body weight. Behavior recorded for 15 min in box, analyzed with EthoVision XT. Seizure-related behaviors scored manually.
