Juxtacellular Recordings in Ventral tegmental area and Locus coeruleus
By Julia Julia Leonard onThis protocol describes juxtacellular recordings in the VTA and LC of adult mice, involving anesthesia, precise brain targeting, and neuron labeling. Using specialized electrodes and a dual-channel amplifier, neuronal activity is recorded and modulated with current injections. Following recordings, brains are perfused, fixed, and sectioned for detailed morphological analysis.
Systemic inflammation triggers long-lasting neuroinflammation and accelerates neurodegeneration in a rat model of Parkinson’s disease overexpressing human alpha-synuclein
By Julia Julia Leonard onIncreasing efforts have been made to elucidate how genetic and environmental factors interact in Parkinson's disease (PD). In the present study, the authors assessed the development of PD-like symptoms on a PD rat model overexpressing human α-synuclein at a presymptomatic age, exposed to a pro-inflammatory insult by injection of lipopolysaccharide.
Sucrose preference test
By Julia Julia Leonard onThe sucrose preference test assesses the animal’s interest in a sweet-tasting sucrose solution relative to unsweetened water.
Immunohistochemistry free-floating rat brain cryosections
By Julia Julia Leonard onProtocol for immunohistochemistry on rat brain cryosections.
Immunofluorescence free-floating rat brain cryosections
By Julia Julia Leonard onProtocol for immunofluorescence on rat brain cryosections.
Optical densitometry of tyrosine hydroxylase fibers
By Julia Julia Leonard onProtocol for quantifying TH+ fibers in rat striatum and substantia nigra pars reticulata (SNpr).
Constant potential amperometry in vitro
By Julia Julia Leonard onProtocol for constant potential amperometry recordings in vitro of evoked dopamine release in the rat striatum.
Rotarod test
By Julia Julia Leonard onProtocol to assess motor coordination and balance by using an accelerating rod.
Open field test
By Julia Julia Leonard onThe open field test is a simple test to evaluate general motor and explorative behavior in rodents.
pAAV_hSyn_GCaMP6m_sWPRE_spA_mDLX_nls_mRuby3_ssWPRE_spA
By Julia Julia Leonard onA viral construct designed to express GCaMP6m pan-neuronally and express a red fluorophore (mRuby) in the nucleus of cortical interneurons. This construct is ideal for dual-imaging experiments to capture activity of two neuronal populations and to identify interneurons with red fluorescence in parallel or post-mortem.
pAAV_Ef1a_DIO_MyrPalm_mScarlet_WT_IRES_mNeon_minBack
By Julia Julia Leonard onBright green (mNeon) and red (mRuby) fluorescence proteins are expressed, with the former being cytoplasmic and the latter membrane-targeted. This expression is ideal for tracking long diffuse axons in cortical structures in vivo or for slice histology and cleared brain tissues post-mortem.
pAAV_EF1a_DIO_jGCaMP8f_nano_ribo_WPRE
By Julia Julia Leonard onThis links GCaMP8f with a ribosome-tethered nanobody that restricts GCaMP to the soma of neurons. Particularly useful in very dense nuclei with strong neuropil, which can make post-hoc cell segmentation (i.e., independent component analysis (ICA) or constrained nonnegative matrix factorization (CMNF)) difficult.
pAAV_Ef1a_DIO_hTyrNull_HATag
By Julia Julia Leonard onCre-dependent expression of a truncated human tyrosinase is inactive and does not lead to the accumulation of neuromelanin in catecholaminergic neurons (control virus). Strong expression of Ef1a promoter, HA tag for histological verification of expression.
pAAV_Ef1a_DIO_hTyrHA_minBack
By Julia Julia Leonard onCre-dependent expression of human tyrosinase induces the formation of neuromelanin in catecholaminergic neurons. Expression under strong Ef1a promoter.
pAAV_Cbh_DIO_hTyr
By Julia Julia Leonard onCre-dependent expression of human tyrosinase induces the formation of neuromelanin in catecholaminergic neurons. The CbH promoter is a strong, pan-neuronal, non-silencable promoter.
pAAV_CbH_SIO_SyGCaMP8s_p2a_mRuby
By Julia Julia Leonard onThis construct links GCaMP8s to Synaptophysin and directs the fusion protein to presynaptic sites. It also produces a cytoplasmic red fluorescence protein (mRuby) via the self-splicing peptide 2A. It is mainly used to longitudinally image synaptic activity with 2-Photon at 980nm and visualize axons at 1080nm in vivo.
Induction of aggregation in alpha-synuclein-expressing cells by treatment with preformed fibrils (PFFs)
By Julia Julia Leonard onThis protocol details how to efficiently produce alpha-synuclein aggregates in cells by templating the misfolding of intracellular alpha-synuclein through treatment with preformed fibrils of alpha-synuclein (PFFs).
Kufor-Rakeb Syndrome-Associated Psychosis: A novel loss-of-function ATP13A2 variant and response to treatment
By Julia Julia Leonard onBiallelic (autosomal recessive) pathogenic variants in ATP13A2 cause a form of juvenile-onset parkinsonism, termed Kufor-Rakeb syndrome. The article describes the treatment response in an individual with Kufor-Rakeb syndrome-associated psychosis.
Source data for “Feed-forward metabotropic signaling by Cav1 Ca2+ channels supports pacemaking in pedunculopontine cholinergic neurons”
By Julia Julia Leonard onSource data and images from the paper: Feed-forward metabotropic signaling by Cav1 Ca2+ channels supports pacemaking in pedunculopontine cholinergic neurons.
