Showing 1 - 3 of 3 Items
Bilateral consequences of chronic unilateral deafferentation in the auditory system of the cricket gryllus bimaculatus
Date: 2011-04-01
Creator: Hadley Wilson Horch, Elizabeth Sheldon, Claire C. Cutting, Claire R. Williams, Dana M., Riker, Hannah R. Peckler, Rohit B. Sangal
Access: Open access
- The auditory system of the cricket has the unusual ability to respond to deafferentation by compensatory growth and synapse formation. Auditory interneurons such as ascending neuron 2 (AN-2) in the cricket Gryllus bimaculatus possess a dendritic arbor that normally grows up to, but not over, the midline of the prothoracic ganglion. After chronic deafferentation throughout larval development, however, the AN-2 dendritic arbor changes dramatically, and medial dendrites sprout across the midline where they form compensatory synapses with the auditory afferents from the contralateral ear. We quantified the extent of the effects of chronic, unilateral deafferentation by measuring several cellular parameters of 3 different neuronal components of the auditory system: the deafferented AN-2, the contralateral (or nondeafferented) AN-2 and the contralateral auditory afferents. Neuronal tracers and confocal microscopy were used to visualize neurons, and double-label experiments were performed to examine the cellular relationship between pairs of cells. Dendritic complexity was quantified using a modified Sholl analysis, and the length and volume of processes and presynaptic varicosities were assessed under control and deafferented conditions. Chronic deafferentation significantly influenced the morphology of all 3 neuronal components examined. The overall dendritic complexity of the deafferented AN-2 dendritic arbor was reduced, while both the contralateral AN-2 dendritic arbor and the remaining, intact, auditory afferents grew longer. We found no significant changes in the volume or density of varicosities after deafferentation. These complex cellular changes after deafferentation are interpreted in the light of the reported differential regulation of vesicle-associated membrane protein and semaphorin 2a. Copyright © 2011 S. Karger AG, Basel.
Destabilization of cortical dendrites and spines by BDNF
Date: 1999-01-01
Creator: Hadley Wilson Horch, Alex Krüttgen, Stuart D. Portbury, Lawrence C. Katz
Access: Open access
- Particle-mediated gene transfer and two-photon microscopy were used to monitor the behavior of dendrites of individual cortical pyramidal neurons coexpressing green fluorescent protein (GFP) and brain-derived neurotrophic factor (BDNF). While the dendrites and spines of neurons expressing GFP alone grew modestly over 24-48 hr, coexpressing BDNF elicited dramatic sprouting of basal dendrites, accompanied by a regression of dendritic spines. Compared to GFP-transfected controls, the newly formed dendrites and spines were highly unstable. Experiments utilizing Trk receptor bodies, K252a, and overexpression of nerve growth factor (NGF) demonstrated that these effects were mediated by secreted BDNF interacting with extracellular TrkB receptors. Thus, BDNF induces structural instability in dendrites and spines, which, when restricted to particular portions of a dendritic arbor, may help translate activity patterns into specific morphological changes.
In situ HCR in non-traditional arthropods
Date: 2024-01-25
Creator: Heather Bruce, Hadley Wilson Horch
Access: Open access
- Visualizing the expression of genes is a fundamental tool in molecular biology. Traditional colorimetric in situ hybridization using long RNA probes has been a staple for visualizing gene expression but has many drawbacks. In situ HCR v3.0, developed by Choi et. al. 2018, offers improvements over traditional in situs in nearly every aspect: probes can simply be ordered rather than painstakingly cloned and transcribed, which also makes them cost-effective; an HCR takes just three days to complete rather than five or more days; HCR is robust and works well for first-time users; and HCR probes can be multiplexed, allowing four to eight genes to be visualized in a single sample. HCR has been used successfully in many arthropods, including insects (Drosophila, Tribolium), crustaceans (Parhyale, Daphnia, Artemia), and chelicerates (Limulus horseshoe crab, Acanthoscurria tarantula). In this demo, you will learn how to design and order HCR probes as well as best practices for experimental design.