Title: High-throughput analysis of synapse number and density in human iPSC-derived neurons.
Legend: Formation of synapses, the connections among nerve cells, is a critical process in brain development. To investigate this in induced pluripotent stem cell-derived neurons, we developed a high-throughput method for quantifying the number and density of synapses on individual neurons. We first label a small subset of neurons using a lentiviral vector encoding a fluorescent protein, mCherry. We then immunolabel cultures with proteins localized at the the pre-synaptic (Syn1) and post-synaptic (PSD95) zones. While previous quantification methods relied on manual counting of each individual fluorescent puncta to assess synapse number, we have employed Imaris software that allows us to digitally reconstruct mCherry-expressing neurons and filter out all puncta that are not associated with the neuron of interest. This pipeline allows us to analyze the morphology and synaptic density of a high volume of neurons accurately and efficiently.
Publication (Poster): Russo, M.L., Crockett, M., Halbe, P., Manders, J., Ardeshiri, M.R., West, N., Zhang, S.C., Sousa, A.M.M., Bhattacharyya, A. (2024). High-resolution analysis of synapse density and number in human trisomy 21 neurons. T21RS 5th International Conference.
Investigator: Anita Bhattacharyya, PhD
About the Lab: Anita Bhattacharyya’s lab examines how brain development is altered in developmental disorders characterized by intellectual impairment. The cerebral cortex is the most complex area of the brain and is responsible for functions unique to humans, such as language and abstract thought. Problems in any of the crucial cerebral cortex formation steps can lead to intellectual impairment.