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Light and electron microscopic imaging of synaptic vesicle endocytosis at mouse hippocampal cultures
Source: ncbi.nlm.nih.gov
Article summary:
1. This article provides a protocol for quantitative measurements of endocytosis at cultured synapses.
2. The protocol includes primary culture of mouse hippocampal neurons, studying endocytosis at neurons transfected with a pH-sensitive synaptophysin-pHluorin2× using fluorescent microscopy, and imaging endocytosis at fixed neurons with electron microscopy.
3. Impairments of endocytosis may contribute to the generation of neurological disorders, such as neurodegeneration and spinocerebellar ataxia 13.
Article analysis:
The article is generally reliable and trustworthy in its presentation of the protocol for quantitative measurements of endocytosis at cultured synapses. It provides detailed instructions on how to perform the experiments, including information about the materials and equipment needed, as well as step-by-step method details for hippocampal neuron culture, digestion and dissociation, cell plating, maintenance of cultured cells, etc. The article also mentions potential risks associated with working with 3,3′-diaminobenzidine (DAB), which is a suspected carcinogen.
However, there are some points that could be improved upon in terms of trustworthiness and reliability. For example, while the article does mention potential neurological disorders that impairments of endocytosis may contribute to (e.g., neurodegeneration and spinocerebellar ataxia 13), it does not provide any evidence or research to back up this claim. Additionally, while the article does provide detailed instructions on how to perform the experiments, it does not discuss any potential pitfalls or errors that could occur during the process or how to troubleshoot them if they do occur. Finally, while the article does mention potential risks associated with working with DAB solution (i.e., wearing lab coats, gloves and protective goggles), it does not discuss any other safety precautions that should be taken when performing these experiments (e.g., proper disposal of hazardous materials).
In conclusion, while this article is generally reliable and trustworthy in its presentation of the protocol for quantitative measurements of endocytosis at cultured synapses, there are some areas where it could be improved upon in terms of trustworthiness and reliability (e.g., providing evidence for claims made about neurological disorders caused by impairments in endocytosis; discussing potential pitfalls or errors that could occur during experimentation; discussing other safety
Topics for further research:
Neurological disorders caused by impairments in endocytosis
Potential pitfalls in quantitative measurements of endocytosis
Troubleshooting errors in endocytosis experiments
Safety precautions for working with 3,3′-diaminobenzidine (DAB)
Proper disposal of hazardous materials in endocytosis experiments
Best practices for hippocampal neuron culture