ABSTRACT
Paraquat (PQ) is a well-known neurotoxin closely associated with neurodegenerative Parkinson's disease (PD). Zebrafish are utilized as a model for PD research because of their well-defined neuropathology and locomotor behavior. Here, we highlight protocols for inducing PD using PQ and analyzing locomotor activity in adult zebrafish. Basic Protocol 1 details the treatment of adult male zebrafish with 60 mg/kg PQ via intraperitoneal injection to induce a PD-like phenotype, followed by the steps to perform a locomotor assay. Basic Protocol 2 provides step-by-step guidance for processing the acquired videos in ToxTrac software to understand the locomotor parameters of 0.9% saline- and 60 mg/kg PQ-injected adult zebrafish. The simplicity of the treatment strategy, low-cost video acquisition setup, and free video processing make these protocols accessible without prior experience. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Development of Parkinson's disease features in adult zebrafish Basic Protocol 2: ToxTrac analysis for locomotor assay.
Subject(s)
Herbicides , Neurodegenerative Diseases , Parkinson Disease , Animals , Male , Paraquat/toxicity , Zebrafish , Herbicides/toxicity , Parkinson Disease/etiology , Parkinson Disease/pathology , Neurodegenerative Diseases/complications , PhenotypeABSTRACT
Tubulins are the highly conserved subunit of microtubules which involve in various fundamental functions including brain development. Microtubules help in neuronal proliferation, migration, differentiation, cargo transport along the axons, synapse formation, and many more. Tubulin gene family consisting of multiple isotypes, their differential expression and varied post translational modifications create a whole new level of complexity and diversity in accomplishing manifold neuronal functions. The studies on the relation between tubulin genes and brain development opened a new avenue to understand the role of each tubulin isotype in neurodevelopment. Mutations in tubulin genes are reported to cause brain development defects especially cortical malformations, referred as tubulinopathies. There is an increased need to understand the molecular correlation between various tubulin mutations and the associated brain pathology. Recently, mutations in tubulin isotypes (TUBA1A, TUBB, TUBB1, TUBB2A, TUBB2B, TUBB3, and TUBG1) have been linked to cause various neurodevelopmental defects like lissencephaly, microcephaly, cortical dysplasia, polymicrogyria, schizencephaly, subcortical band heterotopia, periventricular heterotopia, corpus callosum agenesis, and cerebellar hypoplasia. This review summarizes on the microtubule dynamics, their role in neurodevelopment, tubulin isotypes, post translational modifications, and the role of tubulin mutations in causing specific neurodevelopmental defects. A comprehensive list containing all the reported tubulin pathogenic variants associated with brain developmental defects has been prepared to give a bird's eye view on the broad range of tubulin functions.
Subject(s)
Brain Diseases , Malformations of Cortical Development , Microcephaly , Humans , Tubulin/metabolism , Brain/metabolism , Microcephaly/genetics , Mutation/genetics , Malformations of Cortical Development/genetics , Microtubules/metabolism , Brain Diseases/pathologyABSTRACT
Zebrafish (Danio rerio) is used as a model for studying sensorineural hearing loss. The damage to the hair cells can be assessed by scoring rheotaxis behavior in zebrafish. In this study, we newly designed a rheotaxis behavioral assay protocol capable of quantifying rheotaxis behavior in zebrafish larvae. We chemically induced ototoxicity in the larvae using copper sulfate, a well-known ototoxin, and determined rheotaxis at different flow velocities. The simple, cost-effective, and high-throughput rheotaxis assay system can provide great insights into drug development and other behavioral studies.
