Microplastics (MPs) have emerged as significant environmental pollutants, raising concerns about their impact on aquatic ecosystems and human health. While most research focuses on traditional plastic polymers, rubber-based MPs, primarily derived from tire and road wear particles (TRWPs), constitute a major portion of MP pollution. TRWPs are generated by vehicle tire abrasion, releasing over 10,000 tons annually into the environment. These particles contain toxic substances, including heavy metals and polycyclic aromatic hydrocarbons (PAHs), posing risks to ecosystems and human populations, particularly in urban areas with dense road networks. Despite their prevalence, limited studies have quantified TRWP concentrations or assessed their toxicity in artificial water bodies within cities. This study quantifies and evaluates the toxicity of TRWPs in anthropogenic water bodies in urban Japan. Environmental surveys and exposure experiments were conducted across five metropolitan areas (Kanto, Kinki, Chukyo, Kitakyushu/Fukuoka, and Sapporo), with artificial water bodies serving as sampling sites due to their proximity to human activity. The methodology included (1) selection of sampling sites based on hydrological conditions and accessibility, (2) field sampling of water, (3) quantification and qualitative of TRWP concentrations using FE-SEM, Raman spectroscopy, or FTIR, respectively, and (4) toxicity evaluation through controlled exposure experiments on algae, Daphnia magna, and killifish following OECD guidelines. Preliminary water quality analysis suggests higher contamination levels in urban ponds, with elevated conductivity, total dissolved solids (TDS), total nitrogen (TN), and total phosphorus (TP) in locations such as Ohori Park (Fukuoka Prefecture) and Tokugawa Park (Nagoya Prefecture), indicative of potential TRWP pollution. Future experiments will focus on identifying tire particles in pond water to further assess their presence and environmental impact. The findings will contribute to environmental risk assessments, inform policy measures for mitigating TRWP pollution, and advance global awareness of rubber-based MP contamination. This research may also influence tire material improvements and regulatory actions to address the growing environmental challenges posed by TRWP emissions.