ObjectiveThis study was undertaken to investigate migraine glymphatic and meningeal lymphatic vessel (mLV) functions.MethodsMigraine patients and healthy controls (HCs) were prospectively recruited between 2020 and 2023. Diffusion tensor image analysis along the perivascular space (DTI‐ALPS) index for glymphatics and dynamic contrast‐enhanced magnetic resonance imaging parameters (time to peak [TTP]/enhancement integral [EI]/mean time to enhance [MTE]) for para‐superior sagittal (paraSSS)‐mLV or paratransverse sinus (paraTS)‐mLV in episodic migraine (EM), chronic migraine (CM), and CM with and without medication‐overuse headache (MOH) were analyzed. DTI‐ALPS correlations with clinical parameters (migraine severity [numeric rating scale]/disability [Migraine Disability Assessment (MIDAS)]/bodily pain [Widespread Pain Index]/sleep quality [Pittsburgh Sleep Quality Index (PSQI)]) were examined.ResultsIn total, 175 subjects (112 migraine + 63 HCs) were investigated. DTI‐ALPS values were lower in CM (median [interquartile range] = 0.64 [0.12]) than in EM (0.71 [0.13], p = 0.005) and HCs (0.71 [0.09], p = 0.004). CM with MOH (0.63 [0.07]) had lower DTI‐ALPS values than CM without MOH (0.73 [0.12], p < 0.001). Furthermore, CM had longer TTP (paraSSS‐mLV: 55.8 [12.9] vs 40.0 [7.6], p < 0.001; paraTS‐mLV: 51.2 [8.1] vs 44.0 [3.3], p = 0.002), EI (paraSSS‐mLV: 45.5 [42.0] vs 16.1 [9.2], p < 0.001), and MTE (paraSSS‐mLV: 253.7 [6.7] vs 248.4 [13.8], p < 0.001; paraTS‐mLV: 252.0 [6.2] vs 249.7 [1.2], p < 0.001) than EM patients. The MIDAS (p = 0.002) and PSQI (p = 0.002) were negatively correlated with DTI‐ALPS index after Bonferroni corrections (p < q = 0.01).InterpretationCM patients, particularly those with MOH, have glymphatic and meningeal lymphatic dysfunctions, which are highly clinically relevant and may implicate pathogenesis for migraine chronification. ANN NEUROL 2024;95:583–595