Learning and performing music affect the brain and cognition and have a positive impact on emotional states. Negative emotional states such as stress, anxiety, and depression are harmful for teaching. Alternative certification teachers in particular are extremely vulnerable to and suffer greatly from negative emotional states. In addition, humor and coping humor are cognitive processes depending on neural networks of the brain. Therefore, the relationships between alternative certification pre-service teachers’ musical behaviors, negative emotional states, humor styles, and coping humor were examined in order to produce knowledge that can be used to increase effectiveness in education regarding negative emotional states, humor styles, and humor. The participants were 112 pre-service teachers enrolled in the alternative certification program of a public university in Turkey ( N = 112). Responses to six psychometric scales and a musical behaviors questionnaire were collected. Results demonstrated that having music and instrument education, duration of those educations, playing a musical instrument, and being member of a musical group were related to various emotional states, humor styles, and coping humor. The findings obtained from the analyses of the relationships between 11 musical variables and negative emotional states, humor styles, and coping humor are discussed in light of the related literature.