Melanin is a complex pigment that is found in various fungal species and is associated with a multitude of protective functions against environmental stresses. In Cryptococcus neoformans, melanin is synthesized from exogenous substrate and deposited in the cell wall. Although melanin is often cited as a protector against mechanical stress, there is a paucity of direct experimental data supporting this claim. To probe whether melanin enhances cellular strength, we used ultrasonic cavitation and French cell press pressure to stress cryptococcal cells and then measured changes in cellular morphology and fragmentation for melanized and nonmelanized C. neoformans cells. Melanized yeast cells exhibited lower rates of fragmentation and greater cell areas than did nonmelanized yeast cells after sonication or French press passage. When subjected to French press passage, both melanized and nonmelanized cells exhibited responses that were dependent on their culture age. Our results indicate that melanization protects against some of the morphological changes, such as fragmentation and cellular shrinkage, that are initiated by mechanical energy derived from either sonic cavitation or French press passage, thus supporting the notion that this pigment provides mechanical strength for fungal cell walls. IMPORTANCE Melanin was shown in prior microbiological experiments to be associated with protection against environmental stressors, and it has often been cited as being associated with mechanical stress protection. However, there is a lack of direct experimentation to confirm this claim. We examined the responses of melanized and nonmelanized C. neoformans cells to sonication and French press passage, and we report differences in outcomes depending not only on melanization status but also on culture age. Such findings have important implications for the design and interpretation of laboratory experiments involving C. neoformans. In addition, the elucidation of some of the mechanical properties of melanin promotes further research into fungal melanin applications in health care and industry.
Keywords: Cryptococcus neoformans; French press; mechanical stress; melanin; mycology; sonication; stress response.