Pigmentation in the sensory organs of the ascidian larva is essential for normal behavior.

Free-living animals and their larvae utilize light and gravity as cues to navigate in open space. Detection and response to these environmental stimuli are important for the dispersal and settlement of ascidian larvae. Two pigmented structures in the brain of the ascidian larva, the ocellus and the otolith, have been shown to function as the photoreceptive and gravity sensitive organs, respectively. Here, we show that pigmentation is essential for proper phototactic and geotactic behavior in larvae of the ascidian species Ciona savignyi. Two recessive and complementing mutant lines of C. savignyi, immaculate and spotless, that specifically disrupt the pigmentation of the sensory organs during larval development are described. Homozygous mutant larvae are unable to respond properly to gravity and illumination cues while settling. Genetic analysis shows that spotless is caused by a point mutation within the tyrosinase gene that creates a premature stop codon, while the molecular nature of immaculate is unknown. Although the role of pigmentation in the ocellus of C. savignyi is similar to that in vertebrate visual systems, our results demonstrate a novel use of melanin in geotactic behavior.

Alpha 4 integrins and vascular cell adhesion molecule-1 play a role in sympathetic innervation of the heart.

Sympathetic neurons innervate the heart early in postnatal development, an event that is crucial for proper modulation of blood pressure and cardiac function. However, the axon guidance cues that direct sympathetic neurons to the heart, and the neuronal receptors that recognize those cues, are poorly understood. Here we present evidence that interactions between the alpha4beta1 integrin on sympathetic neurons and vascular cell adhesion molecule-1 (VCAM-1) in the heart plays a role in cardiac innervation. The alpha4 subunit was detected on postnatal rat superior cervical ganglion (SCG) neurons in culture and in cryosections of SCG and heart. VCAM-1 immunoreactivity was detected on cardiac myocytes that associate with invading sympathetic neurons. Purified recombinant soluble VCAM-1 (rsVCAM-1) stimulated SCG neurite outgrowth at levels comparable with laminin 2/4 and fibronectin (Fn), and outgrowth on rs-VCAM-1 and Fn was blocked by antibodies specific for the alpha4 and beta1 integrin subunits. Intrathoracic injection of function-blocking antibodies to alpha4 and VCAM-1, as well as a small molecule inhibitor of alpha4 integrins, significantly reduced sympathetic innervation of the heart. These results indicate that the interaction between alpha4 integrin and VCAM-1 is important for sympathetic innervation of the heart.