Innervation of cercarial tegumentary receptors investigated by the Sevier-Munger method

The investigation of the sensory nature of tegumentary receptors in platyhelminths remains a challenge due to technical difficulties related to nerve tissue exposure and its experimental handling. Neuromorphological studies have been carried out but few demonstrated the association of these receptors with the nervous system. This paper introduces the Sevier-Munger method as an alternative approach to study the innervation of tegumentary receptors in larval flatworms. Twenty heterophyid cercariae were fixed in hot 5% formalin, with all washes performed in tap and distilled water. They were developed in a solution of ammoniacal silver and 2% formalin under the microscope for 10 min, with preparations shaken gently throughout the procedure. In all specimens, nerve cells stained black against a pale gold background. Fine nerve fibers of the subsurface nerve plexus were observed. These fibers sent distal branches from the plexus to the cercarial tegument. The branches became fine nerve endings, projecting as receptors in the cephalic (5CIV(5), 2CV(2), and 2CV(4)), anterior (4AIL, 3AIIL, 2AIIIL), midbody (1ML, 3MV), posterior (1PIL, 1PIIL, and 1PIIIL), and caudal (2UD) regions of the cercaria. These results indicate that the Sevier-Munger method is useful to demonstrate the association of cercarial tegumentary receptors with the subsurface nerve plexus. They also recommend the use of alternative methods to further investigate flatworm nervous systems. Moreover, there is a pressing urge for a standardized protocol, combining a plethora of methods and techniques. Interdisciplinary collaboration aiming at a better understanding of the function of flatworm nervous systems is particularly encouraged.