Abstract

Sihlerl's staining technique is a method for visualizing nerves, while maintaining their positional contexts within organs. The technique stains nervous tissue while rendering other tissues translucent. Initial research assessed the feasibility of this technique on the delicate tissues of bat wings that were preserved by different methods. Specimens of Myotis lucifugus, previously frozen, were fixed in solutions of either 10% formalin or 80% ethanol by volume. After modifications, the technique proved successful in showing nerves down to the finest branches in specimens preserved in either solution. Dissection after staining revealed the dactylopatagial membranes (total area 9.87 cm2 ± 0.96) were supplied by branches of the median nerve, and the plagiopatagium (area 16.97 cm2 ± 1.58) was supplied primarily by the ulnar nerve, along with segmental spinal nerves and at least one nerve originating from the hindlimb. Sensory cell complexes, observed to be associated with sensory hairs on dorsal and ventral wing surfaces, were regularly patterned along rows through the wing. Three 5x5 mm areas were compared for number of primary rows and density of sensory cell complexes. Density was highest along the trailing edge of the dactylopatagium between digits III-IV (7 primary rows, 63 ± 4 cells), intermediate along the trailing edge of plagiopatagium (4 primary rows, 47 ± 6 cells), and lower near the center of the plagiopatagium (2 primary rows, 21± 7 cells). Application of this technique to a sample (n=7) of WNS-positive bats has shown that nerve damage can be seen in some areas where fungal damage was present. This research demonstrated that Sihlerls staining is a useful approach for studying the neuroanatomy of bat wings.

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