Texture and nano-scale internal microstructure of otoliths in the Atlantic molly, Poecilia mexicana: a high-resolution EBSD study.

Otoliths of modern bony fishes are massive polycrystalline structures consisting mainly of calcium carbonate (primarily aragonite), and 1-10% organic residuals. Unlike other biomineralisates like shells, teeth and bones, they are not optimized for mechanical loads but serve the senses of hearing and balance in the inner ear. We examined internal structural variation of otoliths through microstructural and texture analyses. Our study applied the electron backscattered diffraction technique (EBSD) to whole sections of saccular otoliths on cave- and surface-dwelling fish. Application of high spatial resolution EBSD on otoliths of the livebearing fish Poecilia mexicana allowed for an investigation of crystal orientation despite the small size (<150 nm) of aragonite crystallites. Crystallites at the rims of otoliths had a higher structural organization than those situated near the center, where no dominant orientation pattern was discernible. Moreover, the medial (sulcal) face of otoliths, which makes contact with the sensory epithelium, was more structured than the lateral (antisulcal) face.

Toxic hydrogen sulphide and dark caves: pronounced male life-history divergence among locally adapted Poecilia mexicana (Poeciliidae).

Chronic environmental stress is known to induce evolutionary change. Here, we assessed male life-history trait divergence in the neotropical fish Poecilia mexicana from a system that has been described to undergo incipient ecological speciation in adjacent, but reproductively isolated toxic/nontoxic and surface/cave habitats. Examining both field-caught and common garden-reared specimens, we investigated the extent of differentiation and plasticity of life-history strategies employed by male P. mexicana. We found strong site-specific life-history divergence in traits such as fat content, standard length and gonadosomatic index. The majority of site-specific life-history differences were also expressed under common garden-rearing conditions. We propose that apparent conservatism of male life histories is the result of other (genetically based) changes in physiology and behaviour between populations. Together with the results from previous studies, this is strong evidence for local adaptation as a result of ecologically based divergent selection.

Local adaptation and pronounced genetic differentiation in an extremophile fish, Poecilia mexicana, inhabiting a Mexican cave with toxic hydrogen sulphide.

We investigated genetic differentiation and migration patterns in a small livebearing fish, Poecilia mexicana, inhabiting a sulfidic Mexican limestone cave (Cueva del Azufre). We examined fish from three different cave chambers, the sulfidic surface creek draining the cave (El Azufre) and a nearby surface creek without the toxic hydrogen sulphide (Arroyo Cristal). Using microsatellite analysis of 10 unlinked loci, we found pronounced genetic differentiation among the three major habitats: Arroyo Cristal, El Azufre and the cave. Genetic differentiation was also found within the cave between different pools. An estimation of first-generation migrants suggests that (i) migration is unidirectional, out of the cave, and (ii) migration among different cave chambers occurs to some extent. We investigated if the pattern of genetic differentiation is also reflected in a morphological trait, eye size. Relatively large eyes were found in surface habitats, small eyes in the anterior cave chambers, and the smallest eyes were detected in the innermost cave chamber (XIII). This pattern shows some congruence with a previously proposed morphocline in eye size. However, our data do not support the proposed mechanism for this morphocline, namely that it would be maintained by migration from both directions into the middle cave chambers. This would have led to an increased variance in eye size in the middle cave chambers, which we did not find. Restricted gene flow between the cave and the surface can be explained by local adaptations to extreme environmental conditions, namely H2S and absence of light. Within the cave system, habitat properties are patchy, and genetic differentiation between cave chambers despite migration could indicate local adaptation at an even smaller scale.