Francesca Biagi

Mitochondrial DNA variability of the pipefish Syngnathus abaster.

This study provides data on the genetic structuring of the pipefish Syngnathus abaster in the western Mediterranean and Adriatic Seas. A total of 109 specimens were collected in brackish-water biotopes. The control region and three other regions of the mitochondrial genome were analysed. The most relevant result was the high genetic structuring found by Bayesian inference (BI), maximum likelihood (ML) and network analyses, which were consistent in showing three well-separated clusters of S. abaster populations. Furthermore, BI and ML did not support the monophyly of the taxon S. abaster. These results suggest the occurrence of a species complex in the study area, whose differentiation may have occurred since the Pleistocene. The results also show a very high genetic variability at the inter-population level, with no shared haplotypes among sites. Evolutionary forces due to the fragmented nature of the brackish-water habitats may account for the high genetic divergence found among the groups and populations. Finally, although dispersal by rafting over long distances may occasionally occur, this study suggests linear stepping-stone model of colonization to be most likely. The complexity of the results obtained suggests that further studies are needed to elucidate the phylogeny of S. abaster.

Analysis of meristic and mitochondrial DNA variation in Syngnathus abaster (Teleostea: Syngnathidae) from two western Mediterranean lagoons

Syngnathus abaster is a euryhaline pipefish distributed in the Mediterranean, Black Sea, and the north-eastern Atlantic. Although its populations are characterised by high morphological plasticity, neither congruent information about the morphological differentiation of S. abaster populations from the Mediterranean lagoons is available, nor population genetic surveys have been so far performed. In this context, the aims of our study were as follow: i) to describe the variation at nine meristic characters of S. abaster from two western Mediterranean brackish-water areas: the Tunis north lagoon (Tunisia) and the Mauguio lagoon (France); ii) to analyse sequences of four mitochondrial DNA regions in order to evaluate the occurrence of genetic variation between the two areas, if any. The morphological survey revealed a subdivision into two distinct groups: the first included the Tunisian specimens, the second the French ones. Genetic analysis evidenced the occurrence of a sharp genetic structuring with high levels of genetic differentiation between Tunisian and French S. abaster populations. Results suggest that the evolutionary forces driven by the different biogeographical and ecological conditions of the two Mediterranean brackish-water areas have boosted the morphological and genetic divergence of the populations here analysed. The scarce potential of long-distance dispersal of S. abaster may have also enhanced the divergence retrieved.

Neurons within the trigeminal mesencephalic nucleus encode for the kinematic parameters of the whisker pad macrovibrissae

It has been recently shown in rats that spontaneous movements of whisker pad macrovibrissae elicited evoked responses in the trigeminal mesencephalic nucleus (Me5). In the present study, electrophysiological and neuroanatomical experiments were performed in anesthetized rats to evaluate whether, besides the whisker displacement per se, the Me5 neurons are also involved in encoding the kinematic properties of macrovibrissae movements, and also whether, as reported for the trigeminal ganglion, even within the Me5 nucleus exists a neuroanatomical representation of the whisker pad macrovibrissae. Extracellular electrical activity of single Me5 neurons was recorded before, during, and after mechanical deflection of the ipsilateral whisker pad macrovibrissae in different directions, and with different velocities and amplitudes. In several groups of animals, single or multiple injections of the tracer Dil were performed into the whisker pad of one side, in close proximity to the vibrissae follicles, in order to label the peripheral terminals of the Me5 neurons innervating the macrovibrissae (whisking‐neurons), and therefore, the respective perikaria within the nucleus. Results showed that: (1) the whisker pad macrovibrissae were represented in the medial‐caudal part of the Me5 nucleus by a single cluster of cells whose number seemed to match that of the macrovibrissae; (2) macrovibrissae mechanical deflection elicited significant responses in the Me5 whisking‐neurons, which were related to the direction, amplitude, and frequency of the applied deflection. The specific functional role of Me5 neurons involved in encoding proprioceptive information arising from the macrovibrissae movements is discussed within the framework of the whole trigeminal nuclei activities.

The mesencephalic–hypoglossal nuclei loop as a possible central pattern generator for rhythmical whisking in rats

It has been previously demonstrated that the Me5 nucleus is involved in the genesis of reflex activities at whisker pad level. Specific Me5 neurons, which provide sensory innervation of the macrovibrissae, are monosynaptically connected with small hypoglossal neurons innervating the extrinsic muscles that control macrovibrissal movements. Artificial whisking, induced by the electrical stimulation of the peripheral stump of the facial nerve and the electrical stimulation of the XII nucleus or the infraorbital nerve, induced evoked responses in the whisker pad extrinsic motor units, along with a significant increase in the electromyographic activity of the extrinsic pad muscles (Mameli et al. in Acta Oto-Laryngol 126:1334–1338, 2006; in Pfűgers Arch Eur J Physiol 456:1189–1198, 2008; in Brain Res 1283:34–40, 2009; in Exp Brain Res 234:753–761, 2016). In anaesthetized rats, we evaluated the possible involvement of this Me5–XII loop in the genesis of rhythmical whisking. The anatomical findings showed that in addition to the ipsilateral, even the contralateral Me5 nucleus could be retrogradely labeled by the Dil tracer injected into the whisker pad of one side, they, furthermore, showed labeled axons extending across the midline between the two nuclei. The electrophysiological findings agreed with the neuroanatomical results, since the mechanical or artificially induced deflection of the whiskers of one side, evoked in the Me5 contralateral nucleus different patterns of responses. The hypothesis that the Me5–XII loops, along with their cross-linked relationship, could act as a “central generator” responsible for the stereotyped symmetrical pattern of macrovibrissal movements such as rhythmical whisking has been discussed.

Autophagic processes increase during senescence in cultured sheep neurons and astrocytes

A possible response to aging is autophagy, a self-digestion process in which portions of cytoplasm are encapsulated by double-membrane-bound structures and delivered to lysosome for degradation. A previous work of our group showed that astrocytes under starving conditions are characterized by a higher upregulation of the marker of autophagy LC3 II than neurons. Aim of the present work was to evaluate LC3 II expression in an aging model consisting in fetal sheep neurons and astrocytes at 10, 20 and 30 days of culture. Such model has been validated by a remarkable activity of β-galactosidase, commonly used to reveal cell aging. LC3 II immunoreactivity in neurons and astrocytes progressively increased with time but differences were observed on the basis of cell density. Indeed, LC3 II immunoreactivity is higher in clusters of neurons and astrocytes and this may be due to the fact that cell-cell contact would represent a second stress in addition to aging itself. Both cell types displayed a reduction in LC3 II signal in nuclei, and a corresponding strengthening in the cytoplasm with time. This may be due to the need of aged cells to remove damaged cytoplasmic components through autophagic processes. Such variation in LC3 II localization could be caused by migration from the nucleus to cytoplasm as well as possible de novo intracytoplasmic production. The present work based on sheep neural cells in vitro may represent a helpful tool in the studies on aging processes in which autophagy plays a remarkable role.