RESUMO
Marsupials are born very immature yet must be sufficiently autonomous to crawl on the mother's belly, find a teat and attach to it to pursue their development. Sensory inputs are necessary to guide the newborn to a teat and induce attachment. The vestibular system, which perceives gravity and head movements, is one of the senses proposed to guide newborns towards the teats but there are conflicting observations about its functionality at birth (postnatal day (P) 0). To test if the vestibular system of opossum newborns is functional and can influence locomotion, we used two approaches. First, we stimulated the vestibular apparatus in in vitro preparations from opossums aged from P1 to P12 and recorded motor responses: at all ages studied, mechanical pressures applied on the vestibular organs induced spinal roots activity whereas head tilts did not induce forelimb muscle contractions. Second, using immunofluorescence, we assessed the presence of Piezo2, a protein involved in mechanotransduction in vestibular hair cells. Piezo2 labeling was scant in the utricular macula at birth, but observed in all vestibular organs at P7, its intensity increasing up to P14; it seemed to stay the same at P21. Our results indicate that neural pathways from the labyrinth to the spinal cord are already in place around birth but that the vestibular organs are too immature to influence motor activity before the end of the second postnatal week in the opossum. It may be the rule in marsupial species that the vestibular system becomes functional only after birth.
RESUMO
Roughly two-thirds of the proteins annotated as transcription factors in dinoflagellate transcriptomes are cold shock domain-containing proteins (CSPs), an uncommon condition in eukaryotic organisms. However, no functional analysis has ever been reported for a dinoflagellate CSP, and so it is not known if they do in fact act as transcription factors. We describe here some of the properties of two CSPs from the dinoflagellate Lingulodinium polyedrum, LpCSP1 and LpCSP2, which contain a glycine-rich C-terminal domain and an N-terminal cold shock domain phylogenetically related to those in bacteria. However, neither of the two LpCSPs act like the bacterial CSP, since they do not functionally complement the Escherichia coli quadruple cold shock domain protein mutant BX04, and cold shock does not induce LpCSP1 and LpCSP2 to detectable levels, based on two-dimensional gel electrophoresis. Both CSPs bind to RNA and single-stranded DNA in a nonspecific manner in electrophoretic mobility shift assays, and both proteins also bind double-stranded DNA nonspecifically, albeit more weakly. These CSPs are thus unlikely to act alone as sequence-specific transcription factors. IMPORTANCE Dinoflagellate transcriptomes contain cold shock domain proteins as the major component of the proteins annotated as transcription factors. We show here that the major family of cold shock domain proteins in the dinoflagellate Lingulodinium do not bind specific sequences, suggesting that transcriptional control is not a predominant mechanism for regulating gene expression in this group of protists.
