Abstract
Adult neurogenesis is widespread among metazoans, it occurs in animals with a network nervous system, as cnidarians, and in animals with a complex and centralized brain, such as mammals, non-mammalian vertebrates, ecdysozoans, and a lophotrochozoan, Octopus vulgaris. Nevertheless, there are important differences among taxa, especially in the number of the regions involved and in cell proliferation rate during the life-cycle. The comparative evaluation of adult neurogenesis among different brain regions is an arduous task to achieve with only stereological techniques. However, in Octopus vulgaris we recently confirmed the presence of active proliferation in the learning-memory centers, multisensory integration centers, and the motor centers of the adult brain. Here, using a flow cytometry technique, we provide a method to quantify the active proliferation in octopus nervous system using a BrdU in vitro administration without exposing the animals to stress or painful injections usually used. This method is in line with the current animal welfare regulations regarding cephalopods, and the flow cytometry-based technique enabled us to measure adult neurogenesis more quickly and reliably than histological techniques, with the additional advantage of processing multiple samples in parallel. Flow cytometry is thus an appropriate technique for measuring and comparing adult neurogenesis in animals that are in a different physiological and/or environmental contexts. A BrdU immunoreactivity distribution, to define the neurogenic areas, and the effective penetration in vitro of the BrdU is also provided.
Highlights
Adult neurogenesis is a process consisting of proliferation, migration, and differentiation of newborn cells, which become functionally integrated into the existing neural circuitry of the adult brain (Duan et al, 2008; Sun et al, 2011)
No specific labeling was observed, as on randomly picked sections from BrdU treated brains where the primary antibody against BrdU was substituted with plain phosphate buffered saline (PBS) to control secondary unspecific staining
Besides confirming adult neurogenesis in the lophotrochozoan clade, this study has quantified the extent of active cell proliferation in the brain of adult O. vulgaris using a flow cytometry assay
Summary
Adult neurogenesis is a process consisting of proliferation, migration, and differentiation of newborn cells, which become functionally integrated into the existing neural circuitry of the adult brain (Duan et al, 2008; Sun et al, 2011). Adult neurogenesis goes on throughout life, the sophisticated balance of multiple factors, such as growth factors, hormones, neurotransmitters, is altered by senescence (Klempin and Kempermann, 2007; Couillard-Després et al, 2011) The latter results in a reduced number of stem cells and cell precursors (Hamilton et al, 2013), in a decrease of their proliferation rate and in a compromised development of new neurons, which taken together lead to a substantial decline in neurogenesis (Riddle and Lichtenwalner, 2007; Couillard-Després et al, 2011; CapillaGonzalez et al, 2015)
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