Physical activity heterogenously modulates NG2-glia population behavior, and is necessary for cognitive enhancement

NG2-glia is a macroglial population, which constitutes about 5-10% of the total cell population in the mammalian brain. These cells have “stem cell-like” features; for instance, they can proliferate and self-renew and they mostly differentiate into oligodendrocytes, a cell type are of great importance as they myelinate axons in the central nervous system, a process essential for the proper function of vertebrates’ nervous system. Although most myelination happens after birth and completed at a young age, it has been shown that it can also occur during adulthood in mammals. Adult myelination can be modulated by experience, but the exact mechanism of this phenomenon remains unclear. Hence, it is thought that neuronal activity could stop the proliferation and promote the differentiation of NG2-glia, and in turn, newly generated oligodendrocytes could provide the new myelin. However, it is still unclear how neuronal activity could lead to changes in NG2-glia behavior in the adult mouse. In this doctoral thesis, I have used a voluntary physical activity (VPA) mouse model to study the effects of experience on NG2-glia, although other mechanism cannot be discarded. Indeed, our results showed an increase in the proliferation and differentiation of NG2-glia in the cerebral cortical grey matter but not in the corresponding white matter after VPA. We also observed that NG2-glia tend to differentiate with two different modalities, and one of them is preferred during VPA. Furthermore, I performed mass spectrometry of sorted NG2-glia to profile them after VPA, and found that the remaining, non-differentiated NG2-glia show less myelin-related proteins. Interestingly, the results of the proteome analysis correlate with the increase in the number of the GPR17+ subset of NG2-glia, which is characterized by its slow differentiation rate, and I observed that this population remains mostly unaffected by VPA. Finally, for the first time, I found that newly generated oligodendrocytes integrate into the circuitry of the cortex and this myelin remodeling contributes in cognitive enhancement induced by exercise

Optical recording from respiratory pattern generator of foetal mouse brainstem reveals a distributed network

Author Posting. © The Authors, 2005. This is the author's version of the work. It is posted here by permission of Elsevier B. V. for personal use, not for redistribution. The definitive version was published in Neuroscience 137 (2006): 1221-1227, doi:10.1016/j.neuroscience.2005.10.053.Unfailing respiration depends on neural mechanisms already present in mammals before birth. Experiments were made to determine how inspiratory and expiratory neurons are grouped in the brainstem of fetal mice. A further aim was to assess whether rhythmicity arises from a single pacemaker or is generated by multiple sites in the brainstem. To measure neuronal firing, a fluorescent calcium indicator dye was applied to embryonic central nervous systems isolated from mice. While respiratory commands were monitored electrically from third to fifth cervical ventral roots, activity was measured optically over areas containing groups of respiratory neurones, or single neurones, along the medulla from the facial nucleus to the pre-Bötzinger complex. Large optical signals allowed recordings to be made during individual respiratory cycles. Inspiratory and expiratory neurones were intermingled. A novel finding was that bursts of activity arose in a discrete area intermittently, occurring during some breaths, but failing in others. Raised CO2 partial pressure or lowered pH increased the frequency of respiration; neurons then fired reliably with every cycle. Movies of activity revealed patterns of activation of inspiratory and expiratory neurones during successive respiratory cycles; there was no evidence for waves spreading systematically from region to region. Our results suggest that firing of neurons in immature respiratory circuits is a stochastic process, and that the rhythm does not depend on a single pacemaker. Respiratory circuits in fetal mouse brainstem appear to possess a high safety factor for generating rhythmicity, which may or may not persist as development proceeds.Supported by grants from FONDECYT #1010242 and #7010242 to J.E., NIH DC05259 to L.C., the state of FL to K.M., and an Evelyn and Melvin Spiegel Foundation Award for a summer research fellowship at the MBL to J.E

A versatile transcription factor: Multiple roles of orthopedia a (otpa) beyond its restricted localization in dopaminergic systems of developing and adult zebrafish (Danio rerio) brains

Many transcription factors boost neural development and differentiation in specific directions and serve for identifying similar or homologous structures across species. The expression of Orthopedia (Otp) is critical for the development of certain cell groups along the vertebrate neuraxis, for example, the medial amygdala or hypothalamic neurosecretory neurons. Therefore, the primary focus of the present study is the distribution of Orthopedia a (Otpa) in the larval and adult zebrafish (Danio rerio) brain. Since Otpa is also critical for the development of zebrafish basal diencephalic dopaminergic cells, colocalization of Otpa with the catecholamine synthesizing enzyme tyrosine hydroxylase (TH) is studied. Cellular colocalization of Otpa and dopamine is only seen in magnocellular neurons of the periventricular posterior tubercular nucleus and in the posterior tuberal nucleus. Otpa-positive cells occur in many additional structures along the zebrafish neuraxis, from the secondary prosencephalon down to the hindbrain. Furthermore, Otpa expression is studied in shh-GFP and islet1-GFP transgenic zebrafish. Otpa-positive cells only express shh in dopaminergic magnocellular periventricular posterior tubercular cells, and only colocalize with islet1-GFP in the ventral zone and prerecess caudal periventricular hypothalamic zone and the perilemniscal nucleus. The scarcity of cellular colocalization of Otpa in islet1-GFP cells indicates that the Shh-islet1 neurogenetic pathway is not active in most Otpa-expressing domains. Our analysis reveals detailed correspondences between mouse and zebrafish forebrain territories including the zebrafish intermediate nucleus of the ventral telencephalon and the mouse medial amygdala. The zebrafish preoptic Otpa-positive domain represents the neuropeptidergic supraopto-paraventricular region of all tetrapods. Otpa domains in the zebrafish basal plate hypothalamus suggest that the ventral periventricular hypothalamic zone corresponds to the otp-expressing basal hypothalamic tuberal field in the mouse. Furthermore, the mouse otp domain in the mammillary hypothalamus compares partly to our Otpa-positive domain in the prerecess caudal periventricular hypothalamic zone (Hc-a)

Developmental Origin of PreBotzinger Complex Respiratory Neurons

A subset of preBötzinger Complex (preBötC) neurokinin 1 receptor (NK1R) and somatostatin peptide (SST)-expressing neurons are necessary for breathing in adult rats, in vivo. Their developmental origins and relationship to other preBötC glutamatergic neurons are unknown. Here we show, in mice, that the “core” of preBötC SST+/NK1R+/SST 2a receptor+ (SST2aR) neurons, are derived from Dbx1-expressing progenitors. We also show that Dbx1-derived neurons heterogeneously coexpress NK1R and SST2aR within and beyond the borders of preBötC. More striking, we find that nearly all non-catecholaminergic glutamatergic neurons of the ventrolateral medulla (VLM) are also Dbx1 derived. PreBötC SST+ neurons are born between E9.5 and E11.5 in the same proportion as non-SST-expressing neurons. Additionally, preBötC Dbx1 neurons are respiratory modulated and show an early inspiratory phase of firing in rhythmically active slice preparations. Loss of Dbx1 eliminates all glutamatergic neurons from the respiratory VLM including preBötC NK1R+/SST+ neurons. Dbx1 mutant mice do not express any spontaneous respiratory behaviors in vivo. Moreover, they do not generate rhythmic inspiratory activity in isolated en bloc preparations even after acidic or serotonergic stimulation. These data indicate that preBötC core neurons represent a subset of a larger, more heterogeneous population of VLM Dbx1-derived neurons. These data indicate that Dbx1-derived neurons are essential for the expression and, we hypothesize, are responsible for the generation of respiratory behavior both in vitro and in vivo

11 Data figures and text NCOMMS-16-21368C.xlsx

Data used in figures and text in the paper NCOMMS-16-21368C entitled:<div><br></div><div>D-serine released by astrocytes in the brainstem regulates the breathing response to CO₂ levels</div><div><br></div><div><p>Central chemoreception is essential for adjusting breathing to physiological demands, and for maintaining CO₂ and pH homeostasis in the brain. CO₂-induced ATP release from brainstem astrocytes stimulates breathing. NMDA receptor antagonism reduces the CO₂-induced hyperventilation by unknown mechanisms. Here we show that astrocytes in the mouse caudal medullary brainstem can synthesize, store, and release D-serine, an agonist for the glycine-binding site of the NMDAR, in response to elevated CO₂levels. We show that systemic and raphe nucleus D-serine administration to awake, unrestrained mice increase the respiratory frequency (fR). Application of D-serine to brainstem slices also increases fR, which was prevented by NMDAR blockade. Inhibition of D-serine synthesis, enzymatic degradation of D-serine, or the sodium fluoroacetate-induced impairment of astrocyte functions decrease the basal fR and the CO₂-induced respiratory response <i>in vivo</i> and <i>in vitro</i>. Our findings suggest that astrocytic release of D-serine may account for the glutamatergic contribution to central chemoreception.</p><div><br></div><div><br></div></div

In vitro approach to the chemical drive of breathing

Since its introduction two decades ago, the isolated brain stem-spinal cord preparation of neonatal rodents has been the preferred method used to reveal the mystery underlying the genesis of the respiratory rhythm. Little research using this in vitro approach has focused on the study of the central respiratory chemosensitivity. Some unexpected findings obtained with the brain stem-spinal cord preparation have added new questions that challenge our previous theoretic framework. Some of these findings are addressed here

Optical analysis of circuitry for respiratory rhythm in isolated brainstem of foetal mice

Respiratory rhythms arise from neurons situated in the ventral medulla. We are investigating their spatial and functional relationships optically by measuring changes in intracellular calcium using the fluorescent, calcium-sensitive dye Oregon Green 488 BAPTA-1 AM while simultaneously recording the regular firing of motoneurons in the phrenic nerve in isolated brainstem/spinal cord preparations of E17 to E19 mice. Responses of identified cells are associated breath by breath with inspiratory and expiratory phases of respiration and depend on CO2 and pH levels. Optical methods including two-photon microscopy are being developed together with computational analyses. Analysis of the spatial pattern of neuronal activity associated with respiratory rhythm, including cross-correlation analysis, reveals a network distributed in the ventral medulla with intermingling of neurons that are active during separate phases of the rhythm. Our experiments, aimed at testing whether initiation of the respiratory rhythm depends on pacemaker neurons, on networks or a combination of both, suggest an important role for networks