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.2018 Jan 3;4(6):ENEURO.0269-17.2017.
doi: 10.1523/ENEURO.0269-17.2017. eCollection 2017 Nov-Dec.

Homeostatic Changes in GABA and Acetylcholine Muscarinic Receptors on GABAergic Neurons in the Mesencephalic Reticular Formation following Sleep Deprivation

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Homeostatic Changes in GABA and Acetylcholine Muscarinic Receptors on GABAergic Neurons in the Mesencephalic Reticular Formation following Sleep Deprivation

Hanieh Toossi et al. eNeuro..

Abstract

We have examined whether GABAergic neurons in the mesencephalic reticular formation (RFMes), which are believed to inhibit the neurons in the pons that generate paradoxical sleep (PS or REMS), are submitted to homeostatic regulation under conditions of sleep deprivation (SD) by enforced waking during the day in mice. Using immunofluorescence, we investigated first, by staining for c-Fos, whether GABAergic RFMes neurons are active during SD and then, by staining for receptors, whether their activity is associated with homeostatic changes in GABAA or acetylcholine muscarinic type 2 (AChM2) receptors (Rs), which evoke inhibition. We found that a significantly greater proportion of the GABAergic neurons were positively stained for c-Fos after SD (∼27%) as compared to sleep control (SC; ∼1%) and sleep recovery (SR; ∼6%), suggesting that they were more active during waking with SD and less active or inactive during sleep with SC and SR. The density of GABAARs and AChM2Rs on the plasma membrane of the GABAergic neurons was significantly increased after SD and restored to control levels after SR. We conclude that the density of these receptors is increased on RFMes GABAergic neurons during presumed enhanced activity with SD and is restored to control levels during presumed lesser or inactivity with SR. Such increases in GABAAR and AChM2R with sleep deficits would be associated with increased susceptibility of the wake-active GABAergic neurons to inhibition from GABAergic and cholinergic sleep-active neurons and to thus permitting the onset of sleep and PS with muscle atonia.

Keywords: AChM2; GABAA; homeostasis; mice; waking.

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Figures

Figure 1.
Figure 1.
Atlas figure and distribution map of GABAergic neurons expressing c-Fos, GABAARs or AChM2Rs within the RFMes.A, Atlas figure of the pontomesencephalic tegmentum showing the traced contours around the RFMes region in three adjacent sections separated by 100-μm intervals, which were collapsed onto the middle section of this figure (bregma −4.24 mm; Paxinos and Franklin, 2001).B, Distribution map of the GAD+ (blue circle) and c-Fos+/GAD+ (red filled blue circle) neurons in an SC mouse brain (B1), an SD mouse brain (B2), and an SR mouse brain (B3). Note the greater number of c-Fos+/GAD+ neurons in the SD mouse.C, Distribution map of the GABA+ (blue circle) and GABAAR+/GABA+ (red filled blue circle) neurons in an SC mouse brain (C1), an SD mouse brain (C2), and an SR mouse brain (C3). Note the slightly higher number of GABAAR+/GABA+ neurons in the SD mouse.D, Distribution map of the GAD+ (blue circle) and AChM2R+/GAD+ (red filled blue circle) neurons in an SC mouse brain (D1), an SD mouse brain (D2), and an SR mouse brain (D3). Note the greater number of AChM2R+/GAD+ neurons in the SD mouse. Scale bar: 1 mm. 3N, oculomotor nucleus; Aq, aqueduct (Sylvius); DR. dorsal raphe nucleus; mlf, medial longitudinal fasciculus; x, decussation of the superior cerebellar peduncle.
Figure 2.
Figure 2.
Sleep-wake states, c-Fos, GABAA, and AChM2 receptors in RFMes GABAergic neurons across groups.A, The percentage of time spent in wake during the 2 h preceding termination differed significantly across groups, being higher in SD as compared to SC and SR and lower in SR as compared to SC.B, The % of GAD+ neurons that were positively immunostained for c-Fos (+) differed significantly between groups, being greater in SD as compared to SC and SR.C, The % of GABA+ neurons which were positively immunostained for the GABAAR (+) increased insignificantly following SD as compared to SC and SR (C1). The luminance of the GABAAR immunofluorescence on GABAAR+/GABA+ neurons differed significantly, being higher in SD as compared to SC (C2).D, The % of GAD+ neurons which were positively immunostained for AChM2R (+) differed significantly between groups, being higher in SD as compared to SC and SR (D1). The luminance of the AChM2R immunofluorescence on AChM2R+/GAD+ neurons differed significantly, being higher in SD as compared to SC and SR (D2). Note that the changes in GABARs and AChM2Rs on RFMes GABAergic neurons parallel the changes in % Wake and % c-Fos+/GAD+ across groups; * indicates significant difference of SD relative to SC; † indicates significant difference of SD relative to SR; § indicates significant difference of SR relative to SC (p < 0.05), according topost hoc paired comparisons following one-way ANOVA (Table 1).
Figure 3.
Figure 3.
c-Fos in RFMes GABAergic neurons across groups. Fluorescent microscopic images show staining for Nissl with FNS (green;A1,B1,C1), immunostaining for GAD (blue;A2,B2,C2, with positive staining indicated by filled arrowheads), and immunostaining for c-Fos (red;A3,B3,C3, with positive staining indicated by filled arrowhead) along with dual staining for Nissl and c-Fos in merged images (green and red;A4,B4,C4, with positive c-Fos staining indicated by filled arrowhead). Note that c-Fos immunostaining is prominent in the nucleus of a GABAergic neuron from an SD mouse (B3,B4), whereas it is not apparent in images from SC or SR mice (A3,A4 orC3,C4, indicated by open arrowheads). Scale bars: 20 μm. Image thickness: 500 nm in all panels.
Figure 4.
Figure 4.
GABAARs in RFMes GABAergic neurons across groups. Confocal microscopic images show all neurons stained for Nissl with FNS (green;A1,B1,C1), the GABAergic neurons immunostained for GABA (blue;A2,B2,C2, indicated by filled arrowhead), and for the GABAARs in single (red;A3,B3,C3, indicated by filled arrowhead) and merged images (A4,B4,C4, indicated by filled arrowhead). Note that in an SC mouse, the GABAAR immunofluorescence is minimally visible, whereas in an SD mouse, it is prominent and bright. In an SR mouse, it appeared less bright than in the SD mouse. In all cases, the immunostaining is relatively continuous though with nonuniform intensity along the plasma membrane of the GABA+ neurons. Scale bars: 20 μm. Image thickness: 500 nm in all panels.
Figure 5.
Figure 5.
AChM2Rs in RFMes GABAergic neurons across groups. Confocal microscopic images show all neurons stained for Nissl with FNS (green;A1,B1,C1), the GABAergic neurons immunostained for GAD (blue;A2,B2,C2, indicated by filled arrowhead), and for the AChM2Rs in single (red;A3,B3,C3, indicated by filled arrowhead), and merged images (A4,B4,C4, indicated by filled arrowhead). Note that in an SC mouse, the AChM2R immunofluorescence is minimally visible along the plasma membrane, whereas in an SD mouse, the AChM2R staining is bright and clearly visible along the full membrane of the GAD+ neuron. In an SR mouse, the staining appeared less bright than in the SD mouse. Scale bars: 20 μm. Image thickness: 500 nm in all panels.
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References

    1. Aton SJ, Seibt J, Dumoulin M, Jha SK, Steinmetz N, Coleman T, Naidoo N, Frank MG (2009) Mechanisms of sleep-dependent consolidation of cortical plasticity. Neuron 61:454–466. 10.1016/j.neuron.2009.01.007 - DOI - PMC - PubMed
    1. Baghdoyan HA, Lydic R (1999) M2 muscarinic receptor subtype in the feline medial pontine reticular formation modulates the amount of rapid eye movement sleep. Sleep 22:835–847. - PubMed
    1. Baghdoyan HA, Rodrigo-Angulo ML, McCarley RW, Hobson JA (1987) A neuroanatomical gradient in the pontine tegmentum for the cholinoceptive induction of desynchronized sleep signs. Brain Res 414:245–261. - PubMed
    1. Boissard R, Gervasoni D, Schmidt MH, Barbagli B, Fort P, Luppi PH (2002) The rat ponto-medullary network responsible for paradoxical sleep onset and maintenance: a combined microinjection and functional neuroanatomical study. Eur J Neurosci 16:1959–1973. - PubMed
    1. Borbély AA, Tobler I, Hanagasioglu M (1984) Effect of sleep deprivation on sleep and EEG power spectra in the rat. Behav Brain Res 14:171–182. - PubMed

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