Inhibition of glutamatergic interneurons of the lateral parafacial region of mice attenuates the ventilatory response to hypercapnia

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The lateral parafacial region (pFL) generates the expiratory motor activity during hypercapnia. It was demonstrated that this region contains excitatory neurons and interneurons (with local projections), with a population of glutamatergic neurons that are silent under resting conditions, but become active at the end of expiration, simultaneously with active expiration, in response to hypercapnia/acidosis. However, the role of pFL glutamatergic interneurons in regulating pulmonary ventilation during hypercapnia is unclear. Herein, we hypothesized that the inhibition of pFL glutamatergic interneurons attenuates the ventilatory responses to hypercapnia. Therefore, we evaluated the effects of the chemogenetic inhibition of pFL glutamatergic interneurons of non-anesthetized mice on ventilatory parameters during resting and hypercapnia. We used genetically modified mice expressing the enzyme cre recombinase in glutamatergic VGLUT2-positive neurons (CEUA 1143/2022), and injected unilaterally in the pFL region a cre-dependent adeno-associated retrograde virus to express the FLP recombinase, followed by a cre- and FLP-dependent virus to express the inhibitory Designer Receptors Exclusively Activated by Designer Drugs (hM4D-Gi) only in pFL glutamatergic interneurons. Using whole-body plethysmography, we measured respiratory frequency (fR), tidal volume (VT), pulmonary ventilation (VE), duration of respiratory cycle, inspiration (DI) and expiration (DE) at normocapnia and hypercapnia (7% CO2) after activating the hM4D-Gi in the pFL glutamatergic interneurons with JHU 37160 (0.1 mg/kg; i.p.). Data are expressed as mean ± standard deviation. By analyzing the expression of mCherry fluorescent protein, we found a significant number of glutamatergic interneurons in the pFL region (68.22 ± 11.7; n=18). At normocapnia, JHU injection did not change the ventilatory parameters of VGLUT2(cre/cre + hM4D-Gi) (n=18) and VGLUT2(cre/cre) (n=12) mice [(ΔfR: -2.4±51 vs 0.2±48 cpm; p=0.88) (ΔVT: -1.3±4 vs -0.2±1 μL·g−1; p=0.39) (ΔVE: -39 ±1198 vs -71±607 μL·g−1·min−1; p=0.72) (ΔCycle duration: 5.3±41 vs -1.9±45 ms; p=0.65) (ΔDI: 6.6±14 vs 6±20 ms; p=0.92) (ΔDE: -1.3±35 vs -1.6±31 ms; p=0.97)]. On the other hand, JHU injection attenuated the hypercapnia-induced ventilatory responses of VGLUT2(cre/cre + hM4D-Gi) (n=18) mice when compared to VGLUT2(cre/cre) (n=9) [(ΔfR: 67±69 vs 157±41 cpm; p=0.0003) (ΔVT: 3.8±5 vs 11.9±4 μL·g−1; p<0.0001) (ΔVE: 2052±2197 vs 5374±1773 μL·g−1·min−1; p<0.0001) (Δcycle duration: -46±50 vs -107±49 ms; p=0.002) (ΔDI: -3.7±16 vs -21±19 ms; p=0.011) (ΔDE: -42±41 vs -86±36 ms; p=0.01)]. In conclusion, our data demonstrate that the pFL glutamatergic interneurons are important for the ventilatory response of mice to hypercapnia. FAPESP, CNPq and CAPES. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.