Cardiovascular responses to microinjections of GABA or anesthetics into the rostral ventrolateral medulla of conscious and anesthetized rats

The rostral ventrolateral medulla (RVLM) contains neurons involved in tonic and reflex control of arterial pressure. We describe the effects of gamma-aminobutyric acid (GABA) and anesthetics injected into the RVLM of conscious and urethane (1.2 g/kg, iv) anesthetized Wistar rats (300-350 g). In conscious rats, bilateral microinjection of GABA (50 nmol/200 nl) induced a small but significant decrease in blood pressure (from 130 ± 3.6 to 110 ± 5.6 mmHg, N = 7). A similar response was observed with sodium pentobarbital microinjection (24 nmol/200 nl). However, in the same animals, the fall in blood pressure induced by GABA (from 121 ± 8.9 to 76 ± 8.8 mmHg, N = 7) or pentobarbital (from 118 ± 4.5 to 57 ± 11.3 mmHg, N = 6) was significantly increased after urethane anesthesia. In contrast, there was no difference between conscious (from 117 ± 4.1 to 92 ± 5.9 mmHg, N = 7) and anesthetized rats (from 123 ± 6.9 to 87 ± 8.7 mmHg, N = 7) when lidocaine (34 nmol/200 nl) was microinjected into the RVLM. The heart rate variations were not consistent and only eventually reached significance in conscious or anesthetized rats. The right position of pipettes was confirmed by histology and glutamate microinjection into the RVLM. These findings suggest that in conscious animals the RVLM, in association with the other sympathetic premotor neurons, is responsible for the maintenance of sympathetic vasomotor tone during bilateral RVLM inhibition. Activity of one or more of these premotor neurons outside the RVLM can compensate for the effects of RVLM inhibition. In addition, the effects of lidocaine suggest that fibers passing through the RVLM are involved in the maintenance of blood pressure in conscious animals during RVLM inhibition

Evidence for neuronal release of isotopically labelled glycine from the rat ventral medullary surface in vivo

Spontaneous and stimulus-induced release of isotopically labelled glycine was studied in the superfused rat dorsal or ventral medullary surface in vivo. Superfusion of the ventral medullary surface of anesthetized (urethane, 1.2 g/kg, ip) male adult Wistar rats (250-350 g) with high K+ (40 mM) surrogate cerebrospinal fluid (CSF) produced an average increase of 45 per cent over the spontaneous efflux of exogenously applied glycine (N = 5, P<0.01). In experiments in which the calcium of the CSF was replaced by an equimolar amount of magnesium, the increase in glycine efflux in response to high K+ was reduced to 15 per cent, a value not statistically different from that observed in control experiments (N = 6). Veratridine stimulation evoked a large (80 per cent) increase in glycine efflux (N = 5, P<0.001), which was inhibited by tetrodotoxin. High potassium or veratridine failed to modify spontaneous release of glycine on the dorsal medullary surface. Results obtained in control experiments showed that neither high K+ nor veratridine is effective in modifying spontaneous efflux of [(3)H]-leucine or [(3)H]-inulin on the ventral or dorsal medullary surface. These data support the hypothesis that glycine is a neurotransmitter on the ventral medullary surface and that it may be part of neural pathways involved in cardiorespiratory regulation present in this region.

Putative pathways involved in cardiovascular responses evoked from the caudal pressor area

1. The caudal pressor area (CPA) is a recently identified site within the ventrolateral medulla which is involved in cardiovascular regulation. CPA chemical stimulation by L-glutamate produces an increase in arterial blood pressure (ABP) while its inhibition by GABA or glycine evokes marked hypotension. In the present study, we sought to determine the potential neural pathways underlyng these responses. 2. In urethane-anesthetized, paralyzed, artificially ventilated rats, CPA inhibition by bilateral microinjection of the inhibitory amino acid glycine (Gly, 100 nmol 200 nl-1 site-1) produced an average decrease of -38 + or - 4.3 mmHg in ABP (n = 6). Ten min after bilateral microinjection of the broad-spectrum glutamate antagonist kynurenic acid (KYN, 2 nmol 200 nl-1 site-1) into the cauldal ventrolateral medulla (CVLM) depressor responses to CPA inhibition were virtually abolished (-3 + or - 1.7 mmHg, P<0.05). Similar microinjection of KYN into the rostral ventrolateral medulla (RVLM) or into the CPA itself did not modify depressor responses to CPA inhibiton by glycine. 3. CPA stimulation by bilateral microinjection of the excitatory amino acid L-glutamate (L-glu, 50 nmol 200 nl-1 site-1) produced an increase in ABP (+43 + or - 5.4 mmHg, N= 6). Bilateral microinjection of the GABA A antagonist bicuculline methiodide (BIC, 200 pmol 200 nl-1 site-1) into the CVLM markedly reduced pressor responses to CPA stimulation (+6 + or - 2.7 mmHg, P<0.05). Similar application of BIC into the RVLM or CPA did not modify pressor responses to CPA stimulation by glutamic acid

Cardiorespiratory alterations produced by pharmacological agents applied to the ventrolateral medulla (intermediate area) of the cat: effect of exercise condition on glycine-induced inhibition

1. To study the action of the intermediate area (IA), coextensive with the rostral ventrolateral medulla, on the neurophysiological mechanisms involved in the regulation of respiration, in terms of inspiratory drive and respiratory timing, cats were submitted to topical application of sodium pentobarbital (30 mg/ml), leptazol (200 mg/ml), glutamate (50 mg/ml) and glycine (100 and 50 mg/ml) to the IA. The effects of electrically induced exercise on the ventilatory response and oxygen uptake (VO2) obtained by topical application of glycine (50 mg/ml) to the IA were also studied. 2. Leptazol reduced minute ventilation (VE) and inspiratory drive (VT/TI) and changed the timing mechanism. Glutamate only increased tidal volume (VT), VE and VT/TI. Arterial blood pressure (AP) increased and heart rate (HR) did not change with either drug. 3. Sodium pentobarbital reduced VT and changed the timing mechanism. Glycine only reduced VE, VT and VT/TI. AP decreased and HR did not change with either drug. 4. The depressor effects of glycine on respiratory pattern, VO2 and CO2 production (VCO2) tended to be attenuated by exercise. 5. The fall in AP due to glycine application did not differ between resting and exercise conditions. 6. Our results indicate that at least two different nervous structures are involved in the IA: one responsible for the respiratory drive and sensitive to glycine and glutamate, and the other responsible for the regulation of the timing mechanism and sensitive to sodium pentobarbital and leptazol

Role of the rostral ventrolateral medulla in the pressor response to stimulation of the nucleus raphe obscurus

1. Electrical stimulation of the nucleus raphe obscurus (NRO) in urethane-anesthetized rats increases arterial blood pressure (BP) between 20 and 95 mmHg (mean, 61.14 +/- 6.57; N = 30). 2. Unilateral electrolytic destruction of the rostral ventrolateral medulla (RVLM) did not reduce BP or heart rate (HR) but significantly reduced the pressor response to NRO stimulation (control, delta 76.0 +/- 5.4 mmHg; after lesion, delta 26.0 +/- 13.9 mmHg; P < 0.01, N = 5). 3. Bilateral destruction of the RVLM reduced basal BP (control, 104.1 +/- 11.4 mmHg; after lesion, 58.0 +/- 5.7 mmHg; P < 0.01) and the pressor response to NRO stimulation (control, delta 71.6 +/- 7.3; after lesion, delta 12.5 +/- 3.8 mmHg; P < 0.01, N = 6). 4. When topically applied to or microinjected into the RVLM, pentobarbital sodium (200 nl/1 microliters, 10 nmol) decreased BP, HR and the pressor response to NRO stimulation (control, delta 56.2 +/- 6.7 mmHg; after pentobarbital, delta 11.2 +/- 3.1 mmHg; P < 0.01, N = 13). Similar effects were obtained when glycine (200 nl, 50 nmol) was microinjected into RVLM (control, delta 40.5 +/- 5.9 mmHg; after glycine, delta 18.1 +/- 4.9 mmHg; P < 0.01, N = 6). 5. We conclude that RVLM is essential for the pressor response to NRO stimulation

Breathing pattern during pharmacological activation and blockade of the intermediate area of the ventrolateral surface of the cat medulla

The present study analyzes the respiratory pattern of chloralose- (50-60 mg/kg,iv) anesthetized cats treated with Nembutal (NE) (30 mg/ml), glycine (GL) (200 mg/ml) or leptazol (LE) (200 mg/ml) topically applied to the intermediate area of the ventrolateral surface of the medulla oblongata in a volume of 20 micronl. Application of NE and GL produced a decrease in ventilation (-24%) and tidal volume (-25%) suggesting that the intermediate area facilitates respiratory drive and inhibits the inspiratory off-switch mechanism. These results are consistent with the view that intermediate area is necessary for the central chemosensitivity to CO2. The topical application of LE produced an increase in inspiration time (12.5%), expiration time (20.8%) and tidal volume (7%). The increased tidal volume caused by LE is compatible with it action as a GL antagonist