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After placement in the stereotaxic holder and suspension by thoracic and lumbar vertebral clamps, the head of the animal was ventroflexed, the dorsal medulla was exposed by occipital craniotomy, the dura and arachnoid membranes were reflected, and a patch of pia membrane was removed to allow insertion of a glass microelectrode. National Institutes of Health, and approved by the University of Wisconsin Medical School’s Institutional Animal Care and Use Committee. Care and use of animals were in accordance with the Guide for the Care and Use of Laboratory Animals, as adopted and promulgated by the U.S. In experiments where phrenic nerve activity and discharge properties of medullary expiratory neurons were recorded, animals were bilaterally vagotomized and paralyzed with gallamine triethiodide (4 mg/kg iv to start, 4-8 mg/h thereafter) and mechanically ventilated with oxygen-enriched (60% O2) room air. Buy nembutal online in tracheal pressure were measured by connecting the tracheal cannula to a T tube, with one side connected to a pressure transducer and the other open to room air through a variable resistor. This po st w᠎as created by GSA  Con᠎tent Generator D em oversion!

Samples were washed, air dried, and exposed to BioMax MR film for 3-5 days. Surgical procedures involved placement of a catheter in the femoral artery to monitor blood pressure and obtain samples for measurement of blood oxygen and hemoglobin concentration, and catheters in both femoral veins to administer drugs and infuse Ringer lactate solution to maintain tissue hydration. Blood pressure, tracheal pressure, rate of breathing, body temperature, end-tidal CO2 and inspired oxygen were monitored continuously and recorded on chart paper. Anesthesia was allowed to lighten enough during the testing period so that under control conditions, a brief squeeze of the hind paw with a padded hemostat, just firm enough to cause ipsilateral hind limb withdrawal without altering blood pressure, heart rate or breathing, evoked a contralateral EMG discharge recorded with an AC preamplifier (2000 × amplification, 100-3000 Hz bandwith). Supplemental doses of pentobarbital (4-8 mg/kg iv) were administered if symptoms occurred that indicated significant lightening of anesthesia: 1) spontaneous increases of arterial blood pressure and heart rate, 2) irregular breathing or discharges of phrenic nerve activity that decreased in duration and increased in frequency, 3) shivering, and 4) movement and cardio-respiratory changes evoked by surgical procedures. In the second paradigm, single electrical shocks were applied to evoke crossed-extensor reflex discharges and measure the discharge latencies.

One stimulating needle, the cathode, was inserted under the Achilles tendon of the left hindlimb, and another needle, the anode, was inserted under the skin on the medial side of the left hind paw to stimulate the tibial nerve at the ankle and activate the reflex (30). Single shocks (70-90 V, 3-ms pulse duration) that evoked short- and long-latency contralateral reflex discharges of maximal intensity in the contralateral vastus lateralis muscle were delivered every 10 s. Phrenic nerve activity and expiratory neuron membrane potential were displayed on an oscilloscope and registered on magnetic tape (DC, 5 KHz), computer (DC, 10 KHz), and chart recorder (DC, 10 KHz). 8 Eldridge FL. Relationship between phrenic nerve activity and ventilation. The effects of opiates on the respiratory activity of thoracic motoneurons in the anesthetized and decerebrate rabbit. 5 Bedford RF and Wollman H. Postoperative respiratory effects of morphine and halothane anesthesia: a study in patients undergoing cardiac surgery. The animals were initially anesthetized with 5% halothane in 100% oxygen, administered while the animals were in an anesthesia chamber. Animals were mounted in a stereotaxic head holder and suspended by thoracic and lumbar spinal clamps. 1202032) and NIH guidelines for use of animals were followed.


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