Activation of ventrolateral preoptic neurons during sleep

Science

Volumn 271, Issue 5246, 1996, Pages 216-219

  Sherin, J E ^a,b,c   Shiromani, P J ^b   McCarley, R W ^b   Saper, C B ^a,b  

^a BETH ISRAEL DEACONESS MEDICAL CENTER   (United States)

^b HARVARD MEDICAL SCHOOL   (United States)

^c UNIVERSITY OF CHICAGO   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CHOLERA TOXIN B SUBUNIT; TRACER; CHOLERA TOXIN; PROTEIN C FOS;

ANIMAL EXPERIMENT; ANIMAL TISSUE; ANTERIOR HYPOTHALAMUS; AROUSAL; ARTICLE; FOREBRAIN; IMMEDIATE EARLY GENE; IMMUNOCYTOCHEMISTRY; INNERVATION; NERVE FUNCTION; NONHUMAN; PREOPTIC AREA; PRIORITY JOURNAL; RAT; SLEEP; SLEEP WAKING CYCLE; THALAMUS VENTRAL NUCLEUS; ANIMAL; BRAIN LEVEL; CHEMISTRY; CIRCADIAN RHYTHM; CYTOLOGY; IMMUNOHISTOCHEMISTRY; MAMMILLARY BODY; NERVE CELL; NERVE TRACT; PHYSIOLOGY;

ANIMALS; BRAIN CHEMISTRY; CHOLERA TOXIN; CIRCADIAN RHYTHM; IMMUNOHISTOCHEMISTRY; MAMILLARY BODIES; NEURAL PATHWAYS; NEURONS; PREOPTIC AREA; PROTO-ONCOGENE PROTEINS C-FOS; RATS; SLEEP;

ANIMALIA;

EID: 0029671044     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.271.5246.216     Document Type: Article

References (64)

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28. In anesthetized rats (acepromazine, 0.75 mg per kilogram of body weight; ketamine, 44.0 mg/kg; xylazine, 2.5 mg/kg), screw electrodes were implanted in the skull to record the electroencephalogram (EEG), and flexible wires were placed in the nuchal muscles to record the electromyogram (EMG). After attachment to Amphenol connectors, the apparatus was fixed in place with dental cement. Rats were housed individually at 24°C with food and water provided ad libitum and lights on from 07:00 to 19:00 (light cycle) and off from 19:00 to 07:00 (dark cycle). One week after surgery, rats were connected by means of flexible wires to a Grass electroencephalograph (model 79) for recording EEG and EMG. Rats were adapted to the setup for 3 days dunng which sample sleep recordings were obtained. Experimental sessions consisted of connecting animals for recording (as dunng adaptation of three previous days) until they were killed.
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37. Because the IGL, SCNd, and VLPO contained only a few FOS-ir neurons in dark cycle (waking) animals, IB increased staining in light cycle (sleeping) animals in these regions was quite conspicuous Although FOS staining generally decreased during the light cycle in the rest of the brain, some areas, including the cingulate cortex, pinform cortex, paraventricular thalamus, and medial preoptic area, contained FOS-ir cells under all conditions We focused on the SCNd, IGL, and VLPO because they were the only cell groups with a clear-cut increase in FOS staining during the light cycle. We cannot, however, eliminate the possibility that neurons scattered in other cell groups accumulate FOS during the light cycle.
38. The VLPO was demarcated for FOS quantification in a coronal plane approximating that in the Paxmos rat brain atlas. Beginning at the rostral pole of the supraoptic and suprachiasmatic nuclei and continuing rostrally, six 40-μm sections were analyzed bilaterally from a one in two series, such that 12 VLPO sectors were counted and averaged for each brain. Each VLPO sector was demarcated by the base of the brain and extended dorsally 0.3 mm and laterally from 1.0 mm lateral to the midline to the edge of the olfactory tubercle (box in Rg. 1A) Behaviors for the hour before the rat was killed (when FOS protein approaches maximal levels in activated neurons [P. I J. Shiromam et al., Mol. Brain Res. 29, 163 (1995)]} were scored manually by EEG and EMG criteria as percentages of waking, synchronized, and desynchronized sleep. The percentage of time spent in synchronized and desynchronized sleep was combined to obtain percent total sleep time values.
39. Mean (± SD) of percent total sleep time for animals in this study was 63 ± 22 for rats killed at 10:00, 67 ± 24 for rats killed at 16 00, and 16 ± 12 for rats killed at 22:00; these values are within published norms [Z. J. M. Van Hulzen and H. M. L. Coenen, Physiol Behav. 25, 807 (1980)]
40. Pearson correlation coefficients were used to determine the significance of the relation between the number of FOS-ir cells in the VLPO and percent total sleep time [G. W. Snedecor and W. G. Cochran, Statistical Methods (Iowa State Univ. Press, Ames, IA, 1989), pp. 177-195
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53. By means of an air-pressure delivery system, 1% CTB dissolved in 0 9% saline (≃1 nl) was injected stereotaxically through a glass micropipette into the TMN region of chloral hydrate-anesthetized rats (350 mg/ kg) Coordinates from the Paxinos rat atlas were: AP, -4.0 mm (from bregma); DV, -9.0 mm (from cortical surface); and RL, +1 5 mm (from midline).
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64. We thank Q -H. Ha and M. Magner for technical assistance, R. Burstein, J. Elmquist, T. Scammel, M. Greenberg, and J. A. Hobson for helpful discussions, and R. Kellems for ADA antiserum. Supported by USPHS grants NS22835, MH10709, and NS30140 and by a Grant-in-aid from the American Heart Association (94013110) and the Department of Veterans Affairs Medical Research Service.