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Volumn 289, Issue 5479, 2000, Pages 628-630

Altered nociceptive neuronal circuits after neonatal peripheral inflammation

Author keywords

[No Author keywords available]

Indexed keywords

ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ARTICLE; CONTROLLED STUDY; EMBRYO DEVELOPMENT; HEALTH HAZARD; INFLAMMATION; NERVE CELL DIFFERENTIATION; NEWBORN; NOCICEPTION; NONHUMAN; PERINATAL DEVELOPMENT; PRIORITY JOURNAL; RAT; SENSORY NERVE; SENSORY STIMULATION; SPINAL CORD NERVE CELL; TISSUE INJURY;

EID: 0034725740     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.289.5479.628     Document Type: Article
Times cited : (362)

References (21)
  • 4
    • 0005456889 scopus 로고
    • K. J. S. Anand and P. J. McGrath, Eds. Elsevier, New York
    • K. J. S. Anand and P. J. McGrath, in Pain in Neonates, K. J. S. Anand and P. J. McGrath, Eds. (Elsevier, New York, 1993), pp. 1-18.
    • (1993) Pain in Neonates , pp. 1-18
    • Anand, K.J.S.1    McGrath, P.J.2
  • 7
    • 0343842803 scopus 로고    scopus 로고
    • note
    • Sprague-Dawley male rat pups received a single injection of CFA (2: 1, CFA:saline) or saline on either PO, P1, P3, or P14 (25 μl in P0, P1, and P3 rat pups and 50 μl in P14 pups) into the left hind paw, or they were untreated. The animals matured undisturbed. Procedures were approved by the National Institute of Dental and Craniofacial Research (NIDCR) Animal Care and Use Committee.
  • 9
    • 0343842801 scopus 로고    scopus 로고
    • note
    • Adult animals (8 to 12 weeks) were anesthetized with sodium pentobarbital, the sciatic nerves were exposed at midthigh, and WGA-HRP (250 μg) or β-HRP (30 μg) was microinjected bilaterally [C. C. LaMotte, S. E. Kapadia, C. M. Shapiro, J. Comp. Neurol. 311, 546 (1991)]. Forty-eight hours later, they were anesthetized and perfused transcardially with 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4). The spinal cord was removed, postfixed, and transferred to 30% sucrose. Tissue was cut into 30-μm sections on a cryostat and processed with the tetramethylbenzidine method [M. M. Mesulam, J. Histochem. Cytochem. 26, 106 (1978)]. Evaluation of labeling was performed blind. Sections were photographed onto Ektachrome 64T transparency film and scanned into Adobe Photoshop 5.0.
  • 10
    • 0343842802 scopus 로고    scopus 로고
    • Web fig. 1 is available at www.sciencemag.org/ feature/data/1051450.shl.
  • 11
    • 0342537334 scopus 로고    scopus 로고
    • note
    • Tissue sections from the L4-5, L5-6, and L6-S1 spinal segments, six sections each, from neonatal treated (n = 4) and untreated rats (n = 2) were digitized (9) and analyzed with NIH image 1.62. The relative density of labeling was measured in a 300 μm by 200 μm rectangle placed over the dorsal horn beginning at the dorsal root entry zone and continuing medially. The most caudal sections were not included in the analysis as the contralateral dorsal horn was devoid of labeling. To normalize the data, we measured background in lamina VII and subtracted it from the dorsal horn values. Relative density was graphed as the percentage of increase with the use of the formula left minus right side density divided by the right side density multiplied by 100. Data were analyzed with repeated measures analysis of variance (ANOVA).
  • 12
    • 0342971596 scopus 로고    scopus 로고
    • note
    • At 8 weeks of age, neonatal CFA-treated rats (n = 3) were perfused and tissue sectioned (9). Adjacent sections were incubated in a rabbit polyclonal antibody to CGRP (1:200,000) or in biotinylated 1B4 lectin (1 μg/ml). Labeling was visualized with standard avidin-biotin methodology with 0.05% 3,3′-diaminobenizidine tetrahydrochloride as the chromagen.
  • 14
    • 0023924046 scopus 로고    scopus 로고
    • note
    • At 8 to 10 weeks of age, baseline paw withdrawal latency of the left and right hind paw to a radiant heat source [K. Hargreaves, R. Dubner, F. Brown, C. Flores, J. Joris, Pain 32, 77 (1988)] was determined in neonatal treated (n = 9) and untreated (n = 9) rats. Withdrawal latency was tested again 24 hours after an injection of 200 μl of CFA (1:1) into the adult rat left hind paw for five times at intervals of 5 min. Latency was calculated as the mean excluding the first, familiarization trial and presented as means ± SEM. Significance was determined with ANOVA and a t test.
  • 15
    • 0342971595 scopus 로고    scopus 로고
    • note
    • Pain behavior was assessed in adult neonatal treated (n = 10) and untreated (n = 7) rats after intraplantar injection of 1.5% formalin (50 μl) in the left hind paw. Weighted pain scores, representing the time spent licking, lifting, and favoring the injected paw, were quantified every 5 min for 60 min after injection (8). Data were analyzed by repeated measures ANOVA and Tukey post hoc tests. To quantify the shift in the time course of the response, we calculated the median of the late phase of pain behavior [the midpoint of the distribution from the local minimum (interphase) to the terminal observation interval] for each rat and subjected it to ANOVA.
  • 16
    • 0343842800 scopus 로고    scopus 로고
    • note
    • Dorsal horn extracellular single-unit discharges were recorded in the lumbar enlargement of adult neonatal treated (n = 5; 120 neurons) and untreated (n = 5; 147 neurons) rats anesthetized with sodium pentobarbital, immobilized with pancuronium bromide, and mechanically ventilated. Spontaneous activity and activity evoked by nonnoxious brushing with a camel's hair brush and noxious pinch applied with an arterial clamp were assessed. Means of three stimulations (applied for 10 s at 10-s intervals) were calculated with Spike2 software. Data were analyzed by repeated measures ANOVA and Tukey post hoc tests.


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