BEHAVIORAL AND NEUROBIOLOGIC EFFECTS OF NEONATAL PAIN

描述：(根据申请者描述改编)新生儿重症监护室 CARE每年将26万名在美国出生的早产儿暴露在 长期疼痛、压力和不适。早产儿的随访研究 新生儿表现出严重的行为障碍，痛阈值改变，注意力集中 缺陷障碍、严重焦虑、高躯体化分数和学习 赤字。研究人员假设，延长新生儿疼痛和 应激可能导致神经元和神经递质发育的改变 这会导致这些行为障碍。 研究人员建议利用新生儿建立临床疼痛模型。 ，并记录它们的发声和疼痛相关行为 不同的出生后年龄对阵发性或重复性疼痛的反应。 不同年龄段疼痛刺激引起的神经元激活将通过 C-Fos作为跨突触标志物的原位表达 杂交(ISH)和免疫细胞化学(ICC)。C-Fos基因的共表达 而与神经元可塑性相关的基因(GAP43和HSP-70)将有所帮助 探讨痛觉刺激与触觉刺激对儿童生长发育的影响 感觉皮质、海马体、丘脑和下丘脑。核酸酶 保护性分析将用于基因表达的初步估计， 然后用ISH和ICC检测细胞细节和共表达。 炎性或热痛的不同影响，增加的影响 疼痛强度和反复疼痛将在随后的实验中进行测试。 在儿童后期，早产儿表现出与疼痛相关的改变 行为、严重焦虑和学习缺陷。因此，小鼠受到了 反复的疼痛将被抚养到成年期，并由防御者进行测试 戒断试验、喷气惊吓反应、莫里斯游泳迷宫试验、 社会歧视测试和酒精偏好测试。 下丘脑-垂体-肾上腺轴的反应和痛阈值 在基线和约束应力后测量(热板试验)以检查 疼痛行为和应激诱导的镇痛。成人行为反应 将与c-Fos、GAP-43的表达和细胞密度相关 U-S和K-阿片受体在上述区域。健壮 行为模式和神经元基因表达的相关性将 产生机械性假说以研究神经生物学基础 新生儿疼痛/应激后的行为。这些研究可能具有重要的意义 对大量人口的行为和医疗保健问题的影响 前早产儿即将进入青春期和成年期 十年。 压力神经生物学实验室将对阿南德博士进行技术培训 分子神经生物学和帮助进行这些实验。主动型 研究包括：HPA轴的中央调节， HPA反应性、下丘脑控制的早期经验 免疫-内分泌相互作用与神经内分泌功能。这些研究 使用各种行为、神经内分泌和分子技术 在基础研究方面有良好记录的科学家的方向 以及校外资助。

DESCRIPTION: (Adapted from the Applicant's Description) Neonatal Intensive Care exposes 260,000 preterm neonates, born in the U.S. each year, to prolonged pain, stress and discomfort. Follow-up studies of ex-premature neonates show major behavioral disorders, altered pain thresholds, attention deficit disorder, severe anxiety, high somatization scores, and learning deficits. The investigators hypothesize that prolonged neonatal pain and stress may lead to changes in neuronal and neurotransmitter development which result in these behavioral disorders. The investigators propose to develop models of clinical pain using newborn rat pups, and record their vocalizations and pain-related behaviors at different postnatal ages in response to episodic or repetitive pain. Neuronal activation from painful stimuli at different ages will be mapped by the expression of c-Fos as a trans-synaptic marker using in situ hybridization (ISH) and immunocytochemistry (ICC). Co-expression of c-Fos and genes associated with neuronal plasticity (GAP43 and HSP-70) will help to probe the effects of painful versus tactile stimulation on development in the sensory cortex, hippocampus, thalamus and hypothalamus. RNAse protection assays will be used for initial estimation of gene expression, followed by ISH and ICC to examine cellular detail and co-expression. Differential effects of inflammatory or thermal pain, effects of increasing pain intensity and repetitive pain will be tested in subsequent experiments. In later childhood, ex-preterm neonates manifest altered pain-related behaviors, severe anxiety, and learning deficits. Thus, rat pups subjected to repetitive pain will be reared to adulthood and tested by the defensive withdrawal test, air-puff startle response, Morris swim maze test, the social discrimination test, and the alcohol preference test. Hypothalamic-pituitary-adrenal axis responses and pain thresholds will be measured (hot plate test) at baseline and after restraint stress to examine pain behaviors and stress-induced analgesia. Adult behavioral responses will be correlated with the expression of c-Fos, GAP-43, and the density of u-s- and K-opioid receptors in the areas outlined above. Robust correlations between behavior patterns and neuronal gene expression will generate mechanistic hypotheses to investigate the neurobiologic basis for behavior following neonatal pain/stress. These studies may have important implications for the behavior and the health-care problems of large numbers of ex-premature infants approaching adolescence and adulthood in the next decade. The Stress Neurobiology Laboratory will train Dr. Anand in the techniques of molecular neurobiology and help in performing these experiments. Active investigations include: central regulation of the HPA axis, effects of early experience on HPA responsiveness, hypothalamic control of immune-endocrine interactions and neuroendocrine function. These studies use a variety of behavioral, neuroendocrine, and molecular techniques under the direction of scientists with established track-records in basic research and extra-mural funding.