DEVELOPMENTAL NEUROCHEMISTRY OF BUPRENORPHINE

丁丙诺啡的发育神经化学

• 批准号: 2897944
• 负责人: Susan E. Robinson
• 金额: $ 20.42万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2897944
• 关键词: animal developmental psychology; behavioral /social science research tag; buprenorphine; developmental neurobiology; dosage; drug delivery systems; embryo /fetus drug adverse effect; embryo /fetus toxicology; gas chromatography mass spectrometry; laboratory rat; neurochemistry; neurotransmitter metabolism; newborn animals; northern blottings; opioid receptor; prenatal stress; radioimmunoassay

Methadone is used in the management of the pregnant opioid addict. However, methadone maintenance has been associated with neonatal withdrawal as well as with behavioral and neurochemical anomalies in both human and animals. Buprenorphine may be a useful drug in the pregnant addict, as it is not associated with an abstinence syndrome. Furthermore, buprenorphine may be superior to methadone during pregnancy, as it lacks full agonist activity at mu-opioid receptors, and thus may not produce the neurochemical changes observed after prenatal methadone exposure. However, unlike methadone, buprenorphine binds to delta- and kappa-opioid receptors, which may result in a different set of neural and behavioral perturbations in the offspring. This project will study the effects of buprenorphine on the developing organism. Rats will be exposed to various doses of buprenorphine pre- and/or postnatally via maternally-implanted osmotic minipumps. Because the rat central nervous system is relatively immature at birth and continues to develop for several weeks afterwards, rats will be exposed to buprenorphine at different times to determine the effect of the drug at different stages of development. It is important to study multiple doses as buprenorphine has a bell-shaped dose-response curve. Methadone exposed groups will be included as positive controls. Female rats will be implanted with a 28-day osmotic minipump containing water, buprenorphine, or methadone on day 8 of pregnancy. Pups will be cross-fostered to mothers who have been implanted with water or buprenorphine pumps. Levels of buprenorphine and its metabolite will be measured in the brains of fetuses and pups by gas chromatography/mass spectrometry. Effects of perinatal buprenorphine exposure will be determined on behavior, brain regional acetylcholine (ACh), dopamine (DA), norepinephrine (NE), serotonin (5-HT), and their metabolites in neonatal (4 day), weanling (21 day), and adult (90 day) female and male rats. Dose-related effects of perinatal buprenorphine will be studied on brain regional endogenous opioids and their receptors as well as on ACh turnover (TRACh) in weanling and adult rats. Developmental milestones and sexual maturation will be assessed. Antinociception/nociception (tail- flick, hot-plate) in the weanling and adult rats. A gas chromatographic/mass spectrometric method will be used to measure ACh and TRACh. HPLC with electrochemical detection will be used to measure DA, NE, 5-HT, and their metabolites. Beta-Endorphin-like, met- and leu- enkephalin-like, and dynorphin-like immunoreactivity will be measured by radioimmunoassay. The Kd and Bmax for mu, delta, and kappa-opioid receptors will be determined by Scatchard analysis. In the event that changes are noted in opioid receptors, mRNA for the particular receptor will be quantified. In the event that ACh content is reduced by buprenorphine, mRNA for choline acetyltransferase will be quantified by the use of Northern blots. Once the effects of perinatal buprenorphine exposure on neurochemistry and behavior are more completely understood, more efficacious therapeutic management can be developed for pregnant addicts and potentially for prevention of drug abuse in their children.

美沙酮用于管理怀孕的阿片类药物成瘾者。 然而，美沙酮维持治疗与新生儿 以及行为和神经化学异常， 人类和动物。丁丙诺啡可能是一种有用的药物在怀孕 成瘾，因为它与戒断综合征无关。此外，委员会认为， 丁丙诺啡可能是上级美沙酮在怀孕期间，因为它缺乏 μ-阿片受体的完全激动剂活性，因此可能不会产生 产前美沙酮暴露后观察到的神经化学变化。 然而，与美沙酮不同，丁丙诺啡与δ-和κ-阿片样物质结合， 受体，这可能会导致一组不同的神经和行为 在后代中的干扰。该项目将研究 丁丙诺啡对发育中的生物体的影响老鼠将暴露于各种 出生前和/或出生后通过母体植入的丁丙诺啡剂量 渗透微型泵因为老鼠的中枢神经系统 出生时不成熟，并在出生后继续发育数周， 大鼠将在不同时间暴露于丁丙诺啡，以确定 药物在不同发展阶段的作用。重要的是 丁丙诺啡的剂量反应呈钟形， 曲线美沙酮暴露组将作为阳性对照。 雌性大鼠将被植入28天渗透微型泵， 水、丁丙诺啡或美沙酮。小狗们会 与被植入水的母亲交叉寄养， 丁丙诺啡泵。丁丙诺啡及其代谢物的水平将 通过气相色谱/质谱法在胎仔和幼仔脑中测量 光谱法围产期丁丙诺啡暴露的影响将 根据行为、脑局部乙酰胆碱（ACh）、多巴胺 (DA)、去甲肾上腺素（NE）、5-羟色胺（5-HT）及其代谢产物， 新生（4天）、断奶（21天）和成年（90天）雌性和雄性 大鼠将研究围产期丁丙诺啡的剂量相关效应， 脑局部内源性阿片类物质及其受体以及ACh 转换（TRACh）在断奶和成年大鼠。发展里程碑和 将评估性成熟。抗伤害性感受/伤害性感受（尾- 轻弹，热板）在断奶和成年大鼠。气体 色谱/质谱法将用于测量乙酰胆碱和 TRACh.将使用具有电化学检测的HPLC来测量DA， NE、5-HT及其代谢产物。β-内啡肽样，蛋氨酸和亮氨酸- 脑啡肽样和强啡肽样免疫反应性将通过 放射免疫法μ、δ和κ-阿片样物质的Kd和Bmax 受体将通过Scatchard分析确定。的情况下 阿片样物质受体的变化，特定受体的mRNA 将被量化。如果乙酰胆碱含量降低， 丁丙诺啡，胆碱乙酰转移酶的mRNA将通过 使用北方印迹法。一旦围产期丁丙诺啡的影响 暴露对神经化学和行为的影响得到了更全面的了解， 可以开发更有效的治疗管理， 吸毒成瘾者和潜在的预防药物滥用在他们的孩子。