DEVELOPMENTAL NEUROCHEMISTRY OF BUPRENORPHINE

美沙酮用于治疗怀孕的阿片类药物成瘾者。 然而，美沙酮的维持与新生儿有关 戒断以及两者的行为和神经化学异常 人类和动物。丁丙诺啡可能是一种对孕妇有用的药物 成瘾者，因为它与戒断综合症无关。此外， 丁丙诺啡在怀孕期间可能优于美沙酮，因为它缺乏 Mu-阿片受体的全部激动剂活性，因此可能不会产生 产前美沙酮暴露后神经化学变化的观察 然而，与美沙酮不同，丁丙诺啡与β-阿片和卡帕-阿片类药物结合 受体，这可能导致一套不同的神经和行为 在后代中的扰动。该项目将研究以下方面的影响 丁丙诺啡在发育中的生物体上。老鼠会暴露在不同的环境中 丁丙诺啡母体植入前和/或产后剂量 渗透性的小泵。因为大鼠的中枢神经系统相对 在出生时不成熟，并在随后的几周内继续发育， 大鼠在不同时间给予丁丙诺啡，以确定 药物在不同发展阶段的作用。这很重要 丁丙诺啡具有钟形剂量反应的多剂量研究 曲线。美沙酮暴露组将被纳入阳性对照。 雌性大鼠将被植入一个28天的渗透性微泵，其中包含 怀孕第8天服用水、丁丙诺啡或美沙酮。小狗将会是 交叉养育给被植入水或 丁丙诺啡泵。丁丙诺啡及其代谢物的水平将是 用气相色谱/质量法测定胎儿和幼崽的脑组织 光谱分析。围产期暴露丁丙诺啡的影响将是 测定行为、大脑局部乙酰胆碱(ACh)、多巴胺 (DA)、去甲肾上腺素(NE)、5-羟色胺(5-羟色胺)及其代谢物。 新生儿(4天)、断奶(21天)和成年(90天)雌雄 老鼠。将研究丁丙诺啡对围产期的剂量相关效应 脑局部内源性阿片类药物及其受体和ACh 断奶和成年大鼠的周转(TRAACH)发展里程碑和 性成熟程度将被评估。抗伤害感受/伤害感受(尾巴- 在断奶大鼠和成年大鼠中)。一种气体 将采用色/质联用法测定ACh和 特拉奇。将采用高效液相色谱-电化学检测相结合的方法测定DA， NE、5-羟色胺及其代谢物。β-内啡肽，Met-和Leu- 脑啡肽样和强啡肽样免疫反应性将通过 放射免疫分析。Mu、Delta和kappa-阿片类药物的KD和Bmax 受体将通过Scatchard分析确定。在发生以下情况时 注意到阿片受体的变化，即特定受体的mrna 将会被量化。如果ACH含量减少了 丁丙诺啡，胆碱乙酰转移酶的信使核糖核酸将通过 北方印迹的使用。一旦丁丙诺啡对围产期的影响 对神经化学和行为的暴露更全面地了解， 可以为怀孕开发更有效的治疗方法 对吸毒成瘾者进行预防，并有可能预防其子女滥用药物。

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.