Quantitative measurement of local cerebral blood flow in the anesthetized mouse using intraperitoneal [14C]iodoantipyrine injection and final arterial heart blood sampling
Quantitative measurement of local cerebral blood flow in the anesthetized mouse using intraperitoneal [14C]iodoantipyrine injection and final arterial heart blood sampling
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DOI：
10.1097/00004647-200001000-00003
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发表时间：
2000-01-01
影响因子：
6.3
通讯作者：
Hossmann, KA
中科院分区：
文献类型：
--
作者：
Maeda, K;Mies, G;Hossmann, KA
[C-14]碘安替比林(IAP)对小鼠局部脑血流量(CBF)的放射自显影测量受到血管插管困难和重复采血失血的限制。作者修改和验证了用[C-14]IAP测定小鼠局部脑血流量的方法，方法是在实验结束时将示踪剂的应用与心脏的一次采血相结合。实验在雄性SV129小鼠氟烷麻醉下进行。腹腔注射Ci[C-14]IAP 15mU后，重复采集动脉血，将麻醉动物浸入液氮中。此外，从心脏采集冰冻血液，以获得最终血液[C-14]的放射性。采集时间与动脉血[C-14]放射性之间的相关分析表明，[C-14]示踪剂在动脉血中的滞后时间为3.3s+/-0.6s，具有极显著的线性关系(P<；0.001，r=0.978)。[C-14]最终动脉血样本的放射性活度(444+/-263nCI/m L)与心脏血的放射性活度(453+/-242nCI/m L)几乎相等，两种动物之间的绝对差异为3.3+/-4.2%(平均+/-SD)。CBF计算的卷积积分是通过对单个动脉血样的放射性进行积分来确定的，或者通过假设从腹膜内注射[C-14]IAP后的[C-14]示踪剂滞后时间到从冰冻心脏的血液样本中测量的值的线性上升来确定的。两种方法计算的区域流量值相差不到11%(不显著)。这种方法可以在不需要任何血管插管的情况下定量测量麻醉小鼠的局部脑血流量，并将流动研究与分子生物学方法相结合，使突变小鼠成为阐明脑损伤分子机制的更强大的工具。
Autoradiographic measurement of local cerebral blood flow (CBF) with [C-14]iodoantipyrine (IAP) is limited in mice by the difficulty in cannulating vessels and the blood loss for repeated blood sampling. The authors modified and validated the method to measure local CBF with [C-14]IAP in mice by combining intraperitoneal tracer application with a single blood sampling from the heart at the end of the experiment. Experiments were carried out in male SV129 mice under halothane anesthesia. After intraperitoneal administration of 15 mu Ci [C-14]IAP, arterial blood samples were collected repeatedly and anesthetized animals were immersed in liquid nitrogen. In addition, frozen blood from the heart was sampled to obtain the final blood [C-14]radioactivity. Correlation analysis between the sampling time and [C-14] radioactivity of the arterial blood revealed a highly significant linear relationship (P < 0.001, r = 0.978) and a lag time of the [C-14]tracer in arterial blood of 3.3 +/- 0.6 seconds. [C-14]radioactivity of the final arterial blood sample (444 +/- 263 nCi/mL) was almost equal to that of the heart blood (453 +/- 242 nCi/mL), and the absolute difference in each animal was 3.3 +/- 4.2% (mean +/- SD). The convolution integrals for the CBF calculation were determined either by integrating the radioactivity of individual arterial blood samples or by assuming a linear rise from [C-14]tracer lag time after intraperitoneal [C-14]IAP injection to the value measured in the blood sample from the frozen heart. Regional flow values calculated by the two methods differed by less than 11% (not significant). This method allows the quantitative measurement of local CBF in anesthetized mice without any vessel catheterization and will make mutant mice a more powerful tool to elucidate the molecular mechanisms of brain injuries by combining flow studies with molecular-biological methods.