Ueno Tatsuya, Duenes Judith A, Kost Louis J, Sarr Michael G
Department of Surgery and the Gastroenterology Research Unit, Mayo Clinic, Rochester, Minnesota 55905, USA.
J Surg Res. 2004 May 15;118(2):136-43. doi: 10.1016/S0022-4804(03)00334-2.
Interest in genomic modulation experimentally often necessitates use of mouse models.
To characterize and quantitate smooth muscle contractile activity of the mouse small intestine using in vitro techniques. Full-thickness jejunal and ileal muscle strips from mice were cut in the direction of longitudinal muscle, suspended in tissue baths (37 degrees C), and connected to force transducers. Spontaneous contractility and two dose-response curves to the cholinergic agonist bethanechol and adrenergic agonist norepinephrine were quantitated for 6 h.
Total contractile activity increased over 4 to 5 h in jejunum (P < 0.01) but not in ileum. Frequency of contractions (counts/min) in jejunum increased from 16 to 33 (P < 0.01) in the first 4 h, then remained stable; ileal frequency did not change. One hour of cold preservation had no major effect on contractile activity and frequency. Bethanechol increased and norepinephrine decreased contractile activity in dose-dependent fashion. The dose of bethanechol producing 50% increase in maximal response did not differ between the first and second dose-response; in contrast, the concentration of norepinephrine producing 50% decrease in activity for the second dose-response in jejunum was decreased compared to the first dose-response (P < 0.01). Cold preservation had no substantive effect on agonist responses.
Experiments in murine jejunal but not ileal longitudinal muscle in vitro must consider early changes in contractile activity after tissue harvest. These experiments serve as a baseline for comparison of stimuli or genetic modifications on murine contractile activity of longitudinal muscle in vivo.
在实验中对基因组调节的研究常常需要使用小鼠模型。
运用体外技术对小鼠小肠平滑肌收缩活性进行表征和定量分析。从小鼠身上切取全层空肠和回肠肌条,沿纵肌方向切割,悬挂于组织浴(37摄氏度)中,并连接到力传感器。对自发收缩性以及对胆碱能激动剂氨甲酰甲胆碱和肾上腺素能激动剂去甲肾上腺素的两条剂量反应曲线进行6小时的定量分析。
空肠的总收缩活性在4至5小时内增加(P<0.01),但回肠没有。空肠收缩频率(次/分钟)在最初4小时内从16增加到33(P<0.01),然后保持稳定;回肠频率没有变化。1小时的冷藏对收缩活性和频率没有重大影响。氨甲酰甲胆碱以剂量依赖性方式增加收缩活性,而去甲肾上腺素则降低收缩活性。产生最大反应增加50%的氨甲酰甲胆碱剂量在第一次和第二次剂量反应之间没有差异;相比之下,空肠中第二次剂量反应导致活性降低50%的去甲肾上腺素浓度与第一次剂量反应相比有所降低(P<0.01)。冷藏对激动剂反应没有实质性影响。
体外对小鼠空肠而非回肠纵肌进行的实验必须考虑组织取材后收缩活性的早期变化。这些实验可作为比较体内刺激或基因修饰对小鼠纵肌收缩活性影响的基线。