高原鼠兔对高原低氧环境有很强的适应性。研究发现,精子特异性乳酸脱氢酶(LDH-C4)基因在高原鼠兔脑组织中表达,为阐明LDH-C4在高原鼠兔脑组织中的作用,应用荧光定量PCR和Western blot方法,测定了Ldh-c基因在高原鼠兔脑组织中的表达水平;应用对精子特异性乳酸脱氢酶(LDH-C4)特异性的抑制剂(N-isopropyl oxamate),通过肌肉注射后,研究抑制剂对高原鼠兔脑组织中LDH比活力、乳酸和ATP含量的影响。结果表明,在mRNA和蛋白水平,Ldh-c基因在高原鼠兔脑组织中均有表达,相对表达水平分别为0.38±0.05和0.74±0.13;当肌肉注射1 m L 1 mol/L的抑制剂30 min后,血液中抑制剂浓度为0.08 mmol/L;与对照组相比,抑制剂组脑组织中LDH比活力、乳酸和ATP含量显著下降,抑制剂对LDH、乳酸和ATP的抑制率分别为30.78%、46.47%和21.04%。结果表明,精子特异性乳酸脱氢酶基因在高原鼠兔脑组织中表达。LDH-C4通过催化无氧糖酵解过程,为其脑组织生命活动提供至少20%的ATP,可能使高原鼠兔减小在低氧环境中对氧气的依赖,增强对低氧环境的适应能力。
The plateau pika(Ochotona curzoniae) has a strong adaptability to a hypoxic plateau environment. We found that the sperm-specific lactate dehydrogenase(LDH-C4) gene is expressed in plateau pika brain. In order to shed light on the role of LDH-C4 in plateau pika brain,we determined the expression levels of Ldh-c gene in plateau pika brain by real-time PCR and western blot. After injecting N-isopropyl oxamate,a LDH-C4 specific inhibitor,in skeletal muscle of hind legs,the activities of LDH,and the contents of lactic acid and ATP in plateau pika brain were measured. The results indicated that the expression levels of Ldh-c mRNA and protein were 0. 38 ± 0. 05 and 0. 74 ± 0. 13. Thirty minutes after1 m L 1 mol / L N-isopropyl oxamate was injected into the biceps femoris muscle,the concentration of N-isopropyl oxamate in blood was 0. 08 mmol / L; the LDH activities,the contents of lactic acid and ATP in the brains of the inhibitor group were decreased significantly compared with that of the control group,and the inhibition rates of N-isopropyl oxamate to LDH,lactic acid and ATP in plateau pika brain were 30. 78%,46. 47% and 21. 04%,respectively. These results indicated that the sperm-specific lactate dehydrogenase gene is expressed in plateau pika brain,and the pika brain got at least 20% ATP for its life activities by catalyzing anaerobic glycolysis. As a result,the plateau pika may reduce dependence on oxygen and enhance the adaptation to the hypoxic environments.