Blood sampling from the retro-orbital plexus, the saphenous vein and the tail vein in rats: comparative effects on selected behavioural and blood variables

We compared the behaviours of rats, and measured various blood parameters, after three blood sampling techniques: orbital puncture while they were under diethyl-ether anaesthesia, blood collection by tail vein puncture under O2-N2O-halothane anaesthesia and puncture of the saphenous vein without anaesthesia. Twelve rats were subjected to the three treatments according to a Latin square design. After each treatment, the behaviour of the rats was automatically monitored using the so-called LABORAS method, which discriminates between grooming, locomotion and inactivity in rats. Based on excitation scores and urine production, it was found that induction of diethyl-ether anaesthesia combined with orbital puncture caused more distress than did the other two blood sampling techniques. The three techniques had no differential effects on the behaviours of grooming, locomotion and inactivity. Collecting 0.5 ml of blood by orbital puncture was +/-7 times faster than doing so by saphenous vein puncture and +/- 15 times faster than collecting blood by tail vein puncture while the rats were under O2-N2O-halothane anaesthesia. The levels of some haematological and plasma variables differed significantly between the three blood collection techniques. These observations may help to select the most appropriate technique of blood sampling with respect to anticipated discomfort in the animals.     

A simple method for intravenous injection and blood collection in the chinchilla (Chinchilla laniger)

A simple method is outlined for intravenous injection and blood collection from a peripheral vein in the chinchilla. Seven peripheral vein sites were successfully venipunctured in unanaesthetized chinchillas: the femoral, cephalic, auricular, saphenous, dorsalis penis, lateral abdominal and tail veins. The femoral and cephalic veins were found to be the best sites for injection and blood collection.     

Drawing blood from rats through the saphenous vein and by cardiac puncture

Drawing blood from rodents is necessary for a large number of both in vitro and in vivo studies. Sites of blood draws are numerous in rodents: retro-orbital sinus, jugular vein, maxillary vein, saphenous vein, heart. Each technique has its advantages and disadvantages, and some are not approved any more in some countries (e.g., retro-orbital draws in Holland). A discussion of different techniques for drawing blood are available (1-3). Here, we present two techniques for drawing blood from rats, each with its specific applications. Blood draw from the saphenous vein, provided it is done properly, induces minimal distress in animals and does not require anesthesia. This technique allows repeated draws of small amounts of blood, such as needed for pharmacokinetic studies (4,5), determining plasma chemistry, or blood counts (6). Cardiac puncture allows the collection of large amounts of blood from a single animal (up to 10 ml of blood can be drawn from a 150 g rat). This technique is therefore very useful as a terminal procedure when drawing blood from the saphenous would not provide a large enough sample. We use cardiac puncture when we need sufficient amounts of serum from a specific strain of rats to grow T lymphocyte lines in vitro (4-9).     

树[鼠句]的基本实验操作方法

目的对树[鼠句]抓取和保定、采血、灌胃基本实验技术方法进行探讨,逐步规范树[鼠句]实验技术。方法选用成年树[鼠句]进行抓取和保定,分徒于法和器具法（自制捕捉保定袋）,对130只树[鼠句]采用尾静脉、股动（静）脉两种采血方法;采取人用8号胃管可对树[鼠句]绎口灌胃给药。结果所采用徒手、器具的方法抓取和保定树[鼠句]均能有效地控制动物,不会发生动物死亡或很少逃逸;两种方法都顺利采集到所需血量,股动（静）脉单次最大采血量可达2mL而不损伤动物;12只树[鼠句]连续灌胃10d,成功率100％。结论自制的捕捉保定袋经济实用,摸索的几种树[鼠句]实验技术方法具有操作简便、安全、快捷等优点。     

A novel alternative placement site and technique for totally implantable vascular access ports in non‐human primates

Background Two novel approaches to implanting a central venous catheter port in non‐human primates (NHPs) using peripheral insertion are presented and compared. Methods Sixty vascular access port (VAP) implants were attempted in 52 NHPs by saphenous vein puncture (n = 20) or saphenous vein cutdown (n = 40). Results Fifty eight procedures were successful. Eighteen of 20 VAPs were successfully placed using saphenous vein puncture, and 40 of 40 using saphenous vein cutdown. There were no significant differences between procedures. Mean implantation times were similar between groups. At explant or study endpoint, all 58 VAPs were patent. Conclusions Vascular access port implantation by saphenous vein puncture or saphenous vein cutdown is safe and effective in NHPs. It is less invasive than conventional procedures, has fewer complications, provides outstanding patency, and reduces surgery time. Furthermore, it allows for cooperative in‐homecage VAP use, minimizing handling stress. We recommend these refined methods for long‐term vascular access in NHPs.     

The impact of different blood sampling methods on laboratory rats under different types of anaesthesia

Rats with implanted telemetry transponders were blood sampled by jugular puncture, periorbital puncture or tail vein puncture, or sampled by jugular puncture in carbon dioxide (CO2), isoflurane or without anaesthesia in a crossover design. Heart rate, blood pressure and body temperature were registered for three days after sampling. Initially blood pressure increased, but shortly after sampling it decreased, which led to increased heart rate. Sampling induced rapid fluctuations in body temperature, and an increase in body temperature. Generally, rats recovered from sampling within 2-3 h, except for rats sampled from the tail vein, which showed fluctuations in body temperature in excess of 30 h after sampling. Increases in heart rate and blood pressure within the first hours after sampling indicated that periorbital puncture was the method that had the largest acute impact on the rats and that it might take an extra hour to recover from it. CO2 anaesthesia seemed unable to prevent the increase in blood pressure and the fluctuations in body temperature induced by blood sampling, and up to 10 h after sampling, the rats were still affected by CO2 anaesthesia. Rats anaesthetized with isoflurane showed lower increases in blood pressure after, and fewer fluctuations in body temperature during sampling, and the post-anaesthetic effects of isoflurane, if any, seemed to disappear immediately after sampling. It is, therefore, concluded that blood sampling in rats by jugular puncture seems to be the method from which rats most rapidly recover when compared with periorbital puncture and tail vein puncture, and that for anaesthesia, isoflurane is recommended in preference to CO2.     

Impact of blood sampling technique on blood quality and animal welfare in haemophilic mice

Blood collection in mice can be a challenge, in particular for samples used for coagulation analysis as initiation of coagulation during the procedures can influence the results. Blood collection from the retrobulbar venous plexus is commonly used but the method remains controversial. Several alternatives exist but not all are applicable to mice with a compromised coagulation system because of subsequently excessive bleeding. We therefore wanted to explore whether blood collection by puncture of the submandibular vein could replace blood collected from the retrobulbar venous plexus during pharmacokinetic and pharmacodynamic studies in mice lacking coagulation factor VIII (FVIII). The plasma concentrations of recombinant activated factor VII were independent of the blood collection method in a pharmacokinetic study. The same applied to the thromboelastographic profile of mice with normal coagulation in a pharmacodynamic study. However, excessive haemorrhages were observed in all FVIII knockout mice after a single puncture of the submandibular vein and 60% of the mice were euthanized 2-4 h after the blood collection. In contrast, no or only slight haemorrhage was observed in animals subjected to blood collection from the retrobulbar venous plexus. No signs of distress determined by blood glucose level or clinical abnormalities of the eye were observed after puncture of the retrobulbar venous plexus. In conclusion, blood collected by puncture of the submandibular vein and retrobulbar venous plexus has a quality which allows it to be used in coagulation assays. However, because of excessive bleedings, puncture of the submandibular vein is not recommended in mice lacking FVIII.     

Desensitization of the vascular contractile response to cumulative doses of alpha 2-adrenoceptor agonists

Contractile responses to single or cumulative doses of alpha-adrenoceptor agonists were compared in the tail artery and the saphenous vein of the rat. In the rat tail artery, there were no differences in the dose-response relationships to noradrenaline, methoxamine, and KCl whether the agonists were applied as single or cumulative doses. However, the responses to single doses of clonidine and B-HT 920 were significantly larger than similar doses applied cumulatively. In the rat saphenous vein, responses to single doses of noradrenaline, clonidine, and B-HT 920 were also significantly larger than the corresponding cumulative doses. However, there was no difference in the responses to KCl. It was suggested that desensitization of alpha 2-adrenoceptors in these vessels may result in the diminished responses to cumulative doses of the agonists. Desensitization appeared to be specific to alpha 2-adrenoceptors, since the effect was not observed in responses mediated by the alpha 1-adrenoceptors and KCl.     

The effect of cold on adrenergic neurotransmission in canine saphenous arteries and veins

The effect of severe cold (5 to 10 degrees C) on adrenergic neurotransmission was compared in the isolated cutaneous (saphenous) artery and vein of the dog. The vein contracted to sympathetic nerve stimulation at temperatures as low as 10 degrees C; higher temperatures were needed for the artery to contract. Both blood vessels contracted to exogenous norepinephrine at temperatures as low as 5 degrees C. However, the contractile response to exogenous norepinephrine was less in the saphenous artery, and contractions to high K+ solution were depressed by cooling more in the artery than in the vein. During electrical stimulation of the sympathetic nerves in saphenous arteries and veins previously incubated with labeled norepinephrine, progressive cooling from 37 to 5 degrees C caused a sharp decline in overflow of [3H]norepinephrine and its metabolites. However, overflow of labeled norepinephrine in both blood vessels continued at very cold temperatures. Thus the inability of the saphenous artery to contract to sympathetic nerve stimulation at 10 degrees C can be explained by a greater sensitivity of the arterial smooth muscle to the direct depressant effect of cold, rather than to a differential release or metabolism or norepinephrine in the arterial wall or a loss of responsiveness to norepinephrine at very cold temperatures.     

Amplification of sumatriptan-induced contraction in rabbit saphenous vein but not in basilar artery

The modulation of serotonin (5-HT(1B/1D)) receptor-induced vascular contractility by histamine and U-46619 was compared in the rabbit basilar artery and saphenous vein. In the saphenous vein, histamine (5 x 10(-7) M) significantly increased the potency (from pEC(50) of 6.0 to 6.8) and efficacy (from 52.3% to 88.2%) of sumatriptan. Likewise, U-46619 (5 x 10(-9) M) also increased the potency (from pEC(50) of 5.9 to 6.6) and efficacy (from 51.8% to 92.1%) of sumatriptan in the saphenous vein. In contrast, equieffective concentrations of histamine (10(-7) M) and U-46619 (3 x 10(-9) M) failed to amplify contraction to sumatriptan in the basilar artery. Contraction to sumatriptan was inhibited by nitrendipine (10(-7) M) in the basilar artery but not in the saphenous vein, suggesting that different contractile signaling mechanisms are operating in these tissues. Furthermore, U-46619- and thrombin-induced contractility in the basilar artery were also not amplified by histamine, suggesting that lack of amplification of contraction in the basilar artery was not restricted to sumatriptan but was rather a characteristic of this cerebral vessel. These data suggest that in the in vivo plasma milieu sumatriptan will more markedly contract the peripheral saphenous vein than the basilar artery, a cerebral blood vessel.     

一种动态检测小鼠微量血中T细胞受体重排删除环含量的方法

目的:通过检测比较外周血(颈动脉大量采血和微量隐静脉采血)、胸腺组织和脾脏组织等不同成分或组织中T细胞受体重排删除环(T cell receptor rearrangement excision circles,TREC)的含量,并建立一种通过微量外周血PCR预扩增的方法,评估胸腺输出功能。方法:采用C57BL/6小鼠作为实验对象,分成幼年组(5周龄,n=10)和成年组(15周龄,n=10)。经过隐静脉采血及颈动脉采血后处死小鼠,取小鼠胸腺器官和脾脏器官,提取基因组DNA(g DNA),隐静脉微量血g DNA通过PCR预扩增,提纯后再和颈动脉血、胸腺组织和脾脏组织g DNA一起进行实时定量PCR,分析各组织成分TREC的表达水平及差别。结果:幼年组胸腺组织及外周血中TREC的含量比成年组高,且二者趋势基本一致;而脾脏组织结果与其相反;幼年组小鼠胸腺组织中TREC的含量比脾脏组织中高,成年组小鼠结果与之相反;微量外周血经过预扩增后再进行实时定量PCR的结果与直接定量PCR结果一致,且更高效。结论:研究提供了一种新型高效的动态检测T细胞受体重排删除环和检测胸腺输出功能的方法。     

选择性冠状静脉动脉化搭桥术中不同移植物的应用疗效比较

目的:对比选择性冠状静脉动脉化(SCVBG)搭桥治疗弥漫性右冠状动脉狭窄病变中选择乳内动脉和大隐静脉作为桥血管的治疗效果。方法:选择2008年10月到2014年10月在我院行SCVBG搭桥的84例患者资料,其中选择大隐静脉作为桥血管进行冠状静脉动脉化搭桥患者46例(大隐静脉桥组),选择乳内动脉作为桥血管进行冠状静脉动脉化搭桥患者38例(乳内动脉桥组)。随访记录两组患者的生存情况、近期复查超声心动图、冠状动脉CTA及心绞痛复发率。结果:乳内动脉桥组患者总生存率(100%)明显高于大隐静脉桥组(82.6%)(P0.05)。乳内动脉桥组患者桥血管和心中静脉通畅率(100%)明显大于大隐静脉桥组(54.35%)(P0.05)。两组患者左心室射血分数(LVEF)较治疗前明显增加,左心室舒张期末内径(LVEDD)较治疗前明显减小(P0.05)。治疗后,乳内动脉桥组患者心绞痛复发率明显小于大隐静脉桥组(P0.05)。结论:SCVBG搭桥治疗弥漫性右冠状动脉狭窄病变中,选择乳内动脉桥效果优于大隐静脉桥,能明显提高桥血管和心中静脉通畅率,降低心绞痛复发率。     

Mechanisms of vascular adaptation to long-term orthostatic gravitational loading

Frequent symptoms and serious complaints related to orthostatic intolerance are among the important reasons for investigating the long-term control mechanisms of blood vessels especially those of veins. Previously we studied perfused and superfused saphenous vein segments from rats maintained in head-up tilt position for two weeks. It was found that passive lumen capacity and acute pressure induced myogenic response of these vessels increased substantially without measurable change in wall thickness. Sympathetic component of the smooth muscle cell membrane potential determined in vivo was also significantly enhanced in this vein, but no such change was seen in the saphenous artery and in the brachial vessels. In a separate study, rarefaction of microvessels was found in the hind limb oxydative muscles after two-week tilting, while muscular water content was unaltered. These results suggest that long-term gravitational loading may induce adaptive rearrangement of the blood vessel functions. The aim of the present study was to quantitate and compare the density of nerve fiber terminals as well as their synaptic vesicle population in the wall of saphenous vein and artery from tilted rats to those obtained from rats which were maintained in horizontal, control position. It was hypothetized that adaptation of blood vessels to long-term gravitational loading might include also a morphological restructuring of the vascular adrenergic innervation.     

Endoscopic lesser saphenous vein harvesting for coronary artery bypass grafting: first reported case

Endoscopic vessel harvesting has become a widely used modality for harvesting venous and arterial conduits for coronary artery bypass grafting. Specifically, it has been used to harvest the greater saphenous vein, internal thoracic artery, and the radial artery. A case of endoscopic lesser saphenous vein harvesting for coronary artery bypass grafting is reported.     

Heterogeneity of purinergic P2 receptors at the level of the caudal artery of the rat and the saphenous vein of the dog

Phentolamine (10(-5) M) and an inhibitor of the lipoxygenase pathway, nordihydroguaiaretic acid (N. D. G. A.; 8 10(-6) M) antagonized the ATP induced contraction but not antagonized the UTP induced contraction on both rat tail artery and dog saphenous vein. We conclude that the receptors to ATP are distinct from receptors to UTP and that the P2 purinoceptors are an heterogeneous group.     

Distal embolization at the site of a stenotic distal anastomosis of a saphenous vein graft,leading to dye entrapment

This case report describes the entrapment of contrast media after recanalization of a recently occluded saphenous vein graft with balloon predilation, thrombectomy and stent implantation. Recanalization of the respective coronary artery was performed, and the entrapped contrast media within the saphenous vein graft progressed to the left circumflex artery.     

Functional arterio-venous anastomoses between the testicular artery and the pampiniform plexus in the spermatic cord of rams

Blood flow to the testis, haemoglobin oxygen saturation and testosterone concentration in arterial and venous testicular blood vessels were studied in Texel rams in the breeding and non-breeding season. Blood flow in the proximal and distal testicular artery was measured electromagnetically. The mean flow in the proximal testicular artery was 18.5 ml/min and in the distal testicular artery 7.5 ml/min, and there was no detectable seasonal influence. Haemoglobin oxygen saturation and testosterone concentration were measured in the saphenous artery and vein, the distal testicular artery and vein, and in the proximal testicular vein. The haemoglobin oxygen saturation in the proximal testicular vein was significantly higher than in the distal testicular vein in both seasons. The mean testosterone concentration was significantly lower in the proximal testicular vein than in the distal testicular vein in both seasons. Based on haemoglobin oxygen saturation and testosterone data, it was calculated that between 28 and 46% of the testicular arterial blood was bypassing the testis and was directly flowing through arterio-venous anastomoses towards the pampiniform plexus in the spermatic cord of conscious rams. In anaesthetized rams 55 and 64% of the blood was flowing directly from the testicular artery to the pampiniform plexus based on blood flow data. Transfer of testosterone and oxygen by passive diffusion from the testicular artery to the pampiniform plexus and vice versa in the spermatic cord was not detected.     

Coronary anastomoses over intraluminal occluders

A simplified technique with the use of intraluminal vessel occluders to prevent collateral flow of blood and cardioplegic solution during saphenous vein distal coronary artery anastomosis is presented here. Additional advantages of this technique are the stenting of vessels to facilitate vessel approximation and the assurance of anastomotic patency.     

Evidence for nonclassical alpha adrenoceptor blockade by prazosin in isolated rabbit blood vessels

In isolated rabbit ear artery, prazosin shifted the concentration-response curve for norepinephrine (NE) to the right and increased the slope of the curve in the ED₅₀ region. Prazosin caused a smaller shift to the right of the concentration-response curve for NE in rabbit small saphenous vein and had no effect on the slope in the ED₅₀ region. The prazosin-induced increase in the slope of the NE concentration-response curve did not occur in the ear artery when neuronal and extraneuronal uptake were inhibited by desipramine (DMI) and deoxycorticosterone acetate (DOCA). Prazosin concentration ratios were significantly larger in the ear artery than in the small saphenous vein at all prazosin concentrations in the presence or absence of DMI and DOCA. Significant differences were found between prazosin dissociation constants (K_B) determined at three concentrations of prazosin in ear artery. No such differences were found between phentolamine dissociation constants determined in this vessel. Prazosin dissociation constants determined in small saphenous vein were higher than those determined in ear artery. These findings suggest that prazosin behaves as a noncompetitive antagonist in ear artery and that there are differences between the alpha adrenoceptors of the ear artery and those of the small saphenous vein.     

Comparative analysis of ACTH and corticosterone sampling methods in rats

A frequently debated question for studies involving the measurement of stress hormones in rodents is the optimal method for collecting blood with minimal stress to the animal. Some investigators prefer the implantation of indwelling catheters to allow for frequent sampling. Others argue that the implantation of a catheter creates a chronic stress to the animal that confounds stress hormone measures and therefore rely on tail vein sampling. Moreover, some investigators measure hormones in trunk blood samples obtained after anesthesia, a practice that may itself raise hormone levels. To address these controversies, we 1) compared plasma ACTH and corticosterone (Cort) concentrations in pre- and poststress rat blood samples obtained via previously implanted vena cava catheters, tail vein nicks, or clipping the tip off the tail and 2) compared plasma ACTH and Cort in rat blood samples obtained by decapitation with and without anesthesia. Rats sampled via indwelling catheters displayed lower prestress ACTH levels than those sampled by tail vein nick if the time to acquire samples was not limited; however, elevated basal ACTH was not observed in samples obtained by tail clip or tail nick when the samples were obtained within 3 min. Baseline Cort levels were similar in all groups. After restraint stress, the profile of the plasma ACTH and Cort responses was not affected by sampling method. Decapitation with prior administration of CO2 or pentobarbital sodium increased plasma ACTH levels approximately 13- and 2-fold, respectively, when compared with decapitation without anesthesia. These data indicate that tail vein nicking, tail clipping, or indwelling venous catheters can be used for obtaining plasma for ACTH and Cort during acute stress studies without confounding the measurements. However, the elevation in basal ACTH seen in the tail vein nick group at baseline suggests that sampling needs to be completed rapidly (<3 min) to avoid the initiation of the pituitary stress response. Death by CO2 and pentobarbital sodium injection before trunk blood collection cause significant stress to animals, as reflected in the elevated plasma ACTH levels. These results support the use of either chronic vascular cannulas or sampling from a tail vein. However, collection of blood under pentobarbital sodium or CO2 anesthesia is likely to confound the results of stress studies when ACTH is an important endpoint.