A Guide to the Generation of a 6-Hydroxydopamine Mouse Model of Parkinson's Disease for the Study of Non-Motor Symptoms.

In Parkinson's disease (PD), a large number of symptoms affecting the peripheral and central nervous system precede, develop in parallel to, the cardinal motor symptoms of the disease. The study of these conditions, which are often refractory to and may even be exacerbated by standard dopamine replacement therapies, relies on the availability of appropriate animal models. Previous work in rodents showed that injection of the neurotoxin 6-hydroxydopamine (6-OHDA) in discrete brain regions reproduces several non-motor comorbidities commonly associated with PD, including cognitive deficits, depression, anxiety, as well as disruption of olfactory discrimination and circadian rhythm. However, the use of 6-OHDA is frequently associated with significant post-surgical mortality. Here, we describe the generation of a mouse model of PD based on bilateral injection of 6-OHDA in the dorsal striatum. We show that the survival rates of males and females subjected to this lesion differ significantly, with a much higher mortality among males, and provide a protocol of enhanced pre- and post-operative care, which nearly eliminates animal loss. We also briefly discuss the utility of this model for the study of non-motor comorbidities of PD.

Preparation and Monitoring of Small Animals in Renal MRI.

Renal diseases remain devastating illnesses with unacceptably high rates of mortality and morbidity worldwide. Animal models are essential tools to better understand the pathomechanism of kidney-related illnesses and to develop new, successful therapeutic strategies. Magnetic resonance imaging (MRI) has been actively explored in the last decades for assessing renal function, perfusion, tissue oxygenation as well as the degree of fibrosis and inflammation. This chapter aims to provide an overview of the preparation and monitoring of small animals before, during, and after surgical interventions or MR imaging. Standardization of experimental settings such as body temperature or hydration of animals and minimizing pain and distress are essential for diminishing nonexperimental variables as well as for conducting ethical research.This publication is based upon work from the COST Action PARENCHIMA, a community-driven network funded by the European Cooperation in Science and Technology (COST) program of the European Union, which aims to improve the reproducibility and standardization of renal MRI biomarkers.

RatHat: A Self-Targeting Printable Brain Implant System.

There has not been a major change in how neuroscientists approach stereotaxic methods in decades. Here, we present a new stereotaxic method that provides an alternative approach to a traditional u-frame stereotaxic device and reduces costs, surgical time, and aids repeatability. The RatHat brain implantation system is a 3D-printable stereotaxic device for rats that is fabricated prior to surgery and fits to the shape of the skull. RatHat builds are directly implanted into the brain without the need for head-leveling or coordinate-mapping during surgery. The RatHat can be used in conjunction with the traditional u-frame stereotaxic device, but does not require the use of a micromanipulator for successful implantations. Each RatHat contains several primary components including the implant for mounting intracranial components, the surgical stencil for targeting drill sites, and the protective cap for preventing damage from impacts and debris. Each component serves a unique function and can be used together or separately. We demonstrate the feasibility of the RatHat in four different proof-of-principle experiments: (1) a three-pole cannula apparatus, (2) an optrode-electrode assembly, (3) a fixed-electrode array, and (4) a tetrode hyperdrive. Implants were successful, durable, and long-lasting (up to nine months). RatHat print files are easily created, can be modified in computer aided design (CAD) software for a variety of applications, and are easily shared, contributing to open science goals and replications. The RatHat has been adapted to multiple experimental paradigms in our lab and should be a useful new way to conduct stereotaxic implant surgeries in rodents.

Side-to-Side Anastomosis Training Model Using Rat Common Carotid Arteries.

BACKGROUND: The side-to-side anastomosis is one of the difficult bypass configurations that may be used in various complex cerebral vascular and neoplastic cases. Few pure arterial models exist for practicing this bypass subtype. OBJECTIVE: To provide an optimized side-to-side anastomosis training model using rat common carotid arteries (CCA). METHODS: Bilateral CCAs were exposed in the neck of 10 anesthetized Sprague-Dawley rats. The arteries were juxtaposed in parallel, using temporary aneurysm clips applied proximally and distally. CCA caliber and the length of CCA juxtaposition were measured. Side-to-side anastomosis was completed and ischemia time was recorded. Unintended complications were recorded for further analysis. RESULTS: Anastomosis was completed successfully in all animals. The CCAs were approximated in all animals without any difficulty or undue tension. In 2 rats, death occurred prior to completion of anastomosis, which was attributed to injury to the external jugular vein during vessel exposure. Mean ischemia time was 35 min with an average of 22 sutures done to complete the anastomosis. The average CCA caliber was 1.1 ± 0.2 mm and the arteries could be juxtaposed for an average length of 10.2 ± 1.5 mm. CONCLUSION: Full exposure of the cervical segment of the CCAs enables tension-free approximation of adequate length of the vessel for a side-to-side anastomosis. Avoiding complications during exposure helps in prevention of animal death during the ischemia period.

The End-to-Side Anastomosis: A Comparative Analysis of Arterial Models in the Rat.

BACKGROUND: The end-to-side anastomosis is 1 of the most common anastomosis configurations used in cerebrovascular surgery. Although several living practice models have been proposed for this technique, few involve purely arterial vessels. The purpose of the present study was to compare 2 arterial models using common carotid arteries (CCAs) and common iliac arteries (CIAs) in rats. METHODS: The CIAs and CCAs were exposed in 10 anesthetized rats, and their lengths and diameters were measured. Also, the mobilization extent of each vessel along its contralateral counterpart was measured after each artery had been transected at its proximal exposure point. We also studied the technical advantages and disadvantages of each model for practicing end-to-side anastomosis. RESULTS: The average diameters of the CCA and CIA were 1.1 and 1.3 mm, respectively. The average extent of mobilization along the contralateral vessel was 13.9 mm and 10.3 mm for CCA and CIA, respectively. The CCA model had the advantages of greater arterial redundancy (allowing completion of both suture lines extraluminally) and a minimal risk of venous injury. The main disadvantage of the CCA model was the risk of cerebral ischemia. The CIA model was not limited by the ischemic time and provided the technical challenge of microsurgical dissection of the common iliac vein from the CIA, although it had limited CIA redundancy. CONCLUSIONS: Both CCA and CIA models could be efficiently used for practicing the end-to-side anastomosis technique. Each model provides the trainee with a specific set of advantages and disadvantages that could help with the optimal selection of the practice model according to trainee's skill level.

Technical Nuances of Exposing Rat Common Carotid Arteries for Practicing Microsurgical Anastomosis.

BACKGROUND: Animal models are commonly used in training protocols for microsurgical vascular anastomosis. Rat common carotid arteries (CCAs) are frequently used for this purpose. Much attention has been paid to the technical details of various anastomosis configurations using these arteries. However, technical nuances of exposing rat CCAs have been understudied. The purpose of this study is to describe nuances of technique for safely and efficiently exposing rat CCAs in preparation for a vascular anastomosis. METHODS: Bilateral CCAs were exposed and prepared for anastomosis in 10 anesthetized Sprague-Dawley rats through a midline cervical incision. The exposed length of the CCA was measured. Additionally, technical nuances of exposure and surgically relevant anatomic details were recorded. RESULTS: The CCAs were exposed from the sternoclavicular joint to their bifurcation (average length, 19.1 ± 2.8 mm). Tenets important for a safe and efficient exposure of the CCAs included 1) generous subcutaneous dissection to expose the external jugular veins (EJVs), 2) avoiding injury to or compression of the EJVs, 3) superior mobilization of the salivary glands, 4) division of internal jugular veins, 5) opening the carotid sheath at its midlevel and from medial to lateral, and 6) avoiding injury to the vagus nerve or sympathetic trunk. CONCLUSIONS: Using the principles introduced in this study, trainees may safely and efficiently expose rat CCAs in preparation for a bypass.

Transfer of Learning from Practicing Microvascular Anastomosis on Silastic Tubes to Rat Abdominal Aorta.

OBJECTIVE: Learning to perform microvascular anastomosis is difficult. Laboratory practice models using artificial vessels are frequently used for this purpose. However, the efficacy of such practice models has not been objectively assessed for the performance of microvascular anastomosis during live surgical settings. This study was conducted to assess the transfer of learning from practicing microvascular anastomosis on tubes to anastomosing rat abdominal aorta. METHODS: Ten surgeons without any experience in microvascular anastomosis were randomly assigned to an experimental or a control group. Both groups received didactic and visual training on end-to-end microvascular anastomosis. The experimental group received 24 sessions of hands-on training on microanastomosis using Silastic tubes. Next, both groups underwent recall tests on weeks 1, 2, and 8 after training. The recall test consisted of completing an end-to-end anastomosis on the rat's abdominal aorta. Anastomosis score, the time to complete the anastomosis, and the average time to place 1 stitch on the vessel perimeter were compared between the 2 groups. RESULTS: Compared with the control group, the experimental group did significantly better in terms of anastomosis score, total time, and per-stitch time. The measured variables showed stability and did not change significantly between the 3 recall tests. CONCLUSION: The skill of microvascular anastomosis is transferred from practicing on Silastic tubes to rat's abdominal aorta. Considering the relative advantages of Silastic tubes to live rodent surgeries, such as lower cost and absence of ethical issues, our results support the widespread use of Silastic tubes in training programs for microvascular anastomosis.

Microsurgical Bypass Training Rat Model, Part 1: Technical Nuances of Exposure of the Aorta and Iliac Arteries.

BACKGROUND: Animal models using rodents are frequently used for practicing microvascular anastomosis-an essential technique in cerebrovascular surgery. However, safely and efficiently exposing rat's target vessels is technically difficult. Such difficulty may lead to excessive hemorrhage and shorten animal survival. This limits the ability to perform multiple anastomoses on a single animal and may increase the overall training time and costs. We report our model for microsurgical bypass training in rodents in 2 consecutive articles. In part 1, we describe the technical nuances for a safe and efficient exposure of the rat abdominal aorta and common iliac arteries (CIAs) for bypass. METHODS: Over a 2-year period, 50 Sprague-Dawley rats underwent inhalant anesthesia for practicing microvascular anastomosis on the abdominal aorta and CIAs. Lessons learned regarding the technical nuances of vessel exposure were recorded. RESULTS: Several technical nuances were important for avoiding intraoperative bleeding and preventing animal demise while preparing an adequate length of vessels for bypass. The most relevant technical nuances include (1) generous subcutaneous dissection; (2) use of cotton swabs for the blunt dissection of the retroperitoneal fat; (3) combination of sharp and blunt dissection to isolate the aorta and iliac arteries from the accompanying veins; (4) proper control of the posterior branches of the aorta; and (5) efficient division and mobilization of the left renal pedicle. CONCLUSIONS: Applying the aforementioned technical nuances enables safe and efficient preparation of the rat abdominal aorta and CIAs for microvascular anastomosis.

Microsurgical Bypass Training Rat Model: Part 2-Anastomosis Configurations.

BACKGROUND: Mastery of microsurgical anastomosis is key to achieving good outcomes in cerebrovascular bypass procedures. Animal models (especially rodents) provide an optimal preclinical bypass training platform. However, the existing models for practicing different anastomosis configurations have several limitations. OBJECTIVE: We sought to optimize the use of the rat's abdominal aorta and common iliac arteries (CIA) for practicing the 3 main anastomosis configurations commonly used in cerebrovascular surgery. METHODS: Thirteen male Sprague-Dawley rats underwent inhalant anesthesia. The abdominal aorta and the CIAs were exposed. The distances between the major branches of the aorta were measured to find the optimal location for an end-to-end anastomosis. Also, the feasibility of performing side-to-side and end-to-side anastomoses between the CIAs was assessed. RESULTS: All bypass configurations could be performed between the left renal artery and the CIA bifurcation. The longest segments of the aorta without major branches were 1) between the left renal and left iliolumbar arteries (16.9 mm ± 4.6), and 2) between the right iliolumbar artery and the aortic bifurcation (9.7 mm ± 4.7). The CIAs could be juxtaposed for an average length of 7.6 mm ± 1.3, for a side-to-side anastomosis. The left CIA could be successfully reimplanted on to the right CIA at an average distance of 9.1 mm ± 1.6 from the aortic bifurcation. CONCLUSIONS: Our results show that rat's abdominal aorta and CIAs may be effectively used for all the anastomosis configurations used in cerebral revascularization procedures. We also provide technical nuances and anatomic descriptions to plan for practicing each bypass configuration.