Dissection of Rodent Brain Regions: Guided Free-Hand Slicing and Dissection of Frozen Tissue

Spijker, Sabine; Faliagkas, Leonidas; Rao-Ruiz, Priyanka

doi:10.1007/978-1-4939-9662-9_2

Cited by 6 publications

(6 citation statements)

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“…At the end of treatment, the mice were anesthetized and sacrificed by cervical dislocation. The cranial cavity was opened and brain tissues were harvested according to a protocol described previously [49]. The brain samples were fixed in 4% paraformaldehyde and sent to Google Biology (Wuhan, China) for pathological sectioning.…”

Section: Animals and Cellsmentioning

confidence: 99%

Aflatoxin B1 Induces Neurotoxicity through Reactive Oxygen Species Generation, DNA Damage, Apoptosis, and S-Phase Cell Cycle Arrest

Huang

Chen

Wang

et al. 2020

IJMS

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Aflatoxin B1 (AFB1) is a mycotoxin widely distributed in a variety of food commodities and exhibits strong toxicity toward multiple tissues and organs. However, little is known about its neurotoxicity and the associated mechanism. In this study, we observed that brain integrity was markedly damaged in mice after intragastric administration of AFB1 (300 μg/kg/day for 30 days). The toxicity of AFB1 on neuronal cells and the underlying mechanisms were then investigated in the neuroblastoma cell line IMR-32. A cell viability assay showed that the IC50 values of AFB1 on IMR-32 cells were 6.18 μg/mL and 5.87 μg/mL after treatment for 24 h and 48 h, respectively. ROS levels in IMR-32 cells increased significantly in a time- and AFB1 concentration-dependent manner, which was associated with the upregulation of NOX2, and downregulation of OXR1, SOD1, and SOD2. Substantial DNA damage associated with the downregulation of PARP1, BRCA2, and RAD51 was also observed. Furthermore, AFB1 significantly induced S-phase arrest, which is associated with the upregulation of CDKN1A, CDKN2C, and CDKN2D. Finally, AFB1 induced apoptosis involving CASP3 and BAX. Taken together, AFB1 manifests a wide range of cytotoxicity on neuronal cells including ROS accumulation, DNA damage, S-phase arrest, and apoptosis—all of which are key factors for understanding the neurotoxicology of AFB1.

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Section: Animals and Cellsmentioning

confidence: 99%

Aflatoxin B1 Induces Neurotoxicity through Reactive Oxygen Species Generation, DNA Damage, Apoptosis, and S-Phase Cell Cycle Arrest

Huang

Chen

Wang

et al. 2020

IJMS

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“…Spijker et al reported that, of the specialized dissection instruments, it is necessary to use a forceps, standard razor blade, scalpel handle (3-12 cm), tissue puncher, and metal expeller (18). Also, they emphasized the importance of using a straight forceps with larger surface areas without injuring the brain tissue.…”

Section: █ Discussionmentioning

confidence: 99%

“…Also, they emphasized the importance of using a straight forceps with larger surface areas without injuring the brain tissue. Spijker et al also mention that, due to the risk of injury to the adjacent tissues (18), such as the amygdala and ventral tegmental area (VTA), care should be exercised when a tissue puncher is used.…”

Section: █ Discussionmentioning

confidence: 99%

“…In these experiments, hippocampal tissues obtained especially from rats were used (12,23). Moreover, dissection of the brain tissue is an important step in the sample preparation process for proteomic studies (18). Therefore, in this study, the aim was to surgically explain how to prepare tissue samples by resecting the hippocampal tissue of Wistar-Albino male rats.…”

Section: A B C Dmentioning

confidence: 99%

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Practical performance of hippocampal tissue resection in rats in pharmacomolecular research

Yılmaz

Karaarslan²,

Özbek³

2020

Turkish Neurosurgery

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AIM: To explain how to resect hippocampal tissue in rats used as live mammalian subjects for the resection of fresh cerebral tissue in laboratories. MATERIAL and METHODS:Adult male Wistar-Albino rats (n=50) were used for this purpose. RESULTS:The average bodyweight of the rats was 316.4 ± 11.71 g, and the average weight of the resected hippocampal tissues was 1.01 ± 0.03 g; however, there were no statistically significant differences between the body weights and the hippocampal tissue weights (p>0.05). The hippocampal tissues to be used in the study were excised practically by preserving the original anatomical configuration without injury to the tissue. CONCLUSION:This paper elucidates a simple, step-by-step methodology for performance in the laboratory in order to improve the standardization of hippocampal tissue dissection.

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“…Multiple recent publications have given detailed recommendations for sampling (and often processing) the central and/or peripheral nervous systems (CNS and PNS, respectively) in rodent and nonrodent species to facilitate toxicologic pathology evaluation in nonclinical toxicity studies. 1,2,[16][17][18][19][20][21][22][23][24] In terms of nonrodents, these publications have emphasized sampling in dogs and nonhuman primates (NHPs), which are common test species for nonclinical safety assessment, and sometimes mention other species used to assess specific neurotoxicity questions (eg, chickens, pigs). Importantly, therefore, no existing publications specifically address optimal practices for sampling and processing the nervous system in rabbits for toxicologic pathology evaluation.…”

Section: Introductionmentioning

confidence: 99%

Nervous System Sampling for General Toxicity and Neurotoxicity Studies in Rabbits

Pardo

Rao²,

Morrison

et al. 2020

Toxicol Pathol

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Although manuscripts for multiple species recommending nervous system sampling for histopathology evaluation in safety assessment have been published in the past 15 years, none have addressed the laboratory rabbit. Here, we describe 2 trimming schemes for evaluating the rabbit brain in nonclinical toxicity studies. In both schemes, the intact brain is cut in the coronal plane to permit bilateral assessment. The first scheme is recommended for general toxicity studies (tier 1) in screening agents where there is no anticipated neurotoxic potential; this 6-section approach is consistent with the Society of Toxicologic Pathology (STP) “best practice” recommendations for brain sampling in nonrodents ( Toxicol Pathol 41: 1028-1048, 20131). The second trimming scheme is intended for dedicated neurotoxicity studies (tier 2) to characterize known or suspected neurotoxicants where the nervous system is a key target organ. This tier 2 strategy relies on coronal trimming of the whole brain into 3-mm-thick slices and then evaluating 12 sections. Collection of spinal cord, ganglia, and nerve specimens for rabbits during nonclinical studies should follow published STP “best practice” recommendations for sampling the central nervous system1 and peripheral nervous system ( Toxicol Pathol 46: 372-402, 20182).

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Dissection of Rodent Brain Regions: Guided Free-Hand Slicing and Dissection of Frozen Tissue

Cited by 6 publications

References 1 publication

Aflatoxin B1 Induces Neurotoxicity through Reactive Oxygen Species Generation, DNA Damage, Apoptosis, and S-Phase Cell Cycle Arrest

Aflatoxin B1 Induces Neurotoxicity through Reactive Oxygen Species Generation, DNA Damage, Apoptosis, and S-Phase Cell Cycle Arrest

Practical performance of hippocampal tissue resection in rats in pharmacomolecular research

Nervous System Sampling for General Toxicity and Neurotoxicity Studies in Rabbits

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