In JoVE (2)

Other Publications (10)

Articles by Dariusz Orlowski in JoVE

 JoVE Medicine

Basic Surgical Techniques in the Göttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

1Department of Neurosurgery, Aarhus University Hospital, 2Department of Neurobiology, Institute of Anatomy, Faculty of Health Sciences, Aarhus University, 3Positron Emission Tomography (PET) Centre, Aarhus University Hospital


JoVE 2652

Other articles by Dariusz Orlowski on PubMed

Morphological Development of Microglia in the Postnatal Rat Brain. A Quantitative Study

International Journal of Developmental Neuroscience : the Official Journal of the International Society for Developmental Neuroscience. Dec, 2003  |  Pubmed ID: 14659995

Morphological transformation of lectin-positive microglia/macrophages in the developing rat cerebral hemisphere was analysed using quantitative methods. During the first postnatal month, the cells showed increases in their size and fractal dimension accompanied by a simultaneous decrease in their solidity. Regional variations in dynamics of the process indicated the existence of spatio-temporal developmental gradients within the cerebral hemisphere wall which might correspond with regional patterns of neuronal differentiation. Results of the present study prove that the quantitative methods can be the source of reliable data replacing subjective cell typologies.

Autometallographic Enhancement of the Golgi-Cox Staining Enables High Resolution Visualization of Dendrites and Spines

Histochemistry and Cell Biology. Sep, 2009  |  Pubmed ID: 19504264

We present a method for autometallographic (AMG) enhancement of the Golgi-Cox staining enabling high resolution visualization of dendrites and spines. The method is cheaper and more flexible than conventional enhancement procedures performed with commercial photographic developers. The staining procedure is thoroughly described and we demonstrate with qualitative and quantitative data, how histological tissue sectioning, Golgi-Cox immersion time and different AMG enhancement length may influence the staining of dendrites and spines in the rat hippocampus. The described method will be of value for future behavioural-anatomical studies, examining changes in dendrite branching and spine density caused by brain diseases and their subsequent treatment.

Role of Mycorrhizal Colonization in Plant Establishment on an Alkaline Gold Mine Tailing

International Journal of Phytoremediation. Feb, 2011  |  Pubmed ID: 21598786

The potential role of arbuscular mycorrhizal fungi (AMF) in the revegetation of an alkaline gold mine tailing was studied in Barberton, South Africa. The tailing, characterized by a slow spontaneous plant succession, is colonized by the shrub Dodonaea viscosa and the grasses, Andropogon eucomus and Imperata cylindrica, all colonized by AMF. The effectiveness of mycorrhizal colonization in grasses was tested under laboratory conditions using fungal isolates of various origins. Both grasses were highly mycorrhiza dependent, and the presence of mycorrhizal colonization significantly increased their biomass and survival rates. The fungi originating from the gold tailing were better adapted to the special conditions of the tailing than the control isolate. Although the total colonization rate found for native fungi was lower than for fungi from non-polluted sites, they were more vital and more effective in promoting plant growth. The results obtained might serve as a practical approach to the phytostabilization of alkaline gold tailings.

The Effect of Mycorrhiza on the Growth and Elemental Composition of Ni-hyperaccumulating Plant Berkheya Coddii Roessler

Environmental Pollution (Barking, Essex : 1987). Dec, 2011  |  Pubmed ID: 21835516

The effect of arbuscular mycorrhizal fungi (AMF) on growth and element uptake by Ni-hyperaccumulating plant, Berkheya coddii, was studied. Plants were grown under laboratory conditions on ultramafic soil without or with the AM fungi of different origin. The AM colonization, especially with the indigenous strain, significantly enhanced plants growth and their survival. AMF affected also the elemental concentrations that were studied with Particle-induced X-ray emission (PIXE). AMF (i) increased K and Fe in shoots, Zn and Mn in roots, P and Ca both, in roots and shoots; (ii) decreased Mn in shoots, Co and Ni both, in shoots and roots. Due to higher biomass of mycorrhizal plants, total Ni content was up to 20 times higher in mycorrhizal plants compared to the non-mycorrhizal ones. The AMF enhancement of Ni uptake may therefore provide an improvement of a presently used technique of nickel phytomining.

A Simple Reproducible and Time Saving Method of Semi-automatic Dendrite Spine Density Estimation Compared to Manual Spine Counting

Journal of Neuroscience Methods. Jul, 2012  |  Pubmed ID: 22595026

Estimation of spine number and spine density by manual counting under the assumption that all dendrite protrusions equal spines are often used in studies on neuroplasticity occurring during health, brain diseases, and different experimental paradigms. Manual spine counting is, however, time consuming and biased by inter-observer variation. We present accordingly a quick, reproducible and simple non-stereological semi-automatic spine density estimation method based on the irregularity of the dendrite surface. Using the freeware ImageJ program, microphotographs of Golgi impregnated hippocampal dendrites derived from a previously performed study on the impact of chronic restrained stress were binarized, skeletonized, and the skeleton endings assumed to represent spine positions were counted and the spine densities calculated. The results based on 754 dendrite fragments were compared to manual spine counting of the same dendrite fragments using the Bland-Altman method. The results from both methods were correlated (r=0.79, p<0.0001), The semi-automatic counting method gave a statistically higher (approx. 4%) spine density number, but both counting methods showed similar significant differences between the groups in the CA1 area, and no differences between the groups in the CA3 area. In conclusion, the presented semi-automatic spine density estimation method yields consistently a higher spine density number than manual counting resulting in similar significance between groups. The proposed method may therefore be a reproducible time saving and useful non-stereological approach to spine counting in neuroplasticity studies requiring analysis of hundreds of dendrites.

Mycorrhizal Colonization Affects the Elemental Distribution in Roots of Ni-hyperaccumulator Berkheya Coddii Roessler

Environmental Pollution (Barking, Essex : 1987). Apr, 2013  |  Pubmed ID: 23369753

The effect of arbuscular mycorrhizal fungi (AMF) on the distribution and concentration of elements in roots of Ni-hyperaccumulating plant Berkheya coddii was studied. Micro-PIXE (particle-induced X-ray emission) analysis revealed significant differences between AMF-inoculated and non-inoculated plants as well as between main and lateral roots. The accumulation of P, K, Mn and Zn in the cortical layer of lateral roots of inoculated plants confirmed the important role of AMF in uptake and accumulation of these elements. Higher concentration of P, K, Fe, Ni, Cu and Zn in the vascular stele in roots of AMF-inoculated plants than in the non-inoculated ones indicates more efficient translocation of these elements to the aboveground parts of the plant. These findings indicate the necessity of including the influence of AMF in studies on the uptake of elements by plants and in industrial use of B. coddii for Ni extraction from polluted soils.

Potential Roles for Homer1 and Spinophilin in the Preventive Effect of Electroconvulsive Seizures on Stress-induced CA3c Dendritic Retraction in the Hippocampus

European Neuropsychopharmacology : the Journal of the European College of Neuropsychopharmacology. Aug, 2015  |  Pubmed ID: 25935093

Electroconvulsive therapy (ECT) remains the treatment of choice for patients with severe or drug-resistant depressive disorders, yet the mechanism behind its efficacy remains poorly characterized. In the present study, we used electroconvulsive seizures (ECS), an animal model of ECT, to identify proteins possibly involved in the preventive effect of ECS on stress-induced neuronal atrophy in the hippocampus. Rats were stressed daily using the 21-day 6h daily restraint stress paradigm and subjected to sham seizures, a single ECS on the last day of the restraint period or daily repeated seizures for 10 consecutive days during the end of the restraint period. Consistent with previous findings, dendritic atrophy was observed in the CA3c hippocampal region of chronically stressed rats. In addition, we confirmed our recent findings of increased spine density in the CA1 region following chronic restraint stress. The morphological alterations in the CA3c area were prevented by treatment with ECS. On the molecular level, we showed that the synaptic proteins Homer1 and Spinophilin are targeted by ECS. Repeated ECS blocked stress-induced up-regulation of Spinophilin protein levels and further increased the stress-induced up-regulation of Homer1. Given the roles of Spinophilin in the regulation of AMPA receptors and Homer1 in the regulation of metabotropic glutamate receptors (mGluRs), our data imply the existence of a mechanism where ECS regulate cell excitability by modulating AMPA receptor function and mGluR related calcium homeostasis. These molecular changes could potentially contribute to the mechanism induced by ECS which prevents the stress-induced morphological changes in the CA3c region.

The Telencephalon of the Göttingen Minipig, Cytoarchitecture and Cortical Surface Anatomy

Brain Structure & Function. Oct, 2016  |  Pubmed ID: 27778106

During the last 20 years pigs have become increasingly popular in large animal translational neuroscience research as an economical and ethical feasible substitute to non-human primates. The anatomy of the pig telencephalon is, however, not well known. We present, accordingly, a detailed description of the surface anatomy and cytoarchitecture of the Göttingen minipig telencephalon based on macrophotos and consecutive high-power microphotographs of 15 μm thick paraffin embedded Nissl-stained coronal sections. In 1-year-old specimens the formalin perfused brain measures approximately 55 × 47 × 36 mm (length, width, height) and weighs around 69 g. The telencephalic part of the Göttingen minipig cerebrum covers a large surface area, which can be divided into a neocortical gyrencephalic part located dorsal to the rhinal fissure, and a ventral subrhinal part dominated by olfactory, amygdaloid, septal, and hippocampal structures. This part of the telencephalon is named the subrhinal lobe, and based on cytoarchitectural and sulcal anatomy, can be discerned from the remaining dorsally located neocortical perirhinal/insular, pericallosal, frontal, parietal, temporal, and occipital lobes. The inner subcortical structure of the minipig telencephalon is dominated by a prominent ventricular system and large basal ganglia, wherein the putamen and the caudate nucleus posterior and dorsally are separated into two entities by the internal capsule, whereas both structures ventrally fuse into a large accumbens nucleus. The presented anatomical data is accompanied by surface renderings and high-power macrophotographs illustrating the telencephalic sulcal pattern, and the localization of the identified lobes and cytoarchitectonic areas. Additionally, 24 representative Nissl-stained telencephalic coronal sections are presented as supplementary material in atlas form on http://www.cense.dk/minipig_atlas/index.html and referred to as S1-S24 throughout the manuscript.

The Retrograde Connections and Anatomical Segregation of the Göttingen Minipig Nucleus Accumbens

Frontiers in Neuroanatomy. 2016  |  Pubmed ID: 27994542

Nucleus accumbens (NAcc) has been implicated in several psychiatric disorders such as treatment resistant depression (TRD), and obsessive-compulsive disorder (OCD), and has been an ongoing experimental target for deep brain stimulation (DBS) in both rats and humans. In order to translate basic scientific results from rodents to the human setting a large animal model is needed to thoroughly study the effect of such therapeutic interventions. The aim of the study was, accordingly, to describe the basic anatomy of the Göttingen minipig NAcc and its retrograde connections. Tracing was carried out by MRI-guided stereotactic unilateral fluorogold injections in the NAcc of Göttingen minipigs. After 2 weeks the brains were sectioned and subsequently stained with Nissl-, autometallographic (AMG) development of myelin, and DARPP-32 and calbindin immunohistochemistry. The minipig NAcc was divided in a central core and an outer medial, ventral and lateral shell. We confirmed the NAcc to be a large and well-segregated structure toward its medial, ventral and lateral borders. The fluorogold tracing revealed inputs to NAcc from the medial parts of the prefrontal cortex, BA 25 (subgenual cortex), insula bilaterally, amygdala, the CA1-region of hippocampus, entorhinal cortex, subiculum, paraventricular and anterior parts of thalamus, dorsomedial parts of hypothalamus, substantia nigra, ventral tegmental area (VTA), the retrorubral field and the dorsal and median raphe nuclei. In conclusion the Göttingen minipig NAcc is a large ventral striatal structure that can be divided into a core and shell with prominent afferent connections from several subrhinal and infra-/prelimbic brain areas.

Brain Tissue Reaction to Deep Brain Stimulation-A Longitudinal Study of DBS in the Goettingen Minipig

Neuromodulation : Journal of the International Neuromodulation Society. Feb, 2017  |  Pubmed ID: 28220987

The use of Deep Brain Stimulation (DBS) in treatment of various brain disorders is constantly growing; however, the number of studies of the reaction of the brain tissue toward implanted leads is still limited. Therefore, the aim of our study was to analyze the impact of DBS leads on brain tissue in a large animal model using minipigs.

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