basic-surgical-techniques-gttingen-minipig-intubation-bladder

Basic Surgical Techniques in the G&ouml;ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

The emergence of the G&ouml;ttingen minipig in research of topics such as neuroscience, toxicology, diabetes, obesity, and experimental surgery reflects ...

exposure-pig-cns-for-histological-analysis-manual-for-decapitation

Exposure of the Pig CNS for Histological Analysis: A Manual for Decapitation, Skull Opening, and Brain Removal

Pigs have become increasingly popular in large-animal translational neuroscience research as an economically and ethically feasible substitute to ...

Aarhus University

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.

Morphological development of microglia in the postnatal rat brain. A quantitative study.

Morphologische Entwicklung der Mikroglia in der postnatalen Rattenhirn. Eine quantitative Studie.

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.

Autometallographic enhancement of the Golgi-Cox staining enables high resolution visualization of dendrites and spines.

Wir stellen eine Methode zur Verbesserung der Autometallographic (AMG) des Golgi-Cox F&auml;rbung erm&ouml;glicht hochaufl&ouml;sende Visualisierung von Dendriten und Stacheln. Die Methode ist g&uuml;nstiger und flexibler als konventionelle Erweiterung Verfahren mit kommerziellen fotografischen Entwicklern durchgef&uuml;hrt. Die F&auml;rbung ist gr&uuml;ndlich beschrieben und wir demonstrieren mit qualitativen und quantitativen Daten, wie histologischen Gewebes schneiden, Golgi-Cox-F&auml;rbezeit und verschiedenen AMG Verbesserung L&auml;nge beeinflussen k&ouml;nnen die F&auml;rbung von Dendriten und Dornen im Ratte Hippocampus. Die beschriebene Methode wird Wert f&uuml;r zuk&uuml;nftige Verhaltensst&ouml;rungen-Anatomische Studien, &Auml;nderungen in Dendrit-Dichte Verzweigung und Wirbels&auml;ule verursacht durch Erkrankungen des Nervensystems und deren anschlie&szlig;ende Behandlung haben.

Autometallographic-Verbesserung des Golgi-Cox F&auml;rbung erm&ouml;glicht hochaufl&ouml;sende Visualisierung von Dendriten und Stacheln.

Die potenzielle Rolle von Arbuskul&auml;ren Mykorrhizapilzen (AMF) in der Rekultivierung eine Goldmine alkalischen Tailing wurde in Barberton, S&uuml;dafrika untersucht. Tailing, zeichnet sich durch eine Abfolge langsam spontane Pflanze, ist der Strauch Dodonaea Viscosa Gr&auml;ser, Andropogon Eucomus und Imperata Cylindrica, alle kolonisierten AMF kolonisiert. Die Wirksamkeit der Mykorrhiza Besiedlung in Gr&auml;ser wurde unter Labor getestet, die unterschiedlicher Herkunft Bedingungen mit Pilzen isoliert. Beide Gr&auml;ser waren hoch Mykorrhiza angewiesen, und das Vorhandensein von Mykorrhiza Kolonisation deutlich erh&ouml;ht ihre Preise f&uuml;r Biomasse und &uuml;berleben. Die Pilze aus der gold Tailing wurden besser besonderen Anweisungen f&uuml;r das Tailing als Isolat Steuerelement angepasst. Obwohl die totale Kolonisierung-Rate gefunden f&uuml;r native Pilze war niedriger liegt als f&uuml;r Pilze aus Altlasten, wichtiger und effektiver bei der F&ouml;rderung von Pflanzenwachstum. Die erzielten Ergebnisse k&ouml;nnte als eine praktische Anleitung f&uuml;r die Phytostabilization der alkalischen gold Abraum dienen.

Rolle der Mykorrhiza Besiedlung in Pflanze Einrichtung auf eine Goldmine alkalischen Tailing.

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.

Role of mycorrhizal colonization in plant establishment on an alkaline gold mine tailing.

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)Â 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.

The effect of mycorrhiza on the growth and elemental composition of Ni-hyperaccumulating plant Berkheya coddii Roessler.

Die Wirkung von Arbuskul&auml;ren Mykorrhizapilzen (AMF) auf Wachstum und Element-Aufnahme durch Ni-Hyperaccumulating Pflanze, Berkheya Coddii, wurde untersucht. Pflanzen wurden unter Laborbedingungen auf ultramafischen B&ouml;den ohne oder mit AM Pilze unterschiedlicher Herkunft angebaut. Die AM Kolonisation, vor allem mit den indigenen Stamm, verbessert erheblich Pflanzen Wachstum und ihr &Uuml;berleben. AMF beeinflusst auch die elementaren-Konzentrationen, die mit x-ray Partikel-induzierte Emission (PIXE) untersucht wurden. AMF (i) erh&ouml;ht K und Fe in Triebe, Zn und Mn in Wurzeln, P und Ca sowohl in Wurzeln und Triebe; (Ii) verminderte Mn in Triebe, Co und Ni sowohl in Triebe und Wurzeln. Aufgrund der h&ouml;heren Biomasse Mykorrhiza Pflanzen war Ni Gesamtgehalt in Mykorrhiza Pflanzen im Vergleich zu den nicht-Mykorrhiza in bis zu 20 mal h&ouml;her. Die AMF-Verbesserung der Ni-Aufnahme kann daher eine Verbesserung der derzeit verwendeten Technik Nickel Phytomining vorsehen.

Die Auswirkungen der Mykorrhiza auf das Wachstum und die Elementarzusammensetzung der Ni-Hyperaccumulating Pflanze Berkheya Coddii Roessler.

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.

A simple reproducible and time saving method of semi-automatic dendrite spine density estimation compared to manual spine counting.

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.

Mycorrhizal colonization affects the elemental distribution in roots of Ni-hyperaccumulator Berkheya coddii Roessler.

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.

Potential roles for Homer1 and Spinophilin in the preventive effect of electroconvulsive seizures on stress-induced CA3c dendritic retraction in the hippocampus.

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 telencephalon of the Göttingen minipig, cytoarchitecture and cortical surface anatomy.

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.

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

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.

ArticleTotal : 2	Year
Publication title Cited by 46	2011
Publication title Cited by 26	2017

Article	Year
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\| PubMed ID: 14659995	2003
Autometallographic enhancement of the Golgi-Cox staining enables high resolution visualization of dendrites and spines. Histochemistry and cell biology\| PubMed ID: 19504264	2009
Role of mycorrhizal colonization in plant establishment on an alkaline gold mine tailing. International journal of phytoremediation\| PubMed ID: 21598786	2011
The effect of mycorrhiza on the growth and elemental composition of Ni-hyperaccumulating plant Berkheya coddii Roessler. Environmental pollution (Barking, Essex : 1987)\| PubMed ID: 21835516	2011
A simple reproducible and time saving method of semi-automatic dendrite spine density estimation compared to manual spine counting. Journal of neuroscience methods\| PubMed ID: 22595026	2012
Mycorrhizal colonization affects the elemental distribution in roots of Ni-hyperaccumulator Berkheya coddii Roessler. Environmental pollution (Barking, Essex : 1987)\| PubMed ID: 23369753	2013
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\| PubMed ID: 25935093	2015
The telencephalon of the Göttingen minipig, cytoarchitecture and cortical surface anatomy. Brain structure & function\| PubMed ID: 27778106	2016
The Retrograde Connections and Anatomical Segregation of the Göttingen Minipig Nucleus Accumbens. Frontiers in neuroanatomy\| PubMed ID: 27994542	2016
Brain Tissue Reaction to Deep Brain Stimulation-A Longitudinal Study of DBS in the Goettingen Minipig. Neuromodulation : journal of the International Neuromodulation Society\| PubMed ID: 28220987	2017

Dariusz Orlowski

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