Translate this page to:
Hindi
In JoVE (1)
Other Publications (16)
- Hippocampus
- Anatomy and Embryology
- Biological Psychiatry
- BJU International
- Anatomy and Embryology
- Journal of Neuroscience Methods
- Neuroreport
- Acta Neurochirurgica
- Histochemistry and Cell Biology
- Journal of Neuroscience Methods
- NeuroImage
- Journal of Chemical Neuroanatomy
- Stereotactic and Functional Neurosurgery
- Molecular Therapy : the Journal of the American Society of Gene Therapy
- Journal of Neuroscience Methods
- PloS One
Automatic Translation
This translation into Hindi was automatically generated.
English Version | Other Languages
Articles by Carsten R. Bjarkam in JoVE
JoVE Clinical and Translational Medicine
मूल गौटिंगेन Minipig में सर्जिकल तकनीक: इंटुबैषेण, मूत्राशय कैथीटेराइजेशन, ऊरु पोत कैथीटेराइजेशन, और Transcardial छिड़काव
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
विज्ञान के क्षेत्र में घरेलू और लघु सूअरों का उपयोग हाल के वर्षों में काफी वृद्धि हुई है. कैसे इंटुबैषेण, transurethral मूत्राशय कैथीटेराइजेशन, और्विक धमनी और शिरा कैथीटेराइजेशन, के रूप में अच्छी तरह से transcardial छिड़काव करने के लिए प्रदर्शन करके, हम करने के लिए जैव चिकित्सा अनुसंधान में आगे गौटिंगेन minipigs के मूल्य में वृद्धि के उद्देश्य.
Other articles by Carsten R. Bjarkam on PubMed
Distribution and Morphology of Serotonin-immunoreactive Axons in the Hippocampal Region of the New Zealand White Rabbit. I. Area Dentata and Hippocampus
This study provides a detailed light microscopic description of the morphology and distribution of immunohistochemically stained serotonergic axons in the hippocampal region of the New Zealand white rabbit. The serotonergic axons were segregated morphologically into three types: beaded fibers, fine fibers, and stem-axons, respectively. Beaded fibers were thin serotonergic axons with large varicosities, whereas thin axons with small fusiform or granular varicosities were called fine fibers. Finally, thick straight non-varicose axons were called stem-axons. Beaded fibers often formed large conglomerates with numerous boutons (pericellular arrays) in close apposition to the cell-rich layers in the hippocampal region, e.g., the granular and hilar cell layers of the dentate area and the pyramidal cell layer ventrally in CA3. The pericellular arrays in these layers were often encountered in relation to small calbindin-D2BK-positive cells, as shown by immunohistochemical double staining for serotonin and calbindin-D28K. The beaded and fine serotonergic fibers displayed a specific innervation pattern in the hippocampal region and were encountered predominantly within the terminal field of the perforant path, e.g., the stratum moleculare hippocampi and the outer two-thirds of the dentate molecular layer. These fibers were also frequently seen in the deep part of the stratum oriens and the alveus, forming a dense plexus in relation to large multipolar calbindin-D28K-positive cells and their basal extensions. Stem-axons were primarily seen in the fimbria and alveus. This innervation pattern was present throughout the entire hippocampal formation, but there were considerable septotemporal differences in the density of the serotonergic innervation. A high density of innervation prevailed in the ventral/temporal part of the hippocampal formation, whereas the dorsal/septal part received only a moderate to weak serotonergic innervation. These results suggest that the serotonergic system could modulate the internal hippocampal circuitry by way of its innervation in the terminal field of the perforant path, the hilus fasciae dentatae, and ventrally in the zone closely apposed to the mossy fiber layer and the pyramidal cells of CA3. This modulation could be of a dual nature, mediated directly by single serotonergic fibers traversing the hippocampal layers or indirectly by the pericellular arrays and their close relation to the calbindin-D28K-positive cells. The marked septotemporal differences in innervation density point toward a difference between the ventral and dorsal parts of the hippocampal formation with respect to serotonergic function and need for serotonergic modulation.
The Anatomy of the Porcine Subthalamic Nucleus Evaluated with Immunohistochemistry and Design-based Stereology
This study provides a light-microscopic description of the organization, morphology and number of neurons in the subthalamic nucleus (STN) of the Göttingen minipig. It is based on histological material stained with Nissl, Golgi and autometallographic techniques, and employs design-based stereological estimation of the total neuron number. The organization of several neurotransmitters in the STN has been evaluated in histological preparations stained for acetylcholinesterase (AChE) and immunostained for choline acetyltransferase (ChAT), tyrosine hydroxylase (TH), glutamic acid decarboxylase (GAD) and glutamate. In all of the stained preparations the STN appeared as a distinct lens-shaped structure located in the caudal diencephalon, medial to the internal capsule and ventrolateral to the zona incerta. Rostrally, the STN approached the globus pallidus pars interna, whereas caudally the ventromedial part of the STN was adjacent to the rostral part of the substantia nigra pars compacta (SNc), where some of the neurons of the two nuclei merged. The neurons in the STN had medium-sized (25-40 microm) ovoid or fusiform cell bodies, from which three to six large dendrites emanated in a direction predominantly parallel to the long axis of the STN. Immunohistochemistry revealed that most of the subthalamic neurons were glutamatergic and differed significantly in appearance from the large stellate TH-positive cells of the adjacent SNc. Numerous TH-positive bouton-rich fibers traversed the STN. The GAD-staining revealed a large number of terminals within the boundaries of the STN. The STN was highly AChE-positive, reflecting a prominent innervation by ChAT-positive terminals. The total number of subthalamic neurons in one hemisphere was estimated to be approximately 56,000. We conclude that the neuroarchitecture of the porcine STN is similar to primates, including humans, and appears well-suited for further studies examining the role of the STN in movement disorders.
Therapeutic Strategies for Neurodegenerative Disorders: Emerging Clues from Parkinson's Disease
Our knowledge of Parkinson's disease pathophysiology has greatly expanded during the last century, resulting in successful new medical and neurosurgical approaches toward this common neurodegenerative disorder. These approaches might also be useable in the treatment of psychiatric disorders, which often are linked to atrophic and degenerative processes in the brain; however, the successful application of these techniques in psychiatry requires thorough elucidation of disease pathophysiology to identify proper intervention sites. Likewise, awareness of the differences between the parkinsonian and psychiatric patient populations in terms of age, disease course, and life expectancy, as well as ethical considerations might in the end determine the appropriateness of these therapeutic strategies in psychiatry.
Stereotactic Electrical Stimulation of the Pontine Micturition Centre in the Pig
To apply stereotactic electrical stimulation of the pig brainstem and thus identify a pontine micturition centre.
Distribution and Morphology of Serotonin-immunoreactive Axons in the Retrohippocampal Areas of the New Zealand White Rabbit
This study provides a detailed light microscopic description of the morphology and distribution of serotonin-immunoreactive axons in the paleocortical retrohippocampal areas, viz. the subiculum, presubiculum, parasubiculum and entorhinal area, and the adjoining neocortical perirhinal and retrosplenial cortices of the New Zealand white rabbit. Serotonergic axons could be segregated into three different fiber types named fine fibers, beaded fibers and stem-axons. Fine fibers were evenly distributed thin axons with small fusiform/granular varicosities. Beaded fibers were thin axons with large varicosities, predominantly located in the retrohippocampal supragranular layers, where they often formed pericellular arrays. Stem-axons were thick straight, nonvaricose axons seen in the white matter of psalterium dorsale, alveus and the plexiform layer. The paleocortical retrohippocampal areas had a dense supragranular innervation with numerous tortuous fine and beaded fibers, intermingled in conglomerates with conspicuous varicosities forming pericellular arrays. In contrast, the neocortical area 17 and the lateral part of the perirhinal cortex (area 36) were innervated by evenly distributed fine fibers with a moderate number of small varicosities and few ramifications, whereas, the retrosplenial cortex (areas 29e, 29ab and 29cd), and the medial part of the perirhinal cortex (area 35) displayed an intermediate innervation pattern, probably reflecting the transitional nature of these areas being located between the paleo- and the neocortex. The described dualistic innervation pattern may functionally enable the serotonergic system to exert a strong influence on the supragranular layers of the retrohippocampal areas and thus on the neural input entering these areas from the perirhinal and neighboring polymodal association neocortices, whereas the innervation pattern in the adjoining neocortical areas points towards a more diffuse and general modulation of neural activity herein.
Deep Brain Stimulation Electrode Anchoring Using BioGlue((R)), a Protective Electrode Covering, and a Titanium Microplate
The authors present an easily applicable deep brain stimulation (DBS) electrode anchoring technique for use in human and experimental animals. The anchoring technique combines the use of fibrin glue, a two-component surgical adhesive (BioGlue), a protective electrode covering, and a titanium microplate. The BioGlue (CryoLife International, Inc., Kennesaw, GA, USA) hinders unwanted electrode movement during the electrode fixation step and seals the burr hole, while the protective electrode covering protects the electrode under the titanium microplate which keeps the electrode in a permanent position. The described technique further has the advantage of being cosmetically acceptable to the human patient, and furthermore it perfectly adapts to the smaller and irregular-shaped skull in experimental animals. The described technique has clinically been used to implant DBS-electrodes in the subthalamic nucleus for Parkinson disease and is the preferred DBS-electrode anchoring technique for our experimental DBS-studies in the Göttingen minipig.
Long-term Meditation is Associated with Increased Gray Matter Density in the Brain Stem
Extensive practice involving sustained attention can lead to changes in brain structure. Here, we report evidence of structural differences in the lower brainstem of participants engaged in the long-term practice of meditation. Using magnetic resonance imaging, we observed higher gray matter density in lower brain stem regions of experienced meditators compared with age-matched nonmeditators. Our findings show that long-term practitioners of meditation have structural differences in brainstem regions concerned with cardiorespiratory control. This could account for some of the cardiorespiratory parasympathetic effects and traits, as well as the cognitive, emotional, and immunoreactive impact reported in several studies of different meditation practices.
Long-term Implantation of Deep Brain Stimulation Electrodes in the Pontine Micturition Centre of the Göttingen Minipig
To implant deep brain stimulation (DBS) electrodes in the porcine pontine micturition centre (PMC) in order to establish a large animal model of PMC-DBS.
Autometallographic Enhancement of the Golgi-Cox Staining Enables High Resolution Visualization of Dendrites and Spines
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.
MRI-guided Stereotaxic Targeting in Pigs Based on a Stereotaxic Localizer Box Fitted with an Isocentric Frame and Use of SurgiPlan Computer-planning Software
We present a stereotaxic procedure enabling MRI-guided isocentric stereotaxy in pigs. The procedure is based on the Leksell stereotaxic arch principle, and a stereotaxic localizer box with an incorporated fiducial marking system (sideplates) defining a stereotaxic space similar to the clinical Leksell system. The obtained MRIs can be imported for 3D-reconstruction and coordinate calculation in the clinical stereotaxic software planning system (Leksell SurgiPlan, Elekta AB, Sweden). After MRI the sideplates are replaced by a modified Leksell arch accommodating clinical standard manipulators for isocentric placement of DBS-electrodes, neural tracers and therapeutics in the calculated target coordinates. The mechanical accuracy of the device was within 0.3-0.5 mm. Stereotaxic MRIs were imported to the stereotaxic software planning system with a mean error of 0.4-0.5 mm and a max error of 0.8-0.9 mm. Application accuracy measured on a phantom and on inserted skull markers in nine pigs was within 1 mm in all planes. The intracerebral application accuracy found after placement of 10 manganese trajectories within the full extent of the intracerebral stereotaxic space in two minipigs was equally randomly distributed and within 0.7+/-0.4; 0.5+/-0.4; and 0.7+/-0.3mm in the X, Y, and Z plane. Injection of neural tracers in the subgenual gyrus of three minipigs and placement of encapsulated gene-modified cells in four minipigs confirmed the accuracy and functionality of the described procedure. We conclude that the devised technique and instrumentation enable high-precision stereotaxic procedures in pigs that may benefit future large animal neuroscience research and outline the technical considerations for a similar stereotaxic methodology in other animals.
Neurite Density from Magnetic Resonance Diffusion Measurements at Ultrahigh Field: Comparison with Light Microscopy and Electron Microscopy
Due to its unique sensitivity to tissue microstructure, diffusion-weighted magnetic resonance imaging (MRI) has found many applications in clinical and fundamental science. With few exceptions, a more precise correspondence between physiological or biophysical properties and the obtained diffusion parameters remain uncertain due to lack of specificity. In this work, we address this problem by comparing diffusion parameters of a recently introduced model for water diffusion in brain matter to light microscopy and quantitative electron microscopy. Specifically, we compare diffusion model predictions of neurite density in rats to optical myelin staining intensity and stereological estimation of neurite volume fraction using electron microscopy. We find that the diffusion model describes data better and that its parameters show stronger correlation with optical and electron microscopy, and thus reflect myelinated neurite density better than the more frequently used diffusion tensor imaging (DTI) and cumulant expansion methods. Furthermore, the estimated neurite orientations capture dendritic architecture more faithfully than DTI diffusion ellipsoids.
The Anatomy of the Göttingen Minipig Hypothalamus
The microscopic organization of the Göttingen minipig (sus scrofa) hypothalamus was studied using Nissl stain, acetylcholinesterase histochemistry, and immunohistochemical staining for calretinin, tyrosin hydroxylase, oxytocin, vasopressin, and orexin A. Mediolaterally the minipig hypothalamus can be divided into three cytoarchitectonic distinct longitudinal zones. The periventricular longitudinal zone comprises the supraoptic, paraventricular, median preoptic, anteroventral periventricular, suprachiasmatic and arcuate nuclei. The medial longitudinal zone includes the prominent medial preoptic, ventromedial, dorsomedial and medial mammillary nuclei. Together with the anterior hypothalamic area, they can be further divided into distinct subregions. The dorsal and posterior hypothalamic areas and the retromammillary and lateral mammillary nuclei are cyto- and chemoarchitectonically distinct but cannot be further divided into subregions. The cell sparse, fiber rich lateral longitudinal zone comprises the lateral preoptic and lateral hypothalamic area as well as the perifornical, lateral tuberal and tuberomammillary nuclei. The findings presented here indicate that the cyto- and chemoarchitecture of the Göttingen minipig hypothalamus is comparable to that of rat, landrace pig, monkey, and human and that the Göttingen minipig may be well suited for future, non-primate, large mammal, hypothalamic research.
Safety and Function of a New Clinical Intracerebral Microinjection Instrument for Stem Cells and Therapeutics Examined in the Göttingen Minipig
A new intracerebral microinjection instrument (IMI) allowing multiple electrophysiologically guided microvolume injections from a single proximal injection path in rats has been adapted to clinical use by coupling the IMI to an FHC microTargeting Manual Drive, designed to be used with standard stereotactic frame-based systems and FHC frameless microTargeting Platforms.
Long-term Delivery of Nerve Growth Factor by Encapsulated Cell Biodelivery in the Göttingen Minipig Basal Forebrain
Nerve growth factor (NGF) prevents cholinergic degeneration in Alzheimer's disease (AD) and improves memory in AD animal models. In humans, the safe delivery of therapeutic doses of NGF is challenging. For clinical use, we have therefore developed an encapsulated cell (EC) biodelivery device, capable of local delivery of NGF. The clinical device, named NsG0202, houses an NGF-secreting cell line (NGC-0295), which is derived from a human retinal pigment epithelial (RPE) cell line, stably genetically modified to secrete NGF. Bioactivity and correct processing of NGF was confirmed in vitro. NsG0202 devices were implanted in the basal forebrain of Göttingen minipigs and the function and retrievability were evaluated after 7 weeks, 6 and 12 months. All devices were implanted and retrieved without associated complications. They were physically intact and contained a high number of viable and NGF-producing NGC-0295 cells after explantation. Increased NGF levels were detected in tissue surrounding the devices. The implants were well tolerated as determined by histopathological brain tissue analysis, blood analysis, and general health status of the pigs. The NsG0202 device represents a promising approach for treating the cognitive decline in AD patients.
A Surgical Device for Minimally Invasive Implantation of Experimental Deep Brain Stimulation Leads in Large Research Animals
Deep brain stimulation (DBS) in experimental animals has promoted new indications and refined existing treatments. Implantation of downscaled clinical DBS leads directly compatible with commercially available implantable pulse generators can however be challenging. Accordingly, we have developed a lead implantation device (LID) and technique for minimally invasive implantation of experimental multicontact DBS leads in large research animals.
Diffusion-weighted MRI and Quantitative Biophysical Modeling of Hippocampal Neurite Loss in Chronic Stress
Chronic stress has detrimental effects on physiology, learning and memory and is involved in the development of anxiety and depressive disorders. Besides changes in synaptic formation and neurogenesis, chronic stress also induces dendritic remodeling in the hippocampus, amygdala and the prefrontal cortex. Investigations of dendritic remodeling during development and treatment of stress are currently limited by the invasive nature of histological and stereological methods. Here we show that high field diffusion-weighted MRI combined with quantitative biophysical modeling of the hippocampal dendritic loss in 21 day restraint stressed rats highly correlates with former histological findings. Our study strongly indicates that diffusion-weighted MRI is sensitive to regional dendritic loss and thus a promising candidate for non-invasive studies of dendritic plasticity in chronic stress and stress-related disorders.
