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1.
Magn Reson Med ; 82(3): 984-999, 2019 09.
Article in English | MEDLINE | ID: mdl-31045280

ABSTRACT

PURPOSE: High-resolution three-dimensional (3D) structural MRI is useful for delineating complex or small structures of the body. However, it requires long acquisition times and high SAR, limiting its clinical use. The purpose of this work is to accelerate the acquisition of high-resolution images by combining compressed sensing and parallel imaging (CSPI) on a 3D-GRASE sequence and to compare it with a (CS)PI 3D-FSE sequence. Several sampling patterns were investigated to assess their influence on image quality. METHODS: The proposed k-space sampling patterns are based on two undersampled k-space grids, variable density (VD) Poisson-disc, and VD pseudo-random Gaussian, and five different trajectories described in the literature. Bloch simulations are performed to obtain the transform point spread function and evaluate the coherence of each sampling pattern. Image resolution was assessed by the full-width at half-maximum (FWHM). Prospective CSPI 3D-GRASE phantom and in vivo experiments in knee and brain are carried out to assess image quality, SNR, SAR, and acquisition time compared to PI 3D-GRASE, PI 3D-FSE, and CSPI 3D-FSE acquisitions. RESULTS: Sampling patterns with VD Poisson-disc obtain the lowest coherence for both PD-weighted and T2 -weighted acquisitions. VD pseudo-random Gaussian obtains lower FWHM, but higher sidelobes than VD Poisson-disc. CSPI 3D-GRASE reduces acquisition time (43% for PD-weighted and 40% for T2 -weighted) and SAR (∼45% for PD-weighted and T2 -weighted) compared to CSPI 3D-FSE. CONCLUSIONS: CSPI 3D-GRASE reduces acquisition time compared to a CSPI 3DFSE acquisition, preserving image quality. The design of the sampling pattern is crucial for image quality in CSPI 3D-GRASE image acquisitions.


Subject(s)
Data Compression/methods , Imaging, Three-Dimensional/methods , Magnetic Resonance Imaging/methods , Algorithms , Brain/diagnostic imaging , Humans , Knee/diagnostic imaging , Phantoms, Imaging
2.
Mol Psychiatry ; 16(3): 293-306, 2011 Mar.
Article in English | MEDLINE | ID: mdl-20048751

ABSTRACT

Strong genetic evidence implicates mutations and polymorphisms in the gene Disrupted-In-Schizophrenia-1 (DISC1) as risk factors for both schizophrenia and mood disorders. Recent studies have shown that DISC1 has important functions in both brain development and adult brain function. We have described earlier a transgenic mouse model of inducible expression of mutant human DISC1 (hDISC1) that acts in a dominant-negative manner to induce the marked neurobehavioral abnormalities. To gain insight into the roles of DISC1 at various stages of neurodevelopment, we examined the effects of mutant hDISC1 expressed during (1) only prenatal period, (2) only postnatal period, or (3) both periods. All periods of expression similarly led to decreased levels of cortical dopamine (DA) and fewer parvalbumin-positive neurons in the cortex. Combined prenatal and postnatal expression produced increased aggression and enhanced response to psychostimulants in male mice along with increased linear density of dendritic spines on neurons of the dentate gyrus of the hippocampus, and lower levels of endogenous DISC1 and LIS1. Prenatal expression only resulted in smaller brain volume, whereas selective postnatal expression gave rise to decreased social behavior in male mice and depression-like responses in female mice as well as enlarged lateral ventricles and decreased DA content in the hippocampus of female mice, and decreased level of endogenous DISC1. Our data show that mutant hDISC1 exerts differential effects on neurobehavioral phenotypes, depending on the stage of development at which the protein is expressed. The multiple and diverse abnormalities detected in mutant DISC1 mice are reminiscent of findings in major mental diseases.


Subject(s)
Brain , Gene Expression Regulation, Developmental/genetics , Mental Disorders/genetics , Mutation/genetics , Nerve Tissue Proteins/metabolism , Age Factors , Amphetamine , Analysis of Variance , Animals , Animals, Newborn , Behavior, Animal , Brain/embryology , Brain/growth & development , Brain/pathology , Brain/ultrastructure , Carrier Proteins/metabolism , Chromatography, High Pressure Liquid/methods , Disease Models, Animal , Dizocilpine Maleate , Dopamine/metabolism , Electrochemical Techniques/methods , Embryo, Mammalian , Exploratory Behavior/physiology , Female , Humans , Locomotion/drug effects , Locomotion/genetics , Magnetic Resonance Imaging , Male , Maze Learning/physiology , Mice , Mice, Transgenic , Nerve Tissue Proteins/genetics , Parvalbumins/metabolism , Phenotype , Pregnancy , Silver Staining/methods
3.
J Comp Neurol ; 518(11): 1892-907, 2010 Jun 01.
Article in English | MEDLINE | ID: mdl-20394049

ABSTRACT

The spatial organization of the mouse cerebellum into transverse zones and parasagittal stripes is reflected during the temporal progression of Purkinje cell death in the Lurcher mutant mouse (+/Lc). Neurodegeneration in the +/Lc mutant is apparent by the second postnatal week and is initially seen in all four transverse zones: the anterior (lobules I-V), central (lobules VI, VII), posterior (lobules VIII, dorsal IX), and nodular (ventral lobule IX and lobule X) zone. However, from postnatal day (P)25-P36, Purkinje cell loss proceeds more rapidly in the anterior zone, followed by the posterior and central zones, and is significantly delayed in the nodular zone. Coronal sections through the +/Lc cerebellum reveal that surviving Purkinje cells are restricted to the paraflocculus/flocculus and the nodular zone and could be detected as late as P146 (approximately 5 months). Within this region, the pattern of preferentially surviving calbindin-immunoreactive Purkinje cells reflects the expression of the constitutively expressed small heat shock protein HSP25 in the wild-type cerebellum. Although the role of constitutively expressed HSP25 in the wild-type cerebellum is not clear, it appears to play a neuroprotective role in the flocculonodular region of the +/Lc mutant cerebellum as the percentage of surviving Purkinje cells that are HSP25-immunopositive significantly increases over time.


Subject(s)
Cell Survival , Heat-Shock Proteins/metabolism , Neoplasm Proteins/metabolism , Nerve Degeneration/metabolism , Purkinje Cells/pathology , Purkinje Cells/physiology , Animals , Calbindins , Cerebellum/pathology , Heat-Shock Proteins/genetics , Mice , Mice, Neurologic Mutants , Molecular Chaperones , Neoplasm Proteins/genetics , Nerve Degeneration/genetics , Nerve Degeneration/pathology , Purkinje Cells/cytology , S100 Calcium Binding Protein G/metabolism
4.
Acta Radiol ; 49(1): 6-18, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18210307

ABSTRACT

The purpose of this article is to explain and illustrate the current status and potential applications of automated and color-coded post-processing techniques for the analysis of dynamic multiphasic gadolinium-enhanced magnetic resonance imaging (MRI) of the liver. Post-processing of these images on dedicated workstations allows the generation of time-intensity curves (TIC) as well as color-coded images, which provides useful information on (neo)-angiogenesis within a liver lesion, if necessary combined with information on enhancement patterns of the surrounding liver parenchyma. Analysis of TIC and color-coded images, which are based on pharmacokinetic modeling, provides an easy-to-interpret schematic presentation of tumor behavior, providing additional characteristics for adequate differential diagnosis. Inclusion of TIC and color-coded images as part of the routine abdominal MRI workup protocol may help to further improve the specificity of MRI findings, but needs to be validated in clinical decision-making situations. In addition, these tools may facilitate the diagnostic workup of disease for detection, characterization, staging, and monitoring of antitumor therapy, and hold incremental value to the widely used tumor response criteria.


Subject(s)
Contrast Media/administration & dosage , Gadolinium , Image Enhancement/methods , Image Processing, Computer-Assisted/methods , Liver Diseases/diagnosis , Liver Neoplasms/diagnosis , Magnetic Resonance Imaging/methods , Adult , Color , Female , Humans , Liver/pathology , Male , Middle Aged , Time Factors
5.
Dev Neurobiol ; 68(3): 332-48, 2008 Feb 15.
Article in English | MEDLINE | ID: mdl-18085563

ABSTRACT

The pro-apoptotic factor BAX has recently been shown to contribute to Purkinje cell (PC) apoptosis induced by the neurotoxic prion-like protein Doppel (Dpl) in the prion-protein-deficient Ngsk Prnp(0/0) (NP(0/0)) mouse. In view of cellular prion protein (PrP(c)) ability to counteract Dpl neurotoxicity and favor neuronal survival like BCL-2, we investigated the effects of the anti-apoptotic factor BCL-2 on Dpl neurotoxicity by studying the progression of PC death in aging NP(0/0)-Hu-bcl-2 double mutant mice overexpressing human BCL-2 (Hu-bcl-2). Quantitative analysis showed that significantly more PCs survived in NP(0/0)-Hu-bcl-2 double mutants compared with the NP(0/0) mutants. However, number of PCs remained inferior to wild-type levels and to the increased number of PCs observed in Hu-bcl-2 mutants. In the NP(0/0) mutants, Dpl-induced PC death occurred preferentially in the aldolase C-negative parasagittal compartments of the cerebellar cortex. Activation of glial cells exclusively in these compartments, which was abolished by the expression of Hu-bcl-2 in the double mutants, suggested that chronic inflammation is an indirect consequence of Dpl-induced PC death. This partial rescue of NP(0/0) PCs by Hu-bcl-2 expression was similar to that observed in NP(0/0):Bax(-/-) double mutants with bax deletion. Taken together, these data strongly support the involvement of BCL-2 family-dependent apoptotic pathways in Dpl neurotoxicity. The capacity of BCL-2 to compensate PrP(c) deficiency by rescuing PCs from Dpl-induced death suggests that the BCL-2-like property of PrP(c) may impair Dpl-like neurotoxic pathways in wild-type neurons.


Subject(s)
Apoptosis/drug effects , Prions/genetics , Proto-Oncogene Proteins c-bcl-2/physiology , Purkinje Cells/drug effects , Age Factors , Analysis of Variance , Animals , Cell Count , Cerebellum/cytology , Fructose-Bisphosphate Aldolase/metabolism , GPI-Linked Proteins , Gene Expression Regulation/drug effects , Gene Expression Regulation/genetics , Glial Fibrillary Acidic Protein/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , Prions/toxicity
6.
Phys Med Biol ; 52(20): N475-84, 2007 Oct 21.
Article in English | MEDLINE | ID: mdl-17921571

ABSTRACT

This note summarizes the characterization of the acoustic properties of four materials intended for the development of tissue, and especially breast tissue, phantoms for the use in photoacoustic and ultrasound imaging. The materials are agar, silicone, polyvinyl alcohol gel (PVA) and polyacrylamide gel (PAA). The acoustical properties, i.e., the speed of sound, impedance and acoustic attenuation, are determined by transmission measurements of sound waves at room temperature under controlled conditions. Although the materials are tested for application such as photoacoustic phantoms, we focus here on the acoustic properties, while the optical properties will be discussed elsewhere. To obtain the acoustic attenuation in a frequency range from 4 MHz to 14 MHz, two ultrasound sources of 5 MHz and 10 MHz core frequencies are used. For preparation, each sample is cast into blocks of three different thicknesses. Agar, PVA and PAA show similar acoustic properties as water. Within silicone polymer, a significantly lower speed of sound and higher acoustical attenuation than in water and human tissue were found. All materials can be cast into arbitrary shapes and are suitable for tissue-mimicking phantoms. Due to its lower speed of sound, silicone is generally less suitable than the other presented materials.


Subject(s)
Acoustics/instrumentation , Phantoms, Imaging , Polymers/chemistry , Ultrasonography/instrumentation , Equipment Design , Equipment Failure Analysis , Materials Testing , Scattering, Radiation
7.
Dev Neurobiol ; 67(5): 670-86, 2007 Apr.
Article in English | MEDLINE | ID: mdl-17443816

ABSTRACT

Research efforts to deduce the function of the prion protein (PrPc) in knock-out mouse mutants have revealed that large deletions in the PrPc genome result in the ectopic neuronal expression of the prion-like protein Doppel (Dpl). In our analysis of one such line of mutant mice, Ngsk Prnp0/0 (NP0/0), we demonstrate that the ectopic expression of Dpl in brain neurons induces significant levels of cerebellar Purkinje cell (PC) death as early as six months after birth. To investigate the involvement of the mitochondrial proapoptotic factor BAX in the Dpl-induced apoptosis of PCs, we have analyzed the progression of PC death in aging NP0/0:Bax-/- double knockout mutants. Quantitative analysis of cell numbers showed that significantly more PCs survived in NP0/0:Bax-/- double mutants than in the NP0/0:Bax+/+ mutants. However, PC numbers were not restored to wildtype levels or to the increased number of PCs observed in Bax-/- mutants. The partial rescue of NP0/0 PCs suggests that the ectopic expression of Dpl induces both BAX-dependent and BAX-independent pathways of cell death. The activation of glial cells that is shown to be associated topographically with Dpl-induced PC death in the NP0/0:Bax+/+ mutants is abolished by the loss of Bax expression in the double mutant mice, suggesting that chronic inflammation is an indirect consequence of Dpl-induced PC death.


Subject(s)
Apoptosis/physiology , Prions/physiology , Purkinje Cells/physiology , bcl-2-Associated X Protein/physiology , Animals , Calcium-Binding Proteins/metabolism , Cell Count , Female , Fluorescent Antibody Technique , GPI-Linked Proteins , Genotype , Glial Fibrillary Acidic Protein/metabolism , Gliosis/pathology , Image Processing, Computer-Assisted , Immunoenzyme Techniques , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Prions/genetics , Prions/metabolism , Purkinje Cells/metabolism , bcl-2-Associated X Protein/genetics
8.
J Comp Neurol ; 491(1): 69-78, 2005 Oct 10.
Article in English | MEDLINE | ID: mdl-16127699

ABSTRACT

Four transverse zones can be distinguished in the adult mouse cerebellar cortex based on differential expression of cell-specific antigens, termination patterns of mossy fiber afferents, and phenotypes of mouse mutants with cerebellar defects: the anterior zone (AZ), central zone (CZ), posterior zone (PZ), and nodular zone (NZ). In the heterozygous Lurcher (Lc/+) mouse a zonally restricted abnormality in Purkinje cell development is seen. The Purkinje cell-specific antigen zebrin II is normally differentially expressed in all four zones of the adult cerebellum, but in the Lc/+ mutant is confined to the PZ and NZ, caudal to a transverse boundary in the dorsal aspect of lobule VIII. In this study we wanted to understand why zebrin II expression is arrested at this boundary and whether the Lc mutation affects the differentiation of additional Purkinje cell antigens in a similar manner. To determine this, we took advantage of the dynamic developmental timetable of another Purkinje cell antigen, the small heat shock protein Hsp25. Using immunohistochemistry we demonstrate that cerebellar maturation anterior to the CZ/PZ transverse boundary appears to be unaffected by the Lc allele, in that initial progression of Hsp25 expression in the Lc/+ cerebellum was similar to controls. Double-labeling experiments with anti-Hsp25 and anti-calbindin suggest that characteristic banding patterns of Hsp25 in Lc/+ cerebellum develop and are preserved despite cell loss. Thus, since simple temporal or spatial models cannot account for the zonal restriction seen during Lc/+ cerebellar development, the abnormality may be zebrin II-specific.


Subject(s)
Cerebellum/growth & development , Heat-Shock Proteins/metabolism , Mice, Neurologic Mutants/metabolism , Neoplasm Proteins/metabolism , Nerve Tissue Proteins/metabolism , Purkinje Cells/metabolism , Animals , Calbindins , Cell Differentiation/genetics , Cell Differentiation/physiology , Cerebellum/cytology , Cerebellum/metabolism , Heterozygote , Immunohistochemistry , Mice , Molecular Chaperones , Purkinje Cells/cytology , S100 Calcium Binding Protein G/metabolism
9.
Article in English | MEDLINE | ID: mdl-16685893

ABSTRACT

Noninvasive temperature measurement is feasible with MRI to monitor changes in thermal therapy. Phase shift based MR thermometry gives an estimate of the relative temperature variation between thermal and baseline images. This technique is limited, however, when applied on targets under inter-frame motion. Simple image registration and subtraction are not adequate to recover the temperature properly since the phase shift due to temperature changes is corrupted by an unwanted phase shift. In this work, the unwanted phase shift is predicted from the raw registered phase shift map itself. To estimate the unwanted phase shift, a thin plate smoothing spline is fitted to the values outside the heated region. The spline value in the heated area serves as an estimate for the offset. The estimation result is applied to correct errors in the temperature maps of an ex-vivo experiment.


Subject(s)
Artifacts , Image Enhancement/methods , Image Interpretation, Computer-Assisted/methods , Information Storage and Retrieval/methods , Magnetic Resonance Imaging, Cine/methods , Movement , Thermography/methods , Algorithms , Animals , Hot Temperature , Hyperthermia, Induced/methods , In Vitro Techniques , Liver/physiology , Liver/radiation effects , Reproducibility of Results , Sensitivity and Specificity , Swine
10.
MAGMA ; 17(3-6): 296-302, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15480945

ABSTRACT

The objective of this study was to evaluate the potential of dynamic contrast-enhanced MRI for quantitative characterization of tumor microvessels and to assess the microvascular changes in response to isolated limb perfusion with TNF-alpha and melphalan. Dynamic contrast-enhanced MRI was performed in an experimental cancer model, using a macromolecular contrast medium, albumin-(Gd-DTPA)45. Small fragments of BN 175, a soft-tissue sarcoma, were implanted in 11 brown Norway (BN) rats. Animals were assigned randomly to a control (Haemaccel) or drug-treated group (TNF-alpha/melphalan). MRI was performed at baseline and 24 h after ILP. The transendothelial permeability (K(PS)) and the fractional plasma volume (fPV) were estimated from the kinetic analysis of MR data using a two-compartment bi-directional model. K(PS) and fPV decreased significantly in the drug-treated group compared to baseline (p<0.05). In addition, K(PS) post therapy was significantly lower (p<0.05) in the drug-treated group than in the control group. There was no significant difference in fPV between the drug-treated and the control group after therapy. Tumor microvascular changes in response to isolated limb perfusion can be determined after 24 h by dynamic contrast-enhanced MRI. The data obtained in this experimental model suggest possible applications in the clinical setting, using the appropriate MR contrast agents.


Subject(s)
Albumins , Contrast Media , Gadolinium DTPA , Image Enhancement/methods , Magnetic Resonance Imaging/methods , Melphalan/administration & dosage , Sarcoma, Experimental/diagnosis , Sarcoma, Experimental/drug therapy , Tumor Necrosis Factor-alpha/administration & dosage , Animals , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Chemotherapy, Cancer, Regional Perfusion/methods , Extremities/blood supply , Macromolecular Substances , Male , Prognosis , Rats , Rats, Inbred BN , Treatment Outcome
11.
Psychopharmacology (Berl) ; 174(2): 177-89, 2004 Jul.
Article in English | MEDLINE | ID: mdl-14985933

ABSTRACT

RATIONALE: Sensorimotor gating disruption is one of many neurocognitive deficits seen in schizophrenia. Disorganized thought is one of the cardinal symptoms associated with sensorimotor gating. In an attempt to model sensorimotor gating deficits in rats relevant to the neurodevelopmental hypothesis for schizophrenia, we have used prenatal injections of the antimitotic drug, cytosine arabinoside (Ara-C) to subtly perturb the development of the rat CNS and disrupt sensorimotor gating. OBJECTIVE: To produce rats with either basal sensorimotor gating deficits or increased vulnerability to the disruption of sensorimotor function by apomorphine or phencyclidine (PCP). Prepulse inhibition (PPI) of the acoustic startle response was used to assess sensorimotor gating. METHODS: Three different cohorts of pregnant Sprague Dawley female rats were injected with Ara-C (30 mg/kg in saline) or saline at embryonic days 19.5 and 20.5. The Ara-C and control rats were tested for acoustic startle response and PPI at preadolescent and post-adolescent ages; postnatal day (Pnd) 35 and 56, respectively. Apomorphine (2.0 mg/kg) or phencyclidine (3.0 mg/kg), was given prior to PPI sessions in order to disrupt PPI. RESULTS: At Pnd 35, Ara-C treatment did not significantly affect acoustic startle amplitudes or PPI. However, at PND 56, Ara-C treated rats had significantly lower acoustic startle amplitudes and significantly diminished sensorimotor gating. Pharmacological challenge with the dopamine agonist apomorphine and the glutamate antagonist PCP significantly disrupted sensorimotor gating in the control subjects. Apomorphine did not further disrupt the existing deficit in the Ara-C treated rats. Ara-C treatment did not cause gross loss of neuronal tissue, although there was a subtle and variable disorganization of the pyramidal cell layer in the hippocampal CA2/3 region. CONCLUSION: The results provide evidence to suggest that late embryonic exposure to Ara-C disrupts the circuitry involved in mediating PPI. While the dopamine agonist apomorphine caused a significant disruption in the control rats it did not further disrupt the existing deficit in the Ara-C treated rats. These data provide evidence to support the contention that modest neurodevelopmental insults can significantly affect sensorimotor gating processes in an adult onset dependent manner.


Subject(s)
Antineoplastic Agents/toxicity , Apomorphine/pharmacology , Brain/growth & development , Cytarabine/toxicity , Dopamine Agonists/pharmacology , Hallucinogens/pharmacology , Phencyclidine/pharmacology , Schizophrenia/chemically induced , Animals , Brain/drug effects , Disease Models, Animal , Drug Interactions , Female , Male , Maternal-Fetal Exchange , Pregnancy , Rats , Rats, Sprague-Dawley , Reflex, Startle/drug effects
12.
Neuroreport ; 12(14): 3039-43, 2001 Oct 08.
Article in English | MEDLINE | ID: mdl-11568633

ABSTRACT

+/Lc Purkinje cells degenerate postnatally because of a gain-of-function mutation in the delta2 glutamate receptor (Grid2) that causes a constitutive Na+ current leak. The effect of the resulting chronic depolarization on Purkinje cell metabolism was investigated by measuring levels of cytochrome oxidase (COX) activity in Purkinje cell dendrites using quantitative densitometry. Analysis of wild type controls and +/Lc mutants at P10, P15 and P25 showed that levels of COX activity were significantly increased above control levels by P15 and continued to increase through P25. The increase in COX activity is likely to reflect an increase in oxidative phosphorylation to accommodate the energy demands of removing excess Na+ and Ca2+ entering the Purkinje cells in response to the Grid2 leak current.


Subject(s)
Electron Transport Complex IV/metabolism , Membrane Potentials/genetics , Mice, Neurologic Mutants/metabolism , Nerve Degeneration/enzymology , Purkinje Cells/enzymology , Receptors, Glutamate/genetics , Up-Regulation/genetics , Aging/genetics , Animals , Animals, Newborn/metabolism , Dendrites/enzymology , Dendrites/pathology , Electron Transport Complex IV/genetics , Energy Metabolism/genetics , Female , Histocytochemistry , Male , Mice , Nerve Degeneration/genetics , Nerve Degeneration/physiopathology , Purkinje Cells/pathology , Receptors, Glutamate/metabolism
13.
J Comp Neurol ; 436(1): 82-91, 2001 Jul 16.
Article in English | MEDLINE | ID: mdl-11413548

ABSTRACT

Cerebellar Purkinje cells and granule cells have been studied extensively as models for investigating neuron-target interactions and the regulation of cell numbers in the developing central nervous system. Recent studies of transgenic mice that overexpress a human Bcl-2 transgene in Purkinje cells suggest both that programmed cell death plays an unexpected role in regulating Purkinje cell number and that Purkinje cells influence the number of granule cells. The role of cell death-related proteins and Purkinje-granule cell interactions in cerebellar development was investigated further in this study by counting the number of Purkinje and granule cells in knockout mutants with a deletion in the proapoptotic gene, Bax. The total number of Purkinje cells was estimated using stereological counting principles in six adult wild type mice, four hemizygous Bax +/- controls, and six Bax -/- knockout mutants. The total number of granule cells per cerebellum was estimated in three adult wild type mice, three hemizygous Bax +/- controls, and three Bax -/- knockout mutants. The number of Purkinje cells increased significantly by over 30% in the Bax -/- knockout mutants compared with wild type and hemizygote controls, whereas the number of granule cells was unchanged in the Bax -/- mutants. There was no change in the volume of the cerebellar cortex or in the size of Purkinje cell bodies in the Bax -/- mutants, implying that Purkinje cell density was increased in the Bax -/- mutants. The increase in Purkinje cell numbers in the Bax -/- knockout mice supports previous evidence that Purkinje cells undergo a period of naturally occurring cell death that is mediated at least in part by the cell death proteins Bcl-2 and Bax. The lack of an effect of Bax gene expression on granule cell numbers indicates that Bax is not an obligate participant in naturally occurring cell death in granule cells.


Subject(s)
Cerebellum/cytology , Neurons/cytology , Proto-Oncogene Proteins c-bcl-2 , Proto-Oncogene Proteins/deficiency , Analysis of Variance , Animals , Cell Count , Cell Size , Cerebellum/metabolism , Dendrites/ultrastructure , Heterozygote , Homozygote , Mice , Mice, Knockout , Neurons/classification , Neurons/metabolism , Proto-Oncogene Proteins/genetics , Purkinje Cells/cytology , Purkinje Cells/metabolism , bcl-2-Associated X Protein
14.
J Neurosci ; 20(14): 5339-45, 2000 Jul 15.
Article in English | MEDLINE | ID: mdl-10884318

ABSTRACT

Lurcher is a gain-of-function mutation in the delta2 glutamate receptor gene (Grid2) that turns the receptor into a leaky ion channel. The expression of the Lurcher gene in heterozygous (Grid2(Lc/+)) mutants induces the death of almost all Purkinje cells starting from the second postnatal week. Ninety percent of the granule cells and 60-75% of the inferior olivary neurons die because of the loss of their target neurons, the Purkinje cells. The apoptotic nature of the neurodegeneration has been demonstrated previously by the presence of activated caspase-3 and DNA fragmentation. Bax, a pro-apoptotic gene of the Bcl-2 family, has been shown to be involved in developmental neuronal death. To study the role of Bax in Grid2(Lc/+) neurodegeneration, double mutants with Grid2(Lc/)+ mice and Bax knock-out mice (Bax-/-) were generated. Bax deletion had no effect on the death of Purkinje cells and inferior olivary neurons, although a temporary rescue of some Purkinje cells could be detected in P15 Grid2(Lc/)+;Bax-/- animals. From postnatal day 15 (P15) to P60, the number of granule cells in Grid2(Lc/)+;Bax-/-mice did not significantly change and was significantly increased compared with the number found in Grid2(Lc/)+;Bax+/+ mice. Granule cell number in P60 Grid2(Lc/)+;Bax-/- mice corresponded to 70% of the number found in wild-type mice. Our results show that Bax inactivation in Grid2(Lc/+) mice does not rescue intrinsic Purkinje cell death or the target-related cell death of olivary neurons, but Bax inactivation does inhibit persistently target-related cell death in cerebellar granule cells.


Subject(s)
Cerebellum/metabolism , Neurons/metabolism , Olivary Nucleus/metabolism , Proto-Oncogene Proteins c-bcl-2 , Proto-Oncogene Proteins/deficiency , Purkinje Cells/metabolism , Animals , Apoptosis/genetics , Cell Count , Cell Death/genetics , Cerebellum/pathology , Female , Gene Deletion , Male , Mice , Mice, Knockout , Mice, Neurologic Mutants , Neurons/pathology , Olivary Nucleus/pathology , Proto-Oncogene Proteins/genetics , Purkinje Cells/pathology , bcl-2-Associated X Protein
15.
J Neurobiol ; 42(3): 323-37, 2000 Feb 15.
Article in English | MEDLINE | ID: mdl-10645972

ABSTRACT

Staggerer (sg/sg) is an autosomal recessive mutation in an orphan nuclear hormone receptor gene, RORalpha, that causes a cell-autonomous, lineage-specific block in the development of the Purkinje cell. Purkinje cell number is reduced by about 75-90% in adult mutants, and many of the surviving cells are small and ectopically positioned. To determine whether Purkinje cell numbers are reduced owing to either agenesis or cell death, cohorts of Purkinje cells were labeled with the birth-date marker bromodeoxyuridine (BrdU) at embryonic day (E) 10.5 or E11.5. The total number of BrdU-labeled profiles was then compared between cerebella from wild-type mice, heterozygous staggerer, and staggerer mutants at E17.5 and postnatal day (P)5. There was no significant difference between sg/sg mutants and +/sg or +/+ controls in the number of BrdU-labeled profiles or in cerebellar volumes in the E17 embryos. By P5, however, cerebellar volume was significantly reduced in the sg/sg mutants compared to controls (p <.005) and the number of BrdU-labeled profiles was reduced by 33% following E11.5 BrdU injections (p <.02). The results suggest that Purkinje cell genesis is not affected by the staggerer mutation and that Purkinje cell loss begins some time after E17. RORalpha is highly expressed in Purkinje cells by E14, so the delay between initial RORalpha expression and sg/sg Purkinje cell loss suggests that the staggerer mutation does not directly cause Purkinje cell death.


Subject(s)
Aging , Cerebellum/abnormalities , Embryonic and Fetal Development , Purkinje Cells/cytology , Animals , Calbindins , Cell Death , Cerebellum/embryology , Cerebellum/growth & development , Genes, Recessive , Heterozygote , Mice , Mice, Inbred Strains , Mice, Neurologic Mutants , Nerve Tissue Proteins/analysis , Nuclear Receptor Subfamily 1, Group F, Member 1 , Purkinje Cells/pathology , Purkinje Cells/physiology , Receptors, Cytoplasmic and Nuclear/genetics , S100 Calcium Binding Protein G/analysis , S100 Calcium Binding Protein G/metabolism , Trans-Activators/genetics
16.
C R Acad Sci III ; 323(11): 967-73, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11144029

ABSTRACT

Intrinsic Purkinje cell death in heterozygous Lurcher (Grid2Lc/+) mice is accompanied by the target-related death of granule cells and olivary neurons. The expression of pro-caspase-3 is increased in Grid2Lc/+ Purkinje cells and activated caspase-3 is detected in all three cell types before their death. Bax inactivation in Grid2Lc/+ mutants rescues granule cells but not Purkinje cells. Here, we show that, while Bax inactivation inhibits caspase-3 activation in both cell types, p53 inactivation does not affect caspase-3 activation and neuronal loss in Grid2Lc/+ mice. The up-regulation of pro-caspase-3 in Grid2Lc/+ Purkinje cells is Bax and p53 independent. These results suggest that Grid2Lc/+ granule cell death is dependent on Bax and caspase-3 activation, whereas several pathways can mediate Grid2Lc/+ Purkinje cell death.


Subject(s)
Brain/pathology , Caspases/genetics , Caspases/metabolism , Nerve Degeneration/physiopathology , Neurons/physiology , Proto-Oncogene Proteins c-bcl-2 , Proto-Oncogene Proteins/physiology , Tumor Suppressor Protein p53/physiology , Animals , Apoptosis , Brain/physiopathology , Caspase 3 , Crosses, Genetic , Gene Expression Regulation, Enzymologic , Mice , Mice, Knockout , Mice, Neurologic Mutants , Nerve Degeneration/genetics , Nerve Degeneration/pathology , Neurons/pathology , Proto-Oncogene Proteins/deficiency , Proto-Oncogene Proteins/genetics , Purkinje Cells/pathology , Purkinje Cells/physiology , Receptors, Glutamate/genetics , Receptors, Glutamate/physiology , Tumor Suppressor Protein p53/deficiency , Tumor Suppressor Protein p53/genetics , bcl-2-Associated X Protein
17.
J Neurosci ; 19(13): 5370-9, 1999 Jul 01.
Article in English | MEDLINE | ID: mdl-10377347

ABSTRACT

To explore the role of Engrailed proteins in development of the cerebellum, Engrailed-2 (En-2) was ectopically expressed in cerebellar Purkinje cells from the late embryonic stage into adulthood. The fundamental organization of Purkinje cell sagittal zones as revealed by the "early onset" markers L7-beta-gal and cadherin-8 was found to be virtually identical to that in wild type. In contrast, "late onset" sagittal banding patterns revealed by Purkinje cell markers zebrin I, zebrin II, and 9-O-acetyl GD3 Ganglioside (P-Path), and the granule cell marker NADPH-diaphorase, were disrupted. In general, although some evidence of banding was still detectable, boundaries defined by the latter markers were poorly defined, and the patterns overall took on a diffuse appearance. In parallel with the changes in late onset markers, anterograde tracing of spinocerebellar axons revealed a general diffusion of the mossy fiber projection pattern in lobule VIII and the anterior lobe. These observations suggest that at least two separate mediolateral boundary systems exist in the cerebellum, and these are differentially affected by ectopic En-2 expression. Alternatively, one boundary system exists that remains primarily intact in the mutant, but recognition of this system by a set of late developmental events is perturbed.


Subject(s)
Cerebellar Cortex/growth & development , Gene Expression , Homeodomain Proteins/physiology , Nerve Tissue Proteins/physiology , Purkinje Cells/metabolism , Animals , Axons/metabolism , Body Patterning , Cadherins/analysis , Cells, Cultured , Cerebellar Cortex/embryology , Gangliosides/analysis , Gene Expression Regulation, Developmental , Guanine Nucleotide Exchange Factors , Homeodomain Proteins/genetics , Mice , Mice, Transgenic , Mossy Fibers, Hippocampal/metabolism , NADPH Dehydrogenase/analysis , Nerve Tissue Proteins/analysis , Nerve Tissue Proteins/genetics , Neural Pathways/metabolism , Neuropeptides/genetics , Promoter Regions, Genetic/genetics , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/physiology
19.
J Neurosci ; 18(1): 319-27, 1998 Jan 01.
Article in English | MEDLINE | ID: mdl-9412510

ABSTRACT

The Lurcher mutant has been extensively studied as a model for cell-autonomous and target-related cell death, yet there are still many unknowns concerning the mechanisms of neuronal degeneration in this mutant. As a key regulator of apoptosis, a bcl-2 transgene has been overexpressed in the heterozygous Lurcher mutant to investigate the effects of BCL-2 on two types of in vivo neuronal cell loss in Lurcher: cell-autonomous Purkinje cell degeneration and target-related olivary neuron death. Six adult +/Lc mutants expressing a human bcl-2 transgene (Hu-bcl-2) were generated by crossing +/Lc mutants with NSE71 Hu-bcl-2 transgenic mice. Analysis of these brains showed that bcl-2 overexpression did not prevent +/Lc Purkinje cell degeneration, but it did rescue most olivary neurons from target-related cell death. Although the number of olivary neurons was equivalent to wild-type numbers, the inferior olive nucleus was significantly shorter in its rostrocaudal extent, suggesting that olivary neurons are atrophied. We propose that Lurcher gene action causes Purkinje cell degeneration independently of a BCL-2-mediated pathway. Furthermore, although bcl-2 overexpression rescues olivary neurons from target-related cell death, it does not prevent the atrophy associated with the loss of target-related trophic support.


Subject(s)
Apoptosis/physiology , Mice, Neurologic Mutants , Olivary Nucleus/cytology , Proto-Oncogene Proteins c-bcl-2/genetics , Purkinje Cells/cytology , Animals , Cerebellum/pathology , Female , Humans , Male , Mice , Mice, Transgenic , Mutagenesis/physiology , Nerve Degeneration/genetics , Purkinje Cells/physiology
20.
J Neurobiol ; 32(5): 502-16, 1997 May.
Article in English | MEDLINE | ID: mdl-9110261

ABSTRACT

Neuron-target interactions during development are critical for determining the final numbers of neurons in the nervous system. To investigate the role of Purkinje cells and programmed cell death in the regulation of afferent neuron numbers, we have counted olivary neurons and granule cells in two lines of transgenic mice (NSE73a and NSE71) that overexpress a human gene for bcl-2 (Hu-bcl-2) in Purkinje cells and olivary neurons, but not in granule cells. Bcl-2 overexpression in vivo reduces naturally occurring neuronal cell death and cell death following axotomy, target removal, or ischemia. Olivary neuron numbers in NSE73a and NSE71 transgenic mice are significantly increased compared to controls by 28% and 27%, respectively, while granule cell numbers are only increased in NSE73a mice (29% above controls). We have previously shown that Purkinje cell number is increased by 43% in NSE73a transgenics and by 23% in NSE71 transgenics. The ratio of Purkinje cells to olivary neurons is not significantly different between the control and transgenic mice, while the ratio of granule cells to Purkinje cells is significantly decreased in the NSE71 transgenic mice compared to controls and NSE73a transgenics. The increased numbers of olivary neurons suggest that bcl-2 overexpression rescues these neurons from programmed cell death. The increase in granule cell number in only one transgenic line is discussed with respect to hypotheses that Purkinje cells regulate both granule cell progenitor proliferation and the survival of differentiated granule cells.


Subject(s)
Cerebellum/cytology , Neurons, Afferent/cytology , Neurons, Afferent/enzymology , Olivary Nucleus/cytology , Proto-Oncogene Proteins c-bcl-2/genetics , Animals , Apoptosis/physiology , Cell Count , Cell Size/physiology , Female , Gene Expression Regulation/physiology , Humans , Male , Mice , Mice, Inbred C57BL , Mice, Transgenic , Neurons, Afferent/chemistry , Phosphopyruvate Hydratase/metabolism , Transgenes/physiology
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