Compressed Sensing 3D-GRASE for faster High-Resolution MRI.

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.

Differential effects of prenatal and postnatal expressions of mutant human DISC1 on neurobehavioral phenotypes in transgenic mice: evidence for neurodevelopmental origin of major psychiatric disorders.

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.

Post-processing of dynamic gadolinium-enhanced magnetic resonance imaging exams of the liver: explanation and potential clinical applications for color-coded qualitative and quantitative analysis.

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.

Acoustical properties of selected tissue phantom materials for ultrasound imaging.

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.

Development of Hsp25 expression compartments is not constrained by Purkinje cell defects in the Lurcher mouse mutant.

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.

Numerical matching in the mammalian CNS: lack of a competitive advantage of early over late-generated cerebellar granule cells.

In this report we use postnatal 3H-thymidine injections to test whether granule cells that are generated early in postnatal cerebellar development and whose axons have access to their Purkinje cell target beginning in the first postnatal week have an advantage over granule cells generated 9 days later in the competition for target-related stabilization. In the wild-type mouse, 3-5% of the adult granule cell population is labeled by injection of 3H-thymidine at either postnatal day 4 (P4) or P13. In the lurcher mutant, however, over 40% of the surviving granule cells are labeled by P4 injection while less than 1% are labeled after a P13 injection. Together, these results suggest that time of target contact is a critical factor in the competition for neuronal survival. The results from the lurcher chimeras, however, reveal that the situation is likely to be more complicated. In all chimeras examined, with target sizes ranging from 3 to 108% of wild type, equivalent numbers of granule cells were labeled at P4 and P13. These data lead to the contradictory conclusion that, in this experimental situation, early generated granule cells do not have a competitive advantage over later-generated granule cells. The results are discussed in terms of various models of target stabilization. We propose that, of the various hypotheses, our results are best explained by postulating two distinct mechanisms for developmental cell death. Supporting evidence for this hypothesis from other neuronal systems is also briefly reviewed.

Topographic spinocerebellar mossy fiber projections are maintained in the lurcher mutant.

A variety of recent studies of cerebellar development have focused attention on the role of Purkinje cells as organizing elements for the topography of afferent fiber connectivity in the cerebellum. We have investigated the involvement of Purkinje and granule cells in the maintenance of topographic spinocerebellar mossy fiber projections by analyzing the distribution of spinocerebellar mossy fiber terminals in lurcher (+/Lc) mutant mice. Purkinje cells in the +/Lc mutant degenerate starting after the first week of postnatal development because of an intrinsic genetic defect. The loss of their Purkinje cell targets also results in the death of 90% of the granule cells. We examined the distribution of spinocerebellar mossy fiber terminals in the juvenile and adult +/Lc mutant to determine how the pattern of afferent projections is affected by the loss of Purkinje cells shortly after innervation of the cerebellum. Labeling of spinocerebellar mossy fiber terminals with WGA-HRP in the P38 and adult +/Lc mutant showed that, despite the loss of almost all Purkinje cells and 90% of the granule cells, spinocerebellar mossy fibers project to the appropriate folia and segregate into relatively normal parasagittal bands. While we cannot rule out the possibility that Purkinje cells may be involved in the initial establishment of topographic maps, our results indicate that Purkinje cells are not necessary for the maintenance of the normal spinocerebellar mossy fiber topographic map.

Elimination of Bax expression in mice increases cerebellar purkinje cell numbers but not the number of granule cells.

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.

Evidence of spinocerebellar mossy fiber segregation in the juvenile staggerer cerebellum.

Developmental and experimental studies of climbing fiber and mossy fiber connectivity in the cerebellum have suggested that Purkinje cells are the critical organizing elements for connectivity patterns. This hypothesis is supported by evidence that spinocerebellar mossy fiber projections are abnormally diffuse in P25 sg/sg mutant mice in which the differentiation of a reduced number of sg/sg Purkinje cells is blocked due to a cell autonomous defect. However, mossy fiber distribution may be disrupted in sg/sg mutants not because of the Purkinje cell deficits, but because of the death of virtually all granule cells following the 4th postnatal week. To test this hypothesis, we have analyzed the distribution of wheat germ agglutinin-horseradish peroxidase (WGA-HRP)-labeled spinocerebellar mossy fiber terminals in sg/sg mutants at the end of the period of granule cell genesis (postnatal day [P] 12-P13) and before massive granule cell death (P16). Two percent WGA-HRP was injected into the lower thoracic/upper lumbar region of the spinal cord of eight homozygous sg/sg mutants (P12-P16) and five controls (+/sg and +/+). We have found that spinocerebellar mossy fibers segregate into distinct terminal fields in the anterior cerebellar lobules of P12 to P16 sg/sg mutants, although the medial-lateral distribution of spinocerebellar mossy fiber projections is different from controls. The results from this study and previous analysis of sg/sg mutants support the hypothesis that topographic cues are expressed in the early postnatal staggerer mutant, but mossy fiber terminals become disorganized or retract as granule cells die in the older staggerer mutant. J. Comp. Neurol. 378:354-362, 1997.

Rapid growth of parallel fibers in the cerebella of normal and staggerer mutant mice.

The growth of cerebellar granule cell axons was examined by placing focal implants of 1,1',dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (DiI) in the cerebella of normal and staggerer mutant mice at a series of developmental ages between postnatal day 2 (P2) and P30. Parallel fibers contacting the implant site were brightly labeled by the fluorescent dye, as were the associated granule cell bodies located principally in the internal granule layer. The extent of parallel fiber labeling in the molecular layer and the distance from the implant to the most extreme labeled granule cells were measured in sectioned material. Two additional measures describing the distribution of labeled granule cells about the implant site suggest length bounds for most parallel fibers. Parallel fiber growth is surprisingly rapid; all measures approached peak values at P3-P5, only a few days after the earliest postmitotic granule cells differentiate and migrate. At intermediate ages (P8 and P10), parallel fiber lengths appeared to decrease transiently. At later ages (P15 and beyond), the measures of fiber length increased to their mature values. These values differed little from lengths measured at P3-P5, suggesting that most parallel fiber growth occurs within a few days of cell birth. At early and intermediate ages, parallel fiber lengths in staggerer mice were comparable to controls, suggesting that an interaction with normal healthy Purkinje cells is not essential for parallel fiber outgrowth.

Increased cerebellar Purkinje cell numbers in mice overexpressing a human bcl-2 transgene.

The Purkinje cell is a primary organizer in the development of the cerebellum. Purkinje cells may provide positional information cues that regulate afferent innervation, and Purkinje cell target size controls the adult number of afferent olivary neurons and granule cells. While Purkinje cells are necessary for the survival of olivary neurons and granule cells during periods of programmed cell death, little is known about the survival requirements of Purkinje cells in vivo. To determine if Purkinje cells are subject to programmed cell death during development we have analyzed Purkinje cell numbers in two lines of transgenic mice that overexpress a human gene for bcl-2 (Hu-bcl-2). Bcl-2 is a protooncogene that inhibits apoptosis in many cell types. Overexpression of bcl-2 in vitro and in vivo rescues neurons from trophic factor deprivation or naturally occurring cell death. In the mice analyzed in this study, transgene expression is driven by the neuron-specific enolase promoter that is first expressed embryonically in most regions of the brain in one line and postnatally in the second line. We have counted Purkinje cells in three adult control mice, five early overexpressing transgenics, and three late expressing transgenics. The number of Purkinje cells in the Hu-bcl-2 transgenic mice is significantly increased above control numbers, with an increase of 43% in the embryonically overexpressing line and an increase of 27% in the postnatally overexpressing line. Because bcl-2 overexpression has been shown to rescue other neurons from programmed cell death, the increase in Purkinje cell numbers in overexpressing bcl-2 transgenics suggests that Purkinje cells undergo a period of cell death during normal development.

Altered prepulse inhibition in rats treated prenatally with the antimitotic Ara-C: an animal model for sensorimotor gating deficits in schizophrenia.

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.

Cytochrome oxidase activity is increased in +/Lc Purkinje cells destined to die.

+/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.

Cerebellar Purkinje cells provide target support over a limited spatial range: evidence from lurcher chimeric mice.

The distribution of Purkinje cells, granule cells, and olivary neurons was quantitatively analyzed in a lurcher +/Lc in equilibrium C3H/HeJ chimera in which the surviving wild type Purkinje cells were unilaterally distributed in the left hemicerebella. The left hemisphere of this mouse contains 7600 Purkinje cells, approximately 10% of the number of Purkinje cells in inbred C3H/HeJ mice. The right hemisphere contains 300 Purkinje cells, all of which are found within 200 microns of the midline. As in other +/Lc in equilibrium wild type chimeras, the ratio of granule cells to Purkinje cells is increased in the left hemisphere, reflecting increased granule cell survival. In the right hemisphere, however, the number of granule cells is reduced to that found in +/Lc mutants. In the inferior olive, almost twice as many neurons are found in the right nucleus as opposed to the left nucleus. As the projections of olivary neurons are crossed, the number of olivary neurons is increased in the nuclei that project to the cerebellar hemisphere containing Purkinje cells compared to the olivary nuclei that project to the cerebellar hemisphere with almost no Purkinje cells. The preferential survival of granule cells and olivary neurons that either occupy or project to the hemicerebellum containing Purkinje cells suggests that the availability of trophic support from target Purkinje cell neurons is spatially restricted.

Spinocerebellar mossy fiber terminal topography in the NR2C/PKC gamma double mutant cerebellum.

The spatiotemporal expression patterns of the NR2C subunit of the NMDA receptor and PKC gamma isoform during cerebellar development suggests that both proteins are involved in the molecular mechanisms of synaptogenesis. However, the topographic distribution of WGA-HRP labeled spinocerebellar mossy fiber terminals in NR2C/PKC gamma double mutants (n = 4) appears similar to controls (n = 3). While the results do not rule out a role for NR2C receptor subunits and the PKC gamma isoform in cerebellar synaptogenesis, they indicate that neither is necessary for the formation or maintenance of normal spinocerebellar mossy fiber afferent maps.

The Engrailed-2 homeobox gene and patterning of spinocerebellar mossy fiber afferents.

The mouse Engrailed-2 gene, En-2, appears to be involved in cerebellar pattern formation. Homozygous null mutants for En-2 have abnormal foliation patterns in the posterior half of the cerebellum and there are changes in Purkinje and granule cell gene expression in some posterior folia, possibly reflecting changes in cell identity. We have examined the distribution of spinocerebellar mossy fiber terminals in homozygous En-2hd null mutants to determine if En-2 is involved in regulating the pattern of afferent connectivity in the cerebellum. Spinocerebellar mossy fiber terminals were labeled following WGA-HRP injections in the lumbar region of 5 homozygous En-2hd mutants and 4 heterozygous controls. The distribution of spinocerebellar mossy fiber terminals was consistently altered in lobules VIII and IX of the En-2hd mutants. The principal changes were a reduction in the number of mossy fiber terminal fields in the dorsal aspect of lobule VIII and the dorsal midline field in lobule IX was fused into a single compartment. The results suggest that the deletion of En-2 expression does not transform lobule identity, at least with respect to afferent fiber positional information cues. However, the changes in foliation and afferent connectivity in the En-2 mutant support a broad role for the En-2 gene in cerebellar patterning.

Heat shock protein 25 expression and preferential Purkinje cell survival in the lurcher mutant mouse cerebellum.

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.

BCL-2 counteracts Doppel-induced apoptosis of prion-protein-deficient Purkinje cells in the Ngsk Prnp(0/0) mouse.

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.

BAX contributes to Doppel-induced apoptosis of prion-protein-deficient Purkinje cells.

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.