Neurological Disorder In Mice Research Articles

Neuronal survival factor TAFA2 suppresses apoptosis through binding to ADGRL1 and activating cAMP/PKA/CREB/BCL2 signaling pathway

AimsTAFA2, a cytokine specifically expressed in the central nervous system, plays a vital role in neuronal cell survival. TAFA2 deficiency has been correlated to various neurological disorders in mice and humans. However, the underlying mechanism remains elusive, especially its membrane-binding receptor through which TAFA2 functions. This study aimed to identify the specific binding receptor responsible for the anti-apoptotic effects of TAFA2. Main methodCo-immunoprecipitation (Co-IP) and quantitative mass spectrometry-based proteomic analysis were employed to identify potential TAFA2 binding proteins in V5 knockin mouse brain lysates. Subsequent validation involved in vitro and in vivo Co-IP and pull-down using specific antibodies. The functional analysis included evaluating the effects of ADGRL1 knockout, overexpression, and Lectin-like domain (Lec) deletion mutant on TAFA2's anti-apoptotic activity and analyzing the intracellular signaling pathways mediated by TAFA2 through ADGRL1. Key findingsOur study identified ADGRL1 as a potential receptor for TAFA2, which directly binds to TAFA2 through its lectin-like domain. Overexpression ADGRL1, but not ADGRL1ΔLec, induced apoptosis, which could be effectively suppressed by recombinant TAFA2 (rTAFA2). In ADGRL1−/− cells or re-introducing with ADGRL1ΔLec, responses to rTAFA2 in suppressing cell apoptosis were compromised. Increased cAMP, p-PKA, p-CREB, and BCL2 levels were also observed in response to rTAFA2 treatment, with these responses attenuated in ADGRL1−/− or ADGRL1ΔLec-expressing cells. SignificanceOur results demonstrated that TAFA2 directly binds to the lectin-like domain of ADGRL1, activating cAMP/PKA/CREB/BCL2 signaling pathway, which is crucial in preventing cell death. These results implicate TAFA2 and its receptor ADGRL1 as potential therapeutic targets for neurological disorders.

Creatine kinase brain-type regulates BCAR1 phosphorylation to facilitate DNA damage repair

Creatine kinase (CK) is an essential metabolic enzyme mediating creatine/phosphocreatine interconversion and shuttle to replenish ATP for energy needs. Ablation of CK causes a deficiency in energy supply that eventually results in reduced muscle burst activity and neurological disorders in mice. Besides the well-established role of CK in energy-buffering, the mechanism underlying the non-metabolic function of CK is poorly understood. Here we demonstrate that creatine kinase brain-type (CKB) may function as a protein kinase to regulate BCAR1 Y327 phosphorylation that enhances the association between BCAR1 and RBBP4. Then the complex of BCAR1 and RPPB4 binds to the promoter region of DNA damage repair gene RAD51 and activates its transcription by modulating histone H4K16 acetylation to ultimately promote DNA damage repair. These findings reveal the possible role of CKB independently of its metabolic function and depict the potential pathway of CKB-BCAR1-RBBP4 operating in DNA damage repair.

Bordetella Dermonecrotic Toxin Is a Neurotropic Virulence Factor That Uses CaV3.1 as the Cell Surface Receptor.

Dermonecrotic toxin (DNT) is one of the representative toxins produced by Bordetella pertussis, but its role in pertussis, B. pertussis infection, remains unknown. In this study, we identified the T-type voltage-gated Ca2+ channel CaV3.1 as the DNT receptor by CRISPR-Cas9-based genome-wide screening. As CaV3.1 is highly expressed in the nervous system, the neurotoxicity of DNT was examined. DNT affected cultured neural cells and caused flaccid paralysis in mice after intracerebral injection. No neurological symptoms were observed by intracerebral injection with the other major virulence factors of the organisms, pertussis toxin and adenylate cyclase toxin. These results indicate that DNT has aspects of the neurotropic virulence factor of B. pertussis The possibility of the involvement of DNT in encephalopathy, which is a complication of pertussis, is also discussed.IMPORTANCEBordetella pertussis, which causes pertussis, a contagious respiratory disease, produces three major protein toxins, pertussis toxin, adenylate cyclase toxin, and dermonecrotic toxin (DNT), for which molecular actions have been elucidated. The former two toxins are known to be involved in the emergence of some clinical symptoms and/or contribute to the establishment of bacterial infection. In contrast, the role of DNT in pertussis remains unclear. Our study shows that DNT affects neural cells through specific binding to the T-type voltage-gated Ca2+ channel that is highly expressed in the central nervous system and leads to neurological disorders in mice after intracerebral injection. These data raise the possibility of DNT as an etiological agent for pertussis encephalopathy, a severe complication of B. pertussis infection.

Effect of betulinic acid and its ionic derivatives on M-MuLV replication

Murine leukemia virus (MuLV) is a retrovirus known causing leukemia and neurological disorders in mice, and its viral life cycle and pathogenesis have been investigated extensively over the past decades. As a natural antiviral agent, betulinic acid is a pentacyclic triterpenoid that can be found in the bark of several species of plants (particularly the white birch). One of the hurdles for betulinic acid to release its antiviral potency is its poor water solubility. In this study, we synthesized more water-soluble ionic derivatives of betulinic acid, and examined their activities against Moloney MuLV (M-MuLV). The mouse fibroblast cells stably infected with M-MuLV, 43D cells, were treated with various doses of betulinic acids and its derivatives, and the viral structural protein Gag in cells and media were detected by western blots. Two ionic derivatives containing the benzalkonium cation were found to inhibit the virus production into media and decreased Gag in cells. However, a cell proliferation assay showed that the benzalkonium cation inhibited the growth of 43D cells, suggesting that our ionic derivatives limited virus production through the inhibition of metabolism in 43D cells. Interestingly, all of these betulinic acid compounds exhibited a minimum impact on the processing and release of Gag from 43D cells, which outlines the differences of viral maturation between MuLV and human immunodeficiency virus.

The Impairment of Trans Fatty Acids on Learning, Memory and Brain Amino Acid Neurotransmitters in Mice.

The aim of the study was to demonstrate the impairment of trans fatty acids (TFAs) in neurological disorders in mice. Forty-eight male Kunming mice were randomly divided into four groups with twelve in each group, namely a control group (corn oil group), and TFA groups with low, middle and high dosages. The tested chemicals were given by gavage, once a day, for 12 wk in total, with the volume of the intragastric liquid as 0.1 mL/10 g of body weight. The mice in the control group were given corn oil only. The mice in the TFA groups were given TFA solution (doses were 25, 50 and 100 mg/kg of body weight, respectively). The Morris circle water maze was used to test learning and memory of mice. The activity of acetylcholinesterase (AChE), nitric oxide synthase (NOS) and the level of brain amino acid neurotransmitters were tested. In the Morris water maze task, and compared to the control group, TFAs showed no obvious effect on learning or memory. TFA intake led to a significant decrease of AChE in all TFA groups, and the increase in levels of NOS in the high-dose group. Meanwhile, intake of TFAs increased the levels of Asp, Glu, Gly, and Tau in all TFA groups. The results suggest that long-term intake of TFAs probably impairs learning, memory and brain neurotransmitters in mice.

A translational approach to capture gait signatures of neurological disorders in mice and humans

A method for capturing gait signatures in neurological conditions that allows comparison of human gait with animal models would be of great value in translational research. However, the velocity dependence of gait parameters and differences between quadruped and biped gait have made this comparison challenging. Here we present an approach that accounts for changes in velocity during walking and allows for translation across species. In mice, we represented spatial and temporal gait parameters as a function of velocity and established regression models that reproducibly capture the signatures of these relationships during walking. In experimental parkinsonism models, regression curves representing these relationships shifted from baseline, implicating changes in gait signatures, but with marked differences between models. Gait parameters in healthy human subjects followed similar strict velocity dependent relationships which were altered in Parkinson’s patients in ways that resemble some but not all mouse models. This novel approach is suitable to quantify qualitative walking abnormalities related to CNS circuit dysfunction across species, identify appropriate animal models, and it provides important translational opportunities.

Quantitative Analysis of Mouse Retinal Layers Using Automated Segmentation of Spectral Domain Optical Coherence Tomography Images.

Quantification of retinal layers using automated segmentation of optical coherence tomography (OCT) images allows for longitudinal studies of retinal and neurological disorders in mice. The purpose of this study was to compare the performance of automated retinal layer segmentation algorithms with data from manual segmentation in mice using the Spectralis OCT. Spectral domain OCT images from 55 mice from three different mouse strains were analyzed in total. The OCT scans from 22 C57Bl/6, 22 BALBc, and 11 C3A.Cg-Pde6b+Prph2Rd2 /J mice were automatically segmented using three commercially available automated retinal segmentation algorithms and compared to manual segmentation. Fully automated segmentation performed well in mice and showed coefficients of variation (CV) of below 5% for the total retinal volume. However, all three automated segmentation algorithms yielded much thicker total retinal thickness values compared to manual segmentation data (P < 0.0001) due to segmentation errors in the basement membrane. Whereas the automated retinal segmentation algorithms performed well for the inner layers, the retinal pigmentation epithelium (RPE) was delineated within the sclera, leading to consistently thicker measurements of the photoreceptor layer and the total retina. The introduction of spectral domain OCT allows for accurate imaging of the mouse retina. Exact quantification of retinal layer thicknesses in mice is important to study layers of interest under various pathological conditions.

Interferon-γ Prevents Death of Bystander Neurons during CD8 T Cell Responses in the Brain

T cells restricted to neurotropic viruses are potentially harmful as their activity may result in the destruction of neurons. In the Borna disease virus (BDV) model, antiviral CD8 T cells entering the brain of infected mice cause neurological disease but no substantial loss of neurons unless the animals lack interferon-gamma (IFN-gamma). We show here that glutamate receptor antagonists failed to prevent BDV-induced neuronal loss in IFN-gamma-deficient mice, suggesting that excitotoxicity resulting from glutamate receptor overstimulation is an unlikely explanation for the neuronal damage. Experiments with IFN-gamma-deficient mice lacking eosinophils indicated that these cells, which specifically accumulate in the infected brains of IFN-gamma-deficient mice, are not responsible for CA1 neuronal death. Interestingly, BDV-induced damage of CA1 neurons was reduced significantly in IFN-gamma-deficient mice lacking perforin, suggesting a key role for CD8 T cells in this pathological process. Specific death of hippocampal CA1 neurons could be triggered by adoptive transfer of BDV-specific CD8 T cells from IFN-gamma-deficient mice into uninfected mice that express transgene-encoded BDV antigen at high level in astrocytes. These results indicate that attack by CD8 T cells that cause the death of CA1 neurons might be directed toward regional astrocytes and that IFN-gamma protects vulnerable CA1 neurons from collateral damage resulting from exposure to potentially toxic substances generated as a result of CD8 T cell-mediated impairment of astrocyte function.

Neurological Disorders in Mice Lacking Glycogenes that are Mainly Expressed in Brain

At the present time, 160 human glycogenes encoding glycosyltransferases and sulfotransferases, which add sulfate to carbohydrates, have been cloned and analyzed for their substrate specificity. Mice have almost all genes orthologous to them, and some are specifically expressed in neuronal tissues. In this review, neurological disorders of mice deficient for the glycogenes that synthesize interesting carbohydrate epitopes in neuronal tissues are described and summarized.

Mild overexpression of MeCP2 causes a progressive neurological disorder in mice.

Mutations in the X-linked methyl-CpG-binding protein 2 (MECP2), encoding a transcriptional repressor, cause Rett syndrome and a variety of related neurodevelopmental disorders. The vast majority of mutations associated with human disease are loss-of-function mutations, but precisely what aspect of MeCP2 function is responsible for these phenotypes remains unknown. We overexpressed wild-type human protein in transgenic mice using a large genomic clone containing the entire human MECP2 locus. Detailed neurobehavioral and electrophysiological studies in transgenic line MeCP2(Tg1), which expresses MeCP2 at approximately 2-fold wild-type levels, demonstrated onset of phenotypes around 10 weeks of age. Surprisingly, these mice displayed enhanced motor and contextual learning and enhanced synaptic plasticity in the hippocampus. After 20 weeks of age, however, these mice developed seizures, became hypoactive and approximately 30% of them died by 1 year of age. These data demonstrate that MeCP2 levels must be tightly regulated in vivo, and that even mild overexpression of this protein is detrimental. Furthermore, these results support the possibility that duplications or gain-of-function mutations in MECP2 might underlie some cases of X-linked delayed-onset neurobehavioral disorders.

Channelopathies of inwardly rectifying potassium channels.

Mutations in genes encoding ion channels have increasingly been identified to cause disease conditions collectively termed channelopathies. Recognizing the molecular basis of an ion channel disease has provided new opportunities for screening, early diagnosis, and therapy of such conditions. This synopsis provides an overview of progress in the identification of molecular defects in inwardly rectifying potassium (Kir) channels. Structurally and functionally distinct from other channel families, Kir channels are ubiquitously expressed and serve functions as diverse as regulation of resting membrane potential, maintenance of K(+) homeostasis, control of heart rate, and hormone secretion. In humans, persistent hyperinsulinemic hypoglycemia of infancy, a disorder affecting the function of pancreatic beta cells, and Bartter's syndrome, characterized by hypokalemic alkalosis, hypercalciuria, increased serum aldosterone, and plasma renin activity, are the two major diseases linked so far to mutations in a Kir channel or associated protein. In addition, the weaver phenotype, a neurological disorder in mice, has also been associated with mutations in a Kir channel subtype. Further genetic linkage analysis and full understanding of the consequence that a defect in a Kir channel would have on disease pathogenesis are among the priorities in this emerging field of molecular medicine.

T cell ignorance in mice to Borna disease virus can be overcome by peripheral expression of the viral nucleoprotein.

Infection of neonates with Borna disease virus (BDV) induces severe meningoencephalitis and neurological disorder in wild-type but not in beta(2)-microglobulin-deficient mice of strain MRL (H-2(k)). Temporary in vivo depletion of CD8(+) T cells delayed BDV-induced disease for several weeks. Depletion of CD4(+) T cells had a similar beneficial effect, indicating that the BDV-induced neurological disorder in mice is a CD4(+) T cell-dependent immunopathological process that is mediated by CD8(+) T cells. Lymphocytes prepared from brains of diseased mice were mainly from the CD8(+) T cell subset. They showed up-regulation of activation markers and exerted strong MHC I-restricted cytotoxic activity against target cells expressing the BDV nucleoprotein p40. Infection of B10.BR (H-2(k)) or congenic C57BL/10 (H-2(b)) mice resulted in symptomless, lifelong persistence of BDV in the brain. Superinfection with a recombinant vaccinia virus expressing BDV p40 but not with other vaccinia viruses induced severe neurological disease and encephalitis in persistently infected B10.BR mice but not in persistently infected C57BL/10 mice, indicating that the disease-inducing T cell response is restricted to the nucleoprotein of BDV in H-2(k) mice. Our results demonstrate that the cellular arm of the immune system may ignore the presence of a replicating virus in the central nervous system until proper antigenic stimulation at a peripheral site triggers the antiviral response.

Borna Disease Virus-Induced Neurological Disorder in Mice: Infection of Neonates Results in Immunopathology

Borna disease virus (BDV) is a neurotropic nonsegmented negative-stranded RNA virus that persistently infects warm-blooded animals. In horses and other natural animal hosts, infections with BDV cause meningoencephalitis and behavioral disturbances. Experimental infection of adult mice takes a nonsymptomatic course, an observation previously believed to indicate that this animal species is not suitable for pathogenesis studies. We now demonstrate that BDV frequently induces severe neurological disease in infected newborn mice. Signs of neurological disease were first observed 4 to 6 weeks after intracerebral infection. They included a characteristic nonphysiological position of the hind limbs at an early stage of the disease and paraparesis at a later stage. Histological examination revealed large numbers of perivascular and meningeal inflammatory cells in brains of diseased mice and, unexpectedly, no increase in immunoreactivity to glial fibrillar acidic protein. The incidence and severity of BDV-induced disease varied dramatically among mouse strains. While only 13% of the infected C57BL/6 mice showed disease symptoms, which were mostly transient, more than 80% of the infected MRL mice developed severe neurological disorder. In spite of these differences in susceptibility to disease, BDV replicated to comparable levels in the brains of mice of the various strains used. Intracerebral infections of newborn beta2-microglobulin-deficient C57BL/6 and MRL mice, which both lack CD8+ T cells, did not result in meningoencephalitis or neurological disease, indicating that the BDV-induced neurological disorder in mice is a cytotoxic T-cell-mediated immunopathological process. With this new animal model it should now be possible to characterize the disease-inducing immune response to BDV in more detail.

Cloning and expression of the proteolipid protein DM20 cDNA from the brain of the rainbow trout, Oncorhynchus mykiss

The myelin sheath in higher vertebrates consists predominantly of proteolipid protein (PLP) and its smaller isoform DM20. Mutations in the PLP gene produces several neurological disorders such as Pelizaeus-Merzbacher disease and the rumpshaker phenotype in mice. This paper describes the cloning and expression of DM20 from the brain of Rainbow trout. We have isolated a nearly full-length cDNA clone containing 1835 bp that codes for a protein of 258 amino acids. Trout DM20 shows extensive homology with DM20 from higher vertebrates and includes the four hydrophobic regions that are believed to span the myelin membrane. The DM20 transcript is expressed throughout the central nervous system of the trout but appears at its highest levels in the spinal cord and medulla oblongata. The transcript is expressed at very low levels on hatching day but increases 179-fold by the 5th week. Contrary to higher vertebrates, there is no switch to the PLP transcript in maturing trout. Moreover, the rsh mutation (186 Thr to Ile) that produces the rumpshaking neurological disorder in mice has no effect in trout.

Factors affecting induction of neurological disorders in mice by paralysis-inducing Friend-related PVC viruses.

Our previous studies showed that the passage of the Friend virus complex through rats generated variant MuLVs, designated PVC111, PVC211, PVC321 and PVC441, that induced neurological disorders associated with tremor and paralysis. In this study, we tested the pathogenicity of four different PVC viruses in mice. Although histopathological studies revealed spongiform degeneration in the spinal cords of NFS mice infected with each PVC virus, only PVC441 frequently induced tremor and paralysis. After a long latency, all of these viruses induced leukemia associated with severe anemia. Further studies with PVC441 revealed dose- and age-dependence for tremor induction. In contrast to NFS mice, BALB/c, DBA/2 and C57BL/6 mice infected with PVC441 virus showed no neurological symptoms, although the virus could be isolated from the tissues of central nervous system. Despite the absence of neurological symptoms, a high degree of neuronal degeneration in the lumbar spinal cord was found in PVC441-infected BALB/c mice. A low degree of neuronal degeneration was found in PVC441-infected DBA/2 or C57BL/6 mice. Genetic crosses of these resistant mice with susceptible NFS mice indicated that resistance to tremor induction by PVC441 was dominant in all mouse strains and suggested that various host genes may control the susceptibility of mice to tremor induction by PVC441 virus.

Influence of ptaquiloside on the development of newborn mice.

We attempted to induce neurological disorders in mice by a single administration of various doses of ptaquiloside (PT) within 24 h after birth. Animals which survived for a long period after treatment showed a reduction in body weight, but did not develop any gait disturbance. On the other hand, in treated animals which died before beginning to walk, karyorrhexis in the external granular layer of the cerebellum and karyorrhexis and rosette formation in the neuroblastic layer of the retina were present. In addition, treated animals showed karyorrhexis and great reduction in the number of the cells in the thymus, suggesting cytotoxic and cytolytic effects of alkylating agents on lymphocytes in this organ.

Abnormal rate of granule cell migration in the cerebellum of “weaver” mutant mice

“Weaver,” a neurological disorder in mice is transmitted by an incompletely dominant gene. Both the heterozygous carrier and homozygous weaver mice show alterations in the cytoarchitectonics of the cerebellum. However, no behavioral abnormalities of cerebellar disorder can be detected in the heterozygous carrier. Studies with autoradiography showed that the alterations in the cytoarchitectonics of both the heterozygous carrier and homozygous weaver cerebellums are associated with a reduced rate of granule cell migration.

The Possible Relationship of the Pleuropneumonialike Organisms to Reiter's Disease, Rheumatoid Arthritis and Ulcerative Colitis

The pleuropneumonia-like group of organisms has long been identified with the problem of chronic arthritis in many animal species. In 1898, Nocard and Roux isolated the first member of this group from cattle suffering from pleurisy, pneumonitis, and in a considerable percentage of the cases, from joint involvement. In 1923, Bridre and Donatien found a similar organism in sheep and goats suffering from agalactia, a syndrome which includes mastitis with glandular atrophy and suppression of lactation, chronic arthritis, keratitis, vesiculo-pustular skin eruptions, and in males, scrotal inflammation. The organism was isolated from the milk, conjunctivae, joint fluid, and blood stream.1 Further members of this group of organisms were first isolated by Klieneberger in 1935 in cultures from rats infected with Streptobacillus moniliformis. In the following few years, many strains were isolated from rats both alone and in association with the streptobacillus. Of these, the L4 strain of Woglom and Warren caused a pyogenic polyarthritis in rats and the L7 of Findlay et al.2 caused a spontaneous as well as transmissible and experimental polyarthritis in rats. This strain could also produce an arthritis when injected into mice. In the course of studies on mouse toxoplasmosis, Sabin3 isolated a new filtrable agent, later shown4,5 to be a pleuropneumonia-like organism (called strain A). This organism caused a neurological disorder in mice, called rolling disease by Findlay et al.,5 but if inoculated into older, more resistant mice, it induced a polyarthritis rather than the cerebellar disease.6 Other strains (B, C, D, E) were subsequently isolated' from the conjunctiva, nasal mucosa, and sometimes the brain, lung, and trachea of normal carrier mice. Of these, strain B,5 on intravenous inoculation, produced a chronic, proliferative, progressive arthritis in mice which clinically and pathologically bears a striking resemblance to human arthritis. Clinically there were typical rheumatoid fusiform swellings of the involved joints which often went on to chronicity and ankylosis. The organisms could be isolated from infected joints as late as 70 days after intravenous injection, though the blood stream was rendered sterile in 2 to 3 days. These various mouse strains all induced antibody responses in the host, but did not cross agglutinate and were not serologically identical. Pleuropneumonia-like microorganisms have likewise been found in dogs and guinea pigs' though not in relation to arthritis and also as saprophytes in sewage and decaying matter.' The close similarity of this experimental infection in mice to human arthritis led several workers8-I to study under controlled