Exposure to Toxicants Affects Everyone, Especially the Very Young.

Toxicology is an incredibly complex and diverse area of biomedical science that includes numerous areas of specialization [...].

Mercury Toxicity and Neurogenesis in the Mammalian Brain.

The mammalian brain is formed from billions of cells that include a wide array of neuronal and glial subtypes. Neural progenitor cells give rise to the vast majority of these cells during embryonic, fetal, and early postnatal developmental periods. The process of embryonic neurogenesis includes proliferation, differentiation, migration, the programmed death of some newly formed cells, and the final integration of differentiated neurons into neural networks. Adult neurogenesis also occurs in the mammalian brain, but adult neurogenesis is beyond the scope of this review. Developing embryonic neurons are particularly susceptible to neurotoxicants and especially mercury toxicity. This review focused on observations concerning how mercury, and in particular, methylmercury, affects neurogenesis in the developing mammalian brain. We summarized information on models used to study developmental mercury toxicity, theories of pathogenesis, and treatments that could be used to reduce the toxic effects of mercury on developing neurons.

Adult neurogenesis in the mammalian dentate gyrus.

Earlier observations in neuroscience suggested that no new neurons form in the mature central nervous system. Evidence now indicates that new neurons do form in the adult mammalian brain. Two regions of the mature mammalian brain generate new neurons: (a) the border of the lateral ventricles of the brain (subventricular zone) and (b) the subgranular zone (SGZ) of the dentate gyrus of the hippocampus. This review focuses only on new neuron formation in the dentate gyrus of the hippocampus. During normal prenatal and early postnatal development, neural stem cells (NSCs) give rise to differentiated neurons. NSCs persist in the dentate gyrus SGZ, undergoing cell division, with some daughter cells differentiating into functional neurons that participate in learning and memory and general cognition through integration into pre-existing neural networks. Axons, which emanate from neurons in the entorhinal cortex, synapse with dendrites of the granule cells (small neurons) of the dentate gyrus. Axons from granule cells synapse with pyramidal cells in the hippocampal CA3 region, which send axons to synapse with CA1 hippocampal pyramidal cells that send their axons out of the hippocampus proper. Adult neurogenesis includes proliferation, differentiation, migration, the death of some newly formed cells and final integration of surviving cells into neural networks. We summarise these processes in adult mammalian hippocampal neurogenesis and discuss the roles of major signalling molecules that influence neurogenesis, including neurotransmitters and some hormones. The recent controversy raised concerning whether or not adult neurogenesis occurs in humans also is discussed.

Morphological and functional comparison of lingual papillae in suckling and adult feral cats: Forensic evidence.

Feral cats are considered as strays and are more likely to hunt in the street. We investigated the effect of environmental adaptations on the structures of lingual papillae in feral cats, which could be used as forensic evidence for their identification. There are no reported studies about the structural comparison of lingual papillae between suckling and adult feral cats. The present study described the lingual papillae of both suckling and adult cats macroscopically and microscopically via light and scanning electron microscopy. A total of nine tongue samples each for suckling and adult feral cats were examined grossly and histologically. Papillae distributions of suckling cats were similar to those observed in adult cats. Meanwhile, the shapes of those papillae were markedly different from that of corresponding papillae in adults. The change in taste bud position and size seemed to be related to the progressive growth of the papillae between adult and suckling cats; absence of taste buds in foliate papillae of feral cats at any stage; and marginal papillae which were a characteristic feature for all suckling cats. All previous elements could be affected by the specific feeding behaviour and mastication mode adaptation in suckling and adult feral cats which might help to identify suckling and adult feral cats among other breeds and animal species. We anticipate these findings may provide promising forensic evidence to discriminate between adult and suckling feral cat remains as well as prediction of environmental harshness and feeding behaviour.

Use of zebrafish ( Danio rerio ) embryos as a model to assess effects of mercury on developing skeletal muscle: a morphometric and immunohistochemical study / Uso de embriones de pez cebra ( Danio rerio ) como modelo para evaluar los efectos del mercurio en el desarrollo del músculo esquelético: un estudio morfométrico e inmunohistoquímico

Exposure to mercury in the environment continues to be a significant worldwide concern, especially for developing embryos and fetuses. While extensive research effort has focused on the effects of mercury on the developing nervous system, much less is known concerning adverse effects of mercury on other organ systems, including the development of skeletal muscle. We exposed developing zebrafish embryos to a range of concentrations of mercuric chloride (100 to 400 µg/liter or ppb) and compared them to control embryos (0 µg/L mercuric chloride). Embryos were examined at 48 hours post fertilization (hpf) for morphometry and morphological deformities of skeletal muscle fibers in the trunk and tail. Embryos exposed to 400 ppb mercuric chloride showed decreased trunk and tail areas compared to control embryos. A dose-dependent reduction in muscle fiber length was observed, and exposure to all concentrations of mercuric chloride used in this study resulted in decreased muscle fiber immunohistochemical staining with anti-myosin antibodies. Irregular muscle fiber diameters, twisted muscle fibers, and degenerated muscle fibers were observed in sections of embryos stained with eosin at the higher exposure concentrations. Evidence presented in this study suggests that exposure to even low concentrations of mercuric chloride adversely affects skeletal muscle fiber development or muscle fiber integrity, or both.

La exposición al mercurio en el medio ambiente sigue siendo una preocupación mundial importante, especialmente para el desarrollo de embriones y fetos. Si bien un amplio esfuerzo de investigación se ha centrado en los efectos del mercurio en el sistema nervioso en desarrollo, se sabe mucho menos sobre los efectos adversos en otros sistemas orgánicos, incluido el desarrollo del músculo esquelético. Expusimos embriones de pez cebra en desarrollo a un rango de concentraciones de cloruro de mercurio (100 a 400 mg / l o ppb) y los comparamos con embriones de control (0 mg / L de cloruro de mercurio). Los embriones se examinaron a las 48 horas después de la fertilización (HPF) pararealizar la morfometría y verificar las deformidades morfológicas de las fibras del músculo esquelético en el tronco y la cola. Los embriones expuestos a 400 ppb de cloruro de mercurio mostraron una disminución de las áreas del tronco y la cola en comparación con los embriones de control. Se observó una reducción dependiente de la dosis en la longitud de la fibra muscular, y la exposición a todas las concentraciones de cloruro de mercurio utilizadas en este estudio, dio como resultado una tinción inmunohistoquímica de fibra muscular disminuida con anticuerpos anti-miosina. Se observaron diámetros irregulares de fibras musculares, fibras musculares retorcidas y fibras musculares degeneradas en secciones de embriones teñidos con eosina en las concentraciones de exposición más altas. La evidencia presentada en este estudio sugiere que la exposición incluso a bajas concentraciones de cloruro mercúrico afecta negativamente el desarrollo de la fibra del músculo esquelético o la integridad de la fibra muscular, o ambas.

Survey of nuclear progesterone receptor expression in the uterus of the cyclic and pregnant camel (Camelus dromedarius).

Progesterone receptors (PR) are necessary to mediate the biological effects of progesterone and are integral to the regulation of a number of different aspects of reproduction in mammals including ovulation of the oocyte, implantation of the conceptus and maintenance of pregnancy. This study investigated the expression and localization of progesterone receptors in the uterine wall of both pregnant and cyclic (nonpregnant) camels. Uterine tissue samples were collected from healthy animals and processed for routine histological and immunohistochemical staining techniques to reveal nuclear PR. Demonstration of PR was performed by indirect immunohistochemical techniques using monoclonal antibodies raised against human PR. Immunolocalization of PR was more intense in all four endometrial zones (I-IV) as well as the myometrium of non pregnant (cyclic) animals (animals with newly formed corpus luteum). In contrast, PR immunostaining in both the endometrium and the myometrium was greatly reduced in pregnancy, particularly in the latest stage examined (approximately 366 days of gestation). In conclusion, a better understanding of the expression of steroid hormones and their receptors, including progesterone and the PR is critical to improving the reproductive health and pregnancy in the domesticated dromedary camel.

Robust Cell Detection for Large-Scale 3D Microscopy Using GPU-Accelerated Iterative Voting.

High-throughput imaging techniques, such as Knife-Edge Scanning Microscopy (KESM),are capable of acquiring three-dimensional whole-organ images at sub-micrometer resolution. These images are challenging to segment since they can exceed several terabytes (TB) in size, requiring extremely fast and fully automated algorithms. Staining techniques are limited to contrast agents that can be applied to large samples and imaged in a single pass. This requires maximizing the number of structures labeled in a single channel, resulting in images that are densely packed with spatial features. In this paper, we propose a three-dimensional approach for locating cells based on iterative voting. Due to the computational complexity of this algorithm, a highly efficient GPU implementation is required to make it practical on large data sets. The proposed algorithm has a limited number of input parameters and is highly parallel.

Effects of methyl mercury exposure on pancreatic beta cell development and function.

Methyl mercury is an environmental contaminant of worldwide concern. Since the discovery of methyl mercury exposure due to eating contaminated fish as the underlying cause of the Minamata disaster, the scientific community has known about the sensitivity of the developing central nervous system to mercury toxicity. Warnings are given to pregnant women and young children to limit consumption of foods containing methyl mercury to protect the embryonic, fetal and postnatally developing central nervous system. However, evidence also suggests that exposure to methyl mercury or various forms of inorganic mercury may also affect development and function of other organs. Numerous reports indicate a worldwide increase in diabetes, particularly type 2 diabetes. Quite recently, methyl mercury has been shown to have adverse effects on pancreatic beta (ß) cell development and function, resulting in insulin resistance and hyperglycemia and may even lead to the development of diabetes. This review discusses possible mechanisms by which methyl mercury exposure may adversely affect pancreatic ß cell development and function, and the role that methyl mercury exposure may have in the reported worldwide increase in diabetes, particularly type 2 diabetes. While additional information is needed regarding associations between mercury exposure and specific mechanisms of the pathogenesis of diabetes in the human population, methyl mercury's adverse effects on the body's natural sources of antioxidants suggest that one possible therapeutic strategy could involve supplementation with antioxidants. Thus, it is important that additional investigation be undertaken into the role of methyl mercury exposure and reduced pancreatic ß cell function. Copyright © 2016 John Wiley & Sons, Ltd.

Transmission electron microscopic evaluation of neuronal changes in methylmercury-exposed zebrafish embryos (Danio rerio).

Our work aimed to elucidate the ultrastructural changes associated with brain neurons in wild-type zebrafish embryos exposed to different concentrations of methylmercury. Zebrafish embryos were exposed to one of five concentrations of methylmercury (0 [negative control], 5, 10, 50, and 80 parts per billion) starting at six hours post fertilization (hpf). At 96 hpf, cells in the zebrafish embryo brains were examined using transmission electron microscopy. The developing neurons of the control embryos sowed normal cellular ultrastructure. Few alterations were observed among the neurons of zebrafish embryos exposed to 5 ppb methylmercury. The cells of the embryos exposed to 10 ppb methylmercury showed slight cellular degeneration as demonstrated by the accumulation of electron dens bodies which were presumably lysosomes in different stages of formation. In embryos exposed to 50 ppb methylmercury, the neuronal cytoplasm conained large electron dense lysosomes and the rough endoplasmic reticulum appeared to be reduced and irregular in shape. Furthermore, the embryonic brain neurons exposed to 80 ppb methylmercury showed the most severe ultrastructural changes, including some that were consistent with different stages of the cell death process. Obvious cellular changes were observed in this highest exposure group included: disrupted or degenerating nuclei; fragmentation or vacuolization of mitochondrial cristae; and loss of mitochondrial matrix density. Based on these observations, we conclude that these different morphological patterns of cellular changes may reflect either different stages of the cell death process or different types of cell death due to 24 hours of exposure to 80 ppb methylmercury.

Neural cell proliferation and survival in the hippocampus of adult CaV 2.1 calcium ion channel mutant mice.

Tottering mutant mice carry a mutation in the pore-forming subunit (α1A) of CaV2.1 (P/Q-type) voltage-gated calcium ion (Ca2+) channels resulting in reduced neuronal Ca2+ current density. We assessed male tottering mice for spatial learning using the Morris water maze. Tottering mice performed worse than wild type mice, suggesting abnormal hippocampal function. Because Ca2+ influx via voltage-dependent Ca2+ channels regulates neuronal survival and function, we assessed hippocampus volume and cell density using hematoxylin and eosin stained serial sections. Adult hippocampal neurogenesis was assessed using 5-bromo-2'-deoxyuridine (BrdU) labeling with fluorescent immunohistochemistry (IHC) and proliferating cell nuclear antigen (PCNA) with diaminobenzidine IHC. We double-labeled neurons using fluorescence IHC with BrdU-neuronal nuclei (Neu-N) or double labeling of astrocytes using BrdU-glial fibrillary protein, respectively, to assess cell proliferation and survival. We assessed numbers of dying cells using fluoro-Jade histochemistry. Decreased hippocampal volume, increased dentate hilar and hippocampal CA1 cell densities were observed in tottering mice compared to wild type mice. Cell proliferation was increased in the hilus and CA2 region of tottering mice compared to wild type mice. Dendritic intersections in Sholl analysis were decreased for tottering mouse CA1 pyramidal neurons compared to wild type mice. The increased regional cell density coincides with increases in cell proliferation in similar, non-neurogenic areas of the hippocampus of tottering mice. Thus, hippocampal alterations observed in adult tottering mice appear to result from changes in neuronal morphology and proliferation in non-neurogenic areas of the hippocampus, and less through altered adult hippocampal neurogenesis or cell death.

Fast submicrometer-scale imaging of whole zebrafish using the knife-edge scanning microscope.

Advances in high-resolution 3D microscopy have enabled the investigation of subcellular microstructures in biological specimen. For a full understanding of the organism's structure and function, it is mandatory to obtain data from the whole animal, not just parts of it. In this paper, we present our work with the Knife-Edge Scanning Microscope (KESM) for imaging a Nissl-stained whole zebrafish larva. KESM combines a diamond microtome and line-scan imaging for simultaneous sectioning and imaging in 3D. We show that using the KESM, a zebrafish, less than 3 mm long and diameter less than 500 µm, can be imaged within 1 hour at a resolution of 0.6 µm × 0.7 µm × 1.0 µm. We also present new results on using a vibrating microtome to improve sectioning and imaging robustness.

Compositional analysis of genetically modified corn events (NK603, MON88017×MON810 and MON89034×MON88017) compared to conventional corn.

Compositional analysis of genetically modified (GM) crops continues to be an important part of the overall evaluation in the safety assessment for these materials. The present study was designed to detect the genetic modifications and investigate the compositional analysis of GM corn containing traits of multiple genes (NK603, MON88017×MON810 and MON89034×MON88017) compared with non-GM corn. Values for most biochemical components assessed for the GM corn samples were similar to those of the non-GM control or were within the literature range. Significant increases were observed in protein, fat, fiber and fatty acids of the GM corn samples. The observed increases may be due to the synergistic effect of new traits introduced into corn varieties. Furthermore, SDS-PAGE analysis showed high similarity among the protein fractions of the investigated corn samples. These data indicate that GM corn samples were compositionally equivalent to, and as nutritious as, non-GM corn.

Pluripotent state induction in mouse embryonic fibroblast using mRNAs of reprogramming factors.

Reprogramming of somatic cells has great potential to provide therapeutic treatments for a number of diseases as well as provide insight into mechanisms underlying early embryonic development. Improvement of induced Pluripotent Stem Cells (iPSCs) generation through mRNA-based methods is currently an area of intense research. This approach provides a number of advantages over previously used methods such as DNA integration and insertional mutagenesis. Using transfection of specifically synthesized mRNAs of various pluripotency factors, we generated iPSCs from mouse embryonic fibroblast (MEF) cells. The genetic, epigenetic and functional properties of the iPSCs were evaluated at different times during the reprogramming process. We successfully introduced synthesized mRNAs, which localized correctly inside the cells and exhibited efficient and stable translation into proteins. Our work demonstrated a robust up-regulation and a gradual promoter de-methylation of the pluripotency markers, including non-transfected factors such as Nanog, SSEA-1 (stage-specific embryonic antigen 1) and Rex-1 (ZFP-42, zinc finger protein 42). Using embryonic stem cells (ESCs) conditions to culture the iPS cells resulted in formation of ES-like colonies after approximately 12 days with only five daily repeated transfections. The colonies were positive for alkaline phosphatase and pluripotency-specific markers associated with ESCs. This study revealed the ability of pluripotency induction and generation of mouse mRNA induced pluripotent stem cells (mRNA iPSCs) using transfection of specifically synthesized mRNAs of various pluripotency factors into mouse embryonic fibroblast (MEF) cells. These generated iPSCs exhibited molecular and functional properties similar to ESCs, which indicate that this method is an efficient and viable alternative to ESCs and can be used for further biological, developmental and therapeutic investigations.

The effect of methylmercury exposure on behavior and cerebellar granule cell physiology in aged mice.

Epidemiology studies have clearly documented that the central nervous system is highly susceptible to methylmercury toxicity, and exposure to this neurotoxicant in humans primarily results from consumption of contaminated fish. While the effects of methylmercury exposure have been studied in great detail, comparatively little is known about the effects of moderate to low dose methylmercury toxicity in the aging central nervous system. We examined the toxic effects of a moderate dose of methylmercury on the aging mouse cerebellum. Male and female C57BL/6 mice at 16-20 months of age were exposed to methylmercury by feeding a total dose of 5.0 mg kg(-1) body weight and assessed using four behavioral tests. Methylmercury-treated aged mice performed significantly worse in open field, footprint analysis and the vertical pole test compared with age-matched control mice. Isolated cerebellar granule cells from methylmercury-treated aged mice exhibited higher levels of reactive oxygen species and reduced mitochondrial membrane potentials, but no differences in basal intracellular calcium ion levels compared with age-matched control mice. When aged mice were exposed to a moderate dose of methylmercury, they exhibited a similar degree of impairment when compared with young adult mice exposed to the same moderate dose of methylmercury, as reported in earlier studies from this laboratory. Thus, at least in mice, exposure of the aged brain to moderate concentrations methylmercury does not pose greater risk compared with the young adult brain exposed to similar concentrations of methylmercury.

Identification of Peptide lv, a novel putative neuropeptide that regulates the expression of L-type voltage-gated calcium channels in photoreceptors.

Neuropeptides are small protein-like signaling molecules with diverse roles in regulating neural functions such as sleep/wake cycles, pain modulation, synaptic plasticity, and learning and memory. Numerous drugs designed to target neuropeptides, their receptors, or relevant pathways have been developed in the past few decades. Hence, the discovery and characterization of new neuropeptides and their functions have received considerable attention from scientific research. Computational bioinformatics coupled with functional assays are powerful tools to address the difficulties in discovering new bioactive peptides. In this study, a new bioinformatic strategy was designed to screen full length human and mouse cDNA databases to search for novel peptides. One was discovered and named peptide Lv because of its ability to enhance L-type voltage-gated calcium channel (L-VGCC) currents in retinal photoreceptors. Using matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), peptide Lv was detected in the culture media, which indicated that it was secreted from 661W cells transfected with the gene. In vitro treatments with either glutathione S-transferase (GST) fusion peptide Lv or synthesized peptide Lv enhanced L-VGCC channel activities in cone photoreceptors. At the molecular level, peptide Lv stimulated cAMP production, enhanced phosphorylation of extracellular signal-regulated kinase (ERK), and increased the protein expression of L-VGCCα1 subunits in cone photoreceptors. Therefore, the biological activities of peptide Lv may be very important in the modulation of L-VGCC dependent neural plasticity.

Changing iron content of the mouse brain during development.

Iron is crucial to many processes in the brain yet the percentages of the major iron-containing species contained therein, and how these percentages change during development, have not been reliably determined. To do this, C57BL/6 mice were enriched in (57)Fe and their brains were examined by Mössbauer, EPR, and electronic absorption spectroscopy; Fe concentrations were evaluated using ICP-MS. Excluding the contribution of residual blood hemoglobin, the three major categories of brain Fe included ferritin (an iron storage protein), mitochondrial iron (consisting primarily of Fe/S clusters and hemes), and mononuclear nonheme high-spin (NHHS) Fe(II) and Fe(III) species. Brains from prenatal and one-week old mice were dominated by ferritin and were deficient in mitochondrial Fe. During the next few weeks of life, the brain grew and experienced a burst of mitochondriogenesis. Overall brain Fe concentration and the concentration of ferritin declined during this burst phase, suggesting that the rate of Fe incorporation was insufficient to accommodate these changes. The slow rate of Fe import and export to/from the brain, relative to other organs, was verified by an isotopic labeling study. Iron levels and ferritin stores replenished in young adult mice. NHHS Fe(II) species were observed in substantial levels in brains of several ages. A stable free-radical species that increased with age was observed by EPR spectroscopy. Brains from mice raised on an Fe-deficient diet showed depleted ferritin iron but normal mitochondrial iron levels.

Initial characterization of mice null for Lphn3, a gene implicated in ADHD and addiction.

The LPHN3 gene has been associated with both attention deficit-hyperactivity disorder (ADHD) and addiction, suggesting that it may play a role in the etiology of these disorders. Unfortunately, almost nothing is known about the normal functions of this gene, which has hampered understanding of its potential pathogenic role. To begin to characterize such normal functions, we utilized a gene-trap embryonic stem cell line to generate mice mutant for the Lphn3 gene. We evaluated differential gene expression in whole mouse brain between mutant and wild type male littermates at postnatal day 0 using TaqMan gene expression assays. Most notably, we found changes in dopamine and serotonin receptors and transporters (Dat1, Drd4, 5Htt, 5Ht2a), changes in neurotransmitter metabolism genes (Th, Gad1), as well as changes in neural developmental genes (Nurr, Ncam). When mice were examined at 4-6 weeks of age, null mutants showed increased levels of dopamine and serotonin in the dorsal striatum. Finally, null mutant mice had a hyperactive phenotype in the open field test, independent of sex, and were more sensitive to the locomotor stimulant effects of cocaine. Considered together, these results suggest that Lphn3 plays a role in development and/or regulation of monoamine signaling. Given the central role for monoamines in ADHD and addiction, it seems likely that the influence of LPHN3 genotype on these disorders is mediated through alterations in monoamine signaling.

Smoking during pregnancy: lessons learned from epidemiological studies and experimental studies using animal models.

Numerous epidemiological studies in the human population clearly indicate that smoking while pregnant has deleterious effects on fetal development as well as long-term adverse consequences on postnatal development and maturation of several organ systems. Low birth weight, sudden infant death syndrome (SIDS), behavioral disorders including attention deficit hyperactivity disorder (ADHD), externalizing and internalizing behavioral problems and conduct disorders in children have all been linked to prenatal exposure to tobacco smoke. The major pharmacologically active chemical found in tobacco smoke is nicotine, and prenatal exposure to nicotine has been shown to have significant effect on the development of multiple organ systems, including the nervous, respiratory, and cardiovascular systems. In this review, we define mainstream and sidestream smoke, summarize the major classes of compounds found in cigarette smoke, and describe how use of laboratory animal models can be used to assess mechanisms of toxicity and risk in the human population in general. We then discuss the association with smoking during pregnancy and the occurrence of reduced lung function, low birth weight, the incidence of congenital structural malformations, SIDS, ADHD, cognitive impairment, and mood disorders in children, and review pertinent experimental studies using a variety of animal models of developmental nicotine exposure, including, rats, mice, monkeys, lambs, and pigs that have increased our understanding of the pathophysiology of these disorders.

Striatal expression of Homer1a is affected by genotype but not dystonic phenotype of tottering mice: a model of spontaneously occurring motor disturbances.

Tottering (tg) mice carry a missense mutation in the gene coding for P/Q-type voltage-dependent Ca(2+) channels (VDCCs). Aberrant functioning of P/Q-type VDCCs results in molecular alterations in Ca(2+) currents and in glutamate and dopamine systems. As a consequence, tottering mice exhibit mild ataxia, spontaneous epilepsy, and paroxysmal dyskinesia. In this study, we evaluated whether the tottering mice genotype (homozygous vs. heterozygous) and abnormal movement phenotype (mice exhibiting paroxysmal dyskinesia vs. mice not exhibiting dyskinesia) may affect the expression of Homer1a. Homer1a is a gene whose expression is modulated by glutamate, dopamine and Ca(2+) concentrations. Over-expression of Homer1a has been described in epilepsy and motor dysfunctions. Thereby, changes in Homer1a expression could take place in tottering mice. Studying the expression profile of this gene may shed light on the molecular events occurring in tottering mice. Moreover, tottering mice may represent a valuable animal model for investigating Homer1a involvement in motor disorders. Homer1a expression was decreased in all striatal subregions, with the exclusion of the dorsolateral caudate-putamen, in heterozygous mice compared to wild-type and homozygous mice. Gene expression was decreased in the core of the accumbens in mice exhibiting paroxysmal dyskinesia compared to wild-type mice and to mice not exhibiting dyskinesia. These results demonstrate that the tottering mouse genotype may affect striatal expression of Homer1a, possibly as a result of imbalance between Ca(2+) channels subtypes or Ca(2+)-related molecules in heterozygous vs. homozygous mice.

Multiscale exploration of mouse brain microstructures using the knife-edge scanning microscope brain atlas.

Connectomics is the study of the full connection matrix of the brain. Recent advances in high-throughput, high-resolution 3D microscopy methods have enabled the imaging of whole small animal brains at a sub-micrometer resolution, potentially opening the road to full-blown connectomics research. One of the first such instruments to achieve whole-brain-scale imaging at sub-micrometer resolution is the Knife-Edge Scanning Microscope (KESM). KESM whole-brain data sets now include Golgi (neuronal circuits), Nissl (soma distribution), and India ink (vascular networks). KESM data can contribute greatly to connectomics research, since they fill the gap between lower resolution, large volume imaging methods (such as diffusion MRI) and higher resolution, small volume methods (e.g., serial sectioning electron microscopy). Furthermore, KESM data are by their nature multiscale, ranging from the subcellular to the whole organ scale. Due to this, visualization alone is a huge challenge, before we even start worrying about quantitative connectivity analysis. To solve this issue, we developed a web-based neuroinformatics framework for efficient visualization and analysis of the multiscale KESM data sets. In this paper, we will first provide an overview of KESM, then discuss in detail the KESM data sets and the web-based neuroinformatics framework, which is called the KESM brain atlas (KESMBA). Finally, we will discuss the relevance of the KESMBA to connectomics research, and identify challenges and future directions.