An Analysis of Genetic Predisposition to Hereditary Catalepsy in a Mouse Model of Neuropsychiatric Disorders Using Whole-Genome Sequencing Data.

Catalepsy is a behavioral condition that is associated with severe psychopathologies, including schizophrenia, depression, and Parkinson's disease. In some mouse strains, catalepsy can be induced by pinching the skin at the scruff of the neck. The main locus of hereditary catalepsy in mice has recently been linked to the 105-115 Mb fragment of mouse chromosome 13 by QTL analysis. We performed whole-genome sequencing of catalepsy-resistant and catalepsy-prone mouse strains in order to pinpoint the putative candidate genes related to hereditary catalepsy in mice. We remapped the previously described main locus for hereditary catalepsy in mice to the chromosome region 103.92-106.16 Mb. A homologous human region on chromosome 5 includes genetic and epigenetic variants associated with schizophrenia. Furthermore, we identified a missense variant in catalepsy-prone strains within the Nln gene. Nln encodes neurolysin, which degrades neurotensin, a peptide reported to induce catalepsy in mice. Our data suggest that Nln is the most probable candidate for the role of major gene of hereditary, pinch-induced catalepsy in mice and point to a shared molecular pathway between catalepsy in mice and human neuropsychiatric disorders.

Quantitative Analysis of L1-Retrotransposons in Alzheimer's Disease and Aging.

LINE1 retrotransposons are members of a class of mobile genetic elements capable of retrotransposition in the genome via a process of reverse transcription. LINE1 repeats, integrating into different chromosomal loci, affect the activity of genes and cause different genomic mutations. Somatic variability of the human genome is linked to the activity of some subfamilies of LINE1, in particular, a high level of LINE1 retrotranspositions has been observed in brain tissues. However, the contribution of LINE1 to genomic variability during normal aging and in age-related neurodegenerative diseases is poorly understood. We conducted quantitative real-time PCR analysis of active subfamilies of LINE1 repeats (aL1) using genomic DNA extracted from brain specimens of Alzheimer's disease (AD) patients and individuals without neuropsychiatric pathologies, as well as DNA extracted from blood specimens of individuals of different ages (healthy and AD subjects). Inter-individual quantitative variations of active families of aL1 repeats in the genome were observed. No significant age-dependent differences were identified. Likewise, no difference of aL1 copy number in brain and blood were indicated between AD patients and the aged-matched control group without dementia. These data imply that aging and the AD-associated neurodegenerative process are not the major factors contributing to the retrotransposition processes of active LINE1 repeats.

Complete mitochondrial genome and evolutionary analysis of Turritopsis dohrnii, the "immortal" jellyfish with a reversible life-cycle.

Turritopsis dohrnii (Cnidaria, Hydrozoa, Hydroidolina, Anthoathecata) is the only known metazoan that is capable of reversing its life cycle via morph rejuvenation from the adult medusa stage to the juvenile polyp stage. Here, we present a complete mitochondrial (mt) genome sequence of T. dohrnii, which harbors genes for 13 proteins, two transfer RNAs, and two ribosomal RNAs. The T. dohrnii mt genome is characterized by typical features of species in the Hydroidolina subclass, such as a high A+T content (71.5%), reversed transcriptional orientation for the large rRNA subunit gene, and paucity of CGN codons. An incomplete complementary duplicate of the cox1 gene was found at the 5' end of the T. dohrnii mt chromosome, as were variable repeat regions flanking the chromosome. We identified species-specific variations (nad5, nad6, cob, and cox1 genes) and putative selective constraints (atp8, nad1, nad2, and nad5 genes) in the mt genes of T. dohrnii, and predicted alterations in tertiary structures of respiratory chain proteins (NADH4, NADH5, and COX1 proteins) of T. dohrnii. Based on comparative analyses of available hydrozoan mt genomes, we also determined the taxonomic relationships of T. dohrnii, recovering Filifera IV as a paraphyletic taxon, and assessed intraspecific diversity of various Hydrozoa species.

Whole exome sequencing links dental tumor to an autosomal-dominant mutation in ANO5 gene associated with gnathodiaphyseal dysplasia and muscle dystrophies.

Tumors of the jaws may represent different human disorders and frequently associate with pathologic bone fractures. In this report, we analyzed two affected siblings from a family of Russian origin, with a history of dental tumors of the jaws, in correspondence to original clinical diagnosis of cementoma consistent with gigantiform cementoma (GC, OMIM: 137575). Whole exome sequencing revealed the heterozygous missense mutation c.1067G > A (p.Cys356Tyr) in ANO5 gene in these patients. To date, autosomal-dominant mutations have been described in the ANO5 gene for gnathodiaphyseal dysplasia (GDD, OMIM: 166260), and multiple recessive mutations have been described in the gene for muscle dystrophies (OMIM: 613319, 611307); the same amino acid (Cys) at the position 356 is mutated in GDD. These genetic data and similar clinical phenotypes demonstrate that the GC and GDD likely represent the same type of bone pathology. Our data illustrate the significance of mutations in single amino-acid position for particular bone tissue pathology. Modifying role of genetic variations in another gene on the severity of the monogenic trait pathology is also suggested. Finally, we propose the model explaining the tissue-specific manifestation of clinically distant bone and muscle diseases linked to mutations in one gene.

[Analysis of clusterin gene (CLU/APOJ) polymorphism in Alzheimer's disease patients and in normal cohorts from Russian populations].

Three genes mutations in which cause familial forms of Alzheimer's disease are known to date:PSEN1, PSEN2 and APP; and APOE gene polymorphism is a strong risk factor for Alzheimer's disease. We have evaluated allele and genotype frequency distribution of rs11136000 polymorphism in clusterin (CLU) gene (or apolipoprotein J, APOJ) in populations of three Russian regions and i nAlzheimhner's diseasepatients. Genome-wideassociation studies in samples from several European populations have recently revealed highly significant association o fCLU gene with AD (p = 8.5 x 10(-10)). We found no differences in allele and genotype frequencies of rs11136000 between populations from Moscow, Ural and Siberia regions. The allele frequencies are close to those in European populations. The genetic association analysis in cohort of Alzheimer's disease patients and normal individuals (>500 individuals ineach group) revealed no significant association of the rs11136000 polymorphism in CLU with Alzheimer's disease in Russian populations. Although our resultsdo not confirm the role of CLU gene as a majorgenetic factor forcommon form of Alzheimer's disease, the data do not rule out the possibility of modest effect of CLU and interaction between CLU and APOE genotypes in etiology of Alzheimer's disease.

Achievements and peculiarities in studies of ancient DNA and DNA from complicated forensic specimens.

Studies of ancient DNA specimens started 25 years ago. At that time short mitochondrial DNA (mtDNA) fragments were the main targets in ancient DNA studies. The last three years were especially productive in the development of new methods of DNA purification and analysis. Complete mtDNA molecules and relatively large fragments of nuclear DNA are the targets of ancient DNA studies today. Ancient DNA studies allowed us to study organisms that went extinct more than ten thousand years ago, to reconstruct their phenotypic traits and evolution. Ancient DNA analyses can help understand the development of ancient human populations and how they migrated. A new evolutionary hypothesis and reconstruction of the biota history have been re-created from recent ancient DNA data. Some peculiarities and problems specific to the study of ancient DNA were revealed, such as very limited amounts of DNA available for study, the short length of the DNA fragments, breaks and chemical modifications in DNA molecules that result in "postmortem" mutations or complete blockage of DNA replication in vitro. The same specific features of DNA analysis were revealed for specimens from complicated forensic cases that result in the lack of experimental data or interpretation problems.. Here, we list the specific features of ancient DNA methodology and describe some achievements in fundamental and applied research of ancient DNA, including our own work in the field.

[Human microRNA in norm and pathology].

MicroRNAs (miRNAs) are a class of RNA which controls gene expression at the posttranscription level. Binding with the target RNA, a miRNA can supress translation and/or induce degradation of mRNA. The studies of miRNAs have already shown that miRNA were essential to switch the programs of gene expression during embryo development as well as to control cell functioning of the adult organism. Alteration of the miRNA expression profile may appear not less important in development of pathology than better known structural variations of the proteins. The role of miRNA has been confirmed for a range of common diseases connected to impaired balance of cell proliferation, differentiation and programmed death. This review discusses specific features of miRNA-mediated regulation of gene expression and its role in normal and pathological development of muscle, immune, and nervous systems. The evidence of miRNA involvent in neurodegenerative disorders and mental disorders is demonstrated.

[Molecular basics of Alzheimer's disease].

Studies of molecular mechanisms for Alzheimer's Disease have led to the two major achievements. First, genes with mutations causing Alzheimer's Disease (presenilin genes PSI, PS2 and APP) or bearing a risk factor polymorphism (ApoE) for Alzheimer's Disease were described. Second, the new type of proteases and mechanisms of regulation of cellular differentiation and development by processes of intramembrane proteolysis were identified. These mechanisms, apparently, are universal for various cell types and organisms. Presenilin is a catalytic component of tetra-protein complex (epsilon-/gamma-secretase) cleaving type I transmembrane proteins. Other recently discovered aspartate proteases, IMPAS/SPP, cleave type II transmembrane proteins. Processing of transmembrane proteins by cellular intramembrane proteases results in production of signal peptides, transcriptional factors and short hydrophobic proteins (fragments of transmembrane domains), which may have a physiological function or play a key role in patogenic events associated with ageing (e.g., beta-amyloid formation in Alzheimer's Disease). To date approximately 160 mutations in PSI gene, more than 10 mutations in PS2 gene and 21 mutations in APP gene were described. Early preclinical diagnostics of some early forms of Alzheimer's Disease became possible. Since patogeneses of early and late onset forms of Alzheimer's Disease are similar, identification of molecular or epigenetic factors affecting primary molecular mechanisms (intramembrane or membrane associated proteolysis) underlying the disease may ultimately contribute to development of rational therapy for Alzheimer's Disease.

Effects of human presenilin 1 isoforms on proliferation and survival of rat pheochromocytoma cell line PC12.

Missense mutations in human presenilin 1 gene (hPS1) cause an autosomal dominant, early onset form of Alzheimer's disease (AD). To study effects of mutant presenilin on processes of cell growth, differentiation, and susceptibility to apoptotic signals, we produced a series of rat pheochromocytoma PC12 poly- and monoclonal cell lines stably expressing wild type hPS1 and hPS1 with mutations in amino (N-) and carboxyl (C-) terminal regions of the PS1 protein. Employing a heterologous rat PC12 cell system, we demonstrated that: 1) AD mutations inhibit, in part, processing of hPS1 holoprotein; 2) negative selection against highly expressed hPS1 may occur in polyclonal cell cultures; 3) expression of N-terminus mutant (M146V) hPS1 increases susceptibility to apoptosis in differentiated neuronal PC12 cells under deprivation conditions; 4) monoclones with hPS1 C-terminal AD mutation (C410Y) have lower proliferation rates than monoclones expressing wild type hPS1 under deprivation conditions and during NGF-induced neuronal differentiation. The data demonstrate deleterious effect of PS1 AD mutations. The effect depends on the level of expression of the hPS1 isoforms, the number of passages, and trophic and differentiation conditions used for growing PC12 cells.

[Loach spermatozoa transfer foreign DNA, which expression is discovered in the early development stages]. / Spermatozoidy v'iuna perenosiat chuzherodnuiu DNK, ékspressiia kotoroi obnaruzhivaetsia na stadiiakh rannego razvitiia.

The transfer of plasmid pcDNA3-lacZ by electrotrasfected sperm cells into loach (Misgurnus fossilis L.) ova has been studied. The lacZ gene has been found to express in 3- to 5-day-old prelarvae.

Novel class of polytopic proteins with domains associated with putative protease activity.

A significant proportion of early onset Alzheimer's disease (AD) is caused by mutations in human genes for amyloid precursor protein (APP), presenilins 1 and 2 (PSEN1,2). AD associated mutations in PSEN1,2 genes alter the gamma-secretase cleavage activity of APP resulting in increased production of amyloidogenic Abeta42. PSEN dependent intramembrane proteolysis was described as an important step required for cleavage of Notch receptors, Notch-dependent signal transduction, and processing of other proteins. It is still unclear whether presenilins are unusual intramembrane proteases or they are necessary cofactors of gamma-secretase cleavage of APP and Notch. Identification of other proteins similar to presenilins may resolve this dilemma. We describe here the identification of novel families of genes encoding polytopic transmembrane proteins of Eukaryotes (IMPASes) and Arachaea(membrases). These proteins have a predicted structure similar to presenilins. The amino acid similarity is significant in domains carrying invariant amino acid residues, which are critical in specific presenilin-regulated endoproteolysis. Many members of the IMPAS family have protease associated domains (PA) typical of proteases. We identified and cloned five human IMPAS genes. Expression analysis of the hIMP1 gene (located on chromosome 20) was performed in human cell tissues and transfected cell cultures. The data demonstrate that a conservative class of putative protease-related polytopic proteins related to presenilins exists in multicellular eukaryotes and microorganisms.

[Expression of the CMV-lacZ- and RSV-lacZ-genes in transgenic fish and mouse embryos]. / Ekspressiia CMV-lacZ- i RSV-lacZ-genov v transgennykh émbryionakh ryb i myshei.

The bacterial gene for beta-galactosidase under the control of LTR from either human cytomegalovirus (CMV-lacZ) or the Rous sarcoma virus (RSV-lacZ) was injected into fertilized eggs of Misgurnus fossilis L. loaches and F1 hybrid mice (CBA x C57Bl). Expression of the CMV-lacZ was observed in almost 100% of the loach embryos and larvae for two months following the first day of embryonic development. In some cases, expression points were located only on either the right or the left side of a fish. The spectrum of tissues expressing CMV-lacZ was decreased during embryonic development: CMV-lacZ was expressed only in fin and body muscles of 6- to 8-week-old loaches. In mice, the RSV-lacZ gene was expressed in ectoderm- and mesoderm-derived tissues of a 13-day-old embryo, and the CMV-lacZ gene was expressed in tissues derived from various blastophylli of a 14-day-old embryo. Distribution of transgene expression is discussed with regard to authors' and published data.

[Biological sorption in immunoneurologic diseases]. / Biologicheskaia sorbtsiia pri immunonevrologicheskikh zabolevaniiakh.

A study is presented of 17 patients--9 with myasthenia, 8 with the Guillain--Barré syndrome. The complex treatment program of intensive therapy and reanimation included in all patients the operation of extracorporal heterospleen. Forty-one perfusions were carried out. The operation of splenic sorption allowed to reduce treatment time under conditions of intensive care and reanimation 1.5 times, lethality--by 33%. Employment of biological sorption in patients with such immunoneurological diseases as myasthenia and the Guillain--Barré syndrome removes the humoral autoallergic factor and, thus, favours restoration of nervous system function.