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Topic:Molecular biology

Molecular biology in horses involves the study of molecular processes and genetic mechanisms that underpin equine physiology and health. This field encompasses the analysis of DNA, RNA, proteins, and other biomolecules to understand gene expression, genetic variation, and cellular functions in horses. Techniques such as genomic sequencing, gene expression profiling, and molecular diagnostics are employed to explore topics like hereditary diseases, performance traits, and immune responses in equines. This page assembles peer-reviewed research studies and scholarly articles that investigate the molecular biology of horses, focusing on genetic research, molecular techniques, and their applications in equine science.
Radiation hybrid mapping of equine CDK2, DGKA, DNAJC14, MMP19, CTSL and GAS1.
Animal genetics    November 19, 2005   Volume 36, Issue 6 536-537 doi: 10.1111/j.1365-2052.2005.01381.x
Wittwer C, Chowdhary BP, Distl O.No abstract available
Differential susceptibility of equine and mouse brain microvascular endothelial cells to equine herpesvirus 1 infection.
Archives of virology    November 17, 2005   Volume 151, Issue 4 775-786 doi: 10.1007/s00705-005-0653-3
Hasebe R, Kimura T, Nakamura K, Ochiai K, Okazaki K, Wada R, Umemura T.Equine herpesvirus 1 (EHV-1) shows endotheliotropism in the central nervous system (CNS) of infected horses. However, infection of endothelial cells has not been observed in the CNS of infected mice. To explore the basis for this difference in endotheliotropism, we compared the susceptibility of equine brain microvascular endothelial cells (EBMECs) and mouse brain microvascular endothelial cells (MBMECs) to EHV-1 infection. The kinetics of viral growth in EBMECs was typical of a fully productive infection whereas viral infection in MBMECs seemed to be nonproductive. Immunofluorescence microsco...
cDNA sequence of the horse (Equus caballus) LAMA3 gene and characterization of two intronic SNP markers.
DNA sequence : the journal of DNA sequencing and mapping    November 17, 2005   Volume 16, Issue 6 468-473 doi: 10.1080/10425170500287674
Milenkovic D, Mata X, Chadi S, Guérin G.Laminins are large heterotrimeric basement membrane glycoproteins composed of alpha, beta and gamma chains. The Laminin 5 isoform has an alpha3beta3gamma2 composition and is essential for the adhesion of basal keratinocytes to the underlying epithelial basement membrane where it is mainly located. Mutations in the genes coding for the 3 chains have been associated with a severe skin blistering disease, Herlitz's junctional epidermolysis bullosa (JEB), observed in different species as man, dog, cat and horse. In this study, we report the sequence of the 5.2 kb horse laminin alpha 3 cDNA (LAMA3)...
Exon skipping in the KIT gene causes a Sabino spotting pattern in horses.
Mammalian genome : official journal of the International Mammalian Genome Society    November 11, 2005   Volume 16, Issue 11 893-902 doi: 10.1007/s00335-005-2472-y
Brooks SA, Bailey E.Sabino (SB) is a white spotting pattern in the horse characterized by white patches on the face, lower legs, or belly, and interspersed white hairs on the midsection. Based on comparable phenotypes in humans and pigs, the KIT gene was investigated as the origin of the Sabino phenotype. In this article we report the genetic basis of one type of Sabino spotting pattern in horses that we call Sabino 1, with the alleles represented by the symbols SB1 and sb1. Transcripts of KIT were characterized by reverse transcriptase polymerase chain reaction (RT-PCR) and sequencing cDNA from horses with the g...
Differences in tumor necrosis factor (TNF)alpha and TNF receptor-1-mediated intracellular signaling factors in normal, inflamed and scar-formed horse tendons.
The Journal of veterinary medical science    November 9, 2005   Volume 67, Issue 10 985-991 doi: 10.1292/jvms.67.985
Hosaka Y, Kirisawa R, Ueda H, Yamaguchi M, Takehana K.Tumor necrosis factor (TNF) receptors (TNF-R)-mediated cell survival or apoptosis has been demonstrated in many cells, but little is known about survival or apoptotic signals via TNF-R1 in tendinocytes. In this study, we focused on four signaling factors, TNFalpha, TNF-R1, TNFR-associated factor2 (TRAF2) and caspase-3, in order to elucidate the signaling events in tendinocytes. Samples were obtained from normal, inflamed and scar-formed equine superficial digital flexor tendons. To detect these signaling factors, samples were subjected to immunohistochemistry and Western blot analysis, and som...
Genomic characterization of MHC class I genes of the horse.
Immunogenetics    November 8, 2005   Volume 57, Issue 10 763-774 doi: 10.1007/s00251-005-0034-9
Tallmadge RL, Lear TL, Antczak DF.The availability of a contig of bacterial artificial chromosome (BAC) clones spanning the equine major histocompatibility complex (MHC) made possible a detailed analysis of horse MHC class I genes. Prior to this study, only a single horse MHC class I gene had been sequenced at the genomic level. Although many ( approximately 60) MHC class I cDNA sequences had been determined and published, from this information, it was not possible to determine how many class I loci are expressed in horses or to assign individual sequences to allelic series. In this study, 15 MHC class I genes were identified ...
Studies of fibronectin-binding proteins of Streptococcus equi.
Infection and immunity    October 22, 2005   Volume 73, Issue 11 7243-7251 doi: 10.1128/IAI.73.11.7243-7251.2005
Lannergård J, Flock M, Johansson S, Flock JI, Guss B.Streptococcus equi subsp. equi is the causative agent of strangles, a disease of the upper respiratory tract in horses. The initiation of S. equi subsp. equi infection is likely to involve cell surface-anchored molecules mediating bacterial adhesion to the epithelium of the host. The present study describes the cloning and characterization of FNEB, a fibronectin-binding protein with cell wall-anchoring motifs. FNEB can thus be predicted as cell surface located, contrary to the two previously characterized fibronectin-binding proteins in S. equi subsp. equi, FNE and SFS. Assays of antibody tite...
Production and characterization of recombinant equine prorelaxin.
Domestic animal endocrinology    October 21, 2005   Volume 31, Issue 2 173-185 doi: 10.1016/j.domaniend.2005.10.001
Neumann JL, Lazaris A, Huang YJ, Karatzas C, Ryan PL, Bagnell CA.Relaxin is a peptide hormone produced by a wide variety of mammals. In the horse, the placenta is the major source of relaxin. Since pure equine relaxin is difficult to obtain to study its role in the pregnant mare, the objectives of this study were to produce recombinant equine prorelaxin and characterize its immunological and biological activity. First, an equine relaxin gene cassette was transfected into immortalized bovine mammary epithelial (MAC-T) cells. Second, immunological activity of media conditioned by transfected MAC-T cells was tested by Western blotting and quantified using a ho...
Misregulated Wnt/beta-catenin signaling leads to ovarian granulosa cell tumor development.
Cancer research    October 19, 2005   Volume 65, Issue 20 9206-9215 doi: 10.1158/0008-5472.CAN-05-1024
Boerboom D, Paquet M, Hsieh M, Liu J, Jamin SP, Behringer RR, Sirois J, Taketo MM, Richards JS.Misregulation of the Wnt/beta-catenin signaling pathway is a hallmark of several forms of cancer. Components of the Wnt/beta-catenin pathway are expressed in ovarian granulosa cells; nevertheless, its potential involvement in granulosa cell tumorigenesis has not been examined. To this end, human (n = 6) and equine (n = 18) granulosa cell tumors (GCT) were analyzed for beta-catenin expression by immunohistochemistry. Unlike granulosa cells of normal ovaries, most (15 of 24) GCT samples showed nuclear localization of beta-catenin, suggesting that activation of the Wnt/beta-catenin pathway plays ...
Mass spectral analysis of domestic and wild equine apoA-I and A-II: detection of unique dimeric forms of apoA-II.
Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology    October 17, 2005   Volume 142, Issue 4 369-373 doi: 10.1016/j.cbpb.2005.08.008
Puppione DL, Whitelegge JP, Yam LM, Bassilian S, Schumaker VN, MacDonald MH.In pigs, humans, chimpanzees and probably other great apes, a cysteine at residue 6 enables apolipoprotein A-II to form a homodimer. However, the apoA-IIs of other primates, lacking a cysteine residue, are monomeric. We have already reported that horse apoA-IIs form homodimers due also to a cysteine at residue 6. In this study, we wanted to determine whether other equine apoA-IIs might be monomeric. The high density lipoproteins were ultracentrifugally isolated from the plasmas of a horse (Equus caballus), a donkey (Equus asinus) and five wild equines: two types of zebras (Equus zebra hartmann...
Real-time reverse transcription PCR for detection and quantitative analysis of equine influenza virus.
Journal of clinical microbiology    October 7, 2005   Volume 43, Issue 10 5055-5057 doi: 10.1128/JCM.43.10.5055-5057.2005
Quinlivan M, Dempsey E, Ryan F, Arkins S, Cullinane A.Equine influenza is a cause of epizootic respiratory disease of the equine. The detection of equine influenza virus using real-time Light Cycler reverse transcription (RT)-PCR technology was evaluated over two influenza seasons with the analysis of 171 samples submitted for viral respiratory disease. Increased sensitivity was found in overall viral detection with this system compared to Directigen Flu A and virus isolation, which were 40% and 23%, respectively, that of the RT-PCR. The assay was also evaluated as a viable replacement for the more traditional methods of quantifying equine influe...
Equine and canine Anaplasma phagocytophilum strains isolated on the island of Sardinia (Italy) are phylogenetically related to pathogenic strains from the United States.
Applied and environmental microbiology    October 6, 2005   Volume 71, Issue 10 6418-6422 doi: 10.1128/AEM.71.10.6418-6422.2005
Alberti A, Zobba R, Chessa B, Addis MF, Sparagano O, Pinna Parpaglia ML, C뻝u T, Pintori G, Pittau M.The presence of Anaplasma phagocytophilum, a tick-transmitted zoonotic pathogen, was investigated in Sardinia using a molecular approach. Phylogenetic analysis revealed that Sardinian strains are genetically distinct from the two lineages previously described in Europe and are closely related to strains isolated in different areas of the United States.
Expression of inhibins, activins, insulin-like growth factor-I and steroidogenic enzymes in the equine placenta.
Domestic animal endocrinology    September 29, 2005   Volume 31, Issue 1 19-34 doi: 10.1016/j.domaniend.2005.09.005
Arai KY, Tanaka Y, Taniyama H, Tsunoda N, Nambo Y, Nagamine N, Watanabe G, Taya K.In this study, the expression patterns of inhibins, activins, insulin-like growth factor-I (IGF-I) and steroidogenic enzymes in equine placentae recovered during the latter two-thirds of gestation were examined. Concentrations of inhibin A and inhibin pro-alphaC in endometrial and fetal placental tissue homogenates were very low during the period examined, whereas these tissues contained high concentrations of activin A. In both maternal endometrial and fetal placental tissues, activin A levels decreased as pregnancy progressed. Expression of inhibin alpha-subunit was not observed in the place...
Genetic structure of an endangered Portuguese semiferal pony breed, the Garrano.
Biochemical genetics    September 28, 2005   Volume 43, Issue 7-8 347-364 doi: 10.1007/s10528-005-6775-1
Morais J, Oom MM, Malta-Vacas J, Luís C.The present study intends to survey the genetic variability of an endangered semiferal Portuguese native pony breed, the Garrano. Thirteen microsatellite markers were examined in 277 animals born in 1998, belonging to eight subpopulations corresponding to eight northern Portuguese geographic regions. Mean heterozygosity (H(o)) in the Garrano breed was 0.732, ranging from 0.531 to 0.857 across subpopulations. Allelic frequencies and diversity differed significantly between regions, suggesting the existence of genetic differentiation within the breed confirmed by the population differentiation e...
Proopiomelanocortin gene expression and beta-endorphin localization in the pituitary, testis, and epididymis of stallion.
Molecular reproduction and development    September 24, 2005   Volume 73, Issue 1 1-8 doi: 10.1002/mrd.20341
Soverchia L, Mosconi G, Ruggeri B, Ballarini P, Catone G, Degl'Innocenti S, Nabissi M, Polzonetti-Magni AM.Proopiomelanocortin (POMC) is a precursor protein that contains the sequences of several bioactive peptides including adrenocorticotropin (ACTH), beta-endorphin (beta-EP), and melanocyte-stimulating-hormone (MSH). POMC is synthesized in the pituitary gland, brain, and many peripheral tissues. Immunoreactive POMC-derived peptides as well as POMC-like mRNA have been evidenced in several nonpituitary tissues, thus suggesting that POMC is actively synthesized by these tissues. The present study was aimed at evaluating if also in the case of stallion POMC-derived peptide, beta-EP, is produced local...
Formation of a cytochrome c-nitrous oxide reductase complex is obligatory for N2O reduction by Paracoccus pantotrophus.
Dalton transactions (Cambridge, England : 2003)    September 23, 2005   Issue 21 3501-3506 doi: 10.1039/b501846c
Rasmussen T, Brittain T, Berks BC, Watmough NJ, Thomson AJ.Nitrous oxide reductase (N2OR) catalyses the final step of bacterial denitrification, the two-electron reduction of nitrous oxide (N2O) to dinitrogen (N2). N2OR contains two metal centers; a binuclear copper center, CuA, that serves to receive electrons from soluble donors, and a tetranuclear copper-sulfide center, CuZ, at the active site. Stopped flow experiments at low ionic strengths reveal rapid electron transfer (kobs=150 s-1) between reduced horse heart (HH) cytochrome c and the CuA center in fully oxidized N2OR. When fully reduced N2OR was mixed with oxidized cytochrome c, a similar rat...
Evaluation of a multiplex polymerase chain reaction assay for simultaneous detection of Rhodococcus equi and the vapA gene.
American journal of veterinary research    September 22, 2005   Volume 66, Issue 8 1380-1385 doi: 10.2460/ajvr.2005.66.1380
Halbert ND, Reitzel RA, Martens RJ, Cohen ND.To evaluate sensitivity and specificity of a multiplex polymerase chain reaction (PCR) assay for simultaneous detection of Rhodococcus equi and differentiation of strains that contain the virulence-associated gene (vapA) from strains that do not. Methods: 187 isolates of R equi from equine and nonequine tissue and environmental specimens and 27 isolates of bacterial species genetically or morphologically similar to R equi. Methods: The multiplex PCR assay included 3 gene targets: a universal 311-bp bacterial 16S ribosomal RNA amplicon (positive internal control), a 959-bp R equi-specific targe...
Deprotonation of the horse liver alcohol dehydrogenase-NAD+ complex controls formation of the ternary complexes.
Biochemistry    September 21, 2005   Volume 44, Issue 38 12797-12808 doi: 10.1021/bi050865v
Kovaleva EG, Plapp BV.Binding of NAD+ to wild-type horse liver alcohol dehydrogenase is strongly pH-dependent and is limited by a unimolecular step, which may be related to a conformational change of the enzyme-NAD+ complex. Deprotonation during binding of NAD+ and inhibitors that trap the enzyme-NAD+ complex was examined by transient kinetics with pH indicators, and formation of complexes was monitored by absorbance and protein fluorescence. Reactions with pyrazole and trifluoroethanol had biphasic proton release, whereas reaction with caprate showed proton release followed by proton uptake. Proton release (200-55...
Direct activation of gelatinase B (MMP-9) by hay dust suspension and different components of organic dust.
Veterinary immunology and immunopathology    September 21, 2005   Volume 109, Issue 3-4 289-295 doi: 10.1016/j.vetimm.2005.08.019
Simonen-Jokinen T, Maisi P, Tervahartiala T, McGorum B, Pirie S, Sorsa T.Matrix metalloproteinases (MMPs) are involved in tissue destruction in allergic airway diseases. We studied the ability of various allergenic substances to directly activate recombinant 92kDa proMMP-9. The substances included hay dust suspension (HDS) and its components (supernatant, particulate matter and wash fluid of particulate matter), storage mite extract and two Aspergillus fumigatus extracts. The allergen suspensions were incubated in vitro with proMMP-9. After incubation the conversion of proMMP-9 to 10kDa lower active forms were studied using gelatin zymography and Western immunoblot...
Assignment of BGLAP, BMP2, CHST4, SLC1A3, SLC4A1, SLC9A5 and SLC20A1 to equine chromosomes by FISH and confirmation by RH mapping.
Animal genetics    September 20, 2005   Volume 36, Issue 5 457-461 doi: 10.1111/j.1365-2052.2005.01347.x
Müller D, Kuiper H, Böneker C, Mömke S, Drögemüller C, Chowdhary BP, Distl O.No abstract available
Molecular characterization and functional expression of equine interleukin-1 type I and type II receptor cDNAs.
Veterinary immunology and immunopathology    September 19, 2005   Volume 109, Issue 3-4 219-231 doi: 10.1016/j.vetimm.2005.08.018
Kirisawa R, Hashimoto N, Tazaki M, Yamanaka H, Ishii R, Hagiwara K, Iwai H.cDNA generated from lipopolysaccharide-stimulated equine peripheral blood mononuclear cells was used to amplify and clone type I and type II equine interleukin-1 receptors (IL-1RI and IL-1RII) using primers derived from semi-conserved regions between human and mouse IL-1RI and IL-1RII sequences, respectively. 5' and 3' terminal sequences of equine IL-1RI and IL-1RII were amplified by 5' and 3' rapid amplification of cDNA ends. The deduced amino acid sequence of equine IL-1RI demonstrated 77, 64 and 63% similarity with human, mouse and rat sequences, respectively. The predicted amino acid seque...
Single nucleotide polymorphisms in four functionally related immune response genes in the horse: CD14,TLR4, Cepsilon, andFcepsilon R1 alpha.
International journal of immunogenetics    September 17, 2005   Volume 32, Issue 5 277-283 doi: 10.1111/j.1744-313X.2005.00522.x
Vychodilova-Krenkova L, Matiasovic J, Horin P.The objective of this study was to identify single nucleotide polymorphisms (SNPs) within four functionally related immune response genes in the horse, and to develop genotyping techniques that could be useful for future genomic studies of horse infectious and allergic diseases. The genes analysed were: the lipopolysaccharide (LPS) receptor gene CD14, the toll-like receptor 4 gene TLR4, the gene Cepsilon encoding the IgE heavy chain molecule and the gene FcepsilonR1 alpha coding for the alpha subunit of the IgE receptor molecule. Horse-specific primers amplifying selected gene regions were des...
Nucleotide sequence of complementary DNA encoding for quaking protein of cow, horse and pig.
DNA sequence : the journal of DNA sequencing and mapping    September 9, 2005   Volume 16, Issue 4 300-303 doi: 10.1080/10425170500136731
Murata T, Yamashiro Y, Kondo T, Nakaichi M, Une S, Taura Y.Complementary DNA (cDNA) for bovine quaking gene (Bqk), equine quaking gene (Eqk) and porcine quaking gene (Pqk), which are homologous to mouse quaking gene (qkI), were isolated, and their nucleotide sequences were determined. cDNA sequences of Bqk, Eqk and Pqk showed very high homology to that of qkI at nucleotide level; 94.2, 95.7 and 95.6%, respectively. Deduced amino acid sequences for Bqk, Eqk and Pqk perfectly matched to that of qkI. These findings suggest that the quaking gene family is highly conserved during mammalian evolution, and that Bqk, Eqk and Pqk are likely to have important b...
A novel method for determination of collagen orientation in cartilage by Fourier transform infrared imaging spectroscopy (FT-IRIS).
Osteoarthritis and cartilage    September 9, 2005   Volume 13, Issue 12 1050-1058 doi: 10.1016/j.joca.2005.07.008
Bi X, Li G, Doty SB, Camacho NP.The orientation of collagen molecules is an important determinant of their functionality in connective tissues. The objective of the current study is to establish a method to determine the alignment of collagen molecules in histological sections of cartilage by polarized Fourier transform infrared imaging spectroscopy (FT-IRIS), a method based on molecular vibrations. Methods: Polarized FT-IRIS data obtained from highly oriented tendon collagen were utilized to calibrate the derived spectral parameters. The ratio of the integrated areas of the collagen amide I/II absorbances was used as an ind...
Polymorphic study of equine antiviral MXA gene.
Biochemical genetics    September 8, 2005   Volume 43, Issue 5-6 299-305 doi: 10.1007/s10528-005-5221-8
Ju LH, Onogi A, Ueda J, Yamada K, Nakatsu Y, Ohe M, Hata H, Sasaki K, Watanabe T.No abstract available
Sex reversal syndrome (64,XY; SRY-positive) in a mare demonstrating masculine behaviour.
Journal of animal breeding and genetics = Zeitschrift fur Tierzuchtung und Zuchtungsbiologie    September 1, 2005   Volume 122 Suppl 1 60-63 doi: 10.1111/j.1439-0388.2005.00510.x
Switonski M, Chmurzynska A, Szczerbal I, Lipczynski A, Yang F, Nowicka-Posłuszna A.A 5-year-old Thoroughbred mare was subjected to cytogenetic and molecular analysis because of infertility and masculine behaviour. Chromosome studies, including painting with the whole X chromosome specific probe, revealed a male chromosome complement (64,XY). The PCR amplification of the SRY and ZFY genes showed the presence of both those genes, while the endocrinological study demonstrated a high level of testosterone (9.7 nmol/l). Sequencing of the SRY gene (1121 bp), comprising also 5'- and 3'-UTRs, did not reveal any differences when compared with the sequence of normal stallions. It was ...
Comparative chromosomal studies of E. caballus (ECA) and E. przewalskii (EPR) in a female F1 hybrid.
Journal of animal breeding and genetics = Zeitschrift fur Tierzuchtung und Zuchtungsbiologie    September 1, 2005   Volume 122 Suppl 1 97-102 doi: 10.1111/j.1439-0388.2005.00494.x
Ahrens E, Stranzinger G.Previous research revealed that the karyotypes of Equus przewalskii (2n = 66) and Equus caballus (2n = 64) differ by one pair of metacentric chromosomes, present in ECA but not in EPR, and two pairs of acrocentric chromosomes found only in the EPR karyotype. The formation of a trivalent during meiosis in a male F1 hybrid and the homologies in G-banding patterns suggest that ECA 5 corresponds to two acrocentric EPR chromosomes resulting from a Robertsonian fusion or fission event. Chromosomal investigations of a female interspecies F1 hybrid including banded karyograms and fluorescence in situ ...
[Inhibition of catalytic activity of butyrylcholinesterase from the horse serum by high concentrations of N-methyl-N-(beta-acetoxyethyl)-piperidinium substrate].
Zhurnal evoliutsionnoi biokhimii i fiziologii    August 30, 2005   Volume 41, Issue 4 339-342 
Zhukovskiĭ IuG, Kuznetsova LP, Sochilina EE.No abstract available
The role of stallion seminal proteins in fertilisation.
Animal reproduction science    August 30, 2005   Volume 89, Issue 1-4 159-170 doi: 10.1016/j.anireprosci.2005.06.018
Töpfer-Petersen E, Ekhlasi-Hundrieser M, Kirchhoff C, Leeb T, Sieme H.Seminal plasma proteins are secretory proteins originating mainly from the epididymis and the accessory sex glands. They are involved in the remodelling of the sperm surface which occurs during sperm transit through the male genital tract and continues later at ejaculation. During this process, collectively called post-testicular sperm maturation, the spermatozoa acquire the ability to fertilise an egg. Seminal plasma proteins have been shown to contribute to early and central steps of the fertilisation sequence, e.g. the establishment of the oviductal sperm reservoir, modulation of capacitati...
Detection of equine herpesvirus type 1 using a real-time polymerase chain reaction.
Journal of virological methods    August 30, 2005   Volume 131, Issue 1 92-98 doi: 10.1016/j.jviromet.2005.07.010
Diallo IS, Hewitson G, Wright L, Rodwell BJ, Corney BG.Equid herpesvirus 1 (EHV1) is a major disease of equids worldwide causing considerable losses to the horse industry. A variety of techniques, including PCR have been used to diagnose EHV1. Some of these PCRs were used in combination with other techniques such as restriction enzyme analysis (REA) or hybridisation, making them cumbersome for routine diagnostic testing and increasing the chances of cross-contamination. Furthermore, they involve the use of suspected carcinogens such as ethidium bromide and ultraviolet light. In this paper, we describe a real-time PCR, which uses minor groove-bindi...