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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.
Characterization of muscarinic receptor subtype mediating contraction and relaxation in equine coronary artery in vitro.
Journal of veterinary pharmacology and therapeutics    June 1, 1994   Volume 17, Issue 3 226-231 doi: 10.1111/j.1365-2885.1994.tb00237.x
Obi T, Kabeyama A, Nishio A.In coronary arterial rings isolated from horses, 10(-8)-10(-6) mol/l acetylcholine (ACh) induced concentration-dependent contractions which were potentiated by the removal of endothelium and by pretreatment with L-nitro-arginine (LNAG) or methylene blue (MB). Relatively lower concentrations of ACh (10(-14)-10(-8) mol/l) induced relaxation when the coronary rings were contracted by phenylephrine (PE). ACh-induced contractions in the coronary rings without endothelium were competitively inhibited by each muscarinic subtype selective antagonist in the following order of potency: 4-diphenylacetoxy...
Structural and functional characterization of elastases from horse neutrophils.
The Biochemical journal    June 1, 1994   Volume 300 ( Pt 2), Issue Pt 2 401-406 doi: 10.1042/bj3000401
Dubin A, Potempa J, Travis J.In order better to understand the pathophysiology of the equine form of emphysema, two elastinolytic enzymes from horse neutrophils, referred to as proteinases 2A and 2B, have been extensively characterized and compared with the human neutrophil proteinases, proteinase-3 and elastase. Specificity studies using both the oxidized insulin B-chain and synthetic peptides revealed that cleavage of peptide bonds with P1 alanine or valine residues was preferred. Further characterization of the two horse elastases by N-terminal sequence and reactive-site analyses indicated that proteinases 2A and 2B ha...
Comparison of M and N gene sequences distinguishes variation amongst equine arteritis virus isolates.
The Journal of general virology    June 1, 1994   Volume 75 ( Pt 6) 1491-1497 doi: 10.1099/0022-1317-75-6-1491
Chirnside ED, Wearing CM, Binns MM, Mumford JA.cDNA copies of the M and N genes of equine arteritis virus (EAV) isolates were synthesized by reverse transcription followed by polymerase chain reaction amplification. The cDNA was subjected to a cycle sequencing strategy using Taq polymerase, and the nucleotide and derived amino acid sequences of 10 virus isolates were compared. The M and N genes of all isolates had the same initiation and termination sites as the prototype Bucyrus strain and the encoded proteins were conserved between viruses. Comparison of nucleotide sequence homologies and phylogenetic tree analysis implied the existence ...
Cloning and sequencing of the equine testicular follitropin receptor.
Biochemical and biophysical research communications    May 30, 1994   Volume 201, Issue 1 201-207 doi: 10.1006/bbrc.1994.1689
Robert P, Amsellem S, Christophe S, Benifla JL, Bellet D, Koman A, Bidart JM.To investigate the possibility that specific structural determinants within the equine follitropin receptor (eFSHR) are critical to the enhanced specificity of this receptor compared to other FSHRs, we used the RACE-PCR technique to clone the eFSHR from equine testis. Sequence analysis revealed that the eFSHR is highly homologous to other mammal FSHRs, but it presents 10 unique amino acid residue replacements in the extracellular domain. Furthermore, a potential N-glycosylation site was detected at a position not encountered in other receptors. Northern blot analysis identified three transcrip...
Isolation, primary structures and metal binding properties of neuronal growth inhibitory factor (GIF) from bovine and equine brain.
FEBS letters    May 30, 1994   Volume 345, Issue 2-3 193-197 doi: 10.1016/0014-5793(94)00452-8
Pountney DL, Fundel SM, Faller P, Birchler NE, Hunziker P, Vasák M.Human neuronal growth inhibitory factor (GIF) impairs the survival of cultured neurons and is deficient in the brains of Alzheimer's disease victims. We have isolated and sequenced analogous proteins from bovine and equine brain. By comparing their primary structures with those of human, mouse and rat GIFs, a consensus GIF sequence was obtained. Although this exhibits ca. 65% similarity with primary structures of mammalian metallothioneins (MTs), some significant differences are expected in the content of helix and turn secondary structures. In contrast to MTs, which usually bind 7 Zn(II) ions...
A versatile synthetic peptide-based ELISA for identifying antibody epitopes.
Journal of immunological methods    May 2, 1994   Volume 171, Issue 1 37-44 doi: 10.1016/0022-1759(94)90226-7
Ball JM, Henry NL, Montelaro RC, Newman MJ.A simple, versatile and very inexpensive procedure for cross-linking synthetic peptides to the polystyrene surfaces of micro-well assay plates for use in ELISA was developed. The method is based on the use of poly-L-lysine (PLL) as the anchor protein for synthetic peptides which were then easily and covalently linked to the PLL using glutaraldehyde. The synthetic peptides used for the study were based on the amino acid sequence of the equine infectious anemia virus (EIAV) envelope sequence and evaluated as antigens in an ELISA designed to detect antibodies in serum of EIAV-infected horses and ...
Purification of a plasminogen activator from Streptococcus uberis.
FEMS microbiology letters    May 1, 1994   Volume 118, Issue 1-2 153-158 doi: 10.1111/j.1574-6968.1994.tb06818.x
Leigh JA.A protein capable of activating bovine, equine and ovine plasminogen, but not that from human or porcine plasma, was purified from culture filtrates of Streptococcus uberis (strain 0140J). Purification was achieved by ammonium sulphate precipitation followed by molecular exclusion chromatography. The elution position of the native molecule was equivalent to a molecular mass of approximately 57 kDa. However, the molecular mass, as determined by SDS-PAGE, was 29 kDa, suggesting the existence of a dimeric structure. Purified immunoglobulin from three out of five monoclonal antibodies raised to th...
Studies on glycoprotein-derived carbohydrates.
Biochemical Society transactions    May 1, 1994   Volume 22, Issue 2 370-373 doi: 10.1042/bst0220370
Vliegenthart JF.This research focuses on the study of glycoproteins, specifically investigating their carbohydrate chains and their various functions in living organisms. The article highlights the challenges in isolating specific carbohydrate chains […]
Species specificity and interspecies relatedness in VP4 genotypes demonstrated by VP4 sequence analysis of equine, feline, and canine rotavirus strains.
Virology    May 1, 1994   Volume 200, Issue 2 390-400 doi: 10.1006/viro.1994.1203
Taniguchi K, Urasawa T, Urasawa S.We determined the nucleotide and deduced amino acid sequences of the VP4 genes of five equine, two feline, and two canine rotavirus strains. A high degree of homology (> 97.0%) was found among the VP4 amino acid sequences of the equine strains H2, FI-14, and FI23. Equine strain L338 has a distinct VP4 amino acid sequence from those of the other equine strains (78.1% or less homology), and the L338 VP4 exhibited more than 17.0% divergence at the amino acid level from those of rotavirus strains published so far. The VP4 amino acid sequence of equine strain H1, which showed low homology with t...
Concentration and molecular weight distribution of hyaluronate in synovial fluid from clinically normal horses and horses with diseased joints.
American journal of veterinary research    May 1, 1994   Volume 55, Issue 5 710-715 
Tulamo RM, Heiskanen T, Salonen M.High molecular weight (MW) hyaluronate (HA) is an integral part of synovial fluid (SF), regulating many important physiologic and pathophysiologic mechanisms. Many of its effects depend on, or are reflected in, the concentration and MW of HA. High-performance liquid chromatography was used to assess simultaneously the concentration and MW of HA in SF obtained from horses with various arthritides: acute traumatic arthritis; chronic traumatic arthritis, including degenerative joint disease (DJD); and infectious arthritis. The size-exclusion column was calibrated, using appropriate HA concentrati...
The transition from inhomogeneous to homogeneous kinetics in CO binding to myoglobin.
Biophysical journal    May 1, 1994   Volume 66, Issue 5 1612-1622 doi: 10.1016/S0006-3495(94)80953-1
Agmon N, Doster W, Post F.Heme proteins react inhomogeneously with ligands at cryogenic temperatures and homogeneously at room temperature. We have identified and characterized a transition from inhomogeneous to homogeneous behavior at intermediate temperatures in the time dependence of CO binding to horse myoglobin. The turnover is attributed to a functionally important tertiary protein relaxation process during which the barrier increases dynamically. This is verified by a combination of theory and multipulse measurements. A likely biological significance of this effect is in the autocatalysis of the ligand release p...
Extracellular ATP can activate autonomic signal transduction pathways in cultured equine sweat gland epithelial cells.
The Journal of experimental biology    May 1, 1994   Volume 190 239-252 doi: 10.1242/jeb.190.1.239
Ko WH, O'Dowd JJ, Pediani JD, Bovell DL, Elder HY, Jenkinson DM, Wilson SM.Changes in intracellular free calcium concentration ([Ca2+]i) were monitored in a cell line that was derived from the equine sweat gland epithelium. ATP and closely related compounds could increase [Ca2+]i with a rank order of potency of UTP > or = ATP > ADP >> AMP = adenosine = alpha,beta-methylene-ATP. The responses to ATP and to UTP were initiated by the release of calcium from an internal store and subsequently sustained by calcium influx. The rise in [Ca2+]i thus seems to be mediated by P2U receptors that are coupled to phosphoinositidase C. Some desensitisation of this respon...
Rapid refolding of native epitopes on the surface of cytochrome c.
Biochemistry    April 5, 1994   Volume 33, Issue 13 3967-3973 doi: 10.1021/bi00179a024
Allen MJ, Jemmerson R, Nall BT.Refolding of surface epitopes on horse cytochrome c has been measured by monoclonal antibody binding. Two antibodies were used to probe re-formation of native-like surface structure: one antibody (2B5) binds to native cytochrome c near a type II turn (residue 44) while the other (5F8) binds to a different epitope on the opposite face of the protein near the amino terminus of an alpha-helical segment (residue 60). The results show that within the first approximately 100 ms of refolding all of the unfolded protein collapses to native-like folding intermediates that contain both antibody binding ...
Molecular cloning and sequencing of equine interleukin 4.
Veterinary immunology and immunopathology    April 1, 1994   Volume 40, Issue 4 379-384 doi: 10.1016/0165-2427(94)90047-7
Vandergrifft EV, Swiderski CE, Horohov DW.We have cloned equine interleukin 4 (IL-4) cDNA using the polymerase chain reaction (PCR) and primers based on the human IL-4 sequence. The cDNA was amplified from mitogen-stimulated equine peripheral blood mononuclear cells (PBMC). The cloned PCR product shares extensive homology ith IL-4 sequences from other species.
An equine tetranucleotide repeat: microsatellite MPZ001.
Animal genetics    April 1, 1994   Volume 25, Issue 2 123 doi: 10.1111/j.1365-2052.1994.tb00096.x
Breen M, Downs P, Irvin Z, Bell K.No abstract available
Six equine dinucleotide repeats: microsatellites MPZ002, 3, 4, 5, 6 and 7.
Animal genetics    April 1, 1994   Volume 25, Issue 2 124 doi: 10.1111/j.1365-2052.1994.tb00097.x
Breen M, Downs P, Irvin Z, Bell K.No abstract available
Detection of African horsesickness virus by reverse transcriptase polymerase chain reaction (RT-PCR) using primers for segment 5 (NS1 gene).
The Journal of veterinary medical science    April 1, 1994   Volume 56, Issue 2 347-352 doi: 10.1292/jvms.56.347
Mizukoshi N, Sakamoto K, Iwata A, Ueda S, Kamada M, Fukusho A.The reverse transcription followed by the polymerase chain reaction (RT-PCR) technique was applied to the detection of African horsesickness virus (AHSV) using primers specific for attenuated AHSV serotype 4 segment 5 (NS1 gene). Total RNA which contains both messenger RNA and genomic dsRNA was extracted by the acid guanidinium-phenol-chloroform method from the AHSV infected Vero cells and was used as templates to optimize the RT-PCR. A pair of primer (NP2-NP32) amplified the product of the expected size from all serotypes of attenuated AHSV when four pairs of primers were tested. Using this p...
Characterisation of proteolytic activity of excretory-secretory products from adult Strongylus vulgaris.
Veterinary parasitology    April 1, 1994   Volume 52, Issue 3-4 285-296 doi: 10.1016/0304-4017(94)90120-1
Caffrey CR, Ryan MF.An excretory-secretory (ES) preparation derived from adult Strongylus vulgaris in vitro was assessed for proteolytic activity using azocasein and synthetic, fluorogenic, peptide substrates. Fractionation was by molecular sieve fast protein liquid chromatography (molecular sieve FPLC) and resolution by gelatin-substrate sodium dodecyl sulphate-polyacrylamide gel electrophoresis (gelatin-substrate SDS-PAGE). The cysteine proteinase activator, dithiothreitol (DTT), enhanced azocaseinolysis and hydrolysis of carbobenzoxy-phenylalanyl-arginine-7-amido-4-methylcoumarin (Z-Phe-Arg-NMec) by the ES pre...
Alternative modes of polymerization distinguish the subunits of equine infectious anemia virus reverse transcriptase.
The Journal of biological chemistry    March 18, 1994   Volume 269, Issue 11 8541-8548 
Wöhrl BM, Howard KJ, Jacques PS, Le Grice SF.A comparative study of recombinant 51- and 66-kDa subunits comprising equine infectious anemia virus reverse transcriptase (EIAV RT) is reported. Both polypeptides sedimented as stable homodimers (molecular mass, 102 and 132 kDa, respectively) when analyzed by rate sedimentation through glycerol gradients. Consistent with their dimer composition, each preparation displayed considerable levels of both RNA- and DNA-dependent DNA polymerase activity on different homopolymeric template/primer combinations. However, a detailed analysis of the polymerization products indicated qualitative difference...
Formation of sulphmyoglobin during expression of horse heart myoglobin in Escherichia coli.
FEBS letters    March 7, 1994   Volume 340, Issue 3 281-286 doi: 10.1016/0014-5793(94)80154-1
Lloyd E, Mauk AG.Expression of recombinant horse heart myoglobin in Escherichia coli has been found to result in the production of both native and variable amounts (approximately 16-17% total) of two sulphmyoglobin isomers. The recombinant sulphmyoglobin produced consists primarily of the A and B isomers as identified by 1H NMR spectroscopy with no evidence for production of the C isomer. Conversion of recombinant sulphmyoglobin to the native protein can be achieved by reconstitution with protohaem IX. The possible relationship of this observation to recombinant expression of other heme proteins is discussed.
Proteolysis and antiproteolysis–a delicate balance.
Equine veterinary journal    March 1, 1994   Volume 26, Issue 2 89-90 doi: 10.1111/j.2042-3306.1994.tb04341.x
Matthews AG.No abstract available
Structure of equine type I and type II collagens.
American journal of veterinary research    March 1, 1994   Volume 55, Issue 3 425-431 
Todhunter RJ, Wootton JA, Lust G, Minor RR.Collagen type I was purified from equine skin and flexor tendon, and type II collagen was purified from equine articular cartilage. The proteoglycans in these tissues were extracted, using guanidine HCl; the collagens were solubilized, using pepsin digestion, then were selectively precipitated with NaCl. Gel electrophoresis indicated that the precipitates contained only type I or type II collagen. Amino acid analysis indicated that collagen constituted > 97% of the total protein in the precipitates. Hydroxylation of proline was 42.0 +/- 0.6% (mean +/- SEM) in alpha 1(I) and alpha 2(I), and was...
Posttranscriptional effector domains in the Rev proteins of feline immunodeficiency virus and equine infectious anemia virus.
Journal of virology    March 1, 1994   Volume 68, Issue 3 1998-2001 doi: 10.1128/JVI.68.3.1998-2001.1994
Mancuso VA, Hope TJ, Zhu L, Derse D, Phillips T, Parslow TG.By systematically dissecting the Rev proteins of feline immunodeficiency virus (FIV) and equine infectious anemia virus (EIAV), we have identified within each a short peptide that is functionally interchangeable with the effector domains found in Rev-like proteins from other retroviruses. The active sequences from FIV and EIAV differ in several respects from other known effectors and may represent a distinct class of effector domain.
Identification of 2 stallion sperm-specific proteins and their autoantibody response.
Equine veterinary journal    March 1, 1994   Volume 26, Issue 2 148-151 doi: 10.1111/j.2042-3306.1994.tb04355.x
Teuscher C, Kenney RM, Cummings MR, Catten M.In this study, 2 stallions were immunised with their own spermatozoa to ascertain whether an antisperm autoantibody response could be mounted. The results demonstrated that the stallion can recognise and respond to sperm autoantigens by producing circulating antisperm antibodies, primarily of the IgG class. Such autoantibodies appeared 2-4 weeks after inoculation and persisted for 6-20 weeks. Immunochemical characterisation by western blot identified two major sperm autoantigens, with molecular weights of 70 kD and 62 kD. Control pony stallions immunised with adjuvants alone failed to exhibit ...
Molecular entrapment of small molecules within the interior of horse spleen ferritin.
Archives of biochemistry and biophysics    February 15, 1994   Volume 309, Issue 1 178-183 doi: 10.1006/abbi.1994.1100
Webb B, Frame J, Zhao Z, Lee ML, Watt GD.A procedure for trapping small molecules inside the interior of horse spleen ferritin (HoSF) and methods for characterizing HoSF and its small entrapped molecules are described. HoSF is first dissociated into subunits by adjustment to pH 2 in the presence of the small molecules to be trapped. The pH of the dissociated HoSF is then increased to 7 at which time the dissociated subunits reassemble reforming the 24-mer HoSF, thereby trapping solvent within its interior. HoSF is then separated from unbound molecules by dialysis, ultrafiltration, and/or ammonium sulfate precipitation. Sephadex G-25 ...
Characterization of seven new horse microsatellites: HMS1, HMS2, HMS3, HMS5, HMS6, HMS7 and HMS8.
Animal genetics    February 1, 1994   Volume 25, Issue 1 62 
Guérin G, Bertaud M, Amigues Y.No abstract available
Detection of Borna disease virus RNA in naturally infected animals by a nested polymerase chain reaction.
Journal of virological methods    February 1, 1994   Volume 46, Issue 2 133-143 doi: 10.1016/0166-0934(94)90098-1
Zimmermann W, Dürrwald R, Ludwig H.Borna disease virus in naturally infected horses, a donkey and sheep was detected for the first time by amplification of viral RNA using PCR. In contrast to a control group of healthy horses, brain tissue was positive by this assay in all animals with neurological symptoms. The use of a second round of PCR with nested primers following Southern hybridization confirmed the specificity and increased the sensitivity of the test. Comparison with conventional methods recommends this technique for monitoring of BDV infections at a molecular level.
Structure determination of the disialylated poly-(N-acetyllactosamine)-containing O-linked carbohydrate chains of equine chorionic gonadotropin.
Glycoconjugate journal    February 1, 1994   Volume 11, Issue 1 35-41 doi: 10.1007/BF00732430
Hokke CH, Roosenboom MJ, Thomas-Oates JE, Kamerling JP, Vliegenthart JF.The disialylated poly-(N-acetyllactosamine)-containing O-linked oligosaccharide alditols, released by alkaline borohydride treatment of the enzymically N-deglycosylated beta-subunit of equine chorionic gonadotropin, were purified by fast protein liquid chromatography (FPLC) on Mono Q and analysed by fast ion bombardment mass spectrometry (FAB-MS) and 1H-NMR spectroscopy. The identified oligosaccharide alditols have the following structure: [Formula: see text]
Preparation and characterization of monoclonal antibodies against equine chondrocytes, osteoblasts and osteocytes.
Zentralblatt fur Veterinarmedizin. Reihe A    February 1, 1994   Volume 41, Issue 1 31-36 doi: 10.1111/j.1439-0442.1994.tb00062.x
Katayama Y, Oikawa M, Kaneko M, Yoshihara T, Yoshikawa H, Yoshikawa T.Three monoclonal antibodies capable of individually recognizing chondrocytes, osteoblasts and osteocytes were prepared. EB-1 reacted with a 55-kDa antigen on the chondrocyte membrane, EB-2 with a 110-kDa antigen on the membrane of osteoblasts and/or partial osteocytes, and EB-3 with a 130-kDa antigen on the membrane of osteocytes. These monoclonal antibodies may be useful probes for studying the differentiation and maturation of osteogenic cells.
The equine herpesvirus type 1 glycoprotein homologous to herpes simplex virus type 1 glycoprotein M is a major constituent of the virus particle.
The Journal of general virology    February 1, 1994   Volume 75 ( Pt 2) 439-442 doi: 10.1099/0022-1317-75-2-439
Pilling A, Davison AJ, Telford EA, Meredith DM.Glycoprotein 45 is a major envelope glycoprotein of equine herpesvirus type 1. The gene encoding this protein is located between map units 0.615 and 0.636 on the virus genome and evidence has suggested that it is encoded by gene 52, one of four genes within this region. Using PCR we have amplified gene 52 and subsequently cloned it into a mammalian expression vector under the control of the human cytomegalovirus immediate early gene promoter. The gene was expressed in COS-7 cells and its product was detected by immunofluorescence and Western blotting. The results indicate that glycoprotein 45 ...