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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.
Polymorphic expression of an equine T lymphocyte and neutrophil subset marker.
Veterinary immunology and immunopathology    July 1, 1994   Volume 42, Issue 1 83-89 doi: 10.1016/0165-2427(94)90091-4
Lunn DP, Holmes MA, Duffus WP.This report describes the further characterization of a group of antibodies which have been assigned to Workshop Cluster 1 by the First International Workshop on Equine Leucocyte Antigens. These antibodies recognize a 22 kDa antigen, which is present on a large subset of T lymphocytes and neutrophils, and on medullary thymocytes. The antigen is polymorphic in its expression, and three equine phenotypes could be identified using the described antibodies. The function and homology of the antigen recognized by these antibodies are unknown.
Initial characterization of equine inhibin.
Biology of reproduction    July 1, 1994   Volume 51, Issue 1 63-71 doi: 10.1095/biolreprod51.1.63
Moore KH, Dunbar BS, Bousfield GR, Ward DN.Inhibin has been characterized from a number of mammals; however, it has not been extensively studied in horses. Western blot analysis was used to examine the size heterogeneity of equine inhibin alpha- and beta-subunits. The distribution of equine inhibin activity from the initial sizing column (S-200, 25 x 94 cm) indicated that the majority of equine inhibin activity was present as larger-molecular-size forms. When the large forms were analyzed by Western blot in nonreducing conditions, alpha-subunit bands were detected at 40,000 M(r), 56,000 M(r), 80,000 M(r), and 90,000 M(r); beta a reacti...
Down-regulation followed by re-expression of equine CD4 molecules in response to phorbol myristate acetate.
Veterinary immunology and immunopathology    July 1, 1994   Volume 42, Issue 1 71-82 doi: 10.1016/0165-2427(94)90090-6
Zhang CH, Grünig G, Davis W, Antczak DF.The regulatory effects of phorbol myristate acetate (PMA) on the expression of the CD4 molecule on horse T cells were investigated. On both peripheral blood lymphocytes and thymocytes, PMA resulted in a rapid and transient down-regulation of equine CD4 expression, but had no such effect on the surface expression of equine CD5, CD8 or major histocompatibility complex (MHC) class I and class II molecules. Over 75% of the surface CD4 molecules per cell were lost after a 4 h exposure to PMA at 37 degrees C. The regulation of equine CD4 expression induced by PMA was temperature dependent and revers...
Correlation between monoclonal antibody reactivity and expression of CD4 and CD8 alpha genes in the horse.
Veterinary immunology and immunopathology    July 1, 1994   Volume 42, Issue 1 61-69 doi: 10.1016/0165-2427(94)90089-2
Grünig G, Barbis DP, Zhang CH, Davis WC, Lunn DP, Antczak DF.Equine peripheral blood lymphocytes (PBL) were enriched by positive selection using panning with a mixture of monoclonal antibodies against putative equine CD4 (Equine Leucocyte Antigen Workshop antibodies WS 1 and WS 72), or CD8 molecules (Workshop antibodies WS 12, WS 49, and WS 74). RNA was extracted from CD4 enriched cells (99% purity), from CD8 enriched cells (69% purity), from peripheral blood lymphocytes, and from neonatal equine thymus. RNA extracted from equine granulocytes and from equine kidney served as negative control. The RNA was electrophoresed in agarose and transferred to nyl...
P2 protamines are phosphorylated in vitro by protein kinase C, whereas P1 protamines prefer cAMP-dependent protein kinase. A comparative study of five mammalian species.
European journal of biochemistry    July 1, 1994   Volume 223, Issue 1 165-169 doi: 10.1111/j.1432-1033.1994.tb18979.x
Pirhonen A, Linnala-Kankkunen A, Mënpää PH.P1 protamines isolated from ejaculated human, stallion, bull, boar and ram spermatozoa and P2 protamines from human and stallion spermatozoa were subjected, after alkaline phosphatase treatment, to in vitro phosphorylation reactions using cAMP-dependent protein kinase (PKA) and protein kinase C (PKC). All P1 protamines were phosphorylated by PKA, whereas P2 protamines were phosphorylated only by PKC. In addition, human, stallion and boar, but not bull and ram, P1 protamines were phosphorylated by PKC. After phosphoamino acid analysis, the protamines showing positive signals for phosphoserine (...
An equine B cell surface antigen defined by a monoclonal antibody.
Veterinary immunology and immunopathology    July 1, 1994   Volume 42, Issue 1 91-102 doi: 10.1016/0165-2427(94)90092-2
Zhang CH, Donaldson WL, Antczak DF.A surface antigen of equine B lymphocytes was identified using the Equine Leucocyte Antigen Workshop antibody WS 65. This marker was expressed on almost all equine B cells, but not on T cells, granulocytes or thymocytes. WS 65 strongly stained cells in the follicular areas of lymph nodes and cells in the splenic nodules when tested on frozen tissue sections by immunohistochemistry. Equine leukemic T cells were not labeled by WS 65, and neither were the cells from a horse with B cell leukemia, although these latter cells carried surface immunoglobulin. Immunoprecipitation of lymphocyte membrane...
FTIR analysis of the interaction of azide with horse heart myoglobin variants.
Biochemistry    June 21, 1994   Volume 33, Issue 24 7600-7608 doi: 10.1021/bi00190a013
Bogumil R, Hunter CL, Maurus R, Tang HL, Lee H, Lloyd E, Brayer GD, Smith M, Mauk AG.The interaction of azide with variants of horse heart myoglobin (Mb) has been characterized by Fourier transform infrared (FTIR), electron paramagnetic resonance (EPR), and UV-VIS absorption spectroscopy and by molecular modeling calculations. Distal histidine variants (His64Thr, His64Ile, His64Lys) and charged surface variants (Val67Arg, Lys45Glu, Lys45Glu/Lys63Glu) were included in this study. All variants, with the exception of Val67Arg, have a lower azide affinity than the wild-type protein. Analysis of the temperature dependence of the FTIR spectra (277-313 K) revealed that the wild-type ...
Sequence of a cDNA encoding horse growth hormone.
Gene    June 10, 1994   Volume 143, Issue 2 299-300 doi: 10.1016/0378-1119(94)90115-5
Ascacio-Martínez JA, Barrera-Saldaña HA.A cDNA encoding horse growth hormone (ecGH) was isolated and sequenced. The coding sequence resembles a typical mammalian GH pre-protein and contains a 3' untranslated region of 101 nucleotides carrying two contiguous polyadenylation signals.
Structure of the equine infectious anemia virus Tat protein.
Science (New York, N.Y.)    June 10, 1994   Volume 264, Issue 5165 1584-1587 doi: 10.1126/science.7515512
Willbold D, Rosin-Arbesfeld R, Sticht H, Frank R, Rösch P.Trans-activator (Tat) proteins regulate the transcription of lentiviral DNA in the host cell genome. These RNA binding proteins participate in the life cycle of all known lentiviruses, such as the human immunodeficiency viruses (HIV) or the equine infectious anemia virus (EIAV). The consensus RNA binding motifs [the trans-activation responsive element (TAR)] of HIV-1 as well as EIAV Tat proteins are well characterized. The structure of the 75-amino acid EIAV Tat protein in solution was determined by two- and three-dimensional nuclear magnetic resonance methods and molecular dynamics calculatio...
A highly polymorphic horse microsatellite locus: VHL20.
Animal genetics    June 1, 1994   Volume 25, Issue 3 207 doi: 10.1111/j.1365-2052.1994.tb00129.x
van Haeringen H, Bowling AT, Stott ML, Lenstra JA, Zwaagstra KA.No abstract available
A rapid method for the analysis of influenza virus genes: application to the reassortment of equine influenza virus genes.
Virus research    June 1, 1994   Volume 32, Issue 3 391-399 doi: 10.1016/0168-1702(94)90087-6
Adeyefa CA, Quayle K, McCauley JW.We describe a rapid method for genetic characterisation of influenza virus genes using reverse transcription and amplification by polymerase chain reaction (RT/PCR) of all virus segments simultaneously (multiplex RT/PCR) using primers based on the conserved terminal sequences. The product has been shown to be suitable for determination of partial nucleotide sequences which can be used to search nucleotide sequence databases and rapidly map the genetic origin of each segment. We illustrate the use of the method by analysing genetic reassortment in H7N7 equine influenza viruses.
Comparative genetic analysis of Swiss and Spanish isolates of Echinococcus granulosus by southern hybridization and Random Amplified Polymorphic DNA technique.
Applied parasitology    June 1, 1994   Volume 35, Issue 2 107-117 
Siles-Lucas M, Felleisen R, Cuesta-Bandera C, Gottstein B, Eckert J.Swiss and Spanish isolates of Echinococcus granulosus were compared using different molecular biological techniques: Genomic DNAs isolated from parasites originating from various intermediate hosts were subjected to Southern hybridization with different probes, the same source of DNA was used for DNA amplification using the Random Amplified Polymorphic DNA (RAPD) technique. With both methods the various isolates (metacestodes) of E. granulosus exhibited characteristic banding patterns which allowed us to assign them to the following groups of homologous profiles: (a) isolates of horse and donk...
Identification of the horse epidermal growth factor (EGF) coding sequence and its use in monitoring EGF gene expression in the endometrium of the pregnant mare.
Journal of molecular endocrinology    June 1, 1994   Volume 12, Issue 3 341-350 doi: 10.1677/jme.0.0120341
Stewart F, Power CA, Lennard SN, Allen WR, Amet L, Edwards RM.The PCR technique and highly degenerate oligonucleotide primers were used to amplify a 282 bp fragment of the horse (Equus caballus) epidermal growth factor (EGF) cDNA. The clone corresponded to 94 amino acids of the EGF precursor molecule. The deduced amino acid sequence of the 53 residue EGF mitogenic peptide within the precursor sequence showed 60-70% identity with five other published EGF sequences. The PCR cDNA fragment hybridized to a 4.9 kb transcript in horse kidney and endometrial RNA which was of a similar size to the mature EGF transcript found in other mammalian species. The horse ...
Comparison of thoroughbred and Arabian horses using RAPD markers.
Animal genetics    June 1, 1994   Volume 25 Suppl 1 105-108 doi: 10.1111/j.1365-2052.1994.tb00414.x
Bailey E, Lear TL.We compared pools of DNA from 10 Thoroughbred horses and 10 Arabian horses for the presence of randomly amplified polymorphic DNA (RAPD) markers which might be useful in distinguishing between the breeds. Using 212 decamer oligonucleotides and our polymerase chain reaction (PCR) conditions, 173 of the primers produced scoreable bands. The number of bands ranged from 0 to 9 with an average of 3.6. In family studies using 11 arbitrarily selected primers, five of the 11 primers produced polymorphic bands which exhibited Mendelian inheritance as dominant markers. When comparing the pooled DNA from...
Serological and genomic characterization of equine rotavirus VP4 proteins identifies three different P serotypes.
Virology    June 1, 1994   Volume 201, Issue 2 364-372 doi: 10.1006/viro.1994.1302
Isa P, Snodgrass DR.A series of viral reassortants was prepared between equine rotaviruses H1 (G5), H2 (G3), and L338 (G13) and human rotavirus ST3 (G4). All contained the VP4 cognate gene segment 4 from the equine parental virus and the VP7 cognate gene segment 9 from ST3. Using these viruses and antisera prepared to them, it was shown that each of the three equine viruses possessed a serologically distinct VP4 or P serotype with a > or = 16-fold difference in reciprocal cross-neutralization titers. H1 VP4 was closely related to that of porcine virus OSU, i.e., P7. L338 gene 4 was sequenced, and the sequence and...
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