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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.
X chromosome detection in an XO mare using a human X paint probe, and PCR detection of SRY and amelogenin genes in 3 XY mares.
Equine veterinary journal    September 18, 2001   Volume 33, Issue 5 527-530 doi: 10.2746/042516401776254844
Mäkinen A, Suojala L, Niini T, Katila T, Tozaki T, Miyake Y, Hasegawa T.No abstract available
Characterization of equine E-selectin.
Immunology    September 1, 2001   Volume 103, Issue 4 498-504 doi: 10.1046/j.1365-2567.2001.01262.x
Hedges JF, Demaula CD, Moore BD, McLaughlin BE, Simon SI, MacLachlan NJ.Expression of E-selectin on activated endothelium is a critical initial step that leads to extravasation of leucocytes during inflammation, yet E-selectin is largely uncharacterized in several animal species including the horse. We have sequenced and compared E-selectin genes derived from activated cultures of purified equine (horse), cervid (black-tailed deer) and ovine (sheep) pulmonary artery endothelial cells (ECs). Phylogenetic and amino acid sequence comparisons indicate that bovine, cervid and ovine E-selectin are similar, whereas human and equine E-selectin are more closely related to ...
Chromosome homologies between man and mountain zebra (Equus zebra hartmannae) and description of a new ancestral synteny involving sequences homologous to human chromosomes 4 and 8.
Cytogenetics and cell genetics    August 31, 2001   Volume 93, Issue 3-4 291-296 doi: 10.1159/000057000
Richard F, Messaoudi C, Lombard M, Dutrillaux B.Using human chromosome painting probes, we looked for homologies between human and mountain zebra (Equus zebra hartmannae, Equidae, Perissodactyla) karyotypes. Except for two very short segments, all euchromatic regions were found to have a human homologous chromosome segment. Conserved syntenies previously described in various mammalian orders were detected. Each synteny corresponded to a chromosomal region homologous to two parts of human chromosomes: HSA3 and HSA21, HSA7 and HSA16, HSA12 and HSA22, and HSA16 and HSA19. Chromosomal segments homologous to a part of HSA11 and HSA19p are found ...
The novel picornavirus Equine rhinitis B virus contains a strong type II internal ribosomal entry site which functions similarly to that of Encephalomyocarditis virus.
The Journal of general virology    August 22, 2001   Volume 82, Issue Pt 9 2257-2269 doi: 10.1099/0022-1317-82-9-2257
Hinton TM, Crabb BS.Equine rhinitis B virus (ERBV) has recently been classified as an Erbovirus, a new genus in the Picornaviridae family. ERBV is distantly related to members of the Cardiovirus and Aphthovirus genera which utilize a type II internal ribosome entry sequence (IRES) to initiate translation. We show that ERBV also possesses the core stem-loop structures (H-L) of a type II IRES. The function of the ERBV IRES was characterized using bicistronic plasmids that were analysed both by transfection into BHK-21 cells and by in vitro transcription and translation in rabbit reticulocyte lysates. In both system...
Characterisation of the enzymatic 4-O-acetylation of sialic acids in microsomes from equine submandibular glands.
Glycoconjugate journal    August 21, 2001   Volume 17, Issue 12 849-858 doi: 10.1023/a:1010965128335
Tiralongo J, Schmid H, Thun R, Iwersen M, Schauer R.Microsomes prepared from equine submandibular glands and incubated with tritium-labelled AcCoA incorporated acid-insoluble radioactivity in a manner dependent on time, protein, membrane integrity and AcCoA concentration, with incorporation being optimal at 37 degrees C and pH 6.6. Under the experimental conditions used a K(M) of 32.1 microM for AcCoA and a V(max) of 1.2 pmol/mg protein x min was obtained. The incorporation of acid-insoluble radioactivity was also inhibited by CoA in a competitive manner (K(i)=240 microM), as well as by para-chloromercuribenzoate, 3'-dephospho-CoA, 5'-IDP, 5'-A...
Two new variants of the Rhodococcus equi virulence plasmid, 90 kb type III and type IV, recovered from a foal in Japan.
Veterinary microbiology    August 17, 2001   Volume 82, Issue 4 373-381 doi: 10.1016/s0378-1135(01)00398-4
Takai S, Murata N, Kudo R, Narematsu N, Kakuda T, Sasaki Y, Tsubaki S.This report describes the discovery of two new virulence plasmid types from a crossbred foal with Rhodococcus equi pneumonia in Kumamoto died with severe R. equi pneumonia and ulcerative enteritis. R. equi was isolated in large numbers and isolates from the foal were investigated for the presence of virulence-associated 15-17 kDa antigens (VapA) by colony blotting, using the monoclonal antibody 10G5, and by gene coding for VapA by PCR. Plasmid DNAs extracted from the isolates were digested with restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII. The digestion patterns that resulted d...
Solution NMR determination of the seating(s) of meso-nitro-etioheme-1 in myoglobin: implications for steric constraints to meso position access in heme degradation by coupled oxidation.
Journal of the American Chemical Society    August 17, 2001   Volume 123, Issue 33 8080-8088 doi: 10.1021/ja010651a
Wang J, Li Y, Ma D, Kalish H, Balch AL, La Mar GN.The highly stereoselective cleavage of hemin in myoglobin by coupled oxidation has been attributed to steric barriers that leave more space near the alpha- than the other meso-positions. The steric barriers near meso positions in myoglobin have been investigated by establishing the thermodynamics and dynamics of possible seatings in the pocket of horse myoglobin of a four-fold symmetric etioheme I modified with a bulky nitro group at a single meso position. The cyanomet complex of this reconstituted myoglobin exhibits three sets of (1)H NMR resonances that are linked dynamically and occur in a...
Cytochrome c reconstituted from two peptide fragments displays native-like redox properties.
European journal of biochemistry    August 15, 2001   Volume 268, Issue 16 4537-4543 doi: 10.1046/j.1432-1327.2001.02373.x
Sinibaldi F, Fiorucci L, Mei G, Ferri T, Desideri A, Ascoli F, Santucci R.Recombination of two fragments of horse cytochrome c (the heme-containing N-fragment, residues 1-56, and the C-fragment, residues 57-104), which are substantially unstructured at neutral pH, gives rise to a 1:1 fragment complex with a compact conformation, in which the alpha helical structure and the native Met80-Fe(III) axial bond are recovered. With respect to the native protein, the ferric complex shows a less rigid atomic packing and a decreased stability [Delta(DeltaG(o))D = 14.7 kJ.mol(-1)], ascribed to perturbations involving the Trp59 microenvironment and, to a lower extent, the heme p...
Cytogenetic assignment and genetic characterization of the horse microsatellites, TKY4-18, TKY20, TKY22-24, TKY30-41 derived from a cosmid library.
Animal genetics    August 9, 2001   Volume 32, Issue 3 160-162 doi: 10.1046/j.1365-2052.2001.0723a.x
Hirota K, Tozaki T, Mashima S, Miura N.No abstract available
Characterization of mutations in the rpoB gene associated with rifampin resistance in Rhodococcus equi isolated from foals.
Journal of clinical microbiology    July 28, 2001   Volume 39, Issue 8 2784-2787 doi: 10.1128/JCM.39.8.2784-2787.2001
Fines M, Pronost S, Maillard K, Taouji S, Leclercq R.Treatment with a combination of erythromycin and rifampin has considerably improved survival rates of foals and immunocompromised patients suffering from severe pneumonia caused by Rhodococcus equi. Frequently, because of monotherapy, emergence of rifampin-resistant strains has been responsible for treatment failure. Using consensus oligonucleotides, we have amplified and sequenced the rifampin resistance (Rif(r))-determining regions of 12 rifampin-resistant R. equi strains isolated from three foals and of mutants selected in vitro from R. equi ATCC 3701, a rifampin-susceptible strain. The ded...
Chromosomal distribution of the telomere sequence (TTAGGG)(n) in the Equidae.
Cytogenetics and cell genetics    July 28, 2001   Volume 93, Issue 1-2 127-130 doi: 10.1159/000056964
Lear TL.Telomeres are a class of repetitive DNA sequences that are located at chromosome termini and that act to stabilize the chromosome ends. The rapid karyotypic evolution of the genus Equus has given rise to ten taxa, all with different diploid chromosome numbers. Using fluorescence in situ hybridization (FISH) we localized the mammalian telomere sequence, (TTAGGG)(n), to the chromosomes of nine equid taxa. TTAGGG signal was located at chromosome termini in all species, however additional signal was seen at interstitial sites on some chromosomes in the Burchell's zebra, Equus quagga burchelli, the...
DNA in human and stallion spermatozoa forms local hexagonal packing with twist and many defects.
Journal of structural biology    July 27, 2001   Volume 134, Issue 1 76-81 doi: 10.1006/jsbi.2001.4365
Sartori Blanc N, Senn A, Leforestier A, Livolant F, Dubochet J.In human and other mammal sperm nuclei, DNA is packed in a highly condensed state, the structure of which remains unsolved. Cryoelectron microscopy of vitrified sections provides a first direct view of the local arrangement of the nucleoprotamine filament. DNA aligns in parallel in layers and its orientation rotates along a single-twist direction as in a cholesteric liquid crystal. The structure contains numerous defects, which introduce locally double-twist configurations. Destruction of the SS bonds with dithiotrehitol relaxes the twist and favors the extension of the hexagonal close packing...
Dynamics of structure and energy of horse carboxymyoglobin after photodissociation of carbon monoxide.
Journal of the American Chemical Society    July 18, 2001   Volume 123, Issue 18 4286-4294 doi: 10.1021/ja9944655
Sakakura M, Yamaguchi S, Hirota N, Terazima M.The energetics and structural volume changes after photodissociation of carboxymyoglobin are quantitatively investigated by laser-induced transient grating (TG) and photoacoustic calorimetric techniques. Various origins of the TG signal are distinguished: the phase grating signals due to temperature change, due to absorption spectrum change, and due to volume change. We found a new kinetics of approximately 700 ns (at room temperature), which was not observed by the flash photolysis technique. This kinetics should be attributed to the intermediate between the geminate pair and the fully dissoc...
Changes in molecular expression of aggrecan and collagen types I, II, and X, insulin-like growth factor-I, and transforming growth factor-beta1 in articular cartilage obtained from horses with naturally acquired osteochondrosis.
American journal of veterinary research    July 17, 2001   Volume 62, Issue 7 1088-1094 doi: 10.2460/ajvr.2001.62.1088
Semevolos SA, Nixon AJ, Brower-Toland BD.To determine molecular changes in the expression of insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 (TGF-beta1) in horses with osteochondrosis, and to characterize expression of matrix aggrecan and collagen types I, II, and X in articular cartilage of affected joints. Methods: Articular cartilage from affected stifle or shoulder joints of 11 horses with naturally acquired osteochondrosis and corresponding joints of 11 clinically normal horses. Methods: Harvested specimens were snap frozen in liquid nitrogen, and total RNA was isolated. Specimens were fixed in 4% paraf...
Comparison of the value of pulsed-field gel electrophoresis, random amplified polymorphic DNA and amplified rDNA restriction analysis for subtyping Taylorella equigenitalis.
Veterinary research communications    July 4, 2001   Volume 25, Issue 4 261-269 doi: 10.1023/a:1010674524428
Kagawa S, Moore JE, Murayama O, Matsuda M.Eight strains of Taylorella equigenitalis were identified by a polymerase chain reaction using a primer pair specific to the 16S rDNA of T equigenitalis. These eight strains were chosen because they had previously been shown to represent eight distinct genotypes by pulsed-field gel electrophoresis analysis after separate digestion of the genomic DNA with ApaI or NotI. The eight strains could be classified into six or seven types by random amplified polymorphic DNA analysis using different kinds of primers. Amplified rDNA restriction analysis after separate digestion with five restriction enzym...
Characterization of equine microsatellite loci, TKY102-TKY112.
Animal genetics    June 26, 2001   Volume 32, Issue 2 117-119 doi: 10.1046/j.1365-2052.2001.0700h.x
Mashima S, Tozaki T, Swinburne J, Kakoi H, Binns M, Miura N.No abstract available
Linked markers exclude KIT as the gene responsible for appaloosa coat colour spotting patterns in horses.
Animal genetics    June 26, 2001   Volume 32, Issue 2 98-101 doi: 10.1046/j.1365-2052.2001.00737.x
Terry RR, Bailey E, Bernoco D, Cothran EG.The appaloosa coat colour pattern of the horse is similar to that caused by the rump-white (Rw) gene in the mouse. In the mouse Rw colour pattern is the result of an inversion involving the proto-oncogene c-kit (KIT). Therefore, we investigated KIT as a candidate gene that encodes the appaloosa coat colour gene (Lp) in horses. KIT plays a critical role in haematopoiesis, gametogenesis, and melanogenesis and encodes a transmembrane tyrosine kinase receptor that belongs to the PDGF/CSF-1/c-KIT receptor subfamily. Half-sib families segregating for Lp were uninformative for a reported polymorphism...
Base substitutions in the sequences flanking microsatellite markers HMS3 and ASB2 interfere with parentage testing in the Lipizzan horse.
Animal genetics    June 23, 2001   Volume 32, Issue 1 52 doi: 10.1046/j.1365-2052.2001.0647k.x
Achmann R, Huber T, Wallner B, Dovc P, Müller M, Brem G.No abstract available
Epidermal growth factor regulation of equine glycoprotein hormone alpha subunit expression in trophoblast cells.
Biology of reproduction    June 23, 2001   Volume 65, Issue 1 197-203 doi: 10.1095/biolreprod65.1.197
Thway TM, Wolfe MW.Primates and equids are the only species known to express the placental glycoprotein hormone, chorionic gonadotropin (CG), a heterodimeric glycoprotein composed of an alpha subunit linked to a hormone-specific beta subunit. The regulatory mechanisms involved in the induction of equine glycoprotein alpha subunit gene expression have not been identified. Epidermal growth factor (EGF) receptor is known to transduce signals that alter a number of different cellular functions (cell proliferation, differentiation, hormone secretion, and gene regulation). In the present study, we investigated the reg...
Physical anchorage and orientation of equine linkage groups by FISH mapping BAC clones containing microsatellite markers.
Animal genetics    June 23, 2001   Volume 32, Issue 1 37-39 doi: 10.1046/j.1365-2052.2001.00715.x
Lindgren G, Swinburne JE, Breen M, Mariat D, Sandberg K, Guérin G, Ellegren H, Binns MM.A horse bacterial artificial chromosome (BAC) library was screened for 19 microsatellite markers from unassigned or non-oriented linkage groups. Clones containing 11 (AHT20, EB2E8, HMS45, LEX005, LEX014, LEX023, LEX044, TKY111, UCDEQ425, UCDEQ464 and VIASH21) of these were found, which were from eight different linkage groups. The BAC clones were used as probes in dual colour FISH to identify their precise chromosomal origin. The microsatellite markers are located on nine different horse chromosomes, four of which (ECA6, ECA25, ECA27 and ECA28) had no previously in situ assigned markers.
Amino acid modifications in canine, equine and porcine pituitary growth hormones, identified by peptide-mass mapping.
Journal of chromatography. B, Biomedical sciences and applications    June 22, 2001   Volume 757, Issue 2 237-245 doi: 10.1016/s0378-4347(01)00154-2
Secchi C, Berrini A, Gaggioli D, Borromeo V.Modified amino acid residues in porcine, canine and equine growth hormones purified from pituitary glands were characterised by tryptic mapping and high-performance liquid chromatography with on-line coupled electrospray ionisation mass spectrometry (HPLC-ESI-MS) detection. Hormones from all three species showed the same changes. Conversion of Asp128 to iso-Asp128 was a component of native hormones, while deamidation of Asn12 and Asn98 to Asp and iso-Asp, oxidation of Met4, and cyclisation to the pyroglutamyl derivative of Gln139, probably occurred in vitro, during isolation, storage or hydrol...
Differential regulation of gelatinases by transforming growth factor beta-1 in normal equine chondrocytes.
Osteoarthritis and cartilage    June 12, 2001   Volume 9, Issue 4 325-331 doi: 10.1053/joca.2000.0392
Thompson CC, Clegg PD, Carter SD.Cartilage destruction in osteoarthritis (OA) is associated with increased levels of several matrix metalloproteinases (MMPs), including the gelatinases MMP-2 and MMP-9. While increases in some MMPs may be destructive, up-regulation of others may result from increases in normal tissue turnover. The production of MMP-2 and MMP-9 by the anabolic transforming growth factor beta-1 (TGF-beta1) in normal equine chondrocytes was investigated. Methods: Equine chondrocytes from clinically normal femoropatellar joints were maintained in alginate beads. After serum deprivation, cells were exposed to TGF-b...
p53 protein expression in conjunctival squamous cell carcinomas of domestic animals.
Veterinary ophthalmology    June 9, 2001   Volume 2, Issue 4 227-231 doi: 10.1046/j.1463-5224.1999.00086.x
Sironi G, Riccaboni P, Mertel L, Cammarata G, Brooks DE.The expression of p53 protein was investigated in eight formalin-fixed, paraffin-embedded conjunctival squamous cell carcinomas of five horses and one cow, dog and cat each by an immunohistochemical procedure in order to evaluate protein overexpression. Anti-human p53 protein mouse monoclonal antibodies known to be cross-reactive with p53 protein of the animal species examined were used. Positive p53 nuclear immunostaining was detected in five equine, one bovine and one feline cases. Conversely, no p53 immunostaining was found in the only canine case examined. These results demonstrate a frequ...
Functional and computer modelling studies of haemoglobin from horse. The haemoglobin system of the Sardinian wild dwarf horse.
European journal of biochemistry    June 8, 2001   Volume 268, Issue 11 3313-3320 doi: 10.1046/j.1432-1327.2001.02235.x
Pellegrini M, Corda M, Manca L, Olianas A, Sanna MT, Fais A, De Rosa MC, Bertonati C, Masala B, Giardina B.A study was made of the haemoglobin (Hb) system from the Sardinian dwarf horse (Equus caballus jara), one of the last surviving wild horse species in Europe. The oxygen binding properties of the whole haemolysate and of the four different horse Hbs, separated by ion-exchange chromatography, were studied with special regard to the effect of chloride, 2,3-diphosphoglycerate and lactate. Results indicate that no significant functional differences exist between the four Hb components of horse haemolysate. Moreover, the molecular basis of the intrinsically low oxygen affinity and of the weak intera...
Protein conformation change of myoglobin upon ligand binding probed by ultraviolet resonance Raman spectroscopy.
Biochemistry    June 8, 2001   Volume 40, Issue 23 6956-6963 doi: 10.1021/bi002640k
Haruta N, Aki M, Ozaki S, Watanabe Y, Kitagawa T.Conformational change of myoglobin (Mb) accompanied by binding of a ligand was investigated with 244 nm excited ultraviolet resonance Raman Spectroscopy (UVRR). The UVRR spectra of native sperm whale (sw) and horse (h) Mbs and W7F and W14F swMb mutants for the deoxy and CO-bound states enabled us to reveal the UVRR spectra of Trp7, Trp14, and Tyr151 residues, separately. The difference spectra between the deoxy and CO-bound states reflected the environmental or structural changes of Trp and Tyr residues upon CO binding. The W3 band of Trp7 near the N-terminus exhibited a change upon CO binding...
DH82 cells: a macrophage cell line for the replication and study of equine infectious anemia virus.
Journal of virological methods    May 30, 2001   Volume 95, Issue 1-2 47-56 doi: 10.1016/s0166-0934(01)00288-9
Hines R, Maury W.In vivo, tissue macrophages have been implicated as an important cell for the replication of equine infectious anemia virus (EIAV). Laboratory investigations of EIAV/macrophage interactions, however, have been hampered by the laborious blood monocyte isolation procedures. In addition, adherent equine macrophage cultures generally have poor long-term viability and are resistant to transfection. This report describes an adherent canine macrophage-like cell line, DH82, that supports the replication of EIAV. This cell line was easily transfectable and supported EIAV Tat transactivation of the LTR....
Characterization of expressed sequence tags generated from skin cDNA clones of Equus caballus by single pass sequencing.
Animal biotechnology    May 24, 2001   Volume 12, Issue 1 87-97 doi: 10.1081/ABIO-100102981
Lieto LD, Cothran EG.A cDNA library was built using RNA extracted from the skin tissue of an adult horse. The library was primed with oligo (dT) and sequences were directionally inserted in order to produce an expression library. The library has 5.8X 10(5) plaque forming units with 99.6% recombinant phage. The average insert size is 1.3 Kbp. Three hundred and thirteen expressed sequence tags (ESTs) were generated from sequencing of the 5 prime end of randomly selected skin cDNA clones. The ESTs were sequenced on an ABI 377 using Big-Dye chemistry. A similarity search was performed on each EST using the NCBI non-re...
Uterocalin, a lipocalin provisioning the preattachment equine conceptus: fatty acid and retinol binding properties, and structural characterization.
The Biochemical journal    May 23, 2001   Volume 356, Issue Pt 2 369-376 doi: 10.1042/0264-6021:3560369
Suire S, Stewart F, Beauchamp J, Kennedy MW.The equine conceptus is surrounded by a fibrous capsule that persists until about day 20 of pregnancy, whereupon the capsule is lost, the conceptus attaches to the endometrium and placentation proceeds. Before attachment, the endometrium secretes in abundance a protein of the lipocalin family, uterocalin. The cessation of secretion coincides with the end of the period during which the conceptus is enclosed in its capsule, suggesting that uterocalin is essential for the support of the embryo before direct contact between maternal and foetal tissues is established. Using recombinant protein and ...
Biochemical characterization and surfactant properties of horse allergens.
European journal of biochemistry    May 19, 2001   Volume 268, Issue 10 3126-3136 doi: 10.1046/j.1432-1327.2001.02217.x
Goubran Botros H, Poncet P, Rabillon J, Fontaine T, Laval JM, David B.A new allergen from horse dander, Equ c 5 has been purified. Its biochemical and biophysical properties have been characterized and compared with those of Equ c 1, Equ c 2 and Equ c 4. Their molecular masses, determined by mass spectrometry, were 22 kDa for Equ c 1, 16 kDa for Equ c 2, 18.7 kDa for Equ c 4 and 16.7 kDa for Equ c 5. Their pI values were between 3.8 and 5.25. Equ c 2 and Equ c 5 are not glycosylated, while Equ c 4 contains a tri-antennary tri-sialylated N-linked glycan. Linkages of terminal N-acetylneuraminic acid to galactose were: alpha-(2-->6) in Equ c 4, and both alpha-(2...
Molecular surveillance of the incidence of Taylorella equigenitalis and Pseudomonas aeruginosa from horses in Ireland by sequence-specific PCR.
Equine veterinary journal    May 16, 2001   Volume 33, Issue 3 319-322 doi: 10.2746/042516401776249750
Moore JE, Buckley TC, Millar BC, Gibson P, Cannon G, Egan C, Cosgrove H, Stanbridge S, Anzai T, Matsuda M, Murphy PG.No abstract available