In situ hybridization (ISH) is a molecular technique used to detect specific nucleic acid sequences within fixed tissues and cells. In equine research, ISH is utilized to study gene expression patterns and the localization of specific RNA or DNA sequences in horse tissues. This technique provides insights into the spatial and temporal aspects of gene activity, helping to elucidate the molecular mechanisms underlying various physiological and pathological processes in horses. ISH can be applied to a range of tissues, including those affected by diseases, to better understand the genetic contributions to equine health and disease. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and findings of in situ hybridization in equine studies.
Haynes SE, Reisner AH.Although no major structural or numerical abnormalities were found in the karyotypes of 12 aborted equine fetuses, two unrelated abortuses each carried a large polymorphism for the amount of heterochromatin in chromosome 1. In both karyotypes this chromosome was shown to be larger than its homolog. To determine the nature of the extra DNA in these chromosomes, equine DNA was isolated and characterized by buoyant density analysis. Equine mainband DNA had a buoyant density in neutral CsCl of 1.699 g/cm3, while the highly repetitive (dG+dC)-rich fraction had a buoyant density of 1.715 g/cm3. A ra...
Ryder OA, Hansen SK.A (G + C)-rich density satellite DNA (rho = 1.713 gm/cc) has been purified from splenic DNA of Przewalski's horse, Equus przewalskii, by successive equilibrium density gradient centrifugations. The purified satellite, which may comprise as much as 29% of the total DNA, renatures rapidly; however, analyses of native, single-stranded, and reassociated molecules by analytical ultracentrifugation and melting properties suggest that some sequence heterogeniety exists in the 1.713 gm/cc satellite. Complementary RNA (cRNA) transcribed from satellite DNA has been utilized for in situ hybridization stu...
Lancaster WD, Theilen GH, Olson C.DNAs from bovine papilloma virus(BPV)-induced hamster tumors and from equine connective tissue tumors of unknown etiology were examined for BPV DNA sequences by molecular hybridization. DNA from two distinct classes of BPV (type 1 and type 2) was labeled in vitro and used as probes. Analysis of DNA-DNA reassociation kinetics indicated that both virus types were capable of tumor induction in the hamster. DNA isolated from 6 of 7 equine tumors accelerated the reassociation of the BPV DNA probes. BPV type 1 or type 2 DNA hybridized extensively to DNA from 3 tumors, while 3 other tumors contained ...
Lancaster WD, Olson C, Meinke W.Four of five spontaneous benign equine connective tissue tumors of unknown etiology and a bovine papilloma virus (BPV)-induced equine tumor contained BPV-specific DNA sequences as determined by DNA-DNA hybridization of DNA from tumors with BPV DNA labeled in vitro. Analysis of the kinetics of reassociation indicated that 20-75% of the BPV genome was present in the various tumors. The number of partial BPV genome equivalents ranged from 60 to 500 copies per diploid quantity of cellular DNA. Thermal denaturation profiles of duplexes formed between labeled BPV DNA and DNA from tumor cells indicat...
Oldenhof H, Schütze S, Wolkers WF, Sieme H.Sperm chromatin structure and condensation determine accessibility for damage, and hence success of fertilization and development. The aim of this study was to reveal characteristic spectral features coinciding with abnormal sperm chromatin packing (i.e., DNA-protein interactions) and decreased fertility, using Fourier transform infrared spectroscopy. Chromatin structure in spermatozoa obtained from different stallions was investigated. Furthermore, spermatozoa were exposed to oxidative stress, or treated with thiol-oxidizing and disulfide-reducing agents, to alter chromatin structure and pack...
Peters-Kennedy J, Löhr CV, Cossic B, Glaser AL, Duhamel GE.Equine herpesvirus-5 (EHV-5) is commonly found in healthy asymptomatic horses worldwide. Although a cause-and-effect relationship has not been thoroughly determined, this virus has been associated with several disease conditions including equine multinodular pulmonary fibrosis (EMPF) and 1 case of interface dermatitis. The authors searched the New York State Animal Health Diagnostic Center database for cases of equine interface dermatitis between 2007 and 2022. Ten cases were identified and scrutinized for viral inclusion bodies which were present in 5 of 10 cases. Two similar cases with inter...
de Heus P, Bagó Z, Weidinger P, Lale D, Trachsel DS, Revilla-Fernández S, Matiasek K, Nowotny N.As evidenced by sero-epidemiological studies, infections of horses with the tick-borne encephalitis virus (TBEV) occur frequently in TBEV-endemic areas. However, there are only very few reports of clinical cases. A possible underreporting may be due to a variety of diagnostic challenges. In this study, ELISA and neutralization tests were applied to serum samples. Brain tissue samples were investigated for the presence of nucleic acids of TBEV, Equid alphaherpesvirus 1, Borna disease virus 1, West Nile and Usutu viruses, rustrela virus, as well as Eastern, Western, and Venezuelan equine encepha...
Bugno-Poniewierska M, Jankowska M, Raudsepp T, Kowalska K, Pawlina-Tyszko K, Szmatola T.Chromosomal abnormalities occur in the equine population at a rate of approximately 2%. The use of molecular cytogenetic techniques allows a more accurate identification of chromosomal abnormalities, especially those with a low rate of abnormal metaphases, demonstrating that the actual incidence in equine populations is higher. Objective: Estimation of the number of carriers of karyotypic abnormalities in a sample from a population of young horses of various breeds, using molecular cytogenetic techniques. Methods: Cross-sectional. Methods: Venous blood samples were collected from 500 young hor...
Valera M, Karlau A, Anaya G, Bugno-Poniewierska M, Molina A, Encina A, Azor PJ, Demyda-Peyrás S.Sex chromosomal abnormalities are a well-established cause of reproductive failure in domestic horses. Because of its difficult diagnosis, the Pura Raza Español breeding program established a routine screening for chromosomal abnormalities in all the horses prior to enrolling in the studbook. This genomic procedure combines an initial assessment based on the results from Short Tandem Repeat (STR) parentage testing followed by a Single-Nucleotide Polymorphism (SNP) based copy number aberration (CNA) confirmative analysis in positive cases. Using this methodology, we identified five new individ...
B B, G M, L G, G A, B B, T F, A G, D B, A K, G T, G S, A B, M F, L R.Equine penile tumors are common in horses and are often related to infection with equine papillomavirus type 2 (EcPV2). This study investigated the immune cell infiltrate (ICI) of these tumors in horses, focusing on the role of EcPV2. Using multiplex immunohistochemistry (mIHC) for CD3, CD20, and IBA-1 and immunohistochemistry (IHC) for FoxP3, 27 horses with papillomas (5/27), in situ carcinomas (CISs) (3/27), and squamous cell carcinomas (SCCs) (19/27) were evaluated. Eighteen cases tested positive for EcPV2 by either or both in situ hybridization (ISH) and polymerase chain reaction (PCR) (18...
Bailey VN, Gilbert BM, Vetter M, Oberhaus EL.The mechanism by which photoperiod influences the hypothalamic-pituitary-gonadal (HPG) axis and regulates seasonal reproduction in horses has yet to be fully elucidated. The hypophyseal pars tuberalis (PT) has been indicated as a critical site for the transduction of melatonin signals through melatonin-responsive, PT-specific cells that produce thyroid-stimulating hormone (TSH) in many mammalian species. However, this has yet to be investigated in horses. The objective of this study was to explore the interaction of melatonin and thyroid-stimulating hormone in the equine HPG axis. Pituitaries ...