A karyotype in horses refers to the complete set of chromosomes present in the cells of a horse, typically organized in a systematic arrangement for analysis. Horses generally have 64 chromosomes, comprising 31 pairs of autosomes and one pair of sex chromosomes. Karyotyping is employed to study chromosomal abnormalities that may affect equine development, fertility, and overall health. It provides insights into genetic disorders, hereditary conditions, and breed-specific traits. This page compiles peer-reviewed research studies and scholarly articles that explore the methodology, findings, and implications of karyotype analysis in equine genetics.
Romagnano A, Richer CL.Cell synchronization was used to obtain an adequate percentage of very long chromosomes in equine mitotic spreads. Reported here is our variation, adapted to horse chromosomes, of a method using excess thymidine followed by bromodeoxyuridine incorporation. This technique routinely yields excellent quality cells, predominantly in prometaphase and prophase. Among other differences with the standard technique, this method does not use Colcemid, which, in addition to inhibiting spindle fiber formation, also increases chromosome contraction resulting in thicker and thus fewer bands. Consequently, h...
Richer CL, Romagnano A.Both dynamic G-banding and cell synchronization produced by bromodeoxyuridine (BrdU), were applied to equine chromosomes. BrdU incorporated during the first half of the S-phase is taken up into the R-bands that are early replicating. These bands, which have incorporated BrdU, cannot contract as usual and remain elongated; only the other regions of the chromosome, i.e., the G-bands, contract normally and are sharply defined. BrdU also can be used for cell synchronization. The addition of BrdU in a high concentration, 15 hours before harvest, and its removal 11 hours later, has two effects: init...
Halnan CR.From the standpoint that cytogenetic screening in mares is seldom necessary as an aid to diagnosis of the gonadal dysgenesis syndrome, a series of double-blind trials were conducted to test the proposal that present practice failed to explore the potential for cytogenetics in clinical practice. It was demonstrated that diagnoses of infertility might be made where mares were found to be of normal phenotype by clinical examination. Such mares were found to be gonosmic mosaics. One stallion had a polymorphism of the X chromosome and had poor conception rates. It was demonstrated that the true val...
Halnan CR.The prognostic use of karyotyping in equine breeding has been recommended since 1976. Specimens used and laboratory methods are described. The system of karyotype evaluation is explained together with a glossary of terms. The principal aberrations in horses are defined. The present state of knowledge in cytogenetics in horses is reviewed dealing with infertility in the mare, where sex chromosomes give clear indication of problems; the situation with respect to anomalies found in stallions and hermaphroditism is explained coupled with recent concepts of chromosomal roles in reproduction. The fu...
Braun U, Förster M, Schams D.A case of intersexuality in an arabo-haflinger horse is described. The external genitalia consisted of mammary gland, vulva and hypertrophic clitoris; the inner genitalia of rudimental testes. The sexual behaviour was typically male-like. The chromosome analysis showed the female karyotype 64,XX. The presence of testes in a genetically female animal is explained by translocation of an Y-chromosome fragment bearing the gene for the H-Y-antigen.
Märki U, Osterhoff DR.A method using methotrexate for horse lymphocyte cell synchronization and thymidine for incorporation into DNA replication is described. This method provides a powerful technique for the study of chromosomal abnormalities and detailed analysis of chromosomal replication patterns. The determination of horse karyotypes with many similar chromosomes needs a special method which reveals the numerous and informative stages of the cell cycle. Horse lymphocyte cell cultures treated with colcemid (20 min) and harvested 6 hours after the release of the 17 hour-block with methotrexate show the best resu...
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...
Dunn HO, Smiley D, Duncan JR, McEntee K.The karyotypes of a Welsh pony and a Standardbred were 64,XX/64,XY and 63,XO/64,XY respectively. Both intersexes were true hermaphrodites with bilateral ovotestes. Neither intersex showed stallion-like behaviour. Each one had an underdeveloped penis, bilateral seminal vesicles and uterine tissue. It would appear that the chimerism in these equine intersexes resulted from double fertilization or fusion of blastocysts. Mosaicism in the Standardbred is a possibility, resulting from loss of a Y chromosome by anaphase lag in an early embryonic XY cell.
Blue MG.The objective of this study was to investigate an hypothesis that chromosome anomalies are an important cause of prenatal loss in the mare. An attempt was made to analyse, cytogenetically, a series of 26 equine abortuses. Cell cultures were prepared from a range of tissues, but failed to grow, and chromosome analysis was therefore not possible for any of these specimens. Consequently, a study was made of the metaphase chromosomes prepared from 22 equine embryos after their surgical removal from mares' uteri. The karyotypes prepared for each specimen were normal. The current findings are discus...
Blue MG.This study provides accumulated data to assist the definition of karyotypes from normal and infertile horses. The normal karyotype of the horse (2n = 64) was characterized following Giemsa staining and C- banding, and 23% aneuploidy was found among chromosome counts of cells prepared from 44 clinically normal horses and 24 equine embryos. These expected variations in chromosome counts are especially important in the evaluation of potential mosaicism. Centromere staining was shown to be a valuable aid for the identification of specific chromosomes, in particular the sex chromosomes. Sex chromat...
Scott IS, Long SE.Meiotic preparations were made from testicular material obtained after surgical castration of eight stallions (Equus caballus) with normal spermatogenesis. The material was examined after conventional Giemsa staining and C-banding. C-banding demonstrated that the Y chromosome at diakinesis associated with the short arm of the X chromosome. In 315 cells at diplotene or diakinesis, 56 (17.7%) had univalents and 51 (16.1%) of these involved the sex chromosomes. The overall mean chiasma number was 54.4 +/- 1.8 SD, and the mean calculated nondisjunction (ND) frequency was 3.4%. These results are di...
Hatami-Monazah H, Pandit RV.The group of Caspian ponies studied contained some animals with 65 chromosomes and others with 64 chromosomes. The morphology and G-banding pattern of the chromosomes resembled those of Equus caballus and E. przewalskii. The karyogram of animals with 65 chromosomes was identical to that of the cross between E. caballus and E. przewalskii. It is suggested that the Caspian pony is the product of natural hybridization between E. caballus and E. prezwalskii. Low reproductive effeciency of the Caspian pony is suggested as the cause of decline in the population of these animals.
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...
Trommershausen-Smith A, Hughes JP, Neely DP.Gonadal dysgenesis in the mare is associated with several different karyotypes, including sex chromosome aneuploidy (63,X; 63,X/64,XX; 63,X/64,XY or 65,XXX), the normal male complement (64,XY) and autosomal deletion (64,XX?del2q-). The 63,X is the most common karyotype found in gonadal dysgenesis. Aneuploid cases probably represent spontaneous chromosome non-disjunction during oogenesis, spermatogenesis or early embryonic development. Cases with XY or autosomal deletion may be inherited defects or of spontaneous origin.
Hughes JP, Trommershausen-Smith A.Reproductive failure was studied in 12 phenotypically normal mares (9 Arabian, 3 Quarter horses, 1 Appaloosa and 1 pony). Karyotyping was performed using lymphocytes isolated from peripheral blood by density gradient procedures, followed by standard culture methods for karyotyping. Nine mares had karyotypes of 63,XO; 1 had 63,XO/64,XX; 1 had 63,XO/64,XY and 1 had 64,XY. All mares had small, firm ovaries that when removed and examined from 4 mares, lacked germ cells and consisted of undifferentiated ovarian stroma.
The Journal of heredityNovember 1, 1976
Volume 67, Issue 6 361-367 doi: 10.1093/oxfordjournals.jhered.a108753
Eldridge F, Blazak WF.Karyotypes of the horse with 64 chromosomes, the ass with 62 chromosomes, and the mule with 63 chromosomes are presented. The chromosome complements of each species and their mule hybrid are analyzed and compared.
Hughes JP, Benirschke K, Kennedy PC, Trommershausen-Smith A.Five phenotypically normal but infertile mares were studied; four had karyotypes of 63XO, and one was a 25,64XX/13,63XO mosaic. The mares exhibited small uteri and has small ovaries that lacked germ cells and consisted primarily of undifferentiated ovarian stroma. These cases demonstrate that chromosome analysis is an important technique for the diagnosis of some forms of equine infertility.
Short RV.The infertility of the mule has proved a continuing challenge to scientific thought. Since the chromosomal differences between the two parental species are so great as to render normal meiosis impossible, it is postulated that all mules and hinnies are sterile. The problem now is to explain how mules and hinnies can occasionally produce spermatozoa or ova. The appearance of the mule was sufficient to persuade the ancients that both parents, not just the male, must contribute to the make-up of the offspring. The mule has also taught us that, when the number of oocytes in the ovary is reduced, t...
Kenney RM, Kent MG, Garcia MC, Hurtgen JP.A total of 174 stallions were subjected to a standard fertility examination and classified as fertile, subfertile or sterile. All stallions were phenotypical males involved in breeding programmes with no detectable abnormalities in their reproductive organs. Fertile stallions had no history of any breeding problem. Subfertile stallions were referred with a history of a breeding problem that was subsequently determined not to be attributable to the mares or infectious diseases. They were divided into chromosomally normal and abnormal groups on the basis of karyotype. The relative DNA content of...
Mäkinen A, Katila T, Kuokkanen MT.Only one X chromosome was found in each of the lymphocyte metaphases studied in an infertile mare. Karyotype analysis was made with the CBG and GTG banding techniques. The most obvious clinical abnormality was gonadal hypoplasia.
Martinez Zuviria S, Ciurkiewicz M, Wohlsein P, Madariaga G, Zuccolilli G.Cystic hygroma (hygroma cysticum) is a malformation that has not yet been described as a cause of early pregnancy loss in equines. The condition is a congenital anomaly occurring during embryogenesis due to a failure in which the primitive lymphatic sac does not reach the venous system at the jugular vein, resulting in a lymphatic stasis that starts in the neck region and continues to the rest of the body. From 2015 to 2020, a total of 5,730 ultrasound examinations were performed in mares from 43 different horse farms and embryo transfer farms when sexing pregnancies. In 12 pregnant mares, a s...
Constant SB, Larsen RE, Asbury AC, Buoen LC, Mayo M.A bilateral cryptorchid stallion with mild development of mammary glands was identified as an XX male by karyotyping. Necropsy revealed underdeveloped accessory sex organs and hypoplastic, inguinally located testes that were deficient of spermatogonia. Evaluation of routine hormonal profiles (without karyotyping) would have failed to diagnose this syndrome.
Pieńkowska-Schelling A, Handler J, Neuhauser S, Schelling C.The present case report describes a 6-year old subfertile pony mare, which became pregnant after the eleventh artificial insemination. The examination of the ovaries and the uterus did not reveal any abnormal clinical findings and the mare showed a regular oestrous cycle. Based on cytogenetic and molecular genetic analyses it became possible to elucidate the observed subfertility. The mosaic karyotype of the mare consisted of 63,X (20%) and 64,XX (80%) cells. A PCR analysis failed to amplify sequences from the equine SRY gene. The observed classic 63,X/64,XX mosaicism is a plausible explanatio...
Deriusheva SE, Loginova IuA, Chiriaeva OG, Iaschak K.A high-resolution cytogenetic map (670 bands per haploid set) of RBA-banded chromosomes has been constructed in the domestic horse Equus caballus. The size and distribution of the replication-based R(G)-bands were analyzed using the computer program VideoTest-Karyo. The obtained data were compared to the results of cytogenetic mapping in other mammalian species and human.
Hughes JP, Kennedy PC.TWo cases of XO-gonadal dysgenesis in the mare are presented. Case No 1 was a pure 63, XO, while Case No 2 was a mosaic with a preponderance of XX cells. The clinical picture was one of phenotypically normal female mares with small uteri and infantile ovaries. The ovaries lacked germ cells, and consisted of stroma only. This study emphasizes the importance of chromosome analysis in providing information concerning the mechanisms involved is some cases of equine infertility.
The Journal of heredityMarch 1, 1997
Volume 88, Issue 2 162-164 doi: 10.1093/oxfordjournals.jhered.a023079
Lear TL, Bailey E.The U2 linkage group of horses includes the genes albumin (ALB), vitamin D binding protein (GC), mitochondrial glutamate oxaloacetate transaminase 2 (GOT2), and haptoglobin (HP) which are found on two human chromosomes, namely, 4 (HSA 4) and 16 (HSA 16). Likewise these genes are also found on two different chromosomes in mice, rats, and cattle. Chromosome painting demonstrated that only horse chromosome 3 (ECA 3) hybridized with whole chromosome paints for both HSA 4 and HSA 16. This indicated that the equine U2 linkage group occurs on ECA 3, spanning the centromere. This technique will be use...
Payan-Carreira R, Quaresma M, Oliveira-Martins T, Silva JR.This report describes a case of blind vagina diagnosed during a pre-breeding evaluation of a 5-year-old crossbred Lusitano mare. This mare was mated twice during the previous breeding season but remained open. Clinical evaluation revealed the existence of follicles in the ovaries and an enlarged uterus. An ultrasound examination showed that a granular free-floating fluid distended the uterus. No connection was detected between the uterus and the vagina and a presumptive diagnosis of congenital vaginal obstruction was raised. On vaginal examination it was noticed that the vagina was short in de...
Stewart-Scott IA, Pearce PD, Burkin DJ.Sex chromosome abnormalities have been detected in a further five mares with clinical histories of small ovaries and absent or irregular oestrous cycles. Three mares had 63,XO karyotypes (X monosomy) and two were sex chromosome mosaics with karyotypes of 63,XO/64,XY and 63,XO/64,XX/64,XY respectively. A sex chromosome abnormality (X monosomy) has also been found in a filly where it was suspected because of her short stature.
Long SE.Nine reproductively normal mares, 25 infertile mares and one set of heterosexual twins were examined cytogenetically using conventional giemsa staining, C-banding and G-banding. It was concluded that G-banding was necessary to identify even gross anomalies. Three (12 per cent) of the infertile mares, but none of the controls, had a chromosomal anomaly. One was 63,XO, one a 63,XO/64,XX mixoploid and one a 64,XY sex reversed male. It is argued that a cytogenetic examination is a useful diagnostic technique but that routine screening of the whole population would be uneconomic.
Power MM.The results of Y chromosome measurements in 31 horses are presented. The Y chromosome was identified using G-, R-, and C-banding techniques. From G-banded metaphase spreads, total X and Y chromosome and separate proximal (P) and distal (D) Y-band measurements were made. Within this group, the Y/X ratio (%) for each animal varied from 18.93 to 43.95, with an overall mean of 34.85 and a coefficient of variation (CV) of 16.12. The overall mean P/X ratio (%) was 23.57 with a CV of 20.57, compared with an overall mean D/X ratio (%) of 11.26 with a CV of 15.18. The group studied included 27 Thorough...
Kútvölgyi G, Brabender K, Andersson M, Javkhlan A, Nagy S, Páble T, Egerszegi I, Hidas A, Soós I, Kovács A.The case of an 8-year-old, sexually active but infertile Przewalski's stallion (Equus ferus przewalskii) was studied. Besides the infertility, the stallion also showed permanent problems with its body condition, being obviously weaker than all the other group members. The horse was kept in a separate place for two years with 12 mares in its harem group (six mares had foals earlier); however, none of the mares covered got pregnant. Andrological and cytogenetic investigations revealed underdeveloped testes, arrested spermatogenesis, azoospermia, and XY/XXY/X0 mosaicism. We classify the case as a...
Milliken JE, Paccamonti DL, Shoemaker S, Green WH.A pseudohermaphrodite horse with aggressive stallion-like behavior and ambiguous external genitalia was gonadectomized. The hypoplastic gonads removed from the abdomen were confirmed by histologic examination to be testes. Examination of blood and fibroblasts revealed a 64,XX karyotype.
Halnan CR.From the standpoint that cytogenetic screening in mares is seldom necessary as an aid to diagnosis of the gonadal dysgenesis syndrome, a series of double-blind trials were conducted to test the proposal that present practice failed to explore the potential for cytogenetics in clinical practice. It was demonstrated that diagnoses of infertility might be made where mares were found to be of normal phenotype by clinical examination. Such mares were found to be gonosmic mosaics. One stallion had a polymorphism of the X chromosome and had poor conception rates. It was demonstrated that the true val...
Sato F, Hirota K, Tozaki T, Ito K, Dhakal P, Taya K, Endo Y, Murase H, Nambo Y.A Thoroughbred colt with ambiguous external genitalia was presented for clinical and histological examinations. The colt had a short penis that faced backward between his hind limbs. The measurements of luteinizing hormone, follicle stimulating hormone, testosterone and ir-inhibin showed a tendency to increase gradually from April. Both the sex-determining region of the Y chromosome and amelogenin gene fragments were detected by the PCR method. A cytogenetic analysis revealed the 63,XO/64,XY mosaic karyotype (ratio 83:17). In autopsy, immature symmetrical subcutaneous testes were found in the ...
Lear TL, Trembicki KA, Ennis RB.Giemsa-11 (G-11) staining and in situ hybridization were used to identify the equine chromosome complement of horse x mouse somatic cell hybrids. The presence of horse chromosomes in somatic cell hybrids was determined by differential G-11 staining. The slides were then destained and hybridized with biotinylated total horse (Equus caballus) genomic DNA without suppression. Fluorescence detection permitted rapid confirmation of horse chromosomal DNA in the hybrid cells.
