DOCSLIB.ORG

Bleaching Melanin in Formalin- Fixed and Paraffin-Embedded Melanoma

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Bleaching Melanin in Formalin- Fixed and Paraffin-Embedded Melanoma 1EJH_2019_04_original.qxp_Hrev_master 31/10/19 09:47 Pagina 214 European Journal of Histochemistry 2019; volume 63:3071 Bleaching melanin in formalin- different and partly unknown molecular fixed and paraffin-embedded features. In general, melanins can be Correspondence: Claudio Pigoli, Department defined as a heterogeneous group of mole- of Veterinary Medicine, University of Milan, melanoma specimens using cules deriving from the oxidation of pheno- Via dell'Università 6, 26900 Lodi, Italy. visible light: A pilot study lic compounds with polymerization of the Tel. +39.02.50334165. resulting chemical products. In animals, E-mail: [email protected] 1 2 melanins are subdivided into eumelanins, Claudio Pigoli, Lucia Rita Gibelli, Key words: Bright-field microscopy; canine; 1 3 black to brown in color, and pheomelanins, Mario Caniatti, Luca Moretti, 1 equine; feline; light radiation; photobleaching; 1 1 reddish to yellowish. A third subtype of swine. Giuseppe Sironi, Chiara Giudice melanin, called neuromelanin and often 1Department of Veterinary Medicine, associated to the afore-mentioned groups, is Contributions: CP, LRG, MC, LM, GS, CG, University of Milan, Lodi composed of a mixture of eumelanins and study design, data analysis and interpretation; 2Istituto Zooprofilattico Sperimentale pheomelanins. In particular, it has been LM, performing of the light source spectral measurement; CP, performing of the photo- della Lombardia e dell’Emilia Romagna, hypothesized that neuromelanin granules consist of a pheomelanin core surrounded bleaching protocol, histochemical and Milan 2 immunohistochemical stains; CP, LRG, MC, 3 by a eumelanin layer. Department of Physics, Politecnico di GS, CG, viewed the histological specimens. In animals, melanin granules are pres- Milano, Milan, Italy All the authors have drafted the manuscript ent in various sites including integumentary and reviewed the work critically, have read system, eye, central nervous system and and approved the final manuscript and agreed inner ear. Given its presence in a wide range to be accountable for all aspects of the work. of tissues, it is not surprising that melanin Abstract performs different tasks ranging from mere- Conflict of interest: the authors declare they In fluorescence microscopy, light radia- ly mechanical ones to those of radical scav- have no competing interests. tion can be used to bleach fluorescent mole- engers. The main role of melanin is to pro- Acknowledgements: We would like to thank cules in formalin-fixed and paraffin-embed- tect living organisms from the harmful Massimo Datteri and Laura Tassielli from the ded (FFPE) samples, in order to increase the effects of electromagnetic radiations. In Istituto Zooprofilattico Sperimentale della ratio between signal of interest and back- particular, it is able to absorb radiations in a Lombardia e dell’Emilia Romagna for the ground autofluorescence. We tested if the wide range of wavelengths from the ultravi- technical support, cutting and staining of his- same principle can be exploited in bright olet to the visible spectrum. The energy tological sections. We would also like to thank field microscopy to bleach pigmented absorbed is then redistributed as heat.1 Stefano Tomasoni from DI.C.S.EL srl for his melanoma FFPE sections together with cell Energy absorption is also followed by redox valuable advices regarding the world of LED morphology maintenance. After dewaxing reactions that ultimately can lead to forma- light sources. and rehydration, serial FFPE sections of a tion of molecules having spectral absorp- Received for publication: 9 September 2019. feline diffuse iris melanoma, a canine dermal tion properties that differ from the original Accepted for publication: 15 October 2019. melanoma, a gray horse dermal melanoma polymer. Since melanins absorb both visible and a swine cutaneous melanoma were irra- and not visible light radiation, their oxida- This work is licensed under a Creative diated with visible light for 1, 2, 3, 4 and 5 tion can be achieved both by high-energy Commons Attribution-NonCommercial 4.0 days, prior to Hematoxylin & Eosin staining. radiation, such as ultraviolet rays, and by International License (CC BY-NC 4.0). Complete bleaching was obtained after 1-day low-energy radiation, such as visible light. treatment in feline and swine melanomas, The phenomenon by which a molecule ©Copyright: the Author(s), 2019 while 2 and 3 days were required in canine undergoes oxidation by means of light irra- Licensee PAGEPress, Italy and equine neoplasms, respectively. In all 3 European Journal of Histochemistry 2019; 63:3071 diation is called photo-oxidation. doi:10.4081/ejh.2019.3071 treated samples, cell morphology was main- Although the exact mechanism of melanin tained. Photo-induced bleaching combined photo-oxidation has not yet been explained with immunohistochemistry was tested after in all its aspects, several studies have shown a 3-day photo-treatment using five different that during ultraviolet and visible light irra- Materials and Methods markers. According to the literature, in all diation, singlet oxygen and superoxide samples neoplastic cells stained positive for anions are produced and rapidly quenched Samples vimentin, S100 and PNL2, while negative by melanin. The oxidation induced by these for FVIII and pancytokeratin. In conclusion, reactive oxygen species is considered to In this study, four FFPE samples of pig- visible light can be effectively exploited to play an important role in the photo-degrada- mented melanocytic neoplasms were used. bleach pigmented melanoma FFPE sections tion of melanin itself.3,4 Visible light is rou- In particular, a grey horse dermal melanoma prior to perform routine histochemical and tinely used, in our laboratory, to damage the and a swine cutaneous melanoma were immunohistochemical stains. molecules responsible for background auto- retrieved from the archives of the Istituto fluorescence, thus making them no longer Zooprofilattico Sperimentale della fluorescent, prior to perform immunofluo- Lombardia e dell’Emilia Romagna rescence assays on formalin-fixed and (IZSLER), while a case of feline diffuse iris paraffin-embedded (FFPE) samples.5-8 In melanoma and a case of canine dermal Introduction the present study, we investigated if the melanoma were obtained from the archives Melanins is a generic term derived from same principle could also be exploited in of the Department of Veterinary Medicine, the Greek word “melanos” meaning dark. bright field microscopy to bleach FFPE University of Milan. For each sample, six 4 Although the term melanin is often used to samples of pigmented melanoma, prior to µm-thick serial sections were cut and identify a single entity, it actually encom- perform routine histochemical and placed on SuperFrost Plus™ adhesion passes a number of pigments widely distrib- immunohistochemical stains, ensuring the slides (Thermo Scientific, Waltham, MA, uted in living beings and characterized by maintenance of cell morphology. USA). [page 214] [European Journal of Histochemistry 2019; 63:3071] 1EJH_2019_04_original.qxp_Hrev_master 31/10/19 09:47 Pagina 215 Original Paper Light source and emission spectrum buffer maintained at 24°C (the temperature different markers were tested: three of them evaluation reached by the sections during photo-treat- (vimentin, S100 and PNL2) are commonly In this study a 300 W ViugreumTM spot- ment) and in the dark inside a thermostat used in the melanocytic neoplasm histolog- light (Guangdong Zhongshan Yilangde incubator (Cavallo srl, Buccinasco, MI, ical identification workflow, while two Illumination CO. Ltd., Zhongshan City, Italy). Every 24 h, for 5 days, a section for (pancytokeratin and FVIII) are usually not China) containing a 594-phosphor white each sample was removed from the Petri expressed by melanoma cells. Sections LED panel with an emission divergence of dishes and stored with the control sections. were dewaxed in xylene and rehydrated 120° and emitting a 36000 lum and 6500 K At the end of the fifth day, to minimize the through graded alcohols; endogenous per- light, was used. Since the manufacturer of possible differences given by a non-con- oxidase activity was blocked with 0.3% the spotlight supplies only the color temper- temporary histochemical staining, all histo- H2O2 in methanol for 30 min. Sections were ature of the light emitted but not the specific logical sections were jointly stained with then dipped in Petri dishes containing emission spectrum, the latter was measured hematoxylin-eosin (H&E). 0.05% sodium azide TRIS buffer and pho- using a HR2000+ spectrometer (Ocean tobleached for 3 days. The duration of the Photobleaching protocol combined photo-treatment was chosen after the evalu- Optics, Largo, Florida), which has a wave- with immunohistochemical stain length range of detection from 190 nm to ation of the results of photobleaching and In the immunohistochemical assay, five 1100 nm. The light of the lamp has been H&E combined protocol: the treatment able coupled to the spectrometer through an optical fiber. Photobleaching apparatus The photobleaching apparatus has been made similarly to the one described by Duong and Ha.6 The spotlight was placed in a room maintained at 4°C and placed so that the LED panel was facing upwards and that 4.5 cm thick reflective spacers could be placed on the spotlight protective glass. With this configuration the distance between the samples and the LED panel was 8.8 cm. Above the spacers, two 12-cm square Petri dishes were placed, filled with TRIS buffer pH 7.6 added with 0.05% sodi- um azide, in order to inhibit bacterial growth (Figure 1). A glass mirror was posi- tioned on the Petri dishes so that the light not directly absorbed by the samples could, at least in part, be redirected towards them. Figure 1. Photobleaching apparatus. The spotlight was placed so that the LED panel was fac- With this configuration of the apparatus, the ing upwards and that reflective spacers could be placed on the spotlight protective glass. Above amount of light impinging directly on the the spacers, two 12-cm square Petri dishes, filled with TRIS buffer added with 0.05% sodium samples from the LEDs has been calculated azide, were placed.
Load more

Recommended publications
  • Coat Color in Horses
    COAT COLOR IN HORSES Tabulation of Color of 42,165 Horses Allows Definite Conclusions to Be Drawn as to Value of Different Factors—Errors in Registry and in Genetic Description of Colors—Connection Between Gray and Roan.1 W. S. ANDERSON Assistant in Horse Husbandry, Kentucky Agricultural Experiment Station, Lexington, Ky. URST, Wilson, Harper, Sturte- brown, black and chestnut. As a rule vant, Anderson and others the variations of each of these colors have published papers on the are not recorded in the stud books. Inheritance of Coat Colors in The gray coat is made up of white Horses. It is the purpose of the writer and black hairs and varies from the to give a summary of all the available almost white to the almost black, and figures on the subject and his interpre- includes a large class of horses whose tation of them. The sources of the coat is of the dappled pattern. When figures collected are the various Stud young, the gray horse exhibits this Books. As a matter of fact these can dappled condition or is what is desig- not be accurate. I, myself, have used nated iron gray, but as age comes on the American Saddle Horse Register. the dapples disappear and white and This Register has been compiled within black hairs are to be found. Later on two or three decades and has been the black may almost be lost and result revised within a decade. I find errors in a white horse. This white, however, in it approximating two percent, for does not seem to be of the same nature color and as great a percent, of errors as the white found on spotted ponies that might be considered of a typo- and some classes of horses.
    [Show full text]
  • I . the Color Gene C
    THE ABC OF COLOR INHERITANCE IN HORSES W. E. CASTLE Division of Genetics, University of California, Berkeley, California Received October, 27, 1947 HE study of color inheritance in horses was begun in the early days of Tgenetics. Indeed many facts concerning it had already been established earlier, by DARWINin his book on “Variation of Animals and Plants under Domestication.” At irregular inteivals since then, new attempts have been made to collect and classify in terms of genetic factors the records contained in stud books concerning the colors of colts in relation to the colors of their sires and dams. A full bibliography is given by CREWand BuCHANAN-SMITH (19301. By such studies, we have acquired very full information as to what color a colt may be expected to have, when the color of its parents and grandparents is known. This knowledge is empirical rather than experimental in nature. For horses being slow breeding and expensive are rarely available for direct experi- mental study, such as can be made with the small laboratory mammals, mice, rats, rabbits and guinea pigs. We have definite information that color inheritance in horses involves the existence of mutant genes similar to those demonstrated by experimental studies to be involved in color inheritance of other mammals. But the horse genes have been given special names, as they were successively discovered, and it is difficult at present to correlate them with the better known names and geneic symbols used by the experimental breeders. The present paper is an attempt to make such a correlation. Just as in morphological studies comparative anatomy was found useful and still is used to establish homologies between systems of organs, so in mammalian genetics, a comparative study of gene action in the production of coat colors and color patterns may also be of value.
    [Show full text]
  • Horse Sale Update
    Jann Parker Billings Livestock Commission Horse Sales Horse Sale Manager HORSE SALE UPDATE August/September 2021 Summer's #1 Show Headlined by performance and speed bred horses, Billings Livestock’s “August Special Catalog Sale” August 27-28 welcomed 746 head of horses and kicked off Friday afternoon with a UBRC “Pistols and Crystals” tour stop barrel race and full performance preview. All horses were sold on premise at Billings Live- as the top two selling draft crosses brought stock with the ShowCase Sale Session entries $12,500 and $12,000. offered to online buyers as well. Megan Wells, Buffalo, WY earned the The top five horses averaged $19,600. fast time for a BLS Sale Horse at the UBRC Gentle ruled the day Barrel Race aboard her con- and gentle he was, Hip 185 “Ima signment Hip 106 “Doc Two Eyed Invader” a 2009 Billings' Triple” a 2011 AQHA Sorrel AQHA Bay Gelding x Kis Battle Gelding sired by Docs Para- Song x Ki Two Eyed offered Loose Market On dise and out of a Triple Chick by Paul Beckstead, Fairview, bred dam. UT achieved top sale position Full Tilt A consistant 1D/ with a $25,000 sale price. 486 Offered Loose 2D barrel horse, the 16 hand The Beckstead’s had gelding also ran poles, and owned him since he was a foal Top Loose $6,800 sold to Frank Welsh, Junction and the kind, willing, all-around 175 Head at $1,000 or City OH for $18,000. gelding was a finished head, better Affordability lives heel, breakaway horse as well at Billings, too, where 69 head as having been used on barrels, 114 Head at $1,500+ of catalog horses brought be- poles, trails, and on the ranch.
    [Show full text]
  • Color Coat Genetics
    Color CAMERoatICAN ≤UARTER Genet HORSE ics Sorrel Chestnut Bay Brown Black Palomino Buckskin Cremello Perlino Red Dun Dun Grullo Red Roan Bay Roan Blue Roan Gray SORREL WHAT ARE THE COLOR GENETICS OF A SORREL? Like CHESTNUT, a SORREL carries TWO copies of the RED gene only (or rather, non-BLACK) meaning it allows for the color RED only. SORREL possesses no other color genes, including BLACK, regardless of parentage. It is completely recessive to all other coat colors. When breeding with a SORREL, any color other than SORREL will come exclusively from the other parent. A SORREL or CHESTNUT bred to a SORREL or CHESTNUT will yield SORREL or CHESTNUT 100 percent of the time. SORREL and CHESTNUT are the most common colors in American Quarter Horses. WHAT DOES A SORREL LOOK LIKE? The most common appearance of SORREL is a red body with a red mane and tail with no black points. But the SORREL can have variations of both body color and mane and tail color, both areas having a base of red. The mature body may be a bright red, deep red, or a darker red appearing almost as CHESTNUT, and any variation in between. The mane and tail are usually the same color as the body but may be blonde or flaxen. In fact, a light SORREL with a blonde or flaxen mane and tail may closely resemble (and is often confused with) a PALOMINO, and if a dorsal stripe is present (which a SORREL may have), it may be confused with a RED DUN.
    [Show full text]
  • Captain of the Gray-Horse Troop, by Hamlin Garland 1
    Captain of the Gray-Horse Troop, by Hamlin Garland 1 Captain of the Gray-Horse Troop, by Hamlin Garland Project Gutenberg's The Captain of the Gray-Horse Troop, by Hamlin Garland This eBook is for the use of anyone anywhere at no cost and with almost no restrictions whatsoever. You may copy it, give it away or re-use it under the terms of the Project Gutenberg License included with this eBook or online at www.gutenberg.org Title: The Captain of the Gray-Horse Troop Author: Hamlin Garland Release Date: August 18, 2010 [EBook #33458] Captain of the Gray-Horse Troop, by Hamlin Garland 2 Language: English Character set encoding: ISO-8859-1 *** START OF THIS PROJECT GUTENBERG EBOOK THE CAPTAIN OF THE *** Produced by Mary Meehan and The Online Distributed Proofreading Team at http://www.pgdp.net (This file was produced from images generously made available by The Internet Archive/American Libraries.) THE CAPTAIN OF THE GRAY-HORSE TROOP By HAMLIN GARLAND SUNSET EDITION HARPER & BROTHERS NEW YORK AND LONDON COPYRIGHT, 1901. BY THE CURTIS PUBLISHING COMPANY COPYRIGHT 1902. BY HAMLIN GARLAND [Illustration] CONTENTS I. A CAMP IN THE SNOW II. THE STREETER GUN-RACK III. CURTIS ASSUMES CHARGE OF THE AGENT IV. THE BEAUTIFUL ELSIE BEE BEE V. CAGED EAGLES Captain of the Gray-Horse Troop, by Hamlin Garland 3 VI. CURTIS SEEKS A TRUCE VII. ELSIE RELENTS A LITTLE VIII. CURTIS WRITES A LONG LETTER IX. CALLED TO WASHINGTON X. CURTIS AT HEADQUARTERS XI. CURTIS GRAPPLES WITH BRISBANE XII. SPRING ON THE ELK XIII. ELSIE PROMISES TO RETURN XIV.
    [Show full text]
  • Unit 9 Te Horse Industry
    Unit 9 Te Horse Industry OBJECTIVES KEY WORDS ¾ Discuss the history of horses and their colt points role today. dorsal stripe pony draft horse stallion ¾ Identify common breeds of horses and equine withers ponies, and their characteristics. feathers ¾ Discuss the use of equine for work and feral recreational uses. filly ¾ Locate the parts of the horse. foal gelding ¾ Identify horse colors and markings. hands light horse mare 113 Many people love horses. But just because people enjoy working with horses, does that mean they are suited for a horse-related career? More than likely, the answer is yes. In fact, an enthusiasm for horses is a tremendous bonus. However, the horse industry is very diverse, and the various jobs in the horse industry require diferent types of education, skills and interests. Some jobs require a college education, but many do not. Also, some jobs require a high level of horsemanship, while other jobs require a better ability to work with people than animals. Te equine industry is a multimillion dollar enterprise. Te business is more than just horses—it encompasses feed, tack and equipment, SAE IDEA publications, veterinary care, advertising, clothing, education, and Exploratory many other fields that are either directly or indirectly afected by the Coordinate and conduct equine industry. a horse safety camp. History of the Horse Industry Horses are, quite literally, the maker of legends. From Alexander the Great’s Bucephalus to Walter Farley’s mythical black stallion, people have seen the horse as the embodiment of freedom, power, strength, beauty, and nobility. Te scientific name for the modern domesticated horse is Equus caballus.
    [Show full text]
  • American Paint Horse Association and What It Can Offer You, Call (817) 834-2742, Extension 788
    07CoatColorGenetics 12/14/07 6:51 PM Page A 07CoatColorGenetics 12/14/07 6:51 PM Page B Contents The Genetic Equation of Paint Horses . .IFC Tobiano . .1 Overo . .1 Tovero . .3 Breeding the Tobiano Paint . .4 Genes . .4 Understanding Simple Dominance . .4 Using the Punnett Square . .4 Understanding genes, simple dominance and the Punnett Square . .4 Breeding the Tobiano Paint . .5 Determining Tobiano Homozygosity . .5 Breeding the Overo Paint . .6 Breeding the Frame Overo . .6 Defining Minimal-White Frame Overo . .6 Breeding the Splashed White Overo . .6 Defining Minimal Splashed-White Overo . .6 Breeding the Sabino Overo . .7 Defining Minimal-White Sabino Overo . .7 Breeding the Tovero . .7 Coat Colors . .8 The Basic Rules of Coat Color Genetics . .9 Overo Lethal White Syndrome . .16 Lethal Whites—Fact Versus Fiction . .16 References . .17 Color Description Guide . .BC For more information on the American Paint Horse Association and what it can offer you, call (817) 834-2742, extension 788. Visit APHA’s official Web site at apha.com. The Genetic Equation of Paint Horses Paint Horses are unique from most other breeds because of their spotted coat patterns. Their base coats are the same colors as those of other breeds, but super- imposed over these colors are a variety of white spotting patterns. The three patterns recognized by APHA are tobiano, overo and tovero. The ability to recognize these patterns and under- stand the genetics behind them is essential for Paint Horse breeders. Being knowledgeable about coat pat- terns helps breeders and owners accurately describe their horses. Understanding the genetics that produce these patterns helps breeders increase the proportion of spotted horses in their foal crops.
    [Show full text]
  • Look Alike Colors 4.06 12/14/11 11:44 AM Page 64
    Look Alike Colors 4.06 12/14/11 11:44 AM Page 64 Look-Alike By Laura Hornick Behning ColorsColors Robbi-Sue’s Cassanova (Equinox Brigham x Robbi-Sue Misalert), a brown buckskin stal- lion (registered as dun), owned by Laura Bunke. When the cream gene is present on a brown base, as it is in this horse, the result is a very dark buckskin that is often mistaken for a non-dilute. dentifying what color a horse is can sometimes be difficult. the Morgan breed, where we have so much of the sooty modifier IThere are many colors that look similar, although they are turning clear coats into a much darker version of their original color, caused by completely different genes. After seeing many examples color identification can be a challenging—and confusing—task. of equine color, the knowledgeable breeder will pick up clues to It is unusual, but occasionally palominos can look like flax- help them differentiate between these “look-alike colors.” en red chestnuts. Some examples in our breed include Mac’s Parentage can also play a part in the discovery process. Since Littlebritches (Mac’s Baby x Golden Judy), a 1989 palomino geld- all of the dilution genes and color modifiers (except flaxen) are ing owned by Cindy Cerrigione of Connecticut, and Northerly dominant, a horse must get its color gene from at least one parent. Llwellyn (Northerly Intrigue x Northerly Gifted), a palomino For example, a dun horse must always have a dun parent and a gray gelding owned by Colleen McNichol of Four Seasons Farm in horse must always have a gray parent; dilution genes and modifiers Minnesota.
    [Show full text]
  • Black Piebald
    Black Black is a hair coat colour of horses in which the entire hair coat is black. Black is a relatively uncommon coat colour, and novices frequently mistake dark chestnuts or bays for black. However, some breeds of horses, such as the Friesian horse are almost exclusively black. Black is also common in the Fell pony, Dales Pony. True black horses have dark brown eyes, black skin, and wholly black hair coats without any areas of permanently reddish or brownish hair. They may have pink skin beneath any white markings under the areas of white hair, and if such white markings include one or both eyes, the eyes may be blue Piebald A piebald or pied animal is one that has a spotting pattern of large unpigmented, usually white, areas of hair, feathers, or scales and normally pigmented patches, generally black. The colour of the animal's skin underneath its coat is also pigmented under the dark patches and unpigmented under the white patches. This alternating colour pattern is irregular and asymmetrical. Some animals also exhibit colouration of the irises of the eye that match the surrounding skin (blue eyes for pink skin, brown for dark Chestnut Chestnut is a hair coat colour of horses consisting of a reddish-to-brown coat with a mane and tail the same or lighter in colour than the coat. Genetically and visually, chestnut is characterized by the absolute absence of true black hairs. It is one of the most common horse coat colours, seen in almost every breed of horse. Chestnut is a very common coat colour but the wide range of shades can cause confusion.
    [Show full text]
  • Guidelines to Coat Color & Coat Characteristics Chestnut – Shades
    Guidelines to Coat Color & Coat Characteristics Chestnut – shades from golden red to dark reddish brown. The mane, tail, and legs are not black, but are the color of the body or shades lighter or darker. Black or dark chestnuts may look basically black with the exception of red hairs on the coronet, pasterns, and/or back of fetlocks. Black – true black without any light areas. Bay – reddish shades from reddish tan to dark mahogany brown. All bay horses have black manes and tails, and black legs below the knees and hocks. Brown – black with light mealy areas at muzzle, eyes, flanks, and inside of legs. Palomino – shades of very pale creamy yellow to golden yellow, with flaxen, silver, or white mane and tail. Palomino is produced by the action of a single cream dilution gene on a chestnut base. Buckskin – tan to yellow coat, with black mane and tail and black on lower legs. Buckskins without the dun gene may have a dorsal line, but this is countershading, not a true dorsal stripe. Buckskin is produced by the action of a single cream dilution gene on a bay base. Smoky black – varying shades of black, especially when weathered or sun-faded. May appear to have a black or brown body and may be difficult to distinguish from black, dun, chestnut, or brown, but all smoky blacks will have at least one parent with a dilute gene. Smoky black is produced by the action of a single cream dilution gene on a black base. The dilution of a single dose of cream may not be apparent, or may be very subtle, on a black base.
    [Show full text]
  • 2018 PDF Catalog
    Selling Approximately 100 Head of AQHA Horses Approximately 15 Riding Horses bar none cowboy church – midway, ar 72651 10 miles north of mountain home, ar REFERENCE SIRES OWN SONS OF: Two ID Bartender ● Mr Gold Bucks ● Docs Gabilan ● Smart Chic Olena ● Joe Jack Honey Bar Dun It Like Lena ● Twice As Shiney ● Hollywood Dun It ● CL Poco Gold Doc Royal Blue Texas (Peptoboonsmal) ● Fuel N Shine (Shining Spark) Cee Booger Red ● Fiddlin Beau Jack ● Bet Hesa Cat (High Brow Cat) ● Mr Red Bartender For Information or Catalog: Kenny McCullough, Pres. 870.895.4026 ● Donnie Perry, Vice Pres. 870.656.2198 Teresa Walker, Treasurer 870.373.0106 Call or Text INDEX TO HORSES BY BREEDER: Killian Quarter Horses: 2, 15, 32, 35, 51, 56, 72, 75, 78, 79, 87, 88, 91, 95, 101, 103 Mike Mahan: 5, 10, 29, 45, 97 Kenny McCullough: 1, 30, 33, 35, 37, 44, 52, 62, 65, 67, 69, 71, 73, 80, 82, 86, 89, 94, 100 Montgomery Performance Horses: 8, 16, 18, 27, 38, 40, 46, 64 Donnie & Georgina Perry: 6, 11, 13, 19, 23, 50, 53, 66, 70, 93, 104 Gemstone H Ranch: 3, 12, 14, 17, 22, 26, 36, 48, 49, 58, 68, 77, 81, 90, 98, 102 Jerry & Diane Walker: 7, 20, 47, 57, 60, 76 Scott & Teresa Walker: 9, 21, 24, 25, 28, 31, 34, 39, 42, 43, 54, 59, 61, 63, 74, 84, 85, 92, 96, 99 Walker 4W Farms: 4, 41, 55, 83 ABOUT OFBA Ozark Foundation Breeder’s Association (OFBA) was founded in 1999 by a group of family and friends with a passion for the American Quarter Horse.
    [Show full text]
  • Equine Science
    Lecture 12- Genetics (including coat color) Chapter 10 Gene – basic unit of inheritance. Genes are carried on chromosomes. Alleles – are the different forms of the same gene located at the same position on the chromosome. E,e E=black e=red Genes of somatic cells are always in pairs. Chromosomes are in the nucleus of all cells Genes are composed of DNA. ◦ The 4 Nitrogen bases are: Adenine (A) and thymine (T) Guanine (G) and cytosine (C) Genome – the complete set of genes for that organism. 64 chromosomes, 32 pairs. (31 somatic and 1 sex) Sex determination – XY in stallions, XX in mares. Sexed semen Alleles – two forms of the gene. ◦ Homozygous (GG, gg) ◦ Heterozygous (Gg) Dominant allele – expressed (G) Recessive allele – unexpressed (g) Genotype – genetic information Phenotype – How the expression of the gene appears ◦ Phenotype = genotype + environmental effects W – white gene obscures action of other genes WW is lethal Ww allele pair White lacks pigment in skin and hair ◦ Skin - pink ◦ Hair - white All non-white horses are ww. G – gray Dominant G ◦ GG or Gg will result in a gray horse. G allele horse can be born any color but gray but will gray out with age. Pigmented skin and eyes. At least one parent must be gray. All non-gray horses are gg. E – black gene EE or Ee is a black horse or at least has black points. ee is a red horse, liver chestnut, dark chestnut, chestnut, sorrel. A – Agouti; distribution of black hair AA, Aa gives black points if E is present aa presents a totally black horse if E is present C – pigment dilution CC : fully pigmented horses CCcr : red is diluted to yellow; black is unaffected.
    [Show full text]