Hanebrink, Earl L.

Johnson, Bob D.

Snow, Lloyd D.

English

UARK (University of Arkansas )

Journal of the Arkansas Academy of Science Volume 23 Article 14 1969 Electrophoretic Patterns of Plasma Proteins and Hemoglobin of the Pigeon Columba liva domestica Lloyd D. Snow Arkansas State University Earl L. Hanebrink Arkansas State University Bob D. Johnson Arkansas State University Follow this and additional works at: https://scholarworks.uark.edu/jaas  Part of the Molecular Biology Commons, and the Ornithology Commons Recommended Citation Snow, Lloyd D.; Hanebrink, Earl L.; and Johnson, Bob D. (1969) "Electrophoretic Patterns of Plasma Proteins and Hemoglobin of the Pigeon Columba liva domestica," Journal of the Arkansas Academy of Science: Vol. 23 , Article 14. Available at: https://scholarworks.uark.edu/jaas/vol23/iss1/14 This article is available for use under the Creative Commons license: Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0). Users are able to read, download, copy, print, distribute, search, link to the full texts of these articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author. This Article is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Journal of the Arkansas Academy of Science by an authorized editor of ScholarWorks@UARK. For more information, please contact scholar@uark.edu. 80Arkansas Academy of Science Proceedings, Vol. 23, 1969ELECTROPHORETIC PATTERNS OF PLASMAPROTEINS AND HEMOGLOBIN OF THE PIGEONCOLUMBA LIVADOMESTICALloyd D. Snow, Earl L. Hanebrink and Bob D. JohnsonArkansas State UniversityINTRODUCTIONElectrophoresis is being used with increasing frequency byvertebrate taxonomists. This technique takes advantage of the dif-ferent migration rates of protein molecules in an electric field.The objectives of this study were to: (1) determine if breedsof pigeons could be distinguished by studying plasma protein com-ponents and (2) determine if hemoglobin is useful by this techniquein separating various breeds of pigeons. Ifso, this technique couldbe useful in studying molecular evolution during the developmentof present pigeon breeds. According to Darwin (1897) the RockPigeon (Columba liva) may be confidently viewed as the commonparent pigeon stock. It is from this parent stock that numerousbreeds and varieties have been developed. For this study three distinctbreeds were used; namely the Modena, English Trumpeter andGerman Beauty Homer (Die Deutsche Schautaube). These breedsdiffer much in body characteristics, origin and time of development,and their "gene pools" have been somewhat isolated for manyyears.The Modena (Fig. 1) is an Italian creation which had itsbeginnings as far back as 1328. The English Trumpeter (Fig. 2)is also an old breed. Aldrovandi in 1603 apparently described it asa variety of the Runt. The breed was well recognized in Englandby 1735 (Levi, 1957). The German Beauty Homer (Fig. 3) is aGerman creation which owes its origin to German breeders whodeveloped a show pigeon from crosses of Racing Homers, Antwerpsand others around 1908 (Weger, 1954).ACKNOWLEDGEMENTSThis study was primarily supported by a grant from theFaculty Research Committee of Arkansas State University and partlyby NIHGrant ES 00275-02. Photographic work was done by JeneaneSaunders.MATERIALS AND METHODSFifty samples of 1.5 ml of whole blood were taken from thewing vein on the inside of the elbow joint from individuals ofkiodenas, English Trumpeters and German Beauty Homers. Thepigeon was held with its back downward and the wing laterallyspread. Removal of a few feathers made the vein visible (Schermer,1967).80Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14Published by Arkansas Academy of Science, 196981OXElectrophoretic Patterns of Plasma ProteinsFig. 1. The Blue Gazzi Modena Pigeon.81Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14https://scholarworks.uark.edu/jaas/vol23/iss1/1482Arkansas Academy of Science ProceedingsFig. 3. The German Beauty Homer (Die Deutsche Schautaube).Fig. 4. Eight Column Electrophoresis Apparatus.82Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14Published by Arkansas Academy of Science, 1969'83Electrophoretic Patterns of Plasma ProteinsWhole blood was drawn from each pigeon by a B-D insulinsyringe needle and put in a solution of 3.8 percent sodium citrate(4 parts blood and 1 part sodium citrate) to prevent clotting. Theblood was centrifuged for 5 minutes. The plasma was removed by atransfer pipette and frozen. The RBC's were washed six times in a0.75 per cent bird physiological saline solution (Humason, 1967).A ml of distilled water was added to the RBC's to cause hemolysis.Hemoglobin was drawn off with a transfer pipette and frozen untilused.Disc electrophoresis procedures closely followed those of Davis(1961). The analyses were performed with an eight column electro-phoresis apparatus (Fig 4) utilizing graphite electrodes and twocircular reservoirs (2 in. deep and 6 in. in diameter) constructedfrom 6 in. acrylic tubing. Acrylamide gels (0.5 cm x 4.8 cm) andTRIS buffer at pH 8.3 were used throughout. Separation of plasmaproteins was conducted at 25°C and at 12 mA for 150 min at 100volts. After separation the staining was accomplished by usingNaphthol Blue Black. The most distinct electrophoretic patternswere obtained with 15 ul of plasma. The gels were then destainedand stored according to procedures outlined by Davis (1961).RESULTS AND DISCUSSIONStudies prior to 1957 on electrophoresis of avian plasma orserum proteins have been limited to a few isolated birds. Wall andSchlumberger (1957) investigated the plasma electrophoretic pat-terns of the shell parakeet (Melopsittacus undulatus). Sibley andJohnsgard (1959) published on the variability in the electrophoreticpatterns of avian serum proteins. Their studies included a variety ofavian species. In this study the Rock Dove (Columba liva) was in-vestigated. The histogram profiles of the serum proteins showed twolarge components for this species. Perkins (1964) published on electro-phoretic patterns of the serum proteins and hemoglobins of the genus(Lams). He showed no more variation between the species thanwithin a single species for this genus even though there is a widevariation in the size and coloration of these birds. Itwas concludedby Baker and Hanson (1965) that species and subspecies of geesecannot be distinguished on the basis of the blood proteins and thatthere are only minor differences between the genus (Anser) and(Branta). Eleven species including subspecies were used in theirstudy. Rylander (1967) studied electrophoretic patterns of the serumproteins of two genera of the family Scolopacidae. He concludedthat intraspecific variation is great and that there is no significantdifference between the species or even genera in this family. Sibleyand Johnsgard (1959) stated that if reliable measurements of aviansera are desired, sample size must be large, and birds must beseparated by age and sex, just as in studies of morphological char-acters. 83Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14https://scholarworks.uark.edu/jaas/vol23/iss1/1484Arkansas Academy of Science ProceedingsThe results from this study by using 1 acrylamide gel electro-phoresis techniques on three distinct breeds of domesticated pigeonsshow minor individual differences within the breed and also dif-ferences between individuals of different breeds. Because of thesevariations there was no one representative electropherogram foreach breed of pigeon. Some samples from individuals within a breedwere remarkably similar. Intrabreed variation is not always easyto explain. These variations may be caused by differences in experi-mental procedures, by sex, age and perhaps health of individualpigeons. In general, there was some uniformity in protein patternsamong the pigeons studied. These patterns are shown in Figs. 5-8.Some protein components did not migrate from the starting zoneof the small pore acrylamide gel. For all three breeds a pronounceddisc appeared at 3 to 4 mm below the starting zone. In the regionof 1 to 15 mm from the starting zone the breeds varied in thenumber of narrow slowly migrating protein discs. In this regionthere were 7 to 8 protein discs for the English Trumpeter; 8 to9 for the German Beauty Homer; and 6 to 8 for the Modena. Forall three breeds two wide rapidly migrating plasma protein discsappeared inin the 15-30 mm region of the gels. In the lower 15-30 mmregion the most significant variation between different breeds wasfound in the number of narrow rapidly migrating discs preceedingeach wide disc. English Trumpeters had 1 or 2 narrow protein com-ponents before the first wide protein disc and sometimes one smalldisc before the second wide disc. The German Beauty Homer usuallyliad one narrow protein disc preceeding each large one. Plasma fromModena's also had 1 or 2 narrow discs preceeding each wide disc.The absence of the narrow disc in certain individuals was probablydue to extremely faint discs caused by low concentrations of aplasma protein component and to masking by the wide fast movingdisc, some of which migrated at different speeds between individualpigeons.Because of technical difficulties hemoglobins of the breedswere not extensively studied by this electrophoretic technique. Hemo-globins from the above mentioned breeds seemed to have exceptional-y low specific gravities. Hemoglobin from the White King breedshowed one large component. Other studies have shown hemoglobino be of no value in identifying bird species by electrophoresis tech-niques.SUMMARYFifty pigeons of three distinct breeds (English Trumpeter,German Beauty Homer and Modena) were used in studying patternsof plasma proteins and hemoglobin with disc electrophoresis tech-niques. Individual variations of protein patterns were observed andescribed. Disc electrophoretic techniques were not useful in study-ng pigeon hemoglobins.84Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14Published by Arkansas Academy of Science, 196985Electrophoretic Patterns of Plasma ProteinsFig. 5. Electrophoreograms Of Three Modenas.Fig. 6. Electrophoreograms Of Three English Trumpeters.85Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14https://scholarworks.uark.edu/jaas/vol23/iss1/1486Arkansas Academy of Science ProceedingsFig. 7. Electrophoreograms Of Three German Beauty Homers.r. 8. Disc Electrophoresis Patterns Of Plasma Proteins OnAcrylamide Gels (From left: English Trumpeters, GermanBeatuy Homers and Modenas).86Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14Published by Arkansas Academy of Science, 1969'87Electrophoretic Patterns of Plasma ProteinsCITED LITERATUREBaker, C. M. Ann and H. C. Hanson. 1966. Molecular genetics ofavian proteins—VI. Evolutionary implications of blood pro-teins of eleven species of geese. Comp. Biochem. Physiol.17: 997-1006.Darwin, Charles. 1897. The variation of animals and plants underdomestication. 2nd. ed., 2 vol. D. Appleton and Co., New York.Davis, B. J. 1964. Disc electrophoresis - II.Method and applicationto human serum proteins. Ann. N. Y. Acad. Sci., 121:404-427.Humason, Gretchen L. 1967. Animal techniques. 2nd. ed., W. H.Freeman and Co., San Francisco. 569 p.Levi, Wendell. 1957. The pigeon. Levi Publ. Co., Inc. Sumter,S. C. 667 p.Perkins, Richard. 1964. The electrophoretic patterns of the serumproteins and hemoglobins of the genus Larus. Murrelet 45 :26-28.Rylander, Michael Kent. 1967. The electrophoretic patterns of theserum proteins of the genera Erolia and Ereunetes (FamilyScolopacidae). Murrelet 48: 39-40.Schermer, Siegmund. 1967. The blood morphology of laboratoryanimals. 3rd. ed. F. A. Davis Co., Philadelphia. 200 p.Sibley, Charles G. and Paul A. Johnsgard. 1959. Variability in theelectrophoretic patterns of avian serum proteins. Condor61: 85-95.Wall, Robert L. and Hans G. Schlumberger. 1957. Electrophoresisof plasma proteins in the parakeet. Science 125: 933-993.Weger, Richard. 1954. Die deutsche schautaube. Verlag DerAllgemeinen Geflugel-Zeitung. Schurmann and Klagges,Bochrum. 47 p.87Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14https://scholarworks.uark.edu/jaas/vol23/iss1/14

Electrophoretic Patterns of Plasma Proteins and Hemoglobin of the Pigeon Columba liva domestica

ScholarWorks@UARK

Journal of the Arkansas Academy of ScienceVolume 23 Article 141969Electrophoretic Patterns of Plasma Proteins andHemoglobin of the Pigeon Columba livadomesticaLloyd D. SnowArkansas State UniversityEarl L. HanebrinkArkansas State UniversityBob D. JohnsonArkansas State UniversityFollow this and additional works at: http://scholarworks.uark.edu/jaasPart of the Molecular Biology Commons, and the Ornithology CommonsThis article is available for use under the Creative Commons license: Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0). Users are able toread, download, copy, print, distribute, search, link to the full texts of these articles, or use them for any other lawful purpose, without asking priorpermission from the publisher or the author.This Article is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Journal of the Arkansas Academyof Science by an authorized editor of ScholarWorks@UARK. For more information, please contact scholar@uark.edu, ccmiddle@uark.edu.Recommended CitationSnow, Lloyd D.; Hanebrink, Earl L.; and Johnson, Bob D. (1969) "Electrophoretic Patterns of Plasma Proteins and Hemoglobin of thePigeon Columba liva domestica," Journal of the Arkansas Academy of Science: Vol. 23 , Article 14.Available at: http://scholarworks.uark.edu/jaas/vol23/iss1/1480Arkansas Academy of Science Proceedings, Vol. 23, 1969ELECTROPHORETIC PATTERNS OF PLASMAPROTEINS AND HEMOGLOBIN OF THE PIGEONCOLUMBA LIVADOMESTICALloyd D. Snow, Earl L. Hanebrink and Bob D. JohnsonArkansas State UniversityINTRODUCTIONElectrophoresis is being used with increasing frequency byvertebrate taxonomists. This technique takes advantage of the dif-ferent migration rates of protein molecules in an electric field.The objectives of this study were to: (1) determine if breedsof pigeons could be distinguished by studying plasma protein com-ponents and (2) determine if hemoglobin is useful by this techniquein separating various breeds of pigeons. Ifso, this technique couldbe useful in studying molecular evolution during the developmentof present pigeon breeds. According to Darwin (1897) the RockPigeon (Columba liva) may be confidently viewed as the commonparent pigeon stock. It is from this parent stock that numerousbreeds and varieties have been developed. For this study three distinctbreeds were used; namely the Modena, English Trumpeter andGerman Beauty Homer (Die Deutsche Schautaube). These breedsdiffer much in body characteristics, origin and time of development,and their "gene pools" have been somewhat isolated for manyyears.The Modena (Fig. 1) is an Italian creation which had itsbeginnings as far back as 1328. The English Trumpeter (Fig. 2)is also an old breed. Aldrovandi in 1603 apparently described it asa variety of the Runt. The breed was well recognized in Englandby 1735 (Levi, 1957). The German Beauty Homer (Fig. 3) is aGerman creation which owes its origin to German breeders whodeveloped a show pigeon from crosses of Racing Homers, Antwerpsand others around 1908 (Weger, 1954).ACKNOWLEDGEMENTSThis study was primarily supported by a grant from theFaculty Research Committee of Arkansas State University and partlyby NIHGrant ES 00275-02. Photographic work was done by JeneaneSaunders.MATERIALS AND METHODSFifty samples of 1.5 ml of whole blood were taken from thewing vein on the inside of the elbow joint from individuals ofkiodenas, English Trumpeters and German Beauty Homers. Thepigeon was held with its back downward and the wing laterallyspread. Removal of a few feathers made the vein visible (Schermer,1967).80Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14Published by Arkansas Academy of Science, 196981OXElectrophoretic Patterns of Plasma ProteinsFig. 1. The Blue Gazzi Modena Pigeon.81Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14http://scholarworks.uark.edu/jaas/vol23/iss1/1482Arkansas Academy of Science ProceedingsFig. 3. The German Beauty Homer (Die Deutsche Schautaube).Fig. 4. Eight Column Electrophoresis Apparatus.82Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14Published by Arkansas Academy of Science, 1969'83Electrophoretic Patterns of Plasma ProteinsWhole blood was drawn from each pigeon by a B-D insulinsyringe needle and put in a solution of 3.8 percent sodium citrate(4 parts blood and 1 part sodium citrate) to prevent clotting. Theblood was centrifuged for 5 minutes. The plasma was removed by atransfer pipette and frozen. The RBC's were washed six times in a0.75 per cent bird physiological saline solution (Humason, 1967).A ml of distilled water was added to the RBC's to cause hemolysis.Hemoglobin was drawn off with a transfer pipette and frozen untilused.Disc electrophoresis procedures closely followed those of Davis(1961). The analyses were performed with an eight column electro-phoresis apparatus (Fig 4) utilizing graphite electrodes and twocircular reservoirs (2 in. deep and 6 in. in diameter) constructedfrom 6 in. acrylic tubing. Acrylamide gels (0.5 cm x 4.8 cm) andTRIS buffer at pH 8.3 were used throughout. Separation of plasmaproteins was conducted at 25°C and at 12 mA for 150 min at 100volts. After separation the staining was accomplished by usingNaphthol Blue Black. The most distinct electrophoretic patternswere obtained with 15 ul of plasma. The gels were then destainedand stored according to procedures outlined by Davis (1961).RESULTS AND DISCUSSIONStudies prior to 1957 on electrophoresis of avian plasma orserum proteins have been limited to a few isolated birds. Wall andSchlumberger (1957) investigated the plasma electrophoretic pat-terns of the shell parakeet (Melopsittacus undulatus). Sibley andJohnsgard (1959) published on the variability in the electrophoreticpatterns of avian serum proteins. Their studies included a variety ofavian species. In this study the Rock Dove (Columba liva) was in-vestigated. The histogram profiles of the serum proteins showed twolarge components for this species. Perkins (1964) published on electro-phoretic patterns of the serum proteins and hemoglobins of the genus(Lams). He showed no more variation between the species thanwithin a single species for this genus even though there is a widevariation in the size and coloration of these birds. Itwas concludedby Baker and Hanson (1965) that species and subspecies of geesecannot be distinguished on the basis of the blood proteins and thatthere are only minor differences between the genus (Anser) and(Branta). Eleven species including subspecies were used in theirstudy. Rylander (1967) studied electrophoretic patterns of the serumproteins of two genera of the family Scolopacidae. He concludedthat intraspecific variation is great and that there is no significantdifference between the species or even genera in this family. Sibleyand Johnsgard (1959) stated that if reliable measurements of aviansera are desired, sample size must be large, and birds must beseparated by age and sex, just as in studies of morphological char-acters. 83Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14http://scholarworks.uark.edu/jaas/vol23/iss1/1484Arkansas Academy of Science ProceedingsThe results from this study by using 1 acrylamide gel electro-phoresis techniques on three distinct breeds of domesticated pigeonsshow minor individual differences within the breed and also dif-ferences between individuals of different breeds. Because of thesevariations there was no one representative electropherogram foreach breed of pigeon. Some samples from individuals within a breedwere remarkably similar. Intrabreed variation is not always easyto explain. These variations may be caused by differences in experi-mental procedures, by sex, age and perhaps health of individualpigeons. In general, there was some uniformity in protein patternsamong the pigeons studied. These patterns are shown in Figs. 5-8.Some protein components did not migrate from the starting zoneof the small pore acrylamide gel. For all three breeds a pronounceddisc appeared at 3 to 4 mm below the starting zone. In the regionof 1 to 15 mm from the starting zone the breeds varied in thenumber of narrow slowly migrating protein discs. In this regionthere were 7 to 8 protein discs for the English Trumpeter; 8 to9 for the German Beauty Homer; and 6 to 8 for the Modena. Forall three breeds two wide rapidly migrating plasma protein discsappeared inin the 15-30 mm region of the gels. In the lower 15-30 mmregion the most significant variation between different breeds wasfound in the number of narrow rapidly migrating discs preceedingeach wide disc. English Trumpeters had 1 or 2 narrow protein com-ponents before the first wide protein disc and sometimes one smalldisc before the second wide disc. The German Beauty Homer usuallyliad one narrow protein disc preceeding each large one. Plasma fromModena's also had 1 or 2 narrow discs preceeding each wide disc.The absence of the narrow disc in certain individuals was probablydue to extremely faint discs caused by low concentrations of aplasma protein component and to masking by the wide fast movingdisc, some of which migrated at different speeds between individualpigeons.Because of technical difficulties hemoglobins of the breedswere not extensively studied by this electrophoretic technique. Hemo-globins from the above mentioned breeds seemed to have exceptional-y low specific gravities. Hemoglobin from the White King breedshowed one large component. Other studies have shown hemoglobino be of no value in identifying bird species by electrophoresis tech-niques.SUMMARYFifty pigeons of three distinct breeds (English Trumpeter,German Beauty Homer and Modena) were used in studying patternsof plasma proteins and hemoglobin with disc electrophoresis tech-niques. Individual variations of protein patterns were observed andescribed. Disc electrophoretic techniques were not useful in study-ng pigeon hemoglobins.84Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14Published by Arkansas Academy of Science, 196985Electrophoretic Patterns of Plasma ProteinsFig. 5. Electrophoreograms Of Three Modenas.Fig. 6. Electrophoreograms Of Three English Trumpeters.85Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14http://scholarworks.uark.edu/jaas/vol23/iss1/1486Arkansas Academy of Science ProceedingsFig. 7. Electrophoreograms Of Three German Beauty Homers.r. 8. Disc Electrophoresis Patterns Of Plasma Proteins OnAcrylamide Gels (From left: English Trumpeters, GermanBeatuy Homers and Modenas).86Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14Published by Arkansas Academy of Science, 1969'87Electrophoretic Patterns of Plasma ProteinsCITED LITERATUREBaker, C. M. Ann and H. C. Hanson. 1966. Molecular genetics ofavian proteins—VI. Evolutionary implications of blood pro-teins of eleven species of geese. Comp. Biochem. Physiol.17: 997-1006.Darwin, Charles. 1897. The variation of animals and plants underdomestication. 2nd. ed., 2 vol. D. Appleton and Co., New York.Davis, B. J. 1964. Disc electrophoresis - II.Method and applicationto human serum proteins. Ann. N. Y. Acad. Sci., 121:404-427.Humason, Gretchen L. 1967. Animal techniques. 2nd. ed., W. H.Freeman and Co., San Francisco. 569 p.Levi, Wendell. 1957. The pigeon. Levi Publ. Co., Inc. Sumter,S. C. 667 p.Perkins, Richard. 1964. The electrophoretic patterns of the serumproteins and hemoglobins of the genus Larus. Murrelet 45 :26-28.Rylander, Michael Kent. 1967. The electrophoretic patterns of theserum proteins of the genera Erolia and Ereunetes (FamilyScolopacidae). Murrelet 48: 39-40.Schermer, Siegmund. 1967. The blood morphology of laboratoryanimals. 3rd. ed. F. A. Davis Co., Philadelphia. 200 p.Sibley, Charles G. and Paul A. Johnsgard. 1959. Variability in theelectrophoretic patterns of avian serum proteins. Condor61: 85-95.Wall, Robert L. and Hans G. Schlumberger. 1957. Electrophoresisof plasma proteins in the parakeet. Science 125: 933-993.Weger, Richard. 1954. Die deutsche schautaube. Verlag DerAllgemeinen Geflugel-Zeitung. Schurmann and Klagges,Bochrum. 47 p.87Journal of the Arkansas Academy of Science, Vol. 23 [1969], Art. 14http://scholarworks.uark.edu/jaas/vol23/iss1/14

https://scholarworks.uark.edu/cgi/viewcontent.cgi?article=3014&context=jaas

Electrophoretic Patterns of Plasma Proteins and Hemoglobin of the Pigeon Columba liva domestica

Abstract

Similar works

Full text

Available Versions

UARK (University of Arkansas )

ScholarWorks@UARK