CROSS-REACTIVE PROPERTIES OF ANTISERA PREPARED IN RABBITS 

 BY STIMULATIONS WITH TELEOST VITELLINS 



By Fred M. Utter. Chemist and George J. Ridgvc ay. Biochemist.^ Bureau of 

 Commercial Fisheries Biological Laboratory. Seattle. Wash.. 98102 



ABSTRACT 



Antisera were prepared by injecting rabbits with egg or 

 serum vitellin preparations from seven teleost species be- 

 longing to different families. The ranges of reactivity of 

 these antisera were tested with sera from mature females of 

 nine teleost families as well as with sera from females of 

 spiny dogfish. Pacific lamprey, and white sturgeon. All of 

 these antisera reacted with vitellins from all species tested 

 from the homologous families. Antisera prepared against 



rockfish and flounder vitellins cross-reacted with sera from 

 mature females of all teleost species tested. A greater anti- 

 genic complexity in the vitellins of more taxonomically 

 advanced species than more primitive species is indicated 

 by the results of the reactions and absorption tests. The 

 results are of practical importance in studies on maturity of 

 fishes and have theoretical implications in the field of 

 systematics. 



Fishery researcher.'^ have used serological 

 techniques with increasing frequency since 

 1950. Much of the work has been directed 

 toward identification of populations either 

 through blood-grouping techniques or studies 

 of variable serum antigens (Gushing, 1964). 

 Ridgway. Klontz, and Matsumoto (1962) ob- 

 served a characteristic antigen in the serum of 

 maturing and mature female sockeye salmon 

 {OncorJiiiiichus iierka) . A review of the litera- 

 ture and subsequent studies by our group has 

 revealed that similar components occur not 

 only in all the teleosts but also in all vertebrate 

 classes were oviparity occurs (Urist and 

 Schjeide, 1961; Drilhon and Fine, 1963), These 

 antigens appear to have considerable practical 

 value in investigations of maturity in female 

 fish because of their connection with the proc- 

 ess of sexual maturation (Ho and Vanstone, 

 1961; Olivereau and Ridgway, 1962; Ridgway, 

 1964; Utter and Ridgway, 1966). 



The function of the blood serum as a trans- 

 porting medium between the site of synthesis 

 in the liver and the site of storage in the ovary 

 appears to explain the presence of yolk com- 

 ponents in the blood (Vanstone. Maw, and 

 Common, 1955), The serum vitellins studied 

 have displayed similar biochemical properties, 

 are characterized as phospholipoproteins, and 



I'revf. 



.Tl.ir,. 



Wfvr l*,ontliti.-H> Harbor. Elaine. 



FISHERY BULLETIN: VOLUME 66, NO, 2 



Published April 1967. 



conform to the classical de.scription of avian 

 vitellin as the water-insoluble fraction of the 

 egg yolk (Jukes and Kay, 1932; Vanstone, 

 Maw, and Common, 1955), 



This report is based on data obtained 

 through testing numerous antisera for cross- 

 reactive properties. The antisera were pre- 

 pared in rabbits against vitellins of teleosts. 

 We intend to bring out points of both practical 

 significance and theoretical interest, 



METHODS AND MATERIALS 



IMMUNOLOGICAL TESTS 



A microslide adaptation of the double dif- 

 fusion method of Ouchterlony as modified by 

 Ridgway, Klontz, and Matsumoto (1962) was 

 used for all serological tests. The agar medium 

 consisted of 1.5 percent Difco agar, 0.72 per- 

 cent sodium chloride, 0,60 percent sodium ci- 

 trate, 0,01 percent merthiolate. and 0,01 per- 

 cent trypan blue. Wells were punched in the 

 agar at 8-mm. intervals and filled to a volume 

 of about 0.01 cc. of reactant. Slides were eval- 

 uated after 24 hours of incubation at 37'- C. 



PRODUCTION OF ANTISERA 



Table 1 lists the antisera used in this study. 

 Egg vitellin preparations were made by blend- 

 ing and centrifuging one part eggs with three 



203 



