116
JOtKN R . CUSHING
staroh gel electrophop& of the acrumg of some rnwine mthropods
that should mrvc as a sourid basis for further investigations using t h h
teohnique. Denuc6 and Cushing (1963) have compared some inveftebrate w u m E by means of immunodectrophoresls.
In retrospect, the information rcviewcd in this section, while not
complek, is sufficient t o show that a number of new techniques are
wsilsble that show groat promhe a9 aids in diacovering genetic
polymorphism in a variety of Inolecular epecies among marine forms.
Thh section conclude8 with t,he notation that the elucidation of
the nature of the evolut.ion of the mechanism of antibody synthesis
and the immune response, with their asgociated genetic considerations,
i 8 a problem that is receiving increasing attention. An introduction t o
thia subject will he found in Cushing and Campbell (1967) and in
publications of Dr. R . A. Good and his arrsociatea (cf. Papermaster
et al., 1963).
IV. METHOD^
A. Inlradwtim
The following eecbiond discuss mcthade of blood grouping that are
particularly useful in the study of marine animals. More detailed
discussions of methods are t o be found in the texh referred to in tho
Introduction and in specific papers referred t o in the various sections.
The reader should note that the choice of a particular method is often
guided by the nature of t h e problem at hand and by the permnal preferences of investigator8 ; also that each pemon usually modifies 60mo
of tho details of any method in order to fit his nee&
B. Collection of samples
Blood samples a m &ken in various ways, E’iA are often bled from
the hemt or gill arch, using hypodermic needles of 18 to 24 gsugcs
and syringes holding up to 50 ml of blood. It i s sometimes convenient
b anesthetize the fLh, for exampIe, by plmieg it in eolution of
tricaiiie methane sulfurlate (MS222, Sandoz Phamaceutieals) at concentrations of I : 20 000 to 1: 8000. It may be n e e c e w to draw
some anticoagulant into the syringe before w e . Antimagulanta that
have boen auccessful include 80diUm citrate, haperin and Alaever’s
Boluticm (of. Campbell et ol. 1063).
Rsh blood C ~ R
&Q
be obtained from the caudal artery of aneathetized fish by levering the taiI and holding the cut surface in a v e d
containing anticoagulant. Care ahould be taken to clean the tail of
BH0888 mucus. This method is particularly useful with amdl fish.
JOtKN R . CUSHING
staroh gel electrophop& of the acrumg of some rnwine mthropods
that should mrvc as a sourid basis for further investigations using t h h
teohnique. Denuc6 and Cushing (1963) have compared some inveftebrate w u m E by means of immunodectrophoresls.
In retrospect, the information rcviewcd in this section, while not
complek, is sufficient t o show that a number of new techniques are
wsilsble that show groat promhe a9 aids in diacovering genetic
polymorphism in a variety of Inolecular epecies among marine forms.
Thh section conclude8 with t,he notation that the elucidation of
the nature of the evolut.ion of the mechanism of antibody synthesis
and the immune response, with their asgociated genetic considerations,
i 8 a problem that is receiving increasing attention. An introduction t o
thia subject will he found in Cushing and Campbell (1967) and in
publications of Dr. R . A. Good and his arrsociatea (cf. Papermaster
et al., 1963).
IV. METHOD^
A. Inlradwtim
The following eecbiond discuss mcthade of blood grouping that are
particularly useful in the study of marine animals. More detailed
discussions of methods are t o be found in the texh referred to in tho
Introduction and in specific papers referred t o in the various sections.
The reader should note that the choice of a particular method is often
guided by the nature of t h e problem at hand and by the permnal preferences of investigator8 ; also that each pemon usually modifies 60mo
of tho details of any method in order to fit his nee&
B. Collection of samples
Blood samples a m &ken in various ways, E’iA are often bled from
the hemt or gill arch, using hypodermic needles of 18 to 24 gsugcs
and syringes holding up to 50 ml of blood. It i s sometimes convenient
b anesthetize the fLh, for exampIe, by plmieg it in eolution of
tricaiiie methane sulfurlate (MS222, Sandoz Phamaceutieals) at concentrations of I : 20 000 to 1: 8000. It may be n e e c e w to draw
some anticoagulant into the syringe before w e . Antimagulanta that
have boen auccessful include 80diUm citrate, haperin and Alaever’s
Boluticm (of. Campbell et ol. 1063).
Rsh blood C ~ R
&Q
be obtained from the caudal artery of aneathetized fish by levering the taiI and holding the cut surface in a v e d
containing anticoagulant. Care ahould be taken to clean the tail of
BH0888 mucus. This method is particularly useful with amdl fish.