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The species-specificity of lens crystallins
ABSTRACTS
302
camera run-up, variable delay between electromagnetically controlled (Iye injection and onset of event, and a variable duration of filming. Fluorescein angiograms are recorded on Kodak R;1R 2498 film using Baird At.omic. l5.l anti infrared absorption angiograms on lCodak lL.4 K %Sl \Vratt,en 1.5 filters; Indocyanine green. film using a Wratten 88A filter. Films arc commerriallp processed.
The Technique
of the Measurement
J. -4. KEIWARD, J.-C’. (!ORNIC. Y. ~~~OTTON AXD J. (CHEVALIER, Prr?'is
The technique using a very results are shown on different demonstrat,ed.
The Crystallin .J. Horr~.
Distribution
.\. \\:I:BP.I~
of the Tensile Cornea1 Wound Strength (exhibit) POI~IJQI~F.~,
precise curves.
P.
l’.i~n
$11. H.
tlynamometer (lnstrom The effect,s of a steroid.
H.sN.\I*II.
dynamomet~er) and the silk
.I. .JA~Y)~.Lu
11:~. 11
is desc,ri bc~l. ‘I‘trl. or nylon sutrln+ ar(’
of the Bovine Lens During Aging (exhibit)
AX,, 0. HOCKWIX.
f107(tl
and quant itiltlvt*l!, The cryst,allin composition of the bovine Icns during agin, or is qualitatively measured by thin-layer isoelectric focusing (Bours, 1971). We used bovine lenses ranging in ag(’ from 0.2 up to approximately 2.5 years. Besides this. also thin-layer isolectric focusing is rnatk of bovine lenses. separated in equator and nnc~leus. Due to the high resolving power of this t,echnique, the c*rystallina c~oultf t)e ~sofbc~~~sr~tl ill it number of components, e.g. one /3-crystallin. 12-14 /3-eryst,allin components and 4-C y-wyrtallin component,s. During aging, the x-crystallin shows a slight inclreasc. the ,8-crystallin components 01 1owr.t isoelectric points show a fast increase during the first five years of life. while the p-rrvst;allirr components of higher isoelectric point,s remain constant dnrinp the aging process. The first and t&d y-crystallin components a.re prominent. throughout aging, especially 111 t!lv embryonic and in the young bovine lens. The first component shows a rapid increase in the first years after birth and a gradual diminishment later to a constant level. The third componrnt shows a rapid decrease in the first years after birth and a gradual diminishment later to a (.ot&ant level. The second and fourth y-cryst,allin camponents show a slight decrease during agitkg. In the embryonic lens, a considerable increase in number and concentration of t,he P-l*rystallltt components of higher isoelectric points is found, while the CX- and y-crystallins show i-1~ saru~~ appearance (Hours and Brahma. 197.3). Regarding the y-crystallin composition. the lens equator shows an csntirc1.v diffcrcnl I~YI~ IIW and nature compared to the lens nucleus (Hours, 1973, 1975).
Hours, Bours, Bours,
J. (1971). J. C”h.romntoyr. 60, 235. J. (1973). E~J. Eye Res. 16, 501. J. (1975). Isoelect,ric focusing of lens crysti;st;tllins. I II Pro~rr.~ I.sotuchophoresis (Ed. Righetti. P. (+.). Elsevler-Excerpta Medim. Bours, J. and Brahma, S. Ii. (1973). l3.r~. Bye IZes. 16, 131.
The Species-specificity A.
~I'EBER,
J.
Bot!rts
of Lens Crystallins ANU 0. HOUKWIK.
i/l isoelectric Fo~usirr!/ II)& North Holland (in ~WSS).
(exhibit)
fiorz,r
Lenx crystallins of various vertebrate species are su bjectetl to thill-layer isoelec+ric ti)vlcsirlg (Bours, 1971). A new classification of species may be defined acc.ording to the iso&+ril, points determined of single crystallin components. A quantification is made by evaluation of densit,ometric measurements of’ st~ainetl isotil(~lls~tl bands. e.g. of a-crystallin and of the single components /3- and y-crystallins following their koelectric points. In this way the tot.al water soluhle lens rxtraets are c.ompared of a numt)t:r ot mammalian species, e.g. man, bovine, pig. rat,, mouse, guinea pig, rabbit, beagle dog, shepherd’s dog and Miinsterliinder dog. For further comparison, lens crystallinn of chicken (Bours. 1974) and fish are used as references. In contrast to the immunologic similarities bet,wcen many vertebrate sprcies (Manski, L\ucrl,a~h and Halbert, 1960), almost no similarity in position of isofocused bands is observed. Even within one race (beagle dog, shepherd’s dog and Miinsterlhnder dog) and between species of t,hr same branch of the evolutionary tree. e.g. in Amphibia (Brahma and McDevitt,, 1974). considerable differences in the number of single @- and y-crystallin components, their isoelectric points and their concentrations are observed.
ABSTR,ACTG
JO3
in relation to experimental cataract.s, records obtained may be of importance in comparison the high species-specificity of lens crystallins in mammals. A marked difference in behaviour each species towards the exposure of combined noxious influences is observed (Hockwin Koch, 1974).
to of and
REFERENCES Bours, J. (1971). J. Chromatogr. 60, 2%. Bours, J. (1974). Docum. Ophthdmol. 37, 1. Brahma, S. K. and McDevitt, D. S. (1974). Exp. Eye lies. 19,379. Horkwin, 0. and Koch, H. R. (1974). Combined noxious influences. In Cataract and Abnor,&ities of the Lens. (Ed. Bellows, J. G.). P. 243. Grune and Stratton, Inc., New York. Manski, IV., Auerbach, T. P. and Halbert, S. P. (1960). Amer. J. Ophthalmol. 50, 985.
A Study of the Change of Proteolytic the Life Span (exhibit) U. H.ra~,
A. d. SWANSON
AND
Enzyme Activity in Bovine Lenses During the Course of
0. HOCKWIR,
Bonn
a&
Charleston
The enzymatic activities of leucine aminopeptidase (E.C.3.4.1.1), e&erase, triglycinpeptidase (E.C.3.4.1.3), protease (E.C.3.4.4.16), and a proteinase were studied. The results show that proteolytic activity can be demonstrated in mammalian lenses and that there is a profound change of activity during the course of the life span. Leucine aminopeptidase, e&erase and triglycinpeptidase show a significant increase in activity after fifteen years of age. The activity of the protease and proteinase increases continuously with increasing age. Most of the activity for the five different prokolytic enzymes was detected in the equator and anterior shell of the lens tissue, whereas lower act.ivity could be demonstrated in the nucleus and posterior shell.
302
camera run-up, variable delay between electromagnetically controlled (Iye injection and onset of event, and a variable duration of filming. Fluorescein angiograms are recorded on Kodak R;1R 2498 film using Baird At.omic. l5.l anti infrared absorption angiograms on lCodak lL.4 K %Sl \Vratt,en 1.5 filters; Indocyanine green. film using a Wratten 88A filter. Films arc commerriallp processed.
The Technique
of the Measurement
J. -4. KEIWARD, J.-C’. (!ORNIC. Y. ~~~OTTON AXD J. (CHEVALIER, Prr?'is
The technique using a very results are shown on different demonstrat,ed.
The Crystallin .J. Horr~.
Distribution
.\. \\:I:BP.I~
of the Tensile Cornea1 Wound Strength (exhibit) POI~IJQI~F.~,
precise curves.
P.
l’.i~n
$11. H.
tlynamometer (lnstrom The effect,s of a steroid.
H.sN.\I*II.
dynamomet~er) and the silk
.I. .JA~Y)~.Lu
11:~. 11
is desc,ri bc~l. ‘I‘trl. or nylon sutrln+ ar(’
of the Bovine Lens During Aging (exhibit)
AX,, 0. HOCKWIX.
f107(tl
and quant itiltlvt*l!, The cryst,allin composition of the bovine Icns during agin, or is qualitatively measured by thin-layer isoelectric focusing (Bours, 1971). We used bovine lenses ranging in ag(’ from 0.2 up to approximately 2.5 years. Besides this. also thin-layer isolectric focusing is rnatk of bovine lenses. separated in equator and nnc~leus. Due to the high resolving power of this t,echnique, the c*rystallina c~oultf t)e ~sofbc~~~sr~tl ill it number of components, e.g. one /3-crystallin. 12-14 /3-eryst,allin components and 4-C y-wyrtallin component,s. During aging, the x-crystallin shows a slight inclreasc. the ,8-crystallin components 01 1owr.t isoelectric points show a fast increase during the first five years of life. while the p-rrvst;allirr components of higher isoelectric point,s remain constant dnrinp the aging process. The first and t&d y-crystallin components a.re prominent. throughout aging, especially 111 t!lv embryonic and in the young bovine lens. The first component shows a rapid increase in the first years after birth and a gradual diminishment later to a constant level. The third componrnt shows a rapid decrease in the first years after birth and a gradual diminishment later to a (.ot&ant level. The second and fourth y-cryst,allin camponents show a slight decrease during agitkg. In the embryonic lens, a considerable increase in number and concentration of t,he P-l*rystallltt components of higher isoelectric points is found, while the CX- and y-crystallins show i-1~ saru~~ appearance (Hours and Brahma. 197.3). Regarding the y-crystallin composition. the lens equator shows an csntirc1.v diffcrcnl I~YI~ IIW and nature compared to the lens nucleus (Hours, 1973, 1975).
Hours, Bours, Bours,
J. (1971). J. C”h.romntoyr. 60, 235. J. (1973). E~J. Eye Res. 16, 501. J. (1975). Isoelect,ric focusing of lens crysti;st;tllins. I II Pro~rr.~ I.sotuchophoresis (Ed. Righetti. P. (+.). Elsevler-Excerpta Medim. Bours, J. and Brahma, S. Ii. (1973). l3.r~. Bye IZes. 16, 131.
The Species-specificity A.
~I'EBER,
J.
Bot!rts
of Lens Crystallins ANU 0. HOUKWIK.
i/l isoelectric Fo~usirr!/ II)& North Holland (in ~WSS).
(exhibit)
fiorz,r
Lenx crystallins of various vertebrate species are su bjectetl to thill-layer isoelec+ric ti)vlcsirlg (Bours, 1971). A new classification of species may be defined acc.ording to the iso&+ril, points determined of single crystallin components. A quantification is made by evaluation of densit,ometric measurements of’ st~ainetl isotil(~lls~tl bands. e.g. of a-crystallin and of the single components /3- and y-crystallins following their koelectric points. In this way the tot.al water soluhle lens rxtraets are c.ompared of a numt)t:r ot mammalian species, e.g. man, bovine, pig. rat,, mouse, guinea pig, rabbit, beagle dog, shepherd’s dog and Miinsterliinder dog. For further comparison, lens crystallinn of chicken (Bours. 1974) and fish are used as references. In contrast to the immunologic similarities bet,wcen many vertebrate sprcies (Manski, L\ucrl,a~h and Halbert, 1960), almost no similarity in position of isofocused bands is observed. Even within one race (beagle dog, shepherd’s dog and Miinsterlhnder dog) and between species of t,hr same branch of the evolutionary tree. e.g. in Amphibia (Brahma and McDevitt,, 1974). considerable differences in the number of single @- and y-crystallin components, their isoelectric points and their concentrations are observed.
ABSTR,ACTG
JO3
in relation to experimental cataract.s, records obtained may be of importance in comparison the high species-specificity of lens crystallins in mammals. A marked difference in behaviour each species towards the exposure of combined noxious influences is observed (Hockwin Koch, 1974).
to of and
REFERENCES Bours, J. (1971). J. Chromatogr. 60, 2%. Bours, J. (1974). Docum. Ophthdmol. 37, 1. Brahma, S. K. and McDevitt, D. S. (1974). Exp. Eye lies. 19,379. Horkwin, 0. and Koch, H. R. (1974). Combined noxious influences. In Cataract and Abnor,&ities of the Lens. (Ed. Bellows, J. G.). P. 243. Grune and Stratton, Inc., New York. Manski, IV., Auerbach, T. P. and Halbert, S. P. (1960). Amer. J. Ophthalmol. 50, 985.
A Study of the Change of Proteolytic the Life Span (exhibit) U. H.ra~,
A. d. SWANSON
AND
Enzyme Activity in Bovine Lenses During the Course of
0. HOCKWIR,
Bonn
a&
Charleston
The enzymatic activities of leucine aminopeptidase (E.C.3.4.1.1), e&erase, triglycinpeptidase (E.C.3.4.1.3), protease (E.C.3.4.4.16), and a proteinase were studied. The results show that proteolytic activity can be demonstrated in mammalian lenses and that there is a profound change of activity during the course of the life span. Leucine aminopeptidase, e&erase and triglycinpeptidase show a significant increase in activity after fifteen years of age. The activity of the protease and proteinase increases continuously with increasing age. Most of the activity for the five different prokolytic enzymes was detected in the equator and anterior shell of the lens tissue, whereas lower act.ivity could be demonstrated in the nucleus and posterior shell.