Progress of Lens Biochemistry Research: Volume in honour of Prof. Dr. med. J. Nordmann

306 Pages·1976·10.78 MB·English

by  A. Brini, M. E. Stoeckel, A. Porte, J. Klethi (auth.), Otto Hockwin (eds.)

Progress of Lens Biochemistry Research
Documenta Ophthalmologica
Proceedings Series volume 8
Editor H. E. Henkes
Dr. W. Junk bv Publishers The Hague 1976
Progress of Lens Biochemistry Research
Volume in honour of
Prof. Dr. med. J. Nordmann
edited by O. Hockwin
Dr. W. Junk bv Publishers The Hague 1976
© Dr W. Junk b.v. Publishers 1976
No part of this book may be reproduced and/or published in any form, by print,
photoprint, microfilm or any other means without written permission from the
publishers.
ISBN-13: 978-90-6193-148-5 e-ISBN-13: 978-94-010-1571-4
001: 10.1007/978-94-010-1571-4
CONTENTS
O. HOCKWIN (Bonn): Preface . . . . . . . . . . .. VII
A. BRINI, M~E. STOECKEL, A. POR TE & J. KLETHI (Stras-
bourg): Intra-Nuclear inclusions in the lens fibres observed in
the galactose-induced cataract in the young rat. . . . . .
T. KUW ABARA & S.OKISAKAA (Bethesda, Boston): Electron
microscopic study of cataractous lenses of diabetic sand rats
(Psammomys obesus) . . . . . . . . . . . . .. 7
D.E.S. TRUMAN, RUTH M. CLAYTON, A.G. GILLIES & H.I.
MACKENZIE (Edinburgh): RNA synthesis in the lenses of
normal chicks and in two strains of chicks with hyperplasia of
the lens epithelium . . . . . . . .. 17
RUTH M. CLA YTON, D.E.S. TRUMAN, J. HUNTER, P.G. ODEI-
GAH & D.1. DE POMERAI (Edinburgh): Protein synthesis and
its regulation in the lenses of normal chicks and in two strains
of chicks with hyperplasia of the lens epithelium . . . .. 27
J. FRANCOIS & V. VICTORIA-TRONCOSO (Ghent): Epithelium
of the adult lens in tissue culture . . . . . . . . .. 39
M. IWIG & D. GLASSER (Halle): On the stability of the differen-
tiated state - Reactivation of lens fiber cells to mitotic growth 47
R.C. BRIGGS, N. WAINWRIGHT & H. ROTHSTEIN (Burling-
ton): On the chromosomal proteins of lens epithelium 57
J.R. REDDAN, N.J. UNAKAR, EV AMARIE KRASICKY & DO-
ROTHY WILSON (Rochester): Triggering of mitosis in the
cultured mammalian lens by proteolytic enzymes . . . .. 67
J. BOURS, KARIN DOEPFMER & O. HOCKWIN (Bonn): Iso-
electric focusing of crystallins from different parts of the
bovine and dog lens in dependence on age . . . .. 75
H. BLOEMENDAL & ANNEKE ZWEERS (Nijmegen): Improved
separation of LM (Low molecular weight) crystallins. . .. 91
H.J. HOENDERS (Nijmegen): Changes in the subunit structure of
a-crystallin from bovine and rabbit eye lenses 105
K. SATOH & M. BANDO (Chiba/Tokyo): A comparative study of
rabbit lens proteins . . . . . . . . . . . . . .. 113
H. MAISEL, J. ALCALA & N. LIESKA (Detroit): The protein
structure of chick lens fiber cell membranes and intracellular
matrix .. ................ 121
A.A. SWANSON, U. HAHN & O. HOCKWIN (Charleston/Bonn):
Investigation of proteolytic activity in bovine lenses with age I.
The exopeptidases . . . . . .. .... 135
W.B. RA THB UN (Minneapolis): Lens glutathione: Metabolism
and possible functions . . . . . .. 145
V.N. REDDY & R.F. HAN (Rochester): Protein-bound gluta-
thione in mammalian lenses and in galactose cataract. . .. 153
H.N. FUKUI, L.O. MEROLA & J.H. KINOSHITA (Bethesda):
The effect of oxidants on the membrane sulfhydryl groups of
the lens. . . . . . . . . .. 161
V
K.l. DILLEY & RUTH VAN HEYNINGEN (Oxford): Some
aspects of human lens metabolism: Glycolysis and protein syn-
thesis . . . . . . 171
C. OHRLOFF, F. BOUS & O. HOCKWIN (Bonn): Competition
for glucose-6-phosphate by enzymes of bovine lenses. . . 177
H.L. KERN, S.A. OSTROVE & C.-K. HO (Bronx): Effect of aging
on transport of amino acids by bovine lens . . . . . .. 185
Y. OBARA, E. COTLlER, R. LINDBERG & 1. HORN (Chicago):
Cholesterol, cholesterol ester, and sphingomyelin complexed
to protein of normal human lens and senile cataracts . 193
INGE KORTF, O. HOCKWIN, F. WINKLER & P. RABE (Bonn):
Studies on the mode of action of I-hydroxy-pyrido-(3,2a)-5-
phenoxazone-3-carboxylic acid in bovine lenses 205
1. KLETHI (Strasbourg): Sodium and potassium in the normal
human lens in relation with age . . . . . . . 213
J.W. PA TTER SON (Farmington): Cell volume regulation in the
lens . . . . . 219
G. DUNCAN & RUTH V AN HEYNINGEN (Oxford): Differences
in the calcium binding capacity of normal and cataractous
lenses . . . .. 229
J.L. RAE & T.R. ST ACEY (Galveston): Intracellular markers in
the crystalline lens of the rat. . . . . . . . . . 233
S. LERMAN (Atlanta): Lens fluorescence in aging and cataract
formation . . . . . . . . . .. 241
J.F.R. KUCK jr. (Atlanta): Effect of long-wave ultraviolet light
on the lens. II. Metabolic inhibitors synergistic with UV in
vitro . . . . . 261
S. ZIGMAN (Rochester): Tryptophan excited states in the lens 267
W. MANSKI, A.T. YORK, G.R. MERRIAM Jr., B.V. WORGUL &
A. SZECHTER (New York): On the independent action of
cataractogenic agents x-rays and galactose 275
H.-R. KOCH, KATRIN DOLDI & O. HOCKWIN (Bonn): Naph-
thalene cataracts in rats. Association of eye pigmentation and
cataract development . . . .. .... 293
S.D. VARMA & J.H. KINOSHITA (Bethesda): Topical treatment
of galactose cataracts . . .. 305
V.E. KINSEY (Rochester): Studies on the Crystalline lens.
XXIV. Bicarbonate content and flux determinations . .. 310
VI
PREFACE
On August 25,1976 Prof. Dr. med. JEAN NORDMANN, Emeritus Professor
of Strasbourg University, will celebrate his 80th anniversary.
The task of paying tribute to Professor NORDMANN's many services to
ophthalmology in a way worthy of the occasion is so difficult that we decided
to concentrate on the focal point of his life's work.
In this issue scientists engaged in lens research from allover the world
offer reports of their latest findings together with their best wishes and
felicitations as a birthday gift to the Nestor of lens research. They thereby
express their gratitude to Professor NORDMANN, who, with the lifelong in-
defatigable enthusiasm of a scientist , has most successfully persued the prob-
lems of lens metabolism.
It is our sincerest hope that we may look forward to many more years in
which to take advantage of Professor NORDMANN's superior knowledge.
Among the contributions to this festive donation we miss the works of
some authors who have always held Professor NORDMANN 's special inter-
est. The editor has received several letters of longstanding friends who would
have liked to submit a manuscript but had been unable to meet the deadline:
Professor NORDMANN may rest assured that they, too , belong to the great
number of congratulators from our field of research.
We have been able to make this dedication thanks to Professor HENKES
who readily agreed to publish our contributions in Documenta Ophthal-
mologica.
Dr. Junk publishers have done their utmost to ensure prompt publication.
OTTO HOCKWlN, Bonn
VII
INTRA-NUCLEAR INCLUSIONS IN THE LENS FIBRES OBSERVED IN
THE GALACTOSE-INDUCED CATARACT IN THE YOUNG RAT
A. BRINI, M.E. STOECKEL, A. PORTE & J. KLETHI
(Strasbourg, France)
ABSTRACT
Electron microscopy of young rat lenses with early changes due to galactose diet point
to an early alteration of the nucleo-plasmatic relationship. Peculiar alterations of the
nuclei are seen in the lens fibres during their maturation.
The galactose-induced cataract can be provoked within 3 to 6 days in the
young rat or rabbit, simply with a galactose diet. The early lesions are
localized along the equator; they are detected with the slit lamp, after
dilatation of the pupil, and usually form a crown of superficial vesicles and
of flat opacities. The latter are proximal to the vesicles and appear later. The
ultrastructure of these lesions is well known (BR INI et al., 1961, 1963;
KUW AB ARA et al., 1969). The vesicle-like appearance is due to a dilatation
of the intercellular spaces (KUWABARA et al., 1969) preceding swelling
and necrosis of fibres. This picture is evidence of an early osmotic alteration
which is generally thought to be related to the accumulation of gala.ctitol -
a result of reduction of abnormally stored galactose - in'the lens (KINO-
SHIT A et al., 1962). This explanation does not account, however, for the
precise localization of the lesions in the fibres at a certain stage of differen-
tiation. The diversified metabolic alterations which were described in several
tissues during human galactosemia and in galactose-fed animals also suggest
a more complex pathogenesis (see SEGAL, 1972).
According to our electron microscopic observations on the young rat on
a galactose diet, an early alteration of the nucleo-plasmatic relationship is
indicated: e.g., peculiar alterations of the nuclei are seen in the lens fibres
during their maturation.
MA TERIAL AND METHODS
Twelve Wistar rats were put on a 50% galactose diet soon after weaning.
Lens alterations were detected with the slit lamp. The lenses were removed
between the third day (when the first lesions were observed) and the tenth
day. They were cut meridionally under fixative (glutaraldehyde 5%, phos-
phate buffer 0.1 M, pH 7.4). Post-fixation with osmic acid was performed in
some cases. The tissue was embedded in araldite-epon. Semi-thin sections
were controlled with light microscopy after toluidine blue, hemalun-eosin or
Feulgen staining. Ultra-thin sections were treated with uranyl-acetate and
lead citrate and studied with a Siemens Elmiskop IA.
Fig. 1. 3 days galactose diet. Semi-thin sagittal section, Hemalun-eosine stained. Equa-
torial vesicular area. X 80.
Fig. 2. 9 days galactose diet. Semi-thin section, hemalun-eosin stained.
A. Same orientation as in fig. I. More advanced stage of cataract (tear in the necrotic
area is an artefact). The nuclei which contain inclusions are proximal to the lesion.
X 80.
B. Detail of fig. 2A (framed area) showing poorly stained spherical intranuclear inclu-
sions. X 500.
Fig. 3. 3 days galactose diet. Equatorial area. Top: superficial layers with normal
appearance : bottom: 'vesicles' by dilatation of the intercellular spaces. X 5,750.
2
RESULTS
On light microscopy control sections, the lesions first appear as an area of
clear vesicles strictly localized along the equator (Fig. I). Electron micro-
scopy shows that these 'vesicles' are actually dilatations of the intercellular
spaces (Fig. 3) at the level of the young fibres, which are still interlinked by
desmosomes and tight junctions, most of the latter being of the gap junction
or nexus-type (PO R TE et aI., 1975). These alterations are not observed in
the epithelium or at a distance from the equator. In the more differentiated
fibres, which are proximal to the vesicle area, only nuclear altelations, as yet
irregular, are observed. Later, the vesicle area presents a downright degener-
ative appearance, with swelling and vacuolization of the fibres and disap-
pearance of the cellular outlines. The alterations of the nuclei in the fibres
beneath the lesions are then obvious and constant (Fig. 2).
They appear as spherical inclusions, which stain slightly with eosin, and
are Feulgen-negative. Several inclusions may be present in the same nucleus
(Fig. 4). With electron microscope (Figs. 3-6), these inclusions appear de-
void of chromatin and constituted of very fine fibrillary material obviously
different from the much coarser material of the fibre cytoplasm. They are
surrounded by chromatin, have no limiting membranes and cannot be con-
sidered as cytoplasmic invaginations. They are independent of. the nucleoles,
which normally appear diversified in this area (fragmentation and mottled
appearance, spherical or annular condensation). Besides the typical inclu-
sions, all the nuclei of the fibres in this part of the lens exhibit diffuse areas
where granular chromatin becomes scarce, and within which fine fibrillar
material accumulates.
Fig. 4. 6 days galactose diet. Nucleus with 3 inclusions. X 7,200.
3