I. 8, No. 9, September 1984
9N 0309-1651
-ELL BIOLOGY
nternational Report:
TBR
i th beGe
\ ( A () : ae
—_ LIBRARY.
iblished for the
TERNATIONAL FEDERATION FOR CELL BIOLOGY
ACADEMIC PRESS
ndon Orlando SanDiego SanFrancisco New York
‘onto Montreal Sydney Tokyo Sao Paulo
,meee e
eee ee ee en ——
Editor
L. M. Franks Department of Cellular Pathology, Imperial Cancer Research Fund, Lincoln’s
Inn Fields, London WC2A 3PX, U.K.
Deputy Editor
B. M. Richards Searle Research Laboratories, High Wycombe, Bucks., U.K.
Editorial Board
J. André Laboratoire de Biologie Cellulaire, Universite de Paris-Sud, Orsay, France.
Dz. Baintgn Department of Pathology, University of California School of Medicine, San
Francisco, U.S.A.
M. Balls University of Nottingham, Nottingham, U.K.
E. A. Barnard Department of Biochemistry, Imperial College, London, U.K.
R. Baserga Temple University School of Medicine, Health Sciences Center, Philadelphia, U.S.A.
O. Behnke Department of Anatomy, University of Copenhagen, Copenhagen, Denmark.
E. L. Benedetti Institut de Biologie Moleculaire, Universite Paris Vil, Paris, France.
M. de Brabander Laboratory of Oncology, Janssen Pharmaceutica, Beerse, Belgium.
G. Cotta-Pereira Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil.
V. J. Cristofalo The Wistar Institute, Philadelphia, U.S.A.
W. W. Franke Institute of Experimental Pathology, Heidelberg, Federal Republic of Germany.
R. Garrett Biostructural Chemistry, Chemistry Department, University of Aarhus, Denmark.
L. A. Ginsel Laboratory for Electron Microscopy, Leiden, The Netherlands.
G. Giudice Instituto di Anatomia Comparata, Universita di Palermo, Palermo, Italy.
D. O. Hall Department of Plant Sciences, King’s College, London, U.K.
S. Inoue Marine Biological Laboratory, Woods Hole, Massachusetts, U.S.A.
L. Jimenez de Asua Department of Biochemistry, St George’s Hospital Medical School, London, U.K.
A. Kane Division of Biology and Medicine, Brown University, Providence, U.S.A.
C. J. Marshall Institute of Cancer Research, Chester Beatty Research Institute, London, U.K.
D. H. Northcote Department of Plant Biochemistry, University of Cambridge, Cambridge, U.K.
Y. Okada Research Institute for Microbial Disease, Osaka University, Japan.
M. L. Pardue Department of Biology, Massachusetts /nstitute of Technology, Cambridge, U.S.A.
H. Passow Max Planck Institut fur Biophysik, Frankfurt am Main, Federal Republic of Germany.
R. Ross Department of Pathology, University of Washington, Seattle, U.S.A.
W. Sachsenmaier Institut fur Biochemie und Experimentel/e Krebsforschung, Universitat Innsbruck,
Innsbruck, Austria.
P. Satir Department of Anatomy, Albert Einstein College of Medicine of Yeshiva University,
1300 Morris Park Avenue, Bronx, New York, U.S.A.
H. G. Schweiger Max Planck Institut fur Zel/biologie, Ladenburg bei Heide/berg, Federal Republic
of Germany.
S. Segal Department of Immunology and Microbiology, Ben Gurion University of the Negev,
Beer Sheva, /srael.
A. S. Spirin Institute of Protein Research, Academy of Science of U.S.S.R., Moscow, U.S.S.R.
J. R. Tata National Institute of Medical Research, London, U.K.
A. Vaheri Department of Virology, University of Helsinki, Finland.
D. F. H. Wallach Division of Radiobiology, Tufts-New England Medical Center, Boston, U.S.A.
E. R. Weibel Anatomisches Institut der Universitat Bern, Bern, Switzerland.
R. Williamson Department of Biochemistry, St Mary’s Hospital, Medical School, London, U.K.
R. E. Williamson Department of Developmental Biology, Australian National University, Canberra,
Australia.
K, E. Wohlfarth Institut fur Cytologie, Universitat Bonn, Bonn, Federal Republic of Germany.
-Bottermann
Copyright © 1984 Academic Press Inc. (London) Ltd. The appearance of the code at the bottom of the
first page of a paper in the journal indicates the copyright owner's consent that copies of the paper may
be made for personal or internal use, or for the personal or internal use of specific clients in the U.S.A.
This consent is given on the condition, within the U.S.A., that the copier pay the stated per-copy fee
through the Copyright Clearance Center,.Inc., 21 Congress Street, Salem, MA 01970, U.S.A. for copying
beyond that permitted by Sections 107 or 108 of the U.S. Copyright Law. This consent does not extend
to other kinds of copying, such as copying for general distribution, for advertising or promotional
purposes, for creating new collective works, for resale or for copying or distributing copies outside the
U.S.A.
Published monthly at 24-28 Oval Road, London NW1, England by Academic Press, Inc. (London)
Limited: Volume 8, inland £66.00 inclusive of postage; abroad $175.00 inclusive of postage.
Subscription orders should be sent to Academic Press Inc. (London) Limited, High Street, Footscray,
Sidcup, Kent DA14 5HP (tel: 01-300-0155), except those Originating in the U.S.A., Canada, and Central
and South America which should be sent to Academic Press, Inc. 111 Fifth Avenue, New York,
NY
10003, U.S.A. Send notices of changes of address to the office of the Publishers at least 6-8 weeks
In
advance. Please include both old and new addresses. Postmaster, send change of address to Cell
Biology International Reports, 111 Fifth Avenue, New York, NY 10003, U.S.A.
saat and mailing in the U.S.A. by Publications Expediting Inc., 200 Meacham Avenue, Elmont,
NY
Second class postage paid at Jamaica, New York 11431, U.S.A.
,Cell Biology International Reports, Vol. 8, No. 9, September 1984 703
Mini-review: Hyaluronidases in early embryonic development
Claudio D. STERN, Department of Anatomy, Downing Street, Cambridge
CB2 3DY, England
In this paper I shall summarise a selection of the most relevant
recent studies which indicate an involvement of the
extracellular matrix and hyaluronidases in the control of
developmental processes in the early embryo. I shall be
concerned principally with processes taking place during
morphogenesis. By this term a shall mean the cellular
rearrangements which involve more or less large fields of the
embryo. To a lesser extent I shall also discuss the relationship
between these morphogenetic processes and cytodifferentiation, a
term which I shall use to refer to the relatively irreversible
change of an individual cell into a "typical" differentiated
member of a histologically, functionally and molecularly
distinct tissue.
Unfortunately, only a few direct investigations of the presence,
dvstribucton- and Wactivitity or hyalwronidases’ in vearly embryos
are available in the literature. Most of the studies to date are
mainly concerned with the presence and characterization of
extracellular materials, and with the incorporation Or
radioactive precursors into these, but very few indeed have
embarked ‘onto a detailed analysis of ‘the turnover of these
compounds during the early stages of embryonic development. Much
of the evidence I shall be considering will perforce be
indirect.
Before embarking on this discussion, it is important to consider
what hyaluronidase is. Far from being a single enzyme, the name
iso "applied’” to Yany -enzyme activity which cleaves the glycosidic
bonds of hyaluronic acid (Merck Index, 4th ed, 1976). The’ Enzyme
Commission currently recognizes at least four distinct enzymes
with "hyaluronidase" activity: sD Gory Se Zh ull soy Baers: 2a. 36
and EUG). 4.529 9141 (Florkin & SiEOuZ , (973
BOGIAA202 041
the existence of many different hyaluronidases has
Furthermore,
demonstrate d aise! many systems. there are “dri tenent
been
in the adult frog from those of the tadpole
hyaluronidases
pye197"); and different activities associated with
Garpson Viet al
from those in the iMmeer Ol Oil Ene ™ same
fibroblast cell surfaces
(Orkin & Toole, 1980a,b). Antibodies raised against rat
cells
hyaluronidase do not Cuoss—Theace with
urinary (renal)
testicular origin and vice-versa (Law & Rowen,
hyaluronidase of
tail fin-derived hyaluronidase shows
1978, 1979, 1981). Tadpole
for hyaluronate aS a substrate over chondroitin
a preference
© 1984 Academic Press Inc. (London) Ltd.
0309-165 1/84/090703-15/$03.00/0
, 704 Cell Biology International Reports, Vol. 8, No. 9, September 1984
4-sulphate, and is unable to degrade chondroitin 6-sulphate or
heparan. Testicular hyaluronidas e, on the other hand, can cleave
both forms of chondroitin sulphate as well as hyaluronate, and
bacterial hyaluronidase cleaves chondroitin sulphate in
preference to hyaluronate (Silbert & DeLuca, 19.7 O)iz-iin the
absence One hydrolysing enzymes, hyaluronate and chondroitin
sulphates are relatively stable in embryos (Derby & Pintar,
TERE,
With these provisos, we can now begin a discussion on the
involvement of hyaluronidases in the regulation of early
developmental processes in the vertebrate embryo.
1. Hyaluronidase activity during morphogenesis
(a) Non-malignant invasion in morphogenesis
of particular interest to» the, clinician sas swell as) tthe
embryologist is the problem of invasion. For the oncologist,
this phenomenon is associated with metastasis and the spatial
spread of malignant tissues into surrounding areas. For the
embryologist, non-malignant invasion 1s associated with normal
morphogenetic events at various times in the development of
embryos, when one tissue displaces another (see Mareel, 1979 and
Vakactcet ab, M980 sfor suecent y yevalewsie Them bese Mexanplesmmo.
this process in embryonic development are perhaps the
implantation of the mammalian blastocyst into the maternal
endometrium, and the process of the formation of the definitive
(gut) endoderm (Stern & Ireland, 1981; \Bellarrsm et tral TeiocioEe
where the ectodermally-derived endoderm tissue displaces and
inserts into the existing endodermal layer (hypoblast). Other
examples include the migration of neural crest cells, initially
into a relatively cell-free but glycosaminoglycan-rich matrix,
and later some of them may move into the mass of mesenchymal
cells forming the somites (Noden, 1978). At the level of single
celiis>, Sfiertilazvation® “itself scouldip bewiconsidercdNasctan einvasiiva
process, where the egg is "invaded" by the sperm.
As tissues and cells are frequently surrounded by extracellular
materials such as basal laminae containing glycosaminoglycans
and proteoglycans, invasive ability, whether malignant or not,
should be accompanied by an ability of the invading cells or
tissues to destroy this Mal Genes Thus one would expect
hyaluronidase activity to be high in invading tissues in cancer
and in non-malignant invasion during embryonic development.
Experimental evidence confirms thass Kolaova (19577) has
demonstrated high levels of hyaluronidase activity in patients
with malignant tumours (see also Coman, 1953 and references
9N 0309-1651
-ELL BIOLOGY
nternational Report:
TBR
i th beGe
\ ( A () : ae
—_ LIBRARY.
iblished for the
TERNATIONAL FEDERATION FOR CELL BIOLOGY
ACADEMIC PRESS
ndon Orlando SanDiego SanFrancisco New York
‘onto Montreal Sydney Tokyo Sao Paulo
,meee e
eee ee ee en ——
Editor
L. M. Franks Department of Cellular Pathology, Imperial Cancer Research Fund, Lincoln’s
Inn Fields, London WC2A 3PX, U.K.
Deputy Editor
B. M. Richards Searle Research Laboratories, High Wycombe, Bucks., U.K.
Editorial Board
J. André Laboratoire de Biologie Cellulaire, Universite de Paris-Sud, Orsay, France.
Dz. Baintgn Department of Pathology, University of California School of Medicine, San
Francisco, U.S.A.
M. Balls University of Nottingham, Nottingham, U.K.
E. A. Barnard Department of Biochemistry, Imperial College, London, U.K.
R. Baserga Temple University School of Medicine, Health Sciences Center, Philadelphia, U.S.A.
O. Behnke Department of Anatomy, University of Copenhagen, Copenhagen, Denmark.
E. L. Benedetti Institut de Biologie Moleculaire, Universite Paris Vil, Paris, France.
M. de Brabander Laboratory of Oncology, Janssen Pharmaceutica, Beerse, Belgium.
G. Cotta-Pereira Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil.
V. J. Cristofalo The Wistar Institute, Philadelphia, U.S.A.
W. W. Franke Institute of Experimental Pathology, Heidelberg, Federal Republic of Germany.
R. Garrett Biostructural Chemistry, Chemistry Department, University of Aarhus, Denmark.
L. A. Ginsel Laboratory for Electron Microscopy, Leiden, The Netherlands.
G. Giudice Instituto di Anatomia Comparata, Universita di Palermo, Palermo, Italy.
D. O. Hall Department of Plant Sciences, King’s College, London, U.K.
S. Inoue Marine Biological Laboratory, Woods Hole, Massachusetts, U.S.A.
L. Jimenez de Asua Department of Biochemistry, St George’s Hospital Medical School, London, U.K.
A. Kane Division of Biology and Medicine, Brown University, Providence, U.S.A.
C. J. Marshall Institute of Cancer Research, Chester Beatty Research Institute, London, U.K.
D. H. Northcote Department of Plant Biochemistry, University of Cambridge, Cambridge, U.K.
Y. Okada Research Institute for Microbial Disease, Osaka University, Japan.
M. L. Pardue Department of Biology, Massachusetts /nstitute of Technology, Cambridge, U.S.A.
H. Passow Max Planck Institut fur Biophysik, Frankfurt am Main, Federal Republic of Germany.
R. Ross Department of Pathology, University of Washington, Seattle, U.S.A.
W. Sachsenmaier Institut fur Biochemie und Experimentel/e Krebsforschung, Universitat Innsbruck,
Innsbruck, Austria.
P. Satir Department of Anatomy, Albert Einstein College of Medicine of Yeshiva University,
1300 Morris Park Avenue, Bronx, New York, U.S.A.
H. G. Schweiger Max Planck Institut fur Zel/biologie, Ladenburg bei Heide/berg, Federal Republic
of Germany.
S. Segal Department of Immunology and Microbiology, Ben Gurion University of the Negev,
Beer Sheva, /srael.
A. S. Spirin Institute of Protein Research, Academy of Science of U.S.S.R., Moscow, U.S.S.R.
J. R. Tata National Institute of Medical Research, London, U.K.
A. Vaheri Department of Virology, University of Helsinki, Finland.
D. F. H. Wallach Division of Radiobiology, Tufts-New England Medical Center, Boston, U.S.A.
E. R. Weibel Anatomisches Institut der Universitat Bern, Bern, Switzerland.
R. Williamson Department of Biochemistry, St Mary’s Hospital, Medical School, London, U.K.
R. E. Williamson Department of Developmental Biology, Australian National University, Canberra,
Australia.
K, E. Wohlfarth Institut fur Cytologie, Universitat Bonn, Bonn, Federal Republic of Germany.
-Bottermann
Copyright © 1984 Academic Press Inc. (London) Ltd. The appearance of the code at the bottom of the
first page of a paper in the journal indicates the copyright owner's consent that copies of the paper may
be made for personal or internal use, or for the personal or internal use of specific clients in the U.S.A.
This consent is given on the condition, within the U.S.A., that the copier pay the stated per-copy fee
through the Copyright Clearance Center,.Inc., 21 Congress Street, Salem, MA 01970, U.S.A. for copying
beyond that permitted by Sections 107 or 108 of the U.S. Copyright Law. This consent does not extend
to other kinds of copying, such as copying for general distribution, for advertising or promotional
purposes, for creating new collective works, for resale or for copying or distributing copies outside the
U.S.A.
Published monthly at 24-28 Oval Road, London NW1, England by Academic Press, Inc. (London)
Limited: Volume 8, inland £66.00 inclusive of postage; abroad $175.00 inclusive of postage.
Subscription orders should be sent to Academic Press Inc. (London) Limited, High Street, Footscray,
Sidcup, Kent DA14 5HP (tel: 01-300-0155), except those Originating in the U.S.A., Canada, and Central
and South America which should be sent to Academic Press, Inc. 111 Fifth Avenue, New York,
NY
10003, U.S.A. Send notices of changes of address to the office of the Publishers at least 6-8 weeks
In
advance. Please include both old and new addresses. Postmaster, send change of address to Cell
Biology International Reports, 111 Fifth Avenue, New York, NY 10003, U.S.A.
saat and mailing in the U.S.A. by Publications Expediting Inc., 200 Meacham Avenue, Elmont,
NY
Second class postage paid at Jamaica, New York 11431, U.S.A.
,Cell Biology International Reports, Vol. 8, No. 9, September 1984 703
Mini-review: Hyaluronidases in early embryonic development
Claudio D. STERN, Department of Anatomy, Downing Street, Cambridge
CB2 3DY, England
In this paper I shall summarise a selection of the most relevant
recent studies which indicate an involvement of the
extracellular matrix and hyaluronidases in the control of
developmental processes in the early embryo. I shall be
concerned principally with processes taking place during
morphogenesis. By this term a shall mean the cellular
rearrangements which involve more or less large fields of the
embryo. To a lesser extent I shall also discuss the relationship
between these morphogenetic processes and cytodifferentiation, a
term which I shall use to refer to the relatively irreversible
change of an individual cell into a "typical" differentiated
member of a histologically, functionally and molecularly
distinct tissue.
Unfortunately, only a few direct investigations of the presence,
dvstribucton- and Wactivitity or hyalwronidases’ in vearly embryos
are available in the literature. Most of the studies to date are
mainly concerned with the presence and characterization of
extracellular materials, and with the incorporation Or
radioactive precursors into these, but very few indeed have
embarked ‘onto a detailed analysis of ‘the turnover of these
compounds during the early stages of embryonic development. Much
of the evidence I shall be considering will perforce be
indirect.
Before embarking on this discussion, it is important to consider
what hyaluronidase is. Far from being a single enzyme, the name
iso "applied’” to Yany -enzyme activity which cleaves the glycosidic
bonds of hyaluronic acid (Merck Index, 4th ed, 1976). The’ Enzyme
Commission currently recognizes at least four distinct enzymes
with "hyaluronidase" activity: sD Gory Se Zh ull soy Baers: 2a. 36
and EUG). 4.529 9141 (Florkin & SiEOuZ , (973
BOGIAA202 041
the existence of many different hyaluronidases has
Furthermore,
demonstrate d aise! many systems. there are “dri tenent
been
in the adult frog from those of the tadpole
hyaluronidases
pye197"); and different activities associated with
Garpson Viet al
from those in the iMmeer Ol Oil Ene ™ same
fibroblast cell surfaces
(Orkin & Toole, 1980a,b). Antibodies raised against rat
cells
hyaluronidase do not Cuoss—Theace with
urinary (renal)
testicular origin and vice-versa (Law & Rowen,
hyaluronidase of
tail fin-derived hyaluronidase shows
1978, 1979, 1981). Tadpole
for hyaluronate aS a substrate over chondroitin
a preference
© 1984 Academic Press Inc. (London) Ltd.
0309-165 1/84/090703-15/$03.00/0
, 704 Cell Biology International Reports, Vol. 8, No. 9, September 1984
4-sulphate, and is unable to degrade chondroitin 6-sulphate or
heparan. Testicular hyaluronidas e, on the other hand, can cleave
both forms of chondroitin sulphate as well as hyaluronate, and
bacterial hyaluronidase cleaves chondroitin sulphate in
preference to hyaluronate (Silbert & DeLuca, 19.7 O)iz-iin the
absence One hydrolysing enzymes, hyaluronate and chondroitin
sulphates are relatively stable in embryos (Derby & Pintar,
TERE,
With these provisos, we can now begin a discussion on the
involvement of hyaluronidases in the regulation of early
developmental processes in the vertebrate embryo.
1. Hyaluronidase activity during morphogenesis
(a) Non-malignant invasion in morphogenesis
of particular interest to» the, clinician sas swell as) tthe
embryologist is the problem of invasion. For the oncologist,
this phenomenon is associated with metastasis and the spatial
spread of malignant tissues into surrounding areas. For the
embryologist, non-malignant invasion 1s associated with normal
morphogenetic events at various times in the development of
embryos, when one tissue displaces another (see Mareel, 1979 and
Vakactcet ab, M980 sfor suecent y yevalewsie Them bese Mexanplesmmo.
this process in embryonic development are perhaps the
implantation of the mammalian blastocyst into the maternal
endometrium, and the process of the formation of the definitive
(gut) endoderm (Stern & Ireland, 1981; \Bellarrsm et tral TeiocioEe
where the ectodermally-derived endoderm tissue displaces and
inserts into the existing endodermal layer (hypoblast). Other
examples include the migration of neural crest cells, initially
into a relatively cell-free but glycosaminoglycan-rich matrix,
and later some of them may move into the mass of mesenchymal
cells forming the somites (Noden, 1978). At the level of single
celiis>, Sfiertilazvation® “itself scouldip bewiconsidercdNasctan einvasiiva
process, where the egg is "invaded" by the sperm.
As tissues and cells are frequently surrounded by extracellular
materials such as basal laminae containing glycosaminoglycans
and proteoglycans, invasive ability, whether malignant or not,
should be accompanied by an ability of the invading cells or
tissues to destroy this Mal Genes Thus one would expect
hyaluronidase activity to be high in invading tissues in cancer
and in non-malignant invasion during embryonic development.
Experimental evidence confirms thass Kolaova (19577) has
demonstrated high levels of hyaluronidase activity in patients
with malignant tumours (see also Coman, 1953 and references
