Embryologists meet 1913 in Rio de Janeiro

From left: J. P. Hill, E. Bresslau and G. S. Sansom 1913

Instituto Oswaldo Cruz

J. P. Hill had made his name studying placentation in Australian marsupials. He returned to England in 1907 as Professor of Zoology at University College London. In October 1913, accompanied by G. S. Sansom he went to Brazil to obtain material from South American marsupials in particular the Brazilian common opossum (Didelphys aurita) (see biography here).

Whilst in Rio de Janeiro he met up with the eminent German zoologist Ernst Bresslau who was there from 1913 until the outbreak of war in the following year (see biography here). Bresslau was interested in the evolutionary origins of the mammary gland. Thus like Hill his research spanned monotremes, marsupials and eutherians. 

Bresslau was then at the University of Strasbourg. Later he was recruited to Cologne where he built up the zoology department. He was dismissed from this post by the Nazi regime in 1934 but went to start up zoology at the newly established University of Sao Paulo. Sadly he died there in the following year.

G. S. Sansom did interesting work on germ layer inversion in the water vole (Arvicola amphibius) and later with Hill on the guinea pig (Cavia porcellus). Sansom was a renowned climber. During World War I he served in the Royal Flying Corps and RAF and was awarded the Military Cross and Distinguished Flying Cross. 

Note on the photo: I have identified Hill and Bresslau by comparison with other photos. The printed text reads, "Prof. Ernest Bresslau - Strasborgo; Prof. J. P. Hill - Londres; Mr G. S. Sansom - Londres"

Hermann von Ihering and polyembryony in armadillos

Uterus of the mulita (Dasypus hybridus) with 9 identical
embryos. From Fernandez Morph Jahrb 1909; 39:302-333

In 1886 Hermann von Ihering opened the uteri of two pregnant armadillos. Both contained 9 fetuses of the same sex. Each fetus had its own amnion but all were enclosed in a common chorion (placenta). He was the first to propose that the embryos were derived from a single fertilized egg with splitting into separate embryos occurring early in development.

Arrangement of fetal membranes in the mulita (Dasypus hybridus).
From Fernandez Morph Jahrb 1909; 39:302-333

Ihering had studied the mulita or Southern Long-Nosed Armadillo (Dasypus hybridus). Later Fernández demonstrated that splitting occurred at the embryonic shield stage in the mulita. Newman & Patterson, working with the nine-banded armadillo (D. novemcinctus), came to a similar conclusion. Fernández, however, was the first to obtain early stages before splitting occurred. More recently, the nine-banded armadillo was the object of elegant studies by Allen Enders (summarized here). Specific polyembryony is known only from Dasypodinae and is thought not to occur in the two other subfamilies of armadillo.

Hermann von Ihering (1850-1930)
CC BY-SA 3.0 Wikimedia Commons

Hermann von Ihering was a German zoologist who relocated to Brazil in reaction to his family's disapproval of his marriage to a widow with a child. This was in 1880. His first years were spent as a collector in the southern state of Rio Grande do Sul, based on an Island known as Ilha do Doutor (Doctor's Island). In 1893 he became the first director of Museu Paulista (State Museum of Sao Paulo) and held this post for 23 years.

Hermann von Ihering´s principal area of expertise was mollusks. He also became an expert on the birds of the State of Sao Paulo, of which he observed 695 species and subspecies. For my Brazilian readers there is an excellent recent biography by Hitoshi Nomura (open access here). It lists 338 of his publications.

His son Rodolpho von Ihering (1883-1939) was also a zoologist. He was appointed vice director of Museu Paulista, which led to the accusation of nepotism that was to force Hermann's resignation. Rodolpho was an expert on fish and is credited with founding Brazilian pisciculture with stations at Porto Alegre in Rio Grande do Sul and Pirassununga, S.P.

References: Biol Zentralblatt 1886; 6:532-9 and Arch Physiol 1886; pp. 443-50

Foetus or fetus?

Human Fetus drawn by Leonardo da Vinci

There was a recent spate of tweeting about the correct spelling in British usage of fetus - or foetus. As the Oxford Dictionary makes clear, the spelling foetus has no etymological basis.

A similar debate 50 years ago was initiated by James Dixon Boyd and William James Hamilton in connection with the first edition of their influential textbook Human Embryology (previous post). This was in the BMJ. Coincidentally Bernard Towers (later Professor of Anatomy and Pediatrics at UCLA) raised the issue in Arch Dis Child. Earlier, Lionel Everard Napier had argued for "fetus" in The Lancet.

The thrust of their arguments was that "fetus" was the only spelling in use until 600 A.D., "foetus" being introduced by Isidorus of Seville on the basis of an erroneous etymology.

Statue of Isidorus of Seville in Madrid
Photo by Luis Garcia CC BY-SA 2.5

Boyd and Hamilton solicited opinion on the subject and the resulting letters fell out 5 to 1 in support of "fetus." Among the supporters was J.H.M. Pinkerton, later Professor of Midwifery and Gynaecology in Belfast. The counter argument, "Foetus is a word of respectable antiquity and lineage," was advanced by Hugh Gault Calwell who is known to have been skilled in Greek and Latin. Sadly, when Human Embryology appeared, it used "foetus" rather than "fetus."

References: BMJ 1967 (5337): 425, (5539): 568, (5540): 631, Arch Dis Child 1967; 42:224, Lancet 1952; 260: 885-6.

Slow incubation of dinosaur eggs: why only birds survived

Cast of Mossospondylus eggs and embryo
Photo from Royal Ontario Museum by Daderot (CC)

Most people have heard that birds are dinosaurs. But that raises an obvious question: why did birds survive the end-Cretaceous mass extinction event that did for their non-avian kin? A paper just published in PNAS suggests this might be explained by different reproductive strategies.

Reconstruction of Protoceratops andrewsi
AntoninJury (Wikimedia Commons) CC BY-SA 4.0

In brief, the authors made CT scans of teeth from fossilized dinosaur embryos. They then counted the von Ebner lines, which reflect the incremental pattern of dentine formation. Applying some quite reasonable assumptions to the data, they estimated the incubation time of the dinosaur eggs as minimum 2.8 months for Protoceratops andrewsi and 5.8 months for Hypachrosaurus stebingeri. The incubation times of modern birds tend to be much shorter though the upper end of the range (11-85 days) overlaps with P. andrewsi.

It is suggested that the relatively long generation times of non-avian dinosaurs put them at a disadvantage in competing with birds, reptiles and mammals during the Cretaceous-Palaeogene transition.

Carnegie Collection of human embryos

Carnegie embryo 8171. Early lacunar stage (Stage 5b)
Courtesy of Dr. Allen C. Enders

An important source for human embryology, including implantation and formation of the placenta, is the Carnegie Collection now housed at the Human Developmental Anatomy Center in Washington D.C. The core of this collection is the carefully dated series of embryos first described by Hertig, Rock and Adams (here).

The Virtual Human Embryo is an online ressource based on the serially sectioned embryos in this collection and includes 3D reconstructions. It covers all 23 Carnegie stages in the first 8 weeks of embryonic development and cannot be too highly recommended.

Carnegie Embryo 7801. Showing extraembryonic coelom (eec)
and secondary yolk sac (sys) (Stage 6)
Courtesy of Dr.Allen C. Enders

Now a group in Amsterdam has used the Carnegie Collection to develop an additional annotated digital atlas of human development (described here). They also utilized material from the Boyd Collection at the Centre for Trophoblast Research in Cambridge.

They make two claims. First that representations in textbooks have become increasingly schematic. This is demonstrably true. Second that the descriptions in standard texts are often based on extrapolation to humans from animal models. It is hard to assess if the latter truly is the case. For example Human Embryology by Hamilton, Boyd and Mossman (previous post) was based on the human embryos in the possession of the three authors. In Germany there was a strong tradition to cover the embryology of all vertebrates, concluding with the human, exemplified by Dietrich Starck's Embryologie.

In physiology, on the other hand, animal data often are presented as if they were human. One example concerns oxygen tensions in various parts of the fetal circulation. Pretty much every textbook of physiology has a large illustration of the fetal circulation with data obtained in sheep by Dawes, Mott and Widdicombe. The figure legends often fail to acknowledge the source or the species or both.

Wombs with a view

ISBN 978-3-319-23567-7

"Illustrations of the Gravid Uterus from the Renaissance through the Nineteenth Century," compiled by Lawrence D. Longo and Lawrence P. Reynolds.

This book contains several iconic images and many that are less well known. Each with a text about the author, artist and engraver as well as an analysis of the influence of the book on contemporary science and midwifery.

Great pains have been taken with reproduction of the images. No doubt many were taken from rare books in Larry Longo's own library. It is a pity he did not live to see the result in print (previous post).

Afterbirth of the sheep with four neat rows of cotyledons
From Girolamo Fabrizio De Formato Foetu 1604

There are plenty of images of the placenta including a few from animals. Anatomists who had dissected the gravid uteri of ruminants and dogs sometimes represented human placenta as cotyledonary or zonary in shape.

I am enjoying this book. It is a pity that the publisher (Springer Nature) did not employ a copy editor. There are many more typos than might be expected in a work of such high quality.

Human development - the first 13 days

Human embryo Carnegie Stage 5c (Carnegie Embryo #7700)
Photomicrograph courtesy of Dr. Allen C. Enders

A system created for cultivating mouse blastocysts has been applied successfully to describe the development of the human embryo for 13 days after in vitro fertilization. This is a step towards opening the black box in our understanding of human embryology (reviewed here). Hitherto we have been confined to interpreting the histological sections of embryos in the Carnegie Collection.

Papers by two groups were just published: Shahbazi et al. in Nature Cell Biology and Deglincerti et al. in Nature. They used appropriate molecular markers to identify epiblast, primitive endoderm (hypoblast) and trophectoderm. In addition they used cytokeratin 7 and human chorionic gonadotrophin as markers for cyto- and syncytiotrophoblast.


Day 13 embryo of the rhesus macaque (Macaca mulatta)
Courtesy of Dr. Allen C. Enders

Both groups showed the appearance of cavities corresponding to the amnion and primary yolk sac as known from studies in the rhesus macaque by Enders, Schlafke and Hendrickx. In the macaque, the yolk sac (at bottom in the figure) is outlined by visceral endoderm (beneath the epiblast) and the more squamous parietal endoderm. These tissues were identified by Shahbazi et al. in human embryos and shown to express the endoderm marker GATA6.  Deglincerti et al. found the GATA6 signal was low in the parietal cells and that they expressed the trophectoderm marker CDX2. This is an interesting observation but hardly justifies them calling these cells "yolk sac trophectoderm." The term was criticized by Janet Rossant in the accompanying News and Views (here)  and it must be hoped it does not gain currency.

As in the macaque, amnion formation was by cavitation. This is nicely described by Shahbazi et al. Unfortunately they use the term pro-amnion, which is appropriate in the mouse but not in primates (contrasted here).

Differentiation of trophectoderm into cytotrophoblast and multinucleated syncytiotrophoblast was confirmed with appearance of lacunae in the latter as appropriate for Carnegie Stage 5c.

Marsupial frogs

Female Brazilian tree frog (Flectonotus pygmaeus) with
brood pouch enclosing the developing embryos
(Mauricio Rivera Correa ShareAlike 2.5)

What happens when frogs abandon their amphibious lifestyle for a more terrestial one? Frogs in the Family Hemiphractidae from South and Central America have evolved some ingenious solutions.

In hemiphractids, the embryo develops on the back of the mother either in a mucous-filled depression or in a closed pouch - as shown above for a Brazilian tree frog (the embryos are under the bumps).

Froglet of a marsupial frog (Gastrotheca ovipera) showing
the external gills. From Nathan 1932 (here)

The embryos may develop into tadpoles and be released to water-filled cavities in plants or skip the tadpole stage and develop directly into froglets. In species of the genus Gastrotheca, embryos have 1-2 pairs of external gills that serve for respiratory gas exchange with maternal tissues in the brood pouch. This would satisfy most people's definition of a placenta. The gills are shed around the time of birth.

Marsupial Frogs by William E. Duellman 2014
Johns Hopkins University Press ISBN 978-1-4214-1676-5

The biology of marsupial frogs is described in painstaking detail in this new book - the destillation of a lifetime's work by William E. Duellman (details here). It is superbly illustrated but at USD 120 a bit pricey. More than half the content comprises species accounts and no doubt it will find a place on the bookshelf of specialists. But it is well worth checking out for its insights into the reproductive biology of frogs. Who knew, for example, that the oocytes of Flectonotus pygmaeus have up to 2000 nuclei reduced during oogenesis to a single one?

For other fun facts on frog reproduction see this video.

Sea squirts, lancelets and acorn worms

A sea squirt (Ciona intestinalis) - a member of Tunicata 
Wikimedia Commons uploaded by perezoso (GFDL)

Genomics has clarified our position in the tree of life. To explain this I need to define some taxonomic terms.

Phylum Chordata comprises three subphyla: Vertebrata (Craniata), Tunicata (Urochordata)and Cephalochordata. Tunicates include sea squirts  such as Ciona (pictured) above. A familiar cephalochordate is the lancelet Branchiostoma lanceolatum better known as Amphioxus (shown below).

Amphioxus or Branchiostoma lanceolatum
(c) Virginia Gewin here (CC-BY-SA 3.0)

Amphioxus has long been used to exemplify the general plan of chordate organization and lancelets used to be regarded as the closest relatives to vertebrates. The genomic evidence, however, has tunicates like the sea squirts as sister group to vertebrates with cephalochordates as a deeper branch. Additional support is given by conserved molecular signatures (here).

Chordates belong in the Superphylum Deuterostomia (brilliantly reviewed by Lowe et al. here) along with Phylum Hemichordata and Phylum Echinodermata. Echinoderms are richly represented in the fossil record and the five extant classes include sea urchins, sea cucumbers and starfish. Hemichordates include the acorn worms for which two genomes just became available (here and here).

Acorn Worms (Hemichordata: Enteropneusta)
from Spengel 1883 (public domain)

One of many findings was a cluster of six genes that are conserved across chordates and implicated in patterning of gill slits. This is significant because gill slits were an innovation in the deuterostome lineage (although secondarily lost in echinoderms).

Relationships between deuterostome phyla were largely worked out through their embryology, an example being the erection of Chordata by Haeckel. Understanding the genes involved in developmental processes remains a focus in working out our evolutionary history (see the review by Lowe et al. mentioned above).

Mouse and human blastocysts compared

Mouse blastocyst with trophectoderm (TE),
epiblast (EPI) and primitive endoderm (PE)  from Selenka 1883

Even before implantation, three cell lineages are apparent in the blastocyst of mouse and human. Outermost is the trophectoderm that will contribute trophoblast to the placenta. The inner cell mass has already differentiated into the epiblast and the primitive endoderm or hypoblast. For mouse, this much has been clear since the pioneering studies of Emil Selenka (here).

Gene expression in these three lineages of mouse placenta has been known for some time. What does this tell us about human preimplantation development? A recent paper in Development (here) suggests less than some might like to think.

• For trophectoderm, key lineages expressed in the mouse (e.g. Id2, Elf5, Eomes) either are not expressed in human trophectoderm or are expressed in alternative lineages.
• There are several genes that are exclusively expressed in human epiblast (e.g.the transcription factor KLF17).
• Expression of some genes in primitive endoderm is conserved between mouse and human (e.g. Foxa2/FOXA2).

These findings support other work indicating that the genes and signalling pathways involved in lineage specification differ between mouse and human blastocysts.

Embryologists then and now

International Institute of Embryology London 2-5 August 1938

The photos in this post are separated by three quarters of a century. We know the identities of the 1938 embryologists because an annotated copy was sent by Fritz Strauss to Harland W. Mossman and is curated in the latter's collection in Madison, Wisconsin.

Top row (left to right): T. Thomson Flynn (Belfast); H. M. W. Woerdeman (Amsterdam); Hans Bluntschli (Bern); Jan Florian (Brno).

Middle row: G.L. Streeter (Carnegie Institution); Karl Peter (Greifswald); Mrs. Katherine Jones Hill (London); E.S. Goodrich (Oxford); Miss E. G. Fraser (London); Warren H. Lewis (Carnegie Institution); A. Celestino da Costa (Lisbon); Paul Gérard (Brussels); H. Woollard (London). 

Bottom row: Otto Grosser (Prague); J.T. Wilson (Cambridge); J. Boeke (Utrecht); Honor B. Fell (Cambridge); Dan de Lange (Utrecht); J. P. Hill (London).

It was my pleasure to attend a recent meeting in Göttingen and find that embryology is still going strong. As in the 1938 Group there are some eminent biologists here. But the most striking difference between the two photos is the inclusion of many postdocs and graduate students with the promise it brings for the future of the field.

Some curiosa

Together with Elliott Smith, J. T. Wilson and J. P. Hill were part of the Fraternity of the Duckmaloi that pioneered research in monotreme and marsupial embryology. Their heirs are Marilyn Renfree (front row fourth from left) and Karen Lychau Hansen (front row third from right).

Theodore Thomson Flynn also worked on marsupials but later turned to fish. He named one species Gibbonsia erroli after his son, who later achieved fame as the swashbuckling film star Errol Flynn.  

One of the few women in the 1938 Photo is Katherine Jones Hill. The daughter of J. P. Hill, she was an embryologist in her own right. She catalogued the Hill Collection now housed in Berlin.