The Mouse Adrenal X-Zone Revisited

 

This photomicrograph (from here) shows the cross section of a female mouse.
The medulla (M), X-zone (XZ) and zona fasciculata (ZF) of the cortex are labelled.
The outer, zona glomerulosa, of the cortex can be seen but is not labelled 

Introduction

For a few months in 1965-66 I worked on the aptly-named X-zone of the mouse adrenal in Hong Kong. In 2018 I wondered what had happened to research on the X-zone. Had its function been discovered? The answers were in fact ‘rather little’ and ‘no’ and so I offered to give a talk at an annual Society for Endocrinology meeting in an attempt to stir up some interest in a problem that has intrigued those interested in the workings of the adrenal gland since the 1920s. This I did in November 2019 at Brighton meeting of the British Endocrine Societies.

I have written below an account of the cellular origins of the X-zone because it now transpires that we were misled in the 1960s by the misinterpretation or misreporting of early findings and that, in fact, the early workers were right about where it came from during development of the embryo. In addition, nobody it seems had spotted an important paper published in 1942 that made our experiments in Hong Kong unnecessary. But first the personal historical background:

Four weeks after we had arrived in Hong Kong, I received an aerogramme from John Phillips he had written on 28 November 1965. He was on his first ‘long leave’ from the University of Sheffield and was spending it back in Sheffield with Ian Chester Jones, where, until December 1962 he had been a lecturer. We had been up to Sheffield several times before leaving for Hong Kong on 1 November and the general idea was that I should have a look to see if steroid hormones found in vertebrates occur in invertebrates as well. I was beginning to see what we had in terms of chromatographic equipment and chemicals in order to make a start when that aerogramme arrived. He wrote:

…I listened to Prof Paul Delost give a lecture on the X Zone last night and I was surprised to hear that he considers the X zone to be under medullary control. He bases this conclusion on the absence of an X zone in an adrenal in which the medullary tissue has been aspirated from the centre of the gland with a needle and vacuum pump—the other gland remaining as a control. The interesting thing about this preparation is that if you castrate the post-pubertal male the X zone reappears in the gland with a medulla but not in the other adrenal without a medulla…But the main criticism of Delost’s approach is that he destroys the vascular bed of the adrenal. This can be overcome by an operation called “enucleation” in which the whole of the adrenal is expressed leaving only the capsule from which a new adrenal cortex regenerates. Will you get some male mice and enucleate adrenals before puberty…

That I did and the results were clear. Given though the techniques available at the time it was difficult to envisage taking the approach further. The caravan moved on.

Cellular Origins of the X-Zone

After its description¹ but misidentification in 1924 by Kiyoshi Masui and Yasushige Tamura of the Imperial University of Tokyo (with a further publication in English in 1926²) and  its naming in 1927 to reflect its unknown function by Evelyn Howard (1904-1999) then at Stanford³, the X-zone of the mouse adrenal excited the interest of pioneering endocrinologists who did not yet describe themselves as such. The structure, which develops after birth between the cortex proper and the medulla, still befits its name; the function of the X-zone remains unknown^4,5. Early work was concerned with its origins and with its hormonal control since it disappears at puberty in males and during first pregnancy in females⁶.

In terms of hormones in the circulation, androgens cause the X zone’s disappearance while LH (luteinising hormone) from the pituitary is necessary for its maintenance^4,7. Although excellent research was done in the early years on the possible cellular origins of the X-zone it is only more recently that cell lineage studies using molecular markers have provided further evidence that it is derived from the fetal or inner adrenal cortex, as opposed to the definitive or outer cortex which forms the well-known zones of the adrenal: glomerulosa, fasciculata, reticularis (references in⁴). Therefore, the X-zone of the mouse appears to be homologous with the human fetal adrenal cortex, which, as its names implies is present only in the developing fetus. Such an origin was suspected by some early workers who referred to the ‘human fetal X-zone’ but strongly denied by others. Other possible homologues are the juxtamedullary zones of various size and appearance observed in some other eutherian mammals (cat, rabbit, voles, hamsters, and shrews)^6,8.

In this short article I first consider whether the more modern findings on the origins of the X-zone are consistent with the early studies since the over-riding impression created in the mind by reading reviews and papers from the latter half of the 20th century is that the X-zone is derived from the inner cells of the zona fasciculata or, in other words, is just another zone of the definitive cortex. Then I review the evidence from little-known perturbative experiments, published, to modern eyes, in obscure places, that throw light on the origins of the X-zone.

That the X-zone is derived from the fetal or inner cortex is entirely consistent with the findings of Harry Waring⁹ who was then working in Liverpool for an M.Sc. The topic was suggested to him by a forgotten promoter of endocrinology in Britain, a famed lecturer in zoology, Ruth Culshaw Bamber (1889-1970) who was always known as Mrs Bisbee.  Horace ‘Harry’ Waring (1910-1980) showed in 1935 that during embryonic development there is—initially—an intermingling of the cortical and medullary elements. Remodelling then concentrates the medulla until there is a clear separation from the cortex. He identified cells, comprising what he called the interlocking zone, between the medulla and cortical elements. These cells became concentrated around the time of birth into a layer around four cells thick. The outer cortex in the meantime was growing and forming the usual zones. But it was that layer of interlocking cells that went on to form the X-zone after birth. Later, as the X-zone degenerated there was left a ‘medullary connective tissue capsule’ or ‘juxtamedullary capsule’ around the medulla formed, it was presumed, from the collapsed stroma⁹.

In 1928, Ruth Deanesly (1901-1997) in London had already observed that after degeneration of the X-zone some of its cells remained around the juxtamedullary capsule¹⁰. Therefore, the key early finding that a secondary X-zone forms after castration of male mice can be explained by growth from these cells, i.e. remnants, capable of division, of an inner (fetal) cortex rather than from a differentiation of cells from the inner zone (z. fasciculata in the mouse) of the outer (definitive) cortex.

I cannot explain why the view prevailed, despite evidence to the contrary, that the X-zone was part of and derived from the outer or definitive cortex. As one example, the following is from the highly influential review written by Helen Wendler Deane (1917-1966) published in 1962⁸, four years before her early death:

These [X-zone] cells differentiate postnatally, at about 2 weeks, from the inner portion of the fasciculata (Whitehead 1933a, Waring 1935).

The problem with this statement is that neither Raymond Whitehead¹¹, working in Manchester, nor Harry Waring⁹ drew any such conclusion. Only Waring of the two studied the origins of the X-zone and his conclusion was, as I have noted above, entirely different.

All the above evidence, even the sophisticated and relatively recent cell lineage studies, have been observational. Those seeking direct, experimental evidence that throws light on the origin of the X-zone would at first sight be discouraged since it all appeared over 50 years ago in obscure publications and/or written in French while one important paper had, I discovered recently, been overlooked entirely.

Until I found that paper, the first experimental work that has a bearing on the problem was thought to have been that in the 1960s from Paul Delost’s laboratory at the University of Clermont-Ferrand in France and in particular that of his student Parviz Chirvan-Nia who later returned to Tehran University of Medical Sciences in Iran. They took advantage of the fact that a secondary X-zone develops after castration in male mice. Their most pertinent finding^12,13 was obtained in mice from which the adrenal medulla had been removed by aspiration through a very fine pipette, a technically difficult procedure. In males in which the entire medulla had been removed, a secondary X-zone failed to develop after castration. By contrast, if even a small piece of medulla remained, an X-zone developed around it. The effect was local; the untreated contralateral adrenal was unaffected.

All of Delost’s work from the 1950s onwards was published in French and had received little attention in the English-speaking world. However, on 27 November 1965 Delost was invited by Ian Chester Jones (1916-1996) to give a seminar in Sheffield, having been one of Chirvan-Nia’s external examiners earlier that year. His old student, John Guest Phillips (1933-1987; FRS 1981), on leave from the University of Hong Kong, was also there and while intrigued by Delost and Chirvan-Nia’s work he and Chester Jones were concerned that, in aspirating the medulla, the venous drainage from the adrenal would have been destroyed. They thought that a complementary approach, that of enucleation, in which end of the adrenal is snipped off and the medulla and most of the cortex squeezed out, would be a useful test. After enucleation the outer cortex redevelops and shows a normal zonation but without a medulla. As a result, John Phillips sent me an aerogramme the next day asking if I, having arrived in Hong Kong four weeks earlier as a PhD student, would take it on. This I did and the results were identical to those obtained by Chirvan-Nia and Delost: a secondary X-zone failed to develop if the medulla had been removed completely; if even a small remnant of medulla remained after incomplete enucleation, an X-zone developed around it¹⁴.

In 2019 while preparing a talk⁵ for the Society of Endocrinology on what has happened to research on the X-zone I discovered our experiments had simply confirmed what had already been published. The same approach, enucleation, had yielded the same results in 1942. The paper, which appears not to have been quoted by anybody working in or reviewing the field, was by Murchie Kilburn McPhail (1907-1987) and his student H.C. Read, of Dalhousie University in Canada¹⁵. I can only assume that it was missed because it appeared in wartime, albeit in a leading journal, when scientists were otherwise occupied. However, another paper from the same authors from the same year was picked up and referred to.

Suggestions as to possible mechanisms as a result of these experimental approaches, for example what would now be termed a paracrine effect of medullary cells on the inner cells of the  cortex against the direction of blood flow, can now be discarded since the removal of the medulla would by any technique result in the extirpation of the inner cortical anlagen formed in and around the juxtamedullary connective tissue capsule after degeneration of the X-zone.

In conclusion, the original observations by Harry Waring on the origins of the X-zone and the experimental evidence on the necessity of the adrenal medulla for the presence of an X-zone are entirely consonant with the X-zone being derived from the fetal or inner cortex and not the definitive or outer adrenal cortex. The recognition that there are two populations of cortical cells only one of which is responsible for the classical zonation of the adult adrenal gland explains many of the false leads followed, blind alleys entered and bold assertions made by those working in the field during the middle decades of the 20th century.

Research over the past 96 years has established the hormonal control of the adrenal X-zone and seemingly settled its cellular origins. Will we also know its function by the time of the centenary of its discovery?

1. Masui K, Yamura Y. 1924. The effects of gonadectomy on the structure of the suprarenal glands of mice, with special reference to the functional relation between this gland and the sex gland of the female (Translated). Nihon Chikusan Gakkaiho 1, 55–79.

2. Tamura Y. 1926. Structural changes in the suprarenal gland of the mouse during pregnancy. Journal of Experimental Biology 4, 81–92.

3. Howard E. 1927. A transitory zone in the adrenal cortex which shows age and sex relationships. American Journal of Anatomy 40, 251-293.

4. Huang C-C J, Kang Y. 2019 The transient cortical zone in the adrenal gland: the mystery of the adrenal X-zone. Journal of Endocrinology 241, R51–R63.

5. Peaker M. 2019. What happened to the adrenal X-zone. Endocrine Abstracts 65 SE 1.1. DOI: 10.1530/endoabs.65.SE1.1.

6. Chester Jones I. 1957. The Adrenal Cortex. Cambridge: Cambridge University Press.

7. Gannon A-L, O’Hara L, Mason JI, Jørgensen A, Frederiksen H, Milne L, Smith S, Mitchell RT, Smith LB. 2019. Androgen receptor signalling in the male adrenal facilitates X-zone regression, cell turnover and protects against adrenal degeneration during ageing. Scientific Reports 9, 10457.

8. Deane, H.W. 1962. The anatomy, chemistry, and physiology of adrenocortical tissue. In The Adrenocortical Hormones Part 1. Handbuch der Experimentellen Pharmakologie, edited by Eichler O & Farah A, subedited by Deane HW, 1-185. Berlin, Springer.

9. Waring H. 1935. The development of the adrenal gland of the mouse. Quarterly Journal of Microscopical Science 78, 329–366.

10. Deanesley R. 1928. A study of the adrenal cortex in the mouse and its relation to the gonads. Proceedings of the Royal Society B 103, 523–536.

11. Whitehead R. 1933. The involution of the transitory cortex of the mouse suprarenal. Journal of Anatomy 67, 387–392.

12. Chirvan-Nia P. 1965. Données nouvelles sar la zone X surrénalienne de las souris. Doctoral Thesis, Université de Clermont.

Brudieux R., Chirvan-Nia P., Delost P. 1966. Sur les relations directes entre la médullo-surrénale et le cortex surrénal. Journal de Physiologie, Paris 58, 213-217.

14. Peaker M, Phillips JG,. Peaker SJ. 1967. A relationship between the medulla and the X-zone of the mouse adrenal. In Proceedings, Third Asia and Oceania Congress of Endocrinology, edited by Litonjua A, vol. 2 317–321. Manila.

15. McPhail MK, Read HC.  1942. Regeneration of adrenal gland following enucleation and transplantation with special reference to X-zone. Endocrinology 31, 486–492.

Frank Evers Beddard: Descriptive zoology at the turn of the 20th century

Frank Evers Beddard

Those browsing in public libraries in the middle of the 20th century would be excused, I hope, since I was one of those browsers, for gaining the impression that F.E. Beddard’s life’s work had been on mammals. Libraries sometimes had on their shelves volumes in the Cambridge Natural History, a series of ten volumes published, out of numerical order, between 1895 and 1909. Beddard wrote volume 10 in 1902; it was on mammals.

It seemed obvious that Beddard was a specialist in mammals; at the time he wrote it he was Prosector to the Zoological Society of London, and London Zoo had lots of dead mammals each week to work on. However, he made his name on an entirely different group of animals, earthworms, the oligochaetes.

Frank Evers Beddard was born in Dudley in 1858, the son of a prosperous businessman. Educated at Harrow and Oxford (graduated 1880) he was appointed to work on the animals collected by the Challenger Expedition of 1872-1876, involving a circumnavigation of the earth for oceanographic research. The job was in Edinburgh and he was assigned the isopods to describe and identify; his results were published in two of the 50 volumes, a marathon of descriptive zoology. In Edinburgh he also began to take an interest in earthworms.

A plate from Beddard's report on isopods
collected by the Challenger Expedition

Beddard was appointed to the prosectorium at London Zoo in 1884. At that time Philip Lutley Sclater was Secretary (i.e. chief executive). It is unfortunate that Beddard’s obituary notice for the Royal Society was written by Sir Peter Chalmers Mitchell, who succeeded Sclater after a bitter, highly political election. In some respects Mitchell was an ideal biographer of Beddard since he had worked in Beddard’s prosectorium and obviously knew him well. However, Mitchell throughout his life seemed hellbent on deriding Sclater and Beddard’s obituary was yet another chance he did not pass over.

Chalmers Mitchell praised Beddard’s working habits in the days of the gentleman scientist:

For a large part of his life he was a harder worker than persons with more ordinary habits could guess. He rose very early, and was usually at his table in the Zoo Prosectorium before 8 a.m. On many days he lunched at the Gardens, taking his notebooks into the restaurant, then worked until 4, when he went home, wrote until an early dinner, again worked after dinner, going to bed at 9. When the routine was broken by his going to a club to lunch, he already had a good morning’s work behind him and could linger in apparent idleness long after the ten-to-four men had ostentatiously hurried away. In later life, naturally, he was easier with himself. 

Beddard did not enjoy a happy life. He married Anne Fletcher in 1886 and had two children but the couple split up. He with the children moved in with his mother in Hampstead. Mitchell continued:

These domestic circumstances are related because they were the first cause of a sense of grievance against life which increased with age, until it became almost a mania of persecution. Apart from the subject of his wrongs, Beddard was a charming companion, with a fund of amusing anecdote that made him a welcome neighbour in clubs or social gatherings. 

It was in his description of Beddard’s work at the Zoo that Mitchell took the opportunity to have a dig at Sclater and remind the world what a great replacement he had been:

During the greater part of Beddard’s tenure of the Prosectorship, there was the unfortunate tradition that the Prosector must always be ready with a paper for the fortnightly Scientific Meetings of the Society. P. L. Sclater, moreover, his official chief, was interested mainly in systematic zoology, and not being an anatomist himself, was over-ready to believe that even a hurried dissection would settle doubtful points in classification. The double pressure led Beddard to produce far too many short papers, and to draw systematic inferences from dissections of single species or even single organs in species. It cannot be said that his output in any part of vertebrate anatomy had a value at all com­mensurate with its bulk. A volume on the ‘Structure and Classification of Birds,’ published in 1895, brought together much useful information ; one on ‘Whales ’ (1900) was pleasantly written; his volume on ‘Mammals in the Cambridge Natural History ’ was a sound and useful compilation. Several others need not be mentioned.

Ouch.

A list of the books he wrote with, in some cases, links to full text sources can be found here.

From Beddards book on animal coloration

Beddard was praised for his work on oligochaetes (which he had done while fulfilling his duties as prosector). Quite how he had time to do this isn’t made clear because he was also lecturer in biology at Guy’s Medical School, as well as external examiner in Oxford and London:

Beddard’s permanent place in Zoology depends on his great ‘Monograph of the Oligochaeta’ (Oxford University Press, 1895) and on the prolonged researches on which it was based. There were already in existence accounts of the group published by F. Vejdovsky in 1884 and L. Vaillant in 1890, but both of these writers had rather neglected the literature on the subject, and subsequent to their volumes very much new information had been obtained by the study of collections from most parts of the world. Beddard had himself dissected and examined nearly all the important types that were known, and had searched the literature carefully. The general account of the anatomy he gave was much in advance of that of any of his predecessors, and his systematic descriptions were clear and well arranged. 

Beddard was elected to the Royal Society in 1892.

The unfortunate taste in the mouth left by Chalmers Mitchell’s account of Beddard’s life—as indeed with much of his writing—extended to reminding the reader, again, of who won the election to succeed Sclater. Beddard, Mitchell wrote, was a candidate but ‘he did not press his claims in face of a general opinion, which he himself frankly shared, that he was not well adapted to the continuous duties of administration of a large organisation’.

Beddard retired from the Zoo, where he was a popular lecturer, in 1915. He died of a heart attack at his house in Hampstead in 1925, aged 67.

Chalmers Mitchell’s comments on the book written by Beddard echo those written by William Plane Pycraft (1868–1942) of the Natural History Museum in an obituary for Nature:

In his books Beddard did himself less than justice. His volume on whales, for example, was good but he could have given us a much better book. The same may be said of his volume on the classification of birds and that on the coloration of animals. In these he seems to have shirked the labour of coming to a decision on the very vexed and controversial points which these two themes presented. He nowhere commits himself to a definite opinion as to whether he does or does not agree with the conclusions arrived at by others, whose views he sets forth without comment. His pages are almost too dispassionate to be helpful.

 

I looked at the book on animal coloration. I found it an excellent introduction for its time, and surely he should have been praised for presenting different views on ‘controversial points’ without necessarily, in the absence of further evidence or informed knowledge, coming down on one side or the other.

Pycraft (who, remember, was one of those duped by the Piltdown hoax) praised the talks that Chalmers Michell so derided; part of the job of the prosector was to keep the amateur and professionals who gathered for Zoological Society meetings content with their membership:

Those who were privileged to listen to his discourses, at the scientific meetings of the Society, will ever remember his extraordinary facility of expression and the clear and rapid way in which he laid abstruse points before his audience. Few, probably, who were listening had ever made the dissections he was describing, yet so admirable was his presentation of the facts he had gleaned, that they could not fail to grasp the essential points laid before them. He had no rival in this regard.

Beddard’s preserved specimens and descriptions live on. I think Beddard is now best described as an exemplar both of what zoology was all about at the end of the 19th and beginning of the 20th centuries and of the sort of man who worked in the field. And I still rather like, dated as it is,  his mammals book.

Pycraft WP (as WPP). 1925. Dr FE Beddard FRS. Nature 116, 215-216.

Mitchell PC (as PCM).1926. Frank Evers Beddard—1858-1925. Proceedings of the Royal Society B 99, xxxvi-xxxvii.

HONG KONG: An Urban Wild Boar

AJP was walking to the Aberdeen MTR Station a couple of weeks ago when, much to his surprise, he spotted a wild boar walking along the footpath towards him.

In the 1960s wild boar were very rare, having previously been abundant. So rare that Patricia Marshall in her Wild Mammals of Hong Kong (1967) wrote of ‘there being possibly only one or two families of not more than five individuals in each. One family on Lantau [island], one in the New Territories’.

Now they are said to be everywhere with a massive increase in the past few years. People feeding them are being blamed for their presence in urban areas. This one was the epitome of nonchalance as it sauntered along the street.

It reminds me of the 1934 temperance song:

Yes, the pig got up and slowly walked away

Slowly walked away, slowly walked away

Yes, the pig got up and he turned and winked at me

As he slowly walked away

The genetic history of these pigs would be interesting, especially since there were thought to be none on Hong Kong island (Aberdeen is on the island for those unfamiliar with the place) by the late 1960s. 

Pallas’s Squirrel in Hong Kong: Feral pets, an introduced or reintroduced species…or even a range expansion?

Fifty years ago the sight of a wild squirrel in Hong Kong would have been unthinkable. But now, Pallas’s Squirrel (Callosciurus erythraeus) can be seen both on Hong Kong Island and in the New Territories. I have no reason to doubt the story that these populations were from the accidental or intentional release of squirrels imported for the pet trade. Well…perhaps a nagging doubt.

Pallas's Squirrel photographed in the New Territories of
Hong Kong by AJP, December 2020. Note the black
'spot' near the end of the tail. In the second photograph
the red belly can be glimpsed.

In the 1960s these squirrels, along with Siberian Chipmunks, were being sold as pets in Hong Kong. The ‘lab boys’ had a couple in a tiny cage in their room in the Zoology Department of the University of Hong Kong, for example. In view of the appearance of flocks of feral Yellow-crested Cockatoos we perhaps should not have been too surprised to have seen squirrels in the trees of Government House on our first return to Hong Kong in 1997.

The standard account of these squirrels in Hong Kong is that the ones on Hong Kong Island are different from those in the New Territories. Those on the island are described as belonging to the subspecies C.e. thai, with those in the New Territories as C.e. styani, the former from Thailand; the latter from Northern China. I am not sure how the feral animals in Hong Kong were assigned to these subspecies and having looked up some of the original papers I am even less sure about the identification of thai other than Thailand is perhaps where the animal dealers said they were imported from. The black hairs near the tip of the tail of the ones from the New Territories do, however, fit the description of styani. The import of the Siberian Chipmunk, Eutamias sibiricus, is also compatible with the importation of C.e. styani, into Hong Kong from the same area of northern China.

In many parts of its range the belly of the squirrel, as its specific name implies, is some shade of red. That is true of those in the New Territories of Hong Kong. That was not the case in at least some of those kept as pets. The ones the lab boys kept were a light yellowish grey—as were those we saw in the gardens of Government House 30 years later.

It does seem odd that a native squirrel had never been reported for Hong Kong. The question is if they were once there and had been extirpated or had never even been part of the native fauna. Two squirrels apparently from near Canton (now Guangzhou)—only 135 km (85 miles) from Hong Kong—were collected by John Reeves between 1812 and 1831; the skins are in the Natural History Museum in London. They are currently assigned to a subspecies, C.e. castaneoventris, with a distribution south of the Pearl River including the island of Hainan. Nothing seems to be known in formal descriptive terms of the Pallas’s Squirrels that occur over much of China (including the mainland north of the Pearl River adjacent to Hong Kong) and this leads me to the first of my nagging questions: is it just possible that the squirrels in the New Territories of Hong Kong are not feral but are native squirrels that have come over the border as Hong Kong became reforested after the devastation of the hillsides for firewood during and shortly after the Japanese occupation?

The whole taxonomy of squirrels of the genus Callosciurus seems to this outsider as a mess, in part due simply to a lack of series of specimens from most parts of the range. Clarity has not be advanced by simplistic molecular phylogenetic studies (using only mitochondrial DNA). That leaves me with a second nagging question: why, if two apparent subspecies were imported for the pet trade in the 1960s has one become feral in mainland Hong Kong and the other on Hong Kong Island? Surely, both would have been sold in the pet shops and bird markets on both sides of the harbour. Did they interbreed on the mainline side and if so does that mean that the ‘styani’ coloration is dominant to the ‘thai’?

Tackling the questions on the origins of the Pallas’s Squirrels would also help settle the obvious corollary: can those in Hong Kong be classified as an ‘introduction’ or a ‘re-introduction’ or even a natural range expansion?

Lurz PWW, Hayssen V, Geissler K, Bertolino S. 2013. Callosciurus erythraeus (Rodentia: Sciuridae). Mammalian Species 45, 60-74.

Moore JC, Tate GHH. 1965. A study of the diurnal squirrels, Sciurinae, of the Indian and Indochinese subregions. Fieldiana 48. Chicago: Chicago Natural History Museum.

A Hong Kong Moth

 AJP found this moth in Hong Kong last week. It seems to be a female Eudocima homaena, noted as being a pest on orange trees in parts of its range.

Do any introduced Painted Frogs still survive in Manchester?

Discoglossus pictus from Sicily
Fabrizio Li Vigni here

In my last post I described how Louis Lantz had bred Painted Frogs (Discoglossus sp.) in Manchester. Because he was unable to rear the young to adulthood he released them into his garden. There grew and bred, and he described how he could then collect them for breeding experiments and to pass on.

The frogs left to fend for themselves in the garden spawned in small open air ponds and spread in neighbouring gardens and garden ponds. This introduction was noted in Deryk Frazer’s Reptiles and Amphibians in Britain (Collins, 1983) with the comment that it was not known whether there were still individuals surviving from Lantz’s releases.

No address was given for Lantz in the 1947 paper in which he discussed his keeping and breeding these frogs and so I suspect that anybody searching the Manchester area would have had no clue as to where to begin looking. The 2017 biographies of Lantz provide two addresses in  the Manchester area where he kept and bred amphibians and reptiles. With the help of genealogy search sites it is possible to pin down the approximate dates he was living at the two locations.

The paper published in 1947 was submitted in March 1946. Given the usual period taken to prepare a final draft we can estimate that Lantz wrote his part at the end of 1945 or in very early 1946. He wrote that he had made the observations ‘during the last 15 years’; in other words around 1930. The first breeding experiments he described were in 1932. At this time Lantz was living at 9 Waterpark Road, Manchester 7. I see from Google Earth that there is a lake in Broughton Park only 130 metres from Lantz’s house. He was still there with his family in 1935. Unfortunately at the time of the 1939 Register, a special census taken in preparation for war, he and his wife were guests at a hotel in Wales but by 1940 he had moved to 2 Kinnaird Road at Withington where the photograph I showed in the last post of his greenhouse was taken. It was at Kinnaird Road that Lantz collected spawn from a neighbour’s pond.

In recent years the single species of Painted Frog has been split into five or six. Lantz explained where he had obtained his frogs and it therefore possible to assign currently recognised species [in square brackets below] to the ones he collected, bought or was given:

Some of the material was collected by the author on Port-Cros and Levant Islands near Hyeres, where the species had not previously been recorded [D. sardus], and later also on Corsica [D. sardus but also possibly D. montalentii]. A few Sardinian [D. sardus] and Portuguese [D. galganoi] specimens were obtained from dealers, but many fine and valuable animals are due to the kindness of Mr J. Armitage (1 specimen from Corsica [D.sardus or D. montalentii]), Gen. M. Berquet (10 specimens from Tunisia [D. pictus]), Mr O. Cyren (3 specimens from Morocco [D. scovazzi]), and Mr R. Maxwell Savage (7 specimens from Port-Cros [D. sardus]), to whom sincerest thanks are expressed. 

In respect of Lantz’s breeding experiments between what were clearly D. sardus (then considered a subspecies, D. pictus sardus) from Sardinia and Port-Cros and D. pictus ( D. p. pictus) from Tunisia, it is interesting to note that these two ‘species’ interbred and back-crossed freely. As in his studies on newts he was interested in the inheritance of skin colour and pattern. What was then pictus came in two dorsal patterns: spotted and striped; sardus spotted only. He found inheritance was of the simple Mendelian type, with ‘striped’ dominant over ‘spotted’.

However, the purpose of this post was not to question whether the current species recognised are good ‘biological’ species, but to point out the existence in the 1930s and 1940s of breeding sites in the Manchester area of introduced Painted Frogs. Although the odds of their continued existence seem long, it is surprising how many gardens in England still have thriving colonies of introduced Midwife Toads. Lantz was not the only herpetologist to introduce deliberately or accidentally Painted Frogs in England. North London in 1960 was the second instance noted by Frazer. Painted Frogs were also commonly imported by dealers, particularly in the earlier decades of the 20th century and numbers must have been released into insecure ponds and vivaria over the years. There are also at present current breeders of one or more species of Discoglossus in Britain.

But back to the original question: has anybody in Manchester had a look or plans to look for Painted Frogs? 

Lantz's plate from the 1947 paper showing his breeding experiment
Original capton wording superimposed on each figure

Distribution of Painted Frogs (Discoglossus)

Bruce HM, Parkes, AS, Lantz LA. 1947. Observations on Discoglossus pictus Otth. Proceedings of the Royal Society B 134, 37-56 (plus two plates).

 

Louis Lantz: Frenchman, Manchester resident, outstanding herpetologist, and important in wider biological circles in 20th century Britain

My first memory of seeing the name Louis Lantz is of being mystified by the appearance in Malcolm Smith’s book, The British Amphibians and Reptiles I had taken out from the local library, of a graph showing growth curves after metamorphosis for three hybrid newts. That graph accompanied the text: ‘Mr L.A. Lantz’s observations on growth shew that the rate depends largely on the quantity of food available and on the length of period during which, because of low temperature or low humidity, little or no food is taken’. I remember being intrigued—who was this Mr Lantz and why did he have hybrids between two species, Triturus cristatus and Triturus marmoratus?*

Only many years later did I begin to learn who Lantz was, why he worked in Manchester, and of his various contributions to herpetology. Biographies published in 2017 in French and In English have attempted to make Lantz and his classical herpetology better known in his native France and in Russia where he had worked. Unfortunately his biographers have largely omitted or underestimated his rôle in the British scientific scene and in the development of genetics and cytogenetics by some of the big players in this then burgeoning field.

Lantz has been described as an amateur herpetologist. That he was in the sense that he was not paid for working on amphibians and reptiles. But he was a professional scientist and technologist who worked in herpetology in his spare time. Lantz was a scientist whether involved in his profession or his ‘professional’ hobby which is why he found collaboration with the major figures in genetics a normal part of scientific discourse. He is also described as having ‘kept salamanders as pets’. I think those of us who have or still do keep reptiles and amphibians would object strongly to use of the word ‘pet’ in such a context.

Louis Amédée Lantz was born in 1886 in Mulhouse, Alsace, now part of northern France but then annexed to Germany. After studying zoology and botany at the University of Montpelier in 1903-4 he moved to the school of industrial chemistry in Mulhouse. There be obtained a diploma in chemical engineering. The school specialised in teaching the use of dyes for the important local textile industry which concentrated on printed fabrics. The school attracted a number of Russian students and Lantz found a job in Russia in 1907. From 1908 he specialised in cotton dyeing and printing at a vast factory in Moscow. Forced to leave in 1918 because of the Russian Revolution—and having married a Russian—he held various jobs in France. Then in 1923 he was appointed Director of Research at the Calico Printers’ Association in Manchester. He stayed there until his retirement in 1951 at the retirement age of 65. Partly, he recorded, because of his wife’s health, he returned to France, intending to live in Paris in order to work at the natural history museum. He died in Lucern, Switzerland in February 1953.

From Ineich et al 2017

   

The laboratory in Manchester, was, for its time, a big one. It employed 20 graduates and 40 technicians seeking to improve and to develop new techniques for printing and dyeing. At the time of formation of the company by merger, Calico Printers was producing 80% of Britain’s output of printed cloth. As well as printing and dyeing, Lantz was also responsible for the development of new, synthetic fibres and coatings. Terylene or PET, later manufactured by ICI, was one outcome in 1941. He and his team had numerous patents for processes and materials they invented.

Having started to keep reptiles and amphibians at home in Alsace, Lantz took every opportunity to study reptiles in the wild, in the Caucasus while he was working in Russia, for example, and to undertake breeding experiments that directly addressed questions in systematics and taxonomy of the time. For example, were some European lizards simply geographical variants (subspecies) or true species? He thus became interested in the whole question of species definition and of genetic reproductive barriers between them. That is why he tried to breed hybrids in captivity and then see if their offspring were fertile.

He came under the influence of the German herpetologist Wilhelm Georg Wolterstorff (1864–1943), a major player in promoting amateur herpetology through the keeping of live animals and in linking amateurs with professional museum scientists, particularly in German-speaking countries. Wolterstorff was particularly interested in newts and salamanders and realised the value of hybridisation experiments. In 1903 he reported that a long-recognised species of newt, Triturus blasii, from western France was in fact a naturally-occurring hybrid between male Great Crested or Warty Newt, Triturus cristatus, and female Marbled Newt, T. marmoratus. Lantz took up the challenge of breeding such hybrids in order to investigate what was going on in terms of genetics in the zone where the northern T. cristatus and the southern T. marmoratus overlap and in which hybridisation sometimes occurs.

I will not describe further Lantz’s work on the lizards of the Caucasus, much of it done in collaboration with the Swede and fellow chemical engineer, Otto Cyrén (1878-1946) and which continued after he left Moscow in 1918. That and other aspects of his work, including keeping and breeding the Common Chameleon (Chamaeleo chamaeleon), have been extensively covered in the 2017 biographies. I shall instead, add to that information by indicating his involvement with the highest levels of research in genetics and cytogenetics in Britain.

A clue to the importance placed on Lantz’s work with newts can be found in the following quotation:

A few people have the capacity for noticing the exceptional very highly developed. I think particularly of my late friend L. A. Lantz, who was secretary of the British Colour Council. This is not, as might be thought, concerned with the rights of immigrants from tropical countries but with textile dyes; and its secretary doubtless had a fine eye for colour differences. He also had a fine eye for lizards and newts. His most remark­ able achievement in this respect was perhaps noticing that he could not tell the sex of a lizard which he saw running about, catching and killing it, and finding that it was an intersex. 

This is from an essay, On Expecting the Unexpected, published in the Rationalist Annual in 1960 and included in the collection, Science and Life, published as a book in 1968. It was written by J.B.S. Haldane. As I said, the top level of evolutionary genetics at the time. But Haldane (and his second wife, Helen Spurway (Haldan [sic] in the 2017 biography in English) was not the only Fellow or future Fellow of the Royal Society that Lantz was supplying with animals and collaborating with. He worked with Michael James Denham White (1910-1983, FRS 1961) then at University College, London, and more significantly with Harold Garnet ‘Mick’ Callan (1917-1993, FRS 1963) on the cytogenetics of his hybrid newts. Newts have large chromosomes and are particularly suitable for studying meiosis; the number of chiasmata between pairs can more easily be counted. After Lantz’s death, Callan and Spurway continued their collaborative work on meiosis in hybrids between different geographical ‘races’ of Triturus cristatus. I have written elsewhere about the people Haldane recruited in the late 1940s to work with Helen at University College London in breeding newts (and fish). The story can be found here. 

Not apparent from the biographies was the many years needed for Lantz’s cross-breeding experiments. From an initial breeding in 1944—in wartime Manchester where Lantz was, as Haldane noted, appointed by de Gaulle as representative of the Free French Forces—the young newts only reached maturity in 1947 and those from back crosses made in that year in 1949-50. Fruit-flies, with a generation time of 1-2 weeks, they were not.

Interest in what determines the skin colour and pattern in amphibians was high in the first half of the 20th century. Paul Kammerer—another protégé of Wolterstorff—had claimed, certainly erroneously and probably fraudulently, that the pattern of stripes and spots on salamanders was determined by the environment. The examples he provided were soon shown to be of salamanders from different parts of the geographical range and that the major differences were genetic in origin. The physiological, i.e. short-term, mechanisms that cause a frog to change colour when moved, say, to a pale substrate, were also becoming clear. Lantz set out to determine what effect a light or dark background would have on larval salamanders and whether changes in colour or pattern were carried into adulthood. He began this work in 1938 on Triturus cristatus. His paper was published posthumously in Haldane’s journal, Journal of Genetics in 1953†.

Lantz also showed that other claims of Kammerer’s, oviparity in viviparous lizards and ovoviviparity in salamanders, were part of the normal plasticity of the species involved in different parts of their geographical range and not novel, acquired, heritable traits.

It was not only his success with newts that attracted attention. He was also breeding the Painted Frog, Discoglossus pictus, at home in Manchester. Alan (Sir Alan from 1968) Parkes (1900-1990, FRS 1933) and Hilda Bruce (1903-1974) (of the famous Bruce Effect in mice) of the National Institute of Medical Research got to know of this and were keen to explore the potential of a species which breeds spontaneously in the summer or can be induced to breed in the winter under laboratory conditions. Lantz supplied them with frogs and he wrote a note as an annex to their paper which described how the adults and tadpoles were kept and bred. Neither Lantz nor Bruce and Parkes succeeded in rearing the metamorphosed young. This problem, common at the time, was almost certainly attributable to deficiencies of calcium and Vitamin D. Lantz did though release young into his garden where they thrived and spread into neighbouring gardens.

Also in touch with Lantz was C.H. Waddington (1905-1975, FRS 1947) who had earlier used Discoglossus pictus for pioneering work in developmental biology. Waddington, then in Edinburgh, was taking Lantz’s advice on breeding the frogs in 1951.

Lantz did not abandon his classical herpetology nor his keeping and breeding of many different species of lizards and amphibians while in Manchester. Far from it. He collected in the Mediterranean region and the Pyrenees. He was clearly in close contact with H. W. Parker (1897-1968) at the Natural History Museum in London; some of his specimens are in the collection.

Thus, while emphasis has been on Lantz’s contributions to classical ‘museum’ herpetology, he was a more significant figure in British science.  Capitalising on his lifelong interest and success in keeping and breeding reptiles and, particularly, amphibians in captivity, he had an important enabling rôle in the research of leading British scientists working on evolutionary genetics and cytogenetics in the mid-20th century.

Louis Lantz was described by Parker in a short obituary for the journal Copeia as one of the outstanding figures in herpetology. I agree.

*There is an error in the text in that the animals are referred to as T. cristatus marmoratus rather than T. cristatus x marmoratus. Smith ought really to have said that he was showing the best available data to make his point on growth rate and the fact they were interspecific hybrids was not germane.

†The results were clear. White or black background (on which the larvae turned lighter or darker respectively) had no effect on the dorsal spot pattern after metamorphosis but the lightness or darkness of the ground colour could be changed with a change in background. For ventral coloration, the results were different. A white background suppressed while a black background promoted the development of black markings. The ventral coloration and patterning could not be reversed by changing the background in adults.

A list of Lantz’s publications is included in the 2017 biographies. However, because there seems to have been confusion when a publication in Proceedings of the Royal Society was listed and scanned for online viewing, a note of his appended to but part of a full paper has been shown as a separate publication. The correct reference is:

Bruce HM, Parkes, AS, Lantz LA. 1947. Observations on Discoglossus pictus Otth. Proceedings of the Royal Society B 134, 37-56 (plus two plates).

These are the two biographies I refer to in the text. The shorter and earlier one is:

Anon. 2007. Lantz, Louis A. (1886-1953). In Contributions to the History of Herpetology Volume 2, edited by Kraig Adler, p 139. Society for the Study of Amphibians and Reptiles.

The 2017 biography has been published twice, once in the USA in English and in a slightly longer form (with an additional author) in France in French:

Ineich I, Doronin I. 2017. Louis Amédée Lantz (1886-1953): The Life and Work of an Alsatian Pioneer of European Herpetology. Herpetological Review 48, 93–108. 

Ineich I, Doronin I, Lescure J. 2017 Vie et œuvre de l’Alsacien Louis Amédée Lantz (1886-1953), pionnier de l’herpétologie européenne. Bull Soc Herp Fr (Bulletin, Société herpétologique de France) 162, 55-106.