Shattered Crystals - Lea's Journey


My sister, Lea, served as President of the Society of Protozoologists in 2001--2002. At the society meeting in June 2003, at Salishan Resort, Oregon, USA, she gave the Past-President’s Address, discussing her journey to that role. The first part of that talk discusses her time as a hidden child and the years that followed after the war, and may be of interest to readers of this website.

The Accidental Protozoologist - My Journey Through the World of Ciliates

Department of Natural Sciences, Baruch College

ABSTRACT. The Past-President’s Address has been the opportunity for the speaker to reminisce about the road traveled to get to this time in life. In this paper, I continue in that tradition. During my journey to the present day, I visited different laboratories, studying the genetics of mating and mating types in Paramecium, Tetrahymena, Blepharisma and Euplotes. I have met and worked with many distinguished scientists, including other Past-Presidents of the Society. I also became an active participant in the Society of Protozoologists. I hope the recounting of my trip will be both entertaining and enlightening.

WHEN I was elected President of the Society, one of my reactions, after the first, of feeling enormous gratitude and pleasure, was that I would have to give a Past-Presidential Address. What would I say?

I first joined the Society of Protozoologists as a graduate student in 1963. I was encouraged to do so by my mentor, Tracy M. Sonneborn. He felt that the meetings were an excellent place for graduate student presentations of their research. He was right. But more than a forum for putting research into context, these meetings also offered opportunities to learn how others were inspired to become scientists. This was particularly true during the Past-Presidents’ addresses. Before presenting either an overview of their research career, or a summary of their most recent research, their talks generally started with reminiscences of how and why he (and occasionally, she) was inspired to study biology in general, and protists in particular. One of the best examples of that genre was given by R. Barclay McGhee (who like me was Secretary before his Presidency) shortly before his death, in 1982 (McGhee 1983). He reminisced about what he called his improbable evolution from a “Tennessee Hillbilly” to a Distinguished Professor. These talks were one of the most wonderful aspects of my early Protozoology meetings. I would like to continue in that tradition.

I needed, as they say in Show Business, “a Hook.” Then I noted that several of my research collaborators had been Presidents or Other Officers of the Society. How did this come to pass? First, I have never been a Principal Investigator, with a laboratory and students of my own. I perforce became a “Visiting Scientist” in the laboratories of Ciliate Geneticists. Second, Ciliate Geneticists were successful candidates for President. So my talk will be about my life, and also some reminiscences about the field of Ciliate Genetics.

As an historical aside (since this is a past-presentation), the reason for the Geneticists being chosen as candidates was to encourage their followers to join the Society. Although Sonneborn was invited to the Chicago meeting that served as the origin of the Society (Corliss 1998), he declined to join. Ciliate geneticists (now calling themselves Ciliate Molecular Biologists) separated and formed their independent group, thus, what might be called, by population geneticists, the founder effect: Sonneborn in 1966 convened their first meeting — at Shelter Island, and they have continued to meet as a separate group. Imagine today if they had joined the Society .... what would we call ourselves, in our occasional and continuing debates about the name of the Society and the Journal?

Let me now start at the beginning of my life. I was born in Halle, Germany, November 9, 1936. My family is Jewish. Exactly on my second birthday, “Kristallnacht”, the destruction of Jewish businesses and the roundup of Jewish men to concentration camps affected us directly. The next year, 1939, my family (just barely) escaped to France as World War II was starting in Europe. We were helped by the underground Resistance and by an organization that ran Children’s Homes for orphaned Jewish children, the OSE (Oeuvre de S;aaecours aux Enfants). My parents volunteered to work in one of the OSE Homes, and my two older sisters and I were able to stay with them. In 1941, just prior to Pearl Harbor, my sisters, Eva and Ruth, aged 10 and 11 respectively, came to America on one of the last ships from Europe in a “Kindertransport” program. Children were placed in foster homes under a program partially helped by the American Friends (Quakers) Service Committee. My mother was reluctant to let me go as I was only 5 years old. Eventually, my parents and I were separated. They were in various detention camps and war factories in occupied France. The story of their survival has been recorded by the Spielberg Foundation and written by my sister Eva as part of a series of published Holocaust Diaries (Kanner and Kugler 1997). I was for a time a so-called “Hidden Child”, living in a convent under a false (non-Jewish) name. I don’t remember much from the years of World War II, except a very strong desire to join my sisters in the United States of America. My wish was granted, and in 1946, my parents and I arrived in New York City and were reunited with my sisters. We moved to Brooklyn, and our one bedroom apartment became the family home for the next 50 years. I was determined to become thoroughly Americanized, and quickly put my German and French behind and learned English.

It is now the 1950’s, and I am attending Midwood High School, and very much interested in Science, especially Biology. I admit that my primary interest was in learning about human structure and function (especially reproductive) as the Biology class seemed to be the best place to become educated in these matters. I also registered for a Physics class, in which I was the only girl. I was made to feel so unwelcome that I ran crying to the dean, who allowed me to drop Physics and take typewriting instead. This turned out to be my most valuable high school class (think computer keyboards!). Like many immigrants, my family was poor, but supportive of education as a way to advance and be successful. My choice of major in college ( I attended Brandeis University on scholarship and work-study) was to continue Science studies, as the most practical way of insuring that I would have a profession. Luckily, I loved my Biology classes. I also managed to get my only A;pl in the Physics class. However, I mostly registered for non-science classes. In college, I took no courses in Protozoology. I did enjoy the Invertebrate Zoology class, especially the text “Animals Without Backbones” ( Buchsbaum 1987). I have never forgotten a photograph of Tarantulas (p 360 in the 3rd edition) , and the caption pointing out that “No one on record has ever objected to these same characteristics ;oblong legs and hairiness;cb in a Wolfhound.” Readers who know me personally will recollect that I ended up the mistress of 3 Irish Wolfhounds, including my favorite, Darwin.

This is a Protozoology presentation, so I now will return to smaller organisms. How did I come to specialize in these wee beasties—and true to form, in their mating habits?

This is due to another aspect of life in the late 1950’s and early l960’s. For most of my college female classmates, it seemed that the college experience would be considered completely successful only if one attained not only a B.A. but a “Mrs.” In 1958, I received both. We moved to New York City, where my husband, Michael A. Bleyman, received his degree as a Philosophy Major and started law school. During those two years, I was a graduate student in the Zoology department of Columbia University. My most memorable accomplishment that first year was that, during the long subway commute from Brooklyn to the Upper West Side of Manhattan, I was able to completely read Tolstoy’s War and Peace. It was at Columbia that I first heard reference to Sonneborn. I took a course with Theodosius Dobzhansky, one of the key geneticists of the so-called “Modern Synthesis”, the others being Ernst Mayr (1942) and George Stebbins (1950). The synthesis referred to a modern theory of evolution based on the combination of new genetic knowledge with the concept of natural selection. Dobzhansky’s book was entitled “Genetics and the Origin of Species” ( 1937). In the preface to the 3rd edition (1951), he pointed out that the fields of genetics, systematics, paleontology, ecology, embryology, and comparative anatomy have been integrated into evolution.. Just two years pre-Watson and Crick (1953), he noted that only the fields of physiology and biochemistry still remained little influenced by the evolutionary approach. He predicted they would be. After all, it was Dobzhansky (1973) who said “Nothing in Biology makes sense except in the light of Evolution.” I completely agree with that statement.

The “Modern Synthesis” left unsolved a precise definition of species. A symposium was held in 1955 entitled “The Species Problem”. Nearly half the pages of the published volume were taken by an article by Sonneborn (1957). It was here that Sonneborn gave fullest expression to his objections to a limited view of species and introduced the term “syngen” for those groups of Ciliates to what we today actually have given Species names (what Mayr called sibling species). A year after the publication of this paper, I sat in Dobzhansky’s Genetics and Evolution class and listened to him lambaste Sonneborn’s concept. This was my introduction to Ciliate Genetics.

I left Columbia after 2 years (M.A. 1961) for the same reason I attended. I was following my husband in his educational migrations. As fate would have it, he decided to return to Philosophy, left New York University Law School and headed for Indiana University, Bloomington, which was a center for Positivist thinking, to study with Michael Scriven. I enrolled in the Biology (then Zoology) Department, completely unsure of anything except a vague interest in “cell biology.” It was a wonderful time to be at Indiana. James D. Watson (Ph.D. ‘50) had recently graduated, leaving behind a reputation as someone who never took notes, but aced everything. H. J. Muller was there, teaching an evolution course so hard that it was acceptable to get a C and still stay in Graduate School. I heard about Friday night seminars at Sonneborn’s house. I learned about Briggs and King’s (1952) transplantation experiments of diploid embryonic nuclei into enucleated frog eggs. I promptly signed up for Sonneborn’s Genetics of Protozoa and Brigg’s Experimental Embryology courses. Then, something happened that made me into a Ciliate Geneticist. All my frog embryos died and all my Paramecia happily mated and reproduced. I went to Sonneborn’s Friday night seminars and my fate was sealed. The seminars were held in the Sonneborns’ house and featured his students presenting their research for what was very pointed criticism and often helpful suggestions. The focus in the Paramecium studies was on nuclear differentiation, nucleo-cytoplasmic interactions, and life cycles. Thus, mating types, serotypes and later, corticotypes were studied. I became part of this group, and was assigned (he worked on the one syngen, one thesis model) my very own Paramecium aurelia syngen: ;ns5. I had to determine the pattern of inheritance of the Odd and Even mating types (labeled IX & X according to the protocol for all the syngens). Most of Sonneborn’s graduate students were in a suite of laboratory rooms. The last room in the suite was the Post-Doc/Visiting Scientist room, which happily for me was next to mine. I became friends with Akio Miyake and Koichi Hiwatashi. It was Hiwatashi who suggested using the vital dye Neutral Red so I could distinguish the two mating types in my cultures. This turned out to be the crucial way to prove that a single cell could change from the odd to the even mating type in the absence of cell division (Bleyman 1967a). This led to “selfing”: intra-clonal mating between cells descended from the same macronucleus and thus genetically identical (Bleyman 1967b). I was not able to explain the mechanism for the mating type change, but the phenomenon of selfing occurs in other ciliate species and it would be a recurring interest of mine.

Working around the corner from me was James D. Berger, the first (although in the future) of Past-Presidents of the Society I have known. He was as intense and dedicated to his research into nuclear differentiation as he continued to be throughout his career studying the cell cycle (Berger 2001) as when he was President of the Society (1998--1999). Down the hall, working with W. J. Van Wagtendonk, in the Biochemistry laboratory was Anthony T. (Tony) Soldo, President of the Society, 1985--1986. Their challenge was to find a completely defined medium for Paramecium, to substitute for the bacterized infusion of either grass (Cerophyl) or baked lettuce in which it was being grown. This was necessary for Sonneborn’s hope that Paramecium aurelia would be the eukaryotic Escherichia coli. By this, he meant that many laboratories would use this one organism to thoroughly explore its genetics and biochemistry. The 1960’s was the decade when molecular genetics became firmly established, and when E. coli was the organism of choice. At that time, I gave a graduate student journal club talk on Marshall Nirenberg’s work deciphering the genetic code (Nirenberg and Matthei 1961). Sonneborn became so excited he promptly invited him, along with other luminaries and future Nobel laureates (Gobind Khorana, Severo Ochoa, Fritz Lipmann, and Seymour Benzer) for a Symposium on RNA coding. I was introduced to Nirenberg as “the little girl who started it all.” Nirenberg was visibly shocked, not realizing that the “it all” was the Symposium. My only contact with Francis Crick was a few years later, when he was doing a Midwest lecture circuit. He came to Illinois after Indiana, and at that time, I was there. I asked him a question, and after he requested that I stand up ;ob”I am standing up, I replied”;cb he listened carefully and said: “Sonneborn asked me that same question in Bloomington!” I would say my training was quite deep, and any influences of Dobzhansky long gone.

We have reached 1963, the year I joined the Society, and presented my first talk (Bleyman 1963). That year, the Society met with AIBS at the University of Massachusetts, in Amherst. In 1964, I left both Indiana and Paramecium, Ph.D. almost completed, (Ph.D. 1966), for the University of Illinois, Urbana. I am still following my husband. He has left Philosophy (I admit partially under my influence, and mostly because H. J. Muller took an interest in him). We both spoke to Sol Spiegelman, who steered Michael to Carl Woese and me to David L. Nanney. Thus did I leave Paramecium, and enter the world of Tetrahymena. I had first heard about this ciliate when I met Joseph Frankel ( President 1989--1990). He came to Indiana and gave a seminar on his cortical studies on the amicronucleate strain of Tetrahymena. Joe Frankel and I may even be distantly related. (When I told Sonneborn this, he replied, “No wonder Joe is so smart” — I was enormously flattered.)

It was not difficult to switch organisms. The only real adjustment I had to make was visual—compared to Tetrahymena, Paramecium are gigantic. Both organisms share the ciliate property of nuclear dimorphism: the genetic and reproductive micronucleus and the somatic and functional macronucleus, both in a common cytoplasm. It is this that makes these organisms excellent for studying nuclear-cytoplasmic interactions and nuclear differentiation.

The University of Illinois was a much more protistological place than Indiana University. John Corliss (President, 1964--1965) was there, before leaving for the University of Maryland. I met Jerome (Jerry) Paulin (President, 1988--1989) who, like both Corliss and myself had also been Secretary of the Society. Their interests were morphological and systematic. The Nanney lab was primarily orientated toward Genetics. Yet systematics could not be escaped. Like Paramecium, Tetrahymena subgroups began as syngens, determined by their mating types. Similarly, with advances in biochemistry and molecular genetics, both these “;auuber species” became individually named species. My old syngen 5 became Paramecium pentaurelia. Tetrahymena thermophila was born. Tetrahymena pyriformis was reserved for the amicronucleate strain (the so-called “immortals”) (Nanney and McCoy 1976 ; Sonneborn 1975).

I spent five wonderful productive (and reproductive: my daughter Anne was born in Illinois) years in Nanney’s laboratory. Nanney also had been Sonneborn’s student, but was determined not to be a Sonneborn clone. After completing his studies at Indiana, he was hired by the University of Michigan, and met Alfred M. Elliott and started studying Tetrahymena. My connection to Elliott, another Past-President (1956--1957) is indirect, through Nanney. In his past president’s address (Elliott 1959), Elliott reviews the early Tetrahymena studies and also discusses “the species problem,” which arose in Tetrahymena when the micronucleate strains were discovered. The approach to studying the biology and genetics of Tetrahymena and Paramecium were similar, and, in my opinion, Nanney not only became more than “Sonneborn redux”, but succeeded in accomplishing Sonneborn’s dream of establishing a ciliate organism which would serve as a “eukaryotic E. coli.” Tetrahymena thermophila ultimately became that organism. This was due, in large part, to two of his students, Eduardo Orias (who studied with Nanney at Michigan) and Peter Bruns who was at Illinois contemporaneously with me. I was a research collaborator with both. I also interacted with Orias when he was President (1997--1998) and I was Secretary. Another reason Tetrahymena became the organism of choice was, and remains, the ease of growing the cells in axenic culture, usually a simple proteose-peptone broth. Telomeres and ribozymes were in the future, but I remind you that these key phenomena were first described in Tetrahymena (see respectively, reviews: Blackburn 1991; Cech 1987.)

My main co-worker in the laboratory was Ellen Simon. Ellen and I studied selfers as a way of getting an insight into nuclear differentiation (Bleyman and Simon 1968; Bleyman et al 1966). One important difference between Paramecium and Tetrahymena mating systems was that in the latter, there was no longer the binary (Odd vs. Even) two-type pattern. Tetrahymena thermophila has a multiple mating type system. The seven possible mating types are specified by the mat locus. Ultimately, the macronucleus is programmed to express one mating type. In some cases, a clone expressed more than one mating type, leading to intra-clonal selfng. The model we worked with was that the macronucleus was organized as sub-units undergoing phenotypic assortment That is, the compound macronucleus can express more than one phenotype, and with subsequent cell divisions , will ultimately give rise to pure (non-selfing) clones. The pattern of assortment becomes a clue as to the number and behavior of the sub-units, and possibly the timing of the determination of mating type. The Macronucleus is 45-ploid, and is now described as containing 250--300 acentromeric chromosome pieces (Orias 2000).

I also became interested in life cycles, since the micronucleate strains of Tetrahymena clearly underwent aging, and had a period of adolescence, prior to becoming sexually mature (Bleyman 1971). Ellen and I discovered an early maturity mutant (Bleyman and Simon 1967), and I studied the phenomenon in Euplotes as well (Dini et al. 1990).

My official position in Nanney’s laboratory was Research Associate. This entitled me to a faculty parking space and almost nothing else. When I asked about maternity leave, I was told that my faculty status did not include that benefit, which was only available to non-faculty staff. The assumption clearly was that faculty members were male and had no need of such leave.

Leave Illinois I eventually did, again following my husband, this time to the University of North Carolina in Chapel Hill. I again found a Research Associate position, but no protozoologists to work with. It was during my time in North Carolina that I divorced Michael. Although this was personally painful, it was professionally liberating. I decided that I would no longer be a Research Associate, but find a tenure track position. My preference was for a return to New York City, where most of my family still lived. At this time, the City University of New York was expanding, and the business school (alias downtown City College) became a full-fledged college: Baruch College. The Biology department was in the market for a Geneticist. It was a perfect match, and I was hired in 1973. There were no research facilities, which meant that although I now had a permanent position, my traveling ciliatologist days were not yet done. That summer, I attended the IVth International Congress of Protozoology in Clermont-Ferrand, France. During my talk, Tracy Sonneborn walked in, and I was so surprised and nervous, that for the first (and last) time in my life, my palms started sweating. Another acquaintance from my Indiana days, Akio Miyake, my suite neighbor was there. He had a laboratory in the Max Planck Institute f;auur Molekular Genetik in Berlin. When he heard I was divorced, and no longer bound to be a dutiful wife, he immediately invited me to spend next summer with him. This meant my learning to work with a new ciliate, Blepharisma, and to return to the country of my birth.

Miyake asked me to bring different strains of Blepharisma for our experiments. He suggested that I get them from Henry I. Hirshfield at New York University. Since I also needed a research space, I asked, and was invited to become a Visiting Research Scientist in his laboratory. I now started to study mating in Blepharisma. They seemed enormous after Tetrahymena, and handily came in albino and red strains, obviating the need for neutral red. In 1974, with 19 strains of 5 different species, I flew to Berlin for the summer. At that time, the city was still divided into sectors and by the Berlin Wall. I introduced myself as an American, and said nothing of my background. I could no longer speak German, but understood quite a lot. Luckily, English is the lingua franca of science, and Akio spoke almost no German as well. We used the large number of strains to classify Blepharisma according to their production of extra-cellular mating substances, called gamones by Miyake. The mating type system is binary and the difference between species was the identity of mt I gamones. All the mt II cells secreted the same molecule. By the time our work (Miyake and Bleyman 1976) was published, Akio had left Berlin for Italy.

My time with Blepharisma was temporary. In terms of investigating the genetics of mating, it was difficult to establish any genetic pattern. I did successfully mate in another sense. I met my second husband, David Minn during my NYU days. The next summer (1975) I was invited to Cornell University to work with Peter Bruns. I now returned to Tetrahymena genetics. Around this time, Orias, who also temporarily had left Tetrahymena as a research subject, returned to this organism, and with Bruns, published a key methods paper (Orias and Bruns 1976). All aspects of the nuclear duality of the organism were now exploited, and I considered myself lucky to be able to participate in the research. I spent the next 10 summers in Ithaca. Peter developed the use of nulllisomic strains, lacking both copies of one or more of the five chromosomes (or in some cases a chromosome arm) in the micronucleus. These strains are useful for genetic mapping ( Bruns and Brussard 1981). The strains are viable because the somatic macronucleus retains and expresses the genetic information. I returned to my first love, mating, to map the mating type locus. Eventually this led to a fruitful collaboration with Orias when we not only mapped the mat locus to chromosome 2, but established meiotic linkage to the ribosomal RNA gene (Bleyman et al, 1992.)

I had now established a pattern of combining the Baruch College teaching duties during the academic year (3--4 courses a semester) and summer research at Cornell University in the Bruns laboratory. However, I kept a connection with Hirshfield which was inadvertently the start of my real involvement with the Society. The Vth International Congress of Protozoology was scheduled for New York City, in 1977, and Hirshfield was invited to be on the local organizing committee. He declined, and sent me instead. Here I was, helping in a small way, and getting to know the other committee members, including William Trager (President, 1960--1961), Seymour Hutner (President, 1961--1962) and the Secretary-General of the Congress, John Lee (President, 1990--1991). When Hutner needed some help with the editing of the proceedings of the Congress, I came to the Haskins Laboratory to help out ( Hutner and Bleyman 1979.) I stayed in his laboratory, and while there, followed up on some ideas for easily keeping Tetrahymena in culture that had been developed independently by Norman Williams (President, 1984--1985) and Jason Wolfe (Williams et al. 1980).

At this time, I was elected to the Executive Committee of the Society (1981--1986) and then Secretary (1991--1997). I also ended my traveling to upstate New York, and traded it in for travel uptown to the Bronx . There I joined Birgit Satir (President, 1987--1988) and continued research on Tetrahymena. We exploited the nullisomic method to map an exocytosis mutant (Bleyman and Satir 1990), which had been isolated when Orias worked in her laboratory (Orias et al. 1983). This exemplifies the great spirit of cooperation and interaction in this field.

In the fall of 1989, I took a sabbatical and went to the University of Camerino, Italy, at the invitation of Piero Luporini. My goal was to use this time to write a review on the Genetics of Ciliates. I was invited to do so by Phyllis Bradbury (President, 1977--1978). She told me that part of the reason she asked me was that she wanted more women represented in the final volume. My review (Bleyman, 1996) summarizes the cellular approach, and will most likely be the final survey of the field of Ciliate Genetics with a non-molecular emphasis. I relied on J. R. Preer, Jr. (President, 1986--1987) for a final reading of the manuscript. This was most appropriate, because Preer, one of Sonneborn’s students, was now his replacement at Indiana, and had successfully combined classical and molecular Genetics in Paramecium (Preer 1986). It was also helpful to me to be able to talk to Geoffrey Beale, who was an annual visitor to Camerino and a pioneer in Paramecium Genetics (Beale 1954) The last part of my time in Italy was spent in Pisa working with Fernando Dini on Euplotes. He and one of his students had found some strains of E. crassus that manifested early maturity. Our analysis of the data showed that unlike other early maturers, inheritance of this trait was transmitted via the cytoplasm, not the nucleus (Dini et al 1990). It was when I was in Italy, on my birthday, November 9, 1989, that the Berlin wall came down. When I returned to Berlin for the IXth International Congress of Protozoology, in 1993 , I was able to travel to Halle, my birthplace. I only knew what my parents had told me, and was struck by the difference between their memory of a vibrant town and the drabness I saw. At least the town square still had the statue of Halle native G. F. Handel, rather than one of Lenin or Stalin. Perhaps those had been toppled.

In 1999 I was elected to serve on the Executive Committee for a new 5-year term, and had looked forward to continuing to serve the Society in that capacity until my retirement. I accepted the nomination for President reluctantly, but was pleased and honored to be elected for the 2001--2002 term. As President-Elect, I organized a symposium on the Tetrahymena Genome. This demonstrated that Tetrahymena Genetics has indeed also traveled far. In March and April of this year, 2003, grants for Tetrahymena macronuclear Genome-Sequencing have been awarded by The National Science Foundation and by the National Institute of General Medical Sciences. As Orias (pers. commun.) put it in his announcement to the Ciliate research community, this “should increase Tetrahymena’s credibility as a model organism.” Although other organisms are in contention as prime eukaryotic models (old standbys like Drosophila and Yeast, and newcomers like Caenorhabditis elegans ;obthe sole survivor of the 2003 Columbia shuttle crash;cb and Arabidopsis thaliana), Tetrahymena has also arrived.

At this time, I have also arrived, or more correctly, ended my journey. The take home lesson of my personal journey illustrates the Yogi Berra saying “If you come to a fork in the road, take it.” To paraphrase another cliche, when the door of opportunity opened, I walked in. I hope all enjoyed following me through.

For more on Lea's time as a hidden child, see this excerpt from Shattered Crystals.

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