In Fall, 1971, I took the University of Wisconsin ­p; Madison Medical
School histology course (Anatomy 710 "Cells and Tissues"), lecture
and lab. Medical school courses had multiple instructors, and one was Allen
W. Clark. Later, he agreed to teach me transmission electron microscopy,
(a lot of work for him and for me!). I wanted to find out if the visual
synapses of two non-phototactic Drosophila mutants (tan and ebony) were
missing visual synapses. I could tell a volume of stories, but I will only
Allen had done electron microscopy (for instance, Clark et al. The ventral
photoreceptor cells of Limulus I The microanatomy, J Gen Physiol, 1969,
54, 289-309). When I worked with him, he was doing electrophysiology on
the effect of black widow spider venom on the neuromuscular junction. When
I sat down at the dissecting microscope to prepare Drosophila heads, there
were the spiders with their hour-glass abdomens, and I hoped they were dead
and their venom glands removed.
Eventually, I obtained sections in an area that I thought might have visual
synapses, but the most conspicuous structures were unknown to me (and to
Allen [and to the departmental chair David Slaughterbach]). I eventually
read some papers and found that these structures were called capitate projections
(Trujillo-Cenoz, 1965, J Ultrastructure Res. 13, 1-33). My many electron
microscope studies, described below, include an entire paper devoted to
capitate projections (Stark and Carlson, Cell and Tissue Research,1986,
246, 481-486). The work with Allen Clark was done while I was a graduate
student, and, while I was a graduate student, I met Stanley D. Carlson who
would be central to my later studies (see "memoirs" in this link).
While I was an Assistant Professor at Hopkins, I published this work with
Clark (Drosophila Information Service vol 50, pp. 105-106, 1973 ). It turned
out that the synapses in tan and ebony were indistinguishable from wild
type. The "article" was an un-refereed research note in a journal
that most researchers did not take seriously. I had been told several times
"Don't bother. Someone in Benzer's lab did that work." I eventually
met that someone, Hansen, who had stopped publishing after getting tenure
While I was an Assistant Professor at Hopkins, I thought it would be useful
to develop EM techniques as part of my program of Research. Tina Ivanyshyn,
an undergraduate, helped. I guess she was into equal rights since one time
she did fixations on female Drosophila instead of males except that the
mutant was maintained across from attached X, so the female was not mutant.
Eventually, she got some nice pictures, and I included one in a grant proposal
submitted in 20 copies, each micrograph carefully produced in the dark room
and carefully taped to the page. A day after submission, the grants officer,
Mr Proctor, called me in and showed me a mangled mailing label. "We
sent in six that day and one was lost in the mail, but we won't know which
one until we get the received notification for the five received and that
can take a month. We've contacted all six grant officers, and they have
agreed to give us an extension." Wouldn't you know it, it was mine,
and I found out a day or two before my family vacation. Rush order in the
dark room and in the production. I drove it from Baltimore to Washington
and plopped it on the NSF desk in person. Eventually, my lost proposal (20?
Copies) came back in a basket mixed with other items of mail.
EM did not get serious for me until 1981. I was at the University of Missouri,
and, by that time, my friend Stan Carlson at Wisconsin had become an expert
in several EM techniques. It is impossible to thank Stan
Carlson as much as he deserves! I applied for and received a Faculty
Development Award for $1,934. I went up there with the goal of looking at
the synapses of the rdgB (retinal degeneration) mutant to see if the post-synaptic
cells degenerated, a question left open after the studies with Bill Harris
(1976, 1977). They do not degenerate (Stark and Carlson, 1982), so the trip
was a success, and Stan pushed the work to publication so quickly. I was
there Aug 2-29, stayed at the campus Y (very cheap) and then at the DeJonghs
(friends from graduate school) when the Y needed space for students after
Wisconsin's semester started. I learned so much! I was scared at all the
tricks I would need to master. Oh, I could tell so many stories! But I will
tell just a few:
Stan's wife, Che Chi, had done her PhD using EM on fly visual system. She
came in one evening (after her work) and showed me how to section to the
correct location, and that eventually became my methodology.
I did some work at the high voltage electron microscope (HVEM). Founded
by Hans Ris, this was an NIH Biotechnology Resource, so they were in the
business of being hospitable (financially) to guests. My travel expenses
for this August trip were complicated, and SM in the deans office helped
with my travel expense voucher. Several years later, rumor had it, she got
in trouble for some creative financial dealings.
Madison, WI is so nice in the summer! Ice cream at Babcock Hall (near where
I worked, the Department of Dairy Husbandry in the Dairy State). Brats at
the Memorial Union next to Lake Mendota with a rock band every evening.
Jogging in Picnic Point or in the Arboretum. (but better bring plenty of
blood since the mosquito is the state airline) Walking up and down State
Street, pretty much like a boardwalk, after work. True I was away from my
family, and my housing was dormitory style, but in later years at places
right next to Lake Mendota. All my subsequent trips, mostly in the summer,
were shorter. I used any excuse to get up there in a collaboration that
lasted about 20 years. So, obviously I need to condense hundreds of stories
to just a few.
Philippa Claude, at the Wisconsin Regional Primate Research Center, had
a freeze fracture apparatus that Stan and I used in 1983 and again later.
My friend from graduate school, Joe Kemnitz, worked at the Primate Center
(and became Director years later [and had become my older son's Godfather
years earlier]). We answered the question whether the diminished photoreceptors
of the outer rhabdomeres absent (ora) mutant had rhodopsin (Stark and Carlson,
1983) [they do not]. I remember one amazing detail about a later trip. We
were ready to work, but there was no liquid nitrogen that morning. For perhaps
the only time in my career, I felt like Superman when it came to ordering,
"OK, Badger welding is leaving a tank at the loading dock at 1 o'clock."
I became an Affiliate Scientist at the Primate Center; they liked my contributions
to their annual reports since I published. Eventually, Philippa had to get
rid of the freeze-fracture apparatus, so she gave it to me. I went up for
a day in January 1991 with my wife and my younger son to make arrangements,
and the highlight was a Badger hockey match.
The HVEM was a way to make microscopy easy since we used thick sections
rather than ultrathin sections, so my initial love affair seemed like cheating.
One time, Missouri gave me about $240 for a plane ticket, and HVEM gave
me about the same amount, so my (cheap) expenses were covered. (I had to
fly the first few years since we had only one car, but later I drove.) The
NIH Biotechnology Resource evolved (and they mothballed the HVEM), became
IMR, Integrated Microscopy Resource, then LOCI
(Laboratory for Optical and Computational Instrumentation). So many people
helped me so much! Eventually, I tried all their tricks but focussed a lot
on confocal microscopy at LOCI with a huge amount of help from Charles Thomas.
By the mid-1980s, I did electron microscopy in my own lab with Randy Sapp
as my grant-salaried technician.
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