A publication of the Center for Reproductive Science
Northwestern University, Evanston and Chicago
Winter 2000 Volume 1 No. 2

From the Director
Seminars
The 1999 Minisymposium
Go-to-Meeting Travel Award for Undergraduates
Storch Scholarship Fund
Focus on Undergraduate Research
CRS Research Notes: "Linking Basic and Clinical Research on Sleep and Circadian Rhythmicity"
Recent Publications

From the Director

We are continuing to work on our web-site, increasing linkages and information. We have begun to receive applications for travel to professional meetings from undergraduates. Let me remind you also that the Storch fund has money to support the research of undergraduates working on ovary-related projects in the laboratory of a center member.

The big fall event was our Minisymposium - described on page 2 .

In our previous issue we listed the faculty members who are members of the Center. In this issue I would like to introduce the three staff members who make the Center possible.

Susan Hall-Perdomo, who has been in the Center since May 1989, is the Center Administrator. She has been a key-player over the years as the grant portfolio expanded from one grant to many. Ms. Hall-Perdomo came to Northwestern University in 1987. She earned her Bachelor of Science degree from Northwestern while working full-time in the Center. She brought to the Center her enthusiasm, academic experience and several years of office experience. She now wears many hats, from hiring personnel to helping with grant proposals. All of the daily activity in the Center is under her responsibility. She has also been a committee member for the Take Our Daughters to Work Day at Northwestern University for the past four years.

Mukhtar Rafiqi has been the secretary in the Center since 1991. Mr. Rafiqi came to Northwestern University after working twenty-one years with the Arabian American Oil Company (Aramco) in Saudi Arabia. He has performed various duties in the Center over the years from accounting to supervising the Minisymposium. Mukhtar has significant computer experience, and he is now in the process of updating the Center's website.

Ila Allen joined the Center in December 1998, as an accounting assistant. She brought with her many years of accounting experience. Ila handles the ordering and paying of invoices for all of the grants, and she makes sure all the work-study students get a paycheck.

Remember- with growing concern about an increasing world population and a deteriorating environment—
reproduction matters!

[Website (developing) http://x.biochem.nwu.edu/CRS/htm] —Neena Schwartz, Director

Seminars

3/13/00 Dr. Ok Kyong Park-Sarge "Steroid receptors: ovulatory overture", 4:00 PM, Frances Searle 3-147, Evanston Campus

3/16/00 Dr. Graeme Bell "Diabetes mellitus: a model for genetic studies of common diseases", 4:00 PM, Prince Faculty Commons, Chicago Campus

4/4/00 Dr. David Albertini "Building better gametes is not just a germ cell thing-interplay of somatic and germ cell lineages in the mammalian ovary", 4:00 PM, Ward 4-075, Chicago Campus

4/6/00 Dr. John Achermann "Steroidogenic factor-1 (SF-1) and DAX-1 in human development and disease", Prince Faculty Commons, 4:00 PM, Chicago Campus

5/1/00 Dr. David Schomberg, "A matter of survival: insulin and IGF-1 action in the ovarian follicle?", Frances Searle 3-147, 4:00 PM, Evanston Campus

5/18/00 Dr. Zena Werb (BMBCB Speaker) "Title TBA", 12:00 Noon, M345 Technological Institute, Evanston Campus

10/16/00 Dr. Jeffrey Rosen, 21st Minisymposium Keynote Speaker. Seminar "Mammary Gland Development and Breast Cancer: Insights from Transgenic and Knockout Mouse Models".

MINISYMPOSIUM - 1999

The 20th Anniversary Minisymposium sponsored by the Center took place on October 18, 1999 in an all-day marathon, beginning with breakfast and ending with wine and cheese at 5:15 p.m. The number of papers presented by trainees in the midwest geographical area has grown from 12 in the first year to 38 this year. Eleven presentations were oral, and the rest were displayed in poster sessions. In addition to presenters from the Chicago and Evanston campuses of Northwestern, papers were also given by trainees from the University of Illinois from the Chicago and Urbana campuses, and from the University of Cincinnati College of Medicine. The day ended with a talk by Dr Phyllis Wise, Professor and Chair of the Department of Physiology at the University of Kentucky College of Medicine on "Neuroendocrine contributions and repercussions of the `menopause': a window to the aging brain". The Symposium was organized by a committee of trainees: Sonali Anand and Glenn Harris of the Department of Neurobiology and Physiology and Shane Cunha and Jennifer Weck of BMBCB, under the supervision of Muhkthar Rafiqi and Susan-Hall Perdomo.

The general consensus after every Minisymposium has been that the papers and posters presented by our trainees, undergraduate, graduate and postdoctoral, equal or exceed in quality those presented at national professional meetings, and serve as a good training ground for our students. Six awards are presented at the end of the meeting: 1st, 2nd 3rd prize for oral presentations and 1st, 2nd and 3rd for posters. The judges are a mix of faculty and postdocs. The awards are named after Constance Campbell, a former faculty member in Neurobiology and Physiology who administered our undergraduate honors program before her death in 1981. This year's winners were as follows: Oral Presentation: 1st- John Achermann, NU Chicago; 2nd- Humphrey H-C Yao, U of IL, Urbana; 3rd- Salil Ginde, U of IL, Chicago. Poster presentation: 1st- Michelle McMullen, Evanston; 2nd- Grace Ma, Evanston; 3rd- Nicole Sleiter, Evanston.

Neena Schwartz (L), Phyllis Wise (R)

TRAINEES: WHAT'S UP?

CRS Go-To-Meeting Travel Award for Undergraduates

A number of undergraduates at Northwestern work in the research laboratories of Center faculty. We have started a fund to enable some of them to attend professional meetings with their preceptors and other lab members. We invite applications for travel for the year 2000 meetings. Undergraduates should submit a letter outlining their research project and a description of the meeting to which they wish to go. They should also estimate the travel cost. Faculty advisors should countersign the documents. Please submit support documents to Dr. Neena Schwartz, Center for Reproductive Science, Hogan Hall, Evanston Campus.

Marcia L. Storch MD Scholarship Fund for Undergraduate Women

Before her death from ovarian cancer Dr. Marcia Storch expressed the wish that contributions be sent to the Center for Reproduction Science at Northwestern University to establish a fund to encourage undergraduate women to study the basic physiology and biochemistry to the ovary. With contributions we have received the Center invites applications from undergraduate women working in the laboratories of Center faculty. A committee will review the applications, which should include a brief description of projected work and a statement from the advisor, and budget for supplies (not to exceed $400). Please submit supporting documents to Dr. Neena Schwartz, Center for Reproductive Sciences, Hogan Building Evanston campus.

Focus on Undergraduate Research

Eva Ma is a junior in the College of Arts and Sciences and is currently pursuing a dual degree in biology and chemistry. Eva recognized that having a solid background in laboratory research is essential to achieving her goal to attend graduate school. Eva was attracted to the concept of applying molecular biology to the broader discipline of physiology, a concept that lies at the heart of the Woodruff laboratory ideology. Eva joined the lab in September of 1999 and is involved in the characterization of a newly discovered inhibin receptor, p120. Her main focus in the lab has been to clone a deletion series of the p120 gene and to use these constructs to gain better understanding of p120 function. In the past six months, Eva has succeeded in cloning just over half of the 13 constructs needed for our study, and has proceeded to analyze some of the constructs in transfection assays. Eva plans to continue her research into her senior year, and her future endeavors will include the use of the p120 deletion constructs for mapping the functional domain of p120 using a variety of techniques.

CRS _ RESEARCH NOTES
Linking Basic and Clinical Research on Sleep and Circadian Rhythmicity
                                            

Two members of the Center for Reproductive Science, Fred W. Turek and Joseph S. Takahashi of the Department of Neurobiology and Physiology in Evanston have worked closely for a number of years with Dr. Phyllis Zee of the Department of Neurology in Chicago to study sleep and circadian rhythms. This collaboration has involved an integration of basic research studies in rodents with the study of human sleep and rhythmicity in both a clinical research setting as well as in the normal work and living environment. Integrated animal/human research projects have focused on, 1) the effects of advanced age on sleep and rhythmicity and on the development of countermeasures for the adverse effects of aging, 2) the mechanisms by which the light-dark cycle entrains the circadian clock, 3) the importance of non-photic stimuli for the regulation of circadian rhythms and sleep, and 4) most recently the genetic basis of the circadian clock itself.

In 1994, Takahashi, Turek and their colleagues at Northwestern reported in Science the discovery of a mutant mouse which did not have a normal 24-hr period to its activity rhythm in constant darkness (DD), but instead when carrying two copies of the mutant gene had a period of about 27-28 hrs. Many of the homozygous mutant mice were also found to become arrhythmic after being exposed to DD for a few weeks, a phenomenon which is essentially never seen in wild type mice. Using genetic mapping, the mutation was soon located to the mid-point of chromosome #5, and then the hard work began. Takahashi led the large team of investigators that won the race to find the first mammalian circadian clock gene when they reported on the cloning of the gene in1997. The gene was appropriately named Clock. This breakthrough discovery was quickly followed by the identification of what appears to be at least eight circadian clock genes (and still counting) in mammals. Clock encodes a novel member of the bHLH-PAS family of transcription factors with features predicting DNA binding, protein dimerization and activation domains. It is now known that Clock dimerizes with another protein, and together they regulate the transcription of other circadian clock genes. It appears that the circadian timing so evident in the whole animal is due to an autoregulatory transcription loop that involves both positive and negative regulatory elements.

The identification of mammalian circadian clock genes now makes it possible to determine if alterations in the expression of these genes may play a role in the aging of the circadian clock system and whether mutations in these genes may underly circadian abnormalities in humans. As the Clock gene story was unfolding, Zee, who is also the head of the Sleep Disorders Clinic at the Northwestern Memorial Medical Center, began searching for families with circadian sleep disorders. Among the most common types of sleep disorders attributed to an alteration of circadian timing are the sleep-wake cycle phase disorders. As the names of these disorders imply, "Delayed Sleep Phase Syndrome" (DSPS) and "Advanced Sleep Phase Syndrome" (ASPS), people with these sleep problems find it difficult to go to asleep and wake up at the normal time. People with ASPS desire to go to sleep long before other people in their environment and then find themselves wide awake in the early morning hours. Conversely, people with DSPS are not able to fall asleep till very late in the night and then find it difficult to get up in the morning to meet the demands of their work/social schedules. Zee has identified a large family with ASPS, where the affected members of the family are about three hours "out of phase" with the unaffected members. The ASPS trait segregates with an autosomal dominant mode of inheritance, and the hunt is on to determine the specific gene responsible for this disorder.

An intriguing tie-in between the Clock gene story and sleep is that Clock mutant animals appear to need less sleep since they sleep 1-2 hrs less than wild-type animals. This finding indicates that the Clock mutation is altering not only the timing of sleep (i.e. the circadian sleep process), but also the amount of sleep the animal experiences (i.e. the homeostatic sleep process). At the present time essentially nothing is known about the genes involved in the "need" for sleep. The finding that a circadian clock gene can influence this need opens up a new avenue for the discovery of sleep genes and may lead to a better understanding of why we need to sleep at all.

There are a number of links between the circadian clock and reproductive systems in mammals. It has been recognized for many years that the circadian clock regulates the timing of ovulation in many mammals, and that the circadian clock is involved in measuring the length of the day; information which is used to time the seasonal reproductive cycle of many vertebrates. Indeed the Clock mutation also affects the reproductive efficiency of female mice. Less understood is how sleep itself impacts the regulatory mechanisms underlying reproductive function. The findings that menstrual cycle abnormalities are more prevalent in shift workers, and that sleep disorders are more prevalent in woman with polycystic ovary syndrome indicate that abnormal sleep and circadian rhythmicity may have adverse consequences for human reproduction. Elucidating the mechanisms and consequences of disrupted sleep/rhythmicity on reproductive function will require the kind of collaboration between clinical and basic researchers that Takahashi, Turek and Zee have established over many years.

References:

Vitaterna MH, King DP, Chang A-M, Kornhauser JM, Lowrey PL, McDonald JD, Dove WF, Pinto LH, Turek FW and Takahashi JS. (1994) Mutagenesis and mapping of a mouse gene, Clock, essential for circadian behavior. Science 264:719-725.

King DP, Zhao Y, Sangoram AM, Wilsbacher LD, Tanaka M, Antoch MP, Steeves TDL, Vitaterna MH, Kornhauser JM, Lowrey PL, Turek FW and Takahashi JS. (1997) Positional cloning of the mouse circadian Clock gene. Cell 89:641-653..

Zee PC and Turek (1999) Introduction to sleep and circadian rhythms, Chapter 1. In: Neurobiology of Sleep and Circadian Rhythms (FW Turek and PC Zee, eds.) Marcel-Dekkar, Inc., New York, pp. 1-18.

Recent Publications

Chen, B., A. Li, D. Wang, M. Wang, L. Zheng and J.R. Bartles (1999) Espin contains an additional actin-binding site in its N terminus and is a major actin-bundling protein of the Sertoli cell-spermatid ectoplasmic specialization junctional plaque. Mol. Biol. Cell. 10:4327-4339.

Sjogren, K., Bohlooly-Y, M., Olsoon, B., Coschigamo, K., Tornell, J., Mohan, S., Isaksson, O.G.P., Baumann, G., Kopchick, J., Ohlsson, C. (2000) Disproportional skeletal growth and markedly decreased bone mineral content in growth hormone receptor -/- mice. Biochem Biophys Res Commun 267:603-608.

Kamat, S.K., Murillo, G., Chatterton, R.T (1999) Hussain, E.A., Amin, D., Mortillaro, E., Peterson, C.T., and Alekel, D.L. (1999) Breast cancer risk factors in two distinct ethnic groups: Indians and Pakisatani caucasian versus American caucasian premenopausal females. Nutr. Cancer 35:16-26.

Fejgin, MD., Pak, SC., Flouret, G., Parsons, NY., and Wilson, L., Jr. (1998) Comparison of the in vivo activity of different oxytocin antagonists in the pregnant baboon. J. Soc. Gynecol. Invest., 5, 251-254.

Xue , J-C and Goldberg, E. (2000)Identification of a novel testis specific protein which is correlated with the meiosis, Biol of Reprod (in press)

Bernard, D.J., I.Y. Merzlyak, T.H. Horton and F.W. Turek (2000). Differential regulation of pituitary gonadotropin subunit messenger ribonucleic acid levels in photostimulated Siberian hamsters. Biol Reprod 62:155-161.

Peters CA, Maizels ET, Hunzicker-Dunn M (1999). Activation of PKC delta in the rat corpus luteum during pregnancy: potential role of prolactin signaling. J Biol Chem 273:37499-37505.

Lee C, Sintich SM, Matthews EP, Shah AH, Kundu SD, Perry KT, Cho JS, Ilio KY, Cronauer MV, Janulis L and Sensibar JA (1999). Transforming growth factor-Beta in benign and malignant prostate. Prostate 39:285-290.

Chappell, P.E. and Levine, J.E. (2000) Stimulation of GnRH surges by estrogen 1: role of hypothalamic progesterone receptors. Endocrinology (in press).

Lin, J. and Linzer, D.I.H. (1999) Induction of megakaryocyte differentiation by a novel pregnancy-specific hormone. J. Biol. Chem. 274:21485-21489.

Ito, M., Park, T., Weck, J., Mayo, K.E. and Jameson, J.L. (2000). Synergic activation of the inhibin a subunit promoter by steroidogenic factor-1 and cAMP. Molecular Endocrinology 14:66-81.

Hayashida, T., Poncelet, A-C., Hubchak, S.C.. Schnaper, H.W. (1999)TGF-b1 activates MAP kinases in human mesangial cells: a possible role in collagen expression. Kidney Int., 56:1710-1720.

Liu, G., Duranteau, L., Carel, J-C., Monroe, J., Doyle, D.A. and Shenker, A. (1999) Leydig-cell tumors caused by an activating mutation of the gene encoding the luteinizing hormone receptor. New Eng J of Med 341:1731-1736.

Bohnsack, B.L., Szabo, M., Kilen, S.M., Tam, D.H.Y. and Schwartz, N.B. (2000) Follistatin suppresses steroid-enhanced follicle-stimulating hormone release in vitro. Biol Reprod 62:636-641.

Rudick, C.N. and Woolley, C.S. Estradiol induces a phasic C-FOS response in the hippocampus of adult female rats. Hippocampus, (in press).

Two members of the Center have chapters in Neuroendocrinology in Physiology and Medicine, Conn and Freeman Eds; Humana Press, 1999.

Jon Levine: Ch. 6. "The Hypothalamus as a Major Integrative Center".
Neena Schwartz: Ch. 8. "Neuroendocrine Regulation of Reproductive Cyclicity".