A PMYR by Joseph G. Kunkel

Biology Department, University of Massachusetts Amherst

The 16 years since my promotion to Full Professor span two sabbaticals and their intervening periods. I will extrapolate from those 16 years of research, teaching and service to my objectives for the future, exploring the places I wish my momentum to take me.

Insect storage protein was my primary research field prior to my 1985 promotion to Full Professor. I became an expert and a reviewer of my field (1, 2); the reviews were widely referenced. I continued my interest in storage proteins with further key publications in major journals (3-10) on various aspects of storage protein synthesis, post-translational modification, transport and utilization by the developing embryo. I have also continued my role as a reviewer of this field collaborating with a founding father (11). During subsequent years I expanded my interests into storage proteins of other groups including marine fish (winter flounder, cod, and tautog) and newly into lobsters. These extensions of my research are based on my conviction that the massive investments of specie's resources flowing through storage proteins, which pass through the bloodstream, can help us predict individual developmental trajectory. My experience with purifying proteins, making antisera against these proteins and using the antisera in protein bioassays is continually reapplied (9, 12). A recently awarded 2-year NOAA/CMER grant is aimed at predicting the time of molting and egg production of lobsters. This grant led to collaboration with Diane Cowan, of The Lobster Conservancy. We just submitted a three year NSF grant proposal which will partner Diane's expertise on lobster behavior with my expertise on storage proteins to extend the objectives of the NOAA/CMER project to a fully funded research program. This lobster project will be the keystone of my future work on storage proteins and will consume at least half of my research energies for the next decade. Our design and code for an online research database (13) developing around our lobster research will be available to future researchers collaborating at a distance.

In 1984 I initiated a new direction in my research, which had its roots in a meeting I had with Lionel Jaffe, who studies pattern formation in development. Using a new technology, non-invasive vibrating probes, he demonstrated patterns of ionic currents surrounding developing oocyte follicles. I suggested that the ionic currents were involved in the massive movements of storage resources; Jaffe insisted the currents were related to polarity development. He invited me to visit his MBL facility to test these hypotheses. After my visit and a full year sabbatical in Berne Switzerland, 1986-87, where I concentrated on the ionic controls of vitellogenic uptake, I returned to Amherst to focus further on this controversy. Whether or not ions have an effect on polarity, I was able to show that there was a definite role of ionic currents in the vitellogenic storage (14-16). I devoted substantial effort since that time in using non-invasive probes to detect correlation between ionic fluxes and cellular processes.

In 1988 I established an NSF funded UMass Vibrating Probe Facility in Amherst. To further the UMass Probe Facility in 1993-94 I did a full year sabbatical at the MBL’s National Vibrating Probe Facility where I learned new ion probe technology and brought it back to Amherst. My UMass Probe Facility became a beta test site for electronics from Applicable Electronics and software from Science Wares, two Massachusetts small businesses. This led to several collaborations with labs focused on particular cellular phenomena such as the growing root tip, the growing pollen tip, the chloride cell and the Xenopus oocyte-expression-system. These efforts have resulted in substantial publications (17-23).

I focus on developing 3-dimensional probe technology, essential to obtaining a total balance sheet of ion flux in and out of a cell. I maintain well-calibrated electronic rigs and work closely with visiting experimenters so that the measurements made can result in calculation of meaningful estimates with associated confidence limits (17, 19, 22). I authored a methodology paper on the intelligent use of buffers in measuring proton flux in and out of tissues (24) which should be referenced by anyone interested in proton flux. We are also developing distance learning approaches to provide training prior to visits to our probe facility.

Furthermore, we recently made progress in application of the non-invasive oxygen and nitric oxide probes. The development of a local facility to use these new micro probes, combined with my interest in 3-D flux measurements for these compounds attracted visitors to the UMass Vibrating Probe Facility in year 2000, including: Margarida Prado, a student of Jose Feijó (Gulbenkian Institute, Portugal), Li-Yih Lee, a student of Pung-Pung Hwang (Taiwan U.) and Petra Sominieski, a Research Associate of Wolfram Nagel (U. Munich, Germany), all leaders in ion regulation research. I plan on spending my upcoming sabbatical in the lab of Wolfram Nagel collaborating to advance the programming of the ASET software, which drives the positioning and measurements by single or multiple voltage- or ion selective- probes.

A prime objective over the next seven years is to extend the momentum gained during this sabbatical to make the simultaneous use of multiple non-invasive probes more automated and convenient for the researcher. This project will be partially funded by continued support from Applicable Electronics, which is committed, on a year by year basis, to provide funding for our probe facility ($24,000+ in 2001). Efforts at our Probe Facility are now partially funded by its visitors and users. For example, I wrote a letter of intent in support of funding a grant to Peter Hepler, pledging my efforts in helping him study pollen tube growth using our ion probe facility. Margarida Prado's visit to our facility was funded by a NATO fellowship. We are talking with the Distance Learning Center to determine if their fee structure can accommodate a workshop fee for training we provide over the internet. I will continue to seek funding for the basic research that we pursue using this instrumentation. I am currently working with Baystate Medical researchers, Margaret Reece and Lucy Bayer, and a Harvard Medical School researcher, former student Anand Iyengar, who are interested in cord blood and embryonic stem cell research. We are seeking startup funding from the Baystate/UMass Biomedical Collaborative and will be applying for external funding based on preliminary findings. Application of ion probe technology to profiling the flux of molecules in and out of stem cells could be a major technological development contributing to their future successful use.

My contributions to service in my profession are vary broadly, from (A) my maintenance of a Cockroach Web Page from which I answer questions posed by citizens about this ancient survivor (25), to (Z) consulting with fellow researchers on biometrical methodology.

A service contribution to my Biology Department has been the institution of a Biology Alumni Magazine and Website (26) which has sought to reconnect alumni with our Departments accomplishments and goals. I will continue as managing editor and web designer for that initiative as long as it is needed. I hope to make the website more interactive and useful to our alumni such that they will think of Biology when they are asked to support their alma mater.

As one of the only active computer programmers on the Biology faculty, I have donated my programming skills to several colleagues' computational projects over the years including a time series analysis by Vincent Dethier years ago. More recently the same program was used in publishing work (18) correlating pollen tube ionic and growth events for Peter Hepler's group. I maintain a webpage that provides access to download these programs (27). I constantly upgrade my programming skills; my current project is developing my ability to access and use large databases using the new high level programming language, J (28).

Service to the University has proceeded via the Faculty Senate University Computer and Electronic Communications Committee. As Secretary for two years I instituted a website (29) which lists agenda and minutes back to 1995 as well as links to relevant materials under current consideration. Now as Committee Chairman I am marshaling discussion of the laptop mandate from our Board of Higher Education and the problem of authentication and communication rights, which are major problems which will affect our future ability to teach and communicate both on and off campus. Issues of information technology continue to confront the university and I intend to stay active on the University committees that deal with IT.

I share my web mastering abilities with the Amherst community by maintaining the Echohill South Association Website (30).

I have taught the Junior Writing Course almost every year since I returned from my sabbatical in 1987. The dramatic changes in communication technology that I am able to apply to this course each year have kept me interested in teaching Junior Writing despite the modest intrinsic intellectual content and repetitive nature of the course. This is a methods course required by students university-wide. I treat Junior Writing as an opportunity to increase the professional communication skills of my students to a level approaching an advanced graduate student. In my most recent presentations of this course I have made it my Science, Technology, Engineering, Math and Technology Educational Collaborative (STEMTEC), Cycle II project. The description of this project and group-based-learning course is to be published in PATHWAYS TO CHANGE edited by Richard Yuretich of the STEMTEC program (31). I am also applying group based learning technology to the Biol 297C, Cell and Molecular Biology Lab, which I teach each spring semester. I am requesting a digital video camera to provide video-taped clips of critical lab demonstrations as well as to video tape student presentations which are part of both Biol 312 and 297C. Digital video technology will also be incorporated in our Distance Learning offerings for the Non-Invasive Probe Facility.

The future of my teaching depends upon my development of a system of teaching-technology and file management, based on Steve Brewer’s BCRC web teaching shells which are made available for all Biology Courses. I consult with George Drake to make sure the procedures I develop are electronically secure. The objective is to make all projects and assignments, submitted for credit in my courses, electronic in nature. This means that every submission is time stamped and I keep an archival copy of the original, my comments and grading of the project. The student gets an electronic copy of the graded project with my highlighted comments. Each year’s coursework is archived onto tape and/or CD for easy access. While easy to describe it is another issue to make it convenient and automated in the modern Internet connected world. I regularly receive assignments either in my UMass office or at home via Email attachment. This results in duplication of storage location and time-stamping of materials with occasion for confusion in the primacy of a copy. A successful IT approach to this security and time budgeting problem within the context of the BCRC teaching shells could be a benefit to all faculty.

This approach is a necessary change in response to an increasingly-technological student body, 80% of which have access to a personal computer. Students become tardy with their projects during the semester and argumentative and even litigious when they do not get their expected grade. Parents may become involved as advocates. These are not isolated observations from one faculty member. I have heard the same analysis from several sources. In order to provide a modern student evaluation system as well as substantiate a reasonably hard line on grades one needs convenient and excellent archival records and statistics to back up grading.

I request affirmation and support in pursuing this technological goal. There are several roadblocks that could be easily removed:

First, though I am one of the principle faculty users of the BCRC and its computers for teaching my courses, my laboratory and office were historically, and remain completely, IBM-PC-based (for good reasons), while the BCRC is completely Macintosh-computer-based. Although this has not prevented me from getting to know the Mac computer, it has made my use of the Macs in the BCRC a reach. When I present a lecture to a BCRC class using my PC laptop, we may all be using Ms Word or Excel, but my implementation is on the PC and theirs is on the Mac (perhaps a different version or revision level); … there is a difference. There have been several instances when my reservation of the BCRC Mac Powerbook followed an earlier reservation, which should have given me the Powerbook in time for my class, but did not. Competing for a single BCRC Powerbook will not do, especially when I need to develop and test my coursework for viewing on a Mac. I request funding for a Mac PowerPC to facilitate my teaching in the BCRC classroom. Obtaining a Mac to experiment with will further allow me to extend my interest in the J computer language, which works cross-platform on PCs, Macs, and Unix machines. I am interested in resuming my teaching of Biometry in our new Internet environment. The last time I taught Biometry we used computer terminals, which were a roadblock to effective learning since students could not access them from their dorms. I would like to try again using the J language as a cross-platform tool. Students have been mandated to have laptops. I am in the forefront of faculty prepared to interact with them; but I need the correct tools for the job.

Second, I respond on-line and in office visits with my students and accommodate their needs in a responsive way by suspending current research work and opening new windows on my computer screen to either demonstrate a procedure or respond to a request that may further require opening several ancillary windows. This was convenient at an earlier time with smaller software footprints and when my desktop computer was the fastest and had (then) large memory. That was four years ago. I find that my current research-grant-funded office computer does not have the speed or memory to rapidly communicate with remote drives; nor does it have several peripherals that are needed to provide flexibility and security of the archival and grading process. This often requires that I shut down some research or service operations in my office computer to accommodate students or move to a lab computer to finish certain operations. Remaining a PC user is a priority since most students own PCs not Macs. I request an upgrade in my office desktop IBM-PC computer to ease this issue. I request a CD writer to provide the necessary IBM and Mac compatible storage and retrieval of archived coursework.

The future may see most faculties using electronic teaching protocols. I need these protocols now. The electronics for accomplishing my goals are available and, with the critical help of colleagues in the Biology Department, I will make these tools work effectively for us.

1. Hagedorn, H. H. and Kunkel, J. G. (1979). Vitellogenin and vitellin in insects. Ann Rev Entomol 24, 475-505.
2. Kunkel, J. G. and Nordin, J. H. (1986). Yolk proteins. Comprehensive Insect Physiology Biochemistry Pharmacology 1, 83-111.
3. Duhamel, R. C. and Kunkel, J. G. (1987). Molting cycle regulation of hemolymph protein clearance in cockroaches: Possible size-dependent mechanism. J Insect Physiol 33, 155-158.
4. Wojchowski, D. M. and Kunkel, J. G. (1987). Purification of two distinct oocyte vitellins and identification of their corresponding vitellogenins in fat body and hemolymph of Blaberus discoidalis. Insect Biochem 17, 189-198.
5. Wojchowski, D. M., Parsons, P., Nordin, J. H. and Kunkel, J. G. (1986). Processing of pro-vitellogenin in insect fat body: A role for high- mannose oligosaccharide. Dev Biol 116, 422-430.
6. Zhang, Y. and Kunkel, J. G. (1991). High abundance calmodulin from Blattella germanica eggs binds to vitellin subunits but disappears during vitellin utilization. Insect Biochem 22, 293-304.
7. Zhang, Y. and Kunkel, J. G. (1994). Most egg calmodulin is a follicle cell contribution to the cytoplasm of the Blattella germanica oocyte. Dev Biol 161, 513-21.
8. Iyengar, A. R. and Kunkel, J. G. (1995). Follicle cell calmodulin in Blattella germanica: Transcript accumulation during vitellogenesis is regulated by juvenile hormone. Dev Biol 170, 314-320.
9. Hartling, R. C. and Kunkel, J. G. (1999). Developmental fate of the yolk protein lipovitellin in embryos and larvae of winter flounder, Pleuronectes americanus. J Exp Zool 284, 686-95.
10. Hartling, R. C., Pereira, J. J. and Kunkel, J. G. (1997). Characterization of a heat-stable fraction of lipovitellin and development of an immunoassay for vitellogenin and yolk protein in winter flounder (Pleuronectes americanus). J Exp Zool 278, 156-66.
11. Telfer, W. H. and Kunkel, J. G. 1991. The function and evolution of insect storage hexamers. Annu Rev Entomol 36, 205-28.
12. Diss, AL, JG Kunkel, ME Montgomery and DE Leonard. 1996. Effects of maternal nutrition and egg provisioning on parameters of larval hatch, survival and dispersal in the gypsy moth, Lymantria dispar L. OECOLOGIA 106, 470-477.
13. Kunkel, J. G., Xu, Y. and Cowan, D. F. (2001). An online lobster research database. URL: http://marlin.bio.umass.edu/biology/kunkel/fish/lobster/.
14. Kunkel, J. G., Koenig, R., Kindle, H. and Lanzrein, B. (1986). Ion Flux during vitellogenesis and patterning of insect oocytes. Adv Invert Reprod 4, 101-108.
15. Kindle, H., Lanzrein, B. and Kunkel, J. G. (1990). The effect of ions, ion channel blockers, and ionophores on uptake of vitellogenin into cockroach follicles. Dev Biol 142, 386-391.
16. Sigel, E., Baur, R., Kunkel, J. G., Kindle, H. and Lanzrein, B. (1990). Demonstration of a voltage dependent calcium current in follicles of the cockroach Nauphoeta cinerea. Invert Reprod Dev 18, 159-164.
17. Kunkel, J. G. and Bowdan, E. (1989). Modeling Currents about vitellogenic oocytes of the cockroach, Blattella germanica IONIC CONTROLS OF DEVELOPMENT II. Biol Bull Woods Hole 176S, 96-102.
18. Kunkel, J. G. and Faszewski, E. (1995). Pattern of potassium ion and proton currents in the ovariole of the cockroach, Periplaneta americana, indicate future embryonic polarity. Biol Bull 189, 197-198.
19. Holdaway-Clarke, T. L., Feijó, J. A., Hackett, G. R., Kunkel, J. G. and Hepler, P. K. (1997). Pollen tube growth and the intracellular cytosolic calcium gradient oscillate in phase while extracellular calcium influx is delayed. Plant Cell 9, 1999-2010.
20. Feijó, J. A., Sainhas, J., Hackett, G. R., Kunkel, J. G. and Hepler, P. K. (1999). Growing pollen tubes possess a constitutive alkaline band in the clear zone and a growth-dependent acidic tip. J Cell Biol 144, 483-96.
21. Feijó, J. A., Sainhas, J., Holdaway-Clarke, T., Cordeiro, M. S., Kunkel, J. G. and Hepler, P. K. (2001). Cellular oscillations and the regulation of growth: the pollen tube paradigm. Bioessays 23, 86-94
22. Cardenas, L., Feijó, J. A., Kunkel, J. G., Sanchez, F., Holdaway-Clarke, T., Hepler, P. K. and Quinto, C. (1999). Rhizobium nod factors induce increases in intracellular free calcium and extracellular calcium influxes in bean root hairs. Plant J 19, 347-52.
23. Cardenas, L., Holdaway-Clarke, T. L., Sanchez, F., Quinto, C., Feijó, J. A., Kunkel, J. G. and Hepler, P. K. (2000). Ion changes in legume root hairs responding to Nod factors. Plant Physiol 123, 443-52.
24. Kunkel, J. G., Lin, L.-Y., Xu, Y., Prado, A. M. M., Feijó, J. A., Hwang, P. P. and Hepler, P. K. (2001). The strategic use of Good buffers to measure proton gradients about growing pollen tubes. In Cell Biology of Plant and Fungal Tip Growth (ed. A. Geitman), pp. 14pp. Amsterdam: IOS Press (in press).
25. Kunkel, J. G. (1995). The Cockroach Home Page. URL: http://marlin.bio.umass.edu/biology/kunkel/blattari.html.
26. Kunkel, J. G. (1999). UMass Biology Alumni Website. URL: http://marlin.bio.umass.edu/biology/alumni/
27. Kunkel, J. G. (2001). Downloadable data and analysis programs in support of publications and general research, URL: http://marlin.bio.umass.edu/biology/kunkel/pub/.
28. Kunkel, J. G. (2001). Joe Kunkel's J Development Project, URL: http://marlin.bio.umass.edu/biology/kunkel/j/.
29. Kunkel, J. G. (1997). Faculty Senate Computer and Electronic Communications Website, URL: http://marlin.bio.umass.edu/biology/kunkel/fsucecc/.
30. Kunkel, J. G. (1998). Echohill South Association webpage, URL: http://www.geocities.com/echohill_2000/.
31. Kunkel, J. G. (2001). Project- and Group-Based Learning of Junior Writing in Biology. In Pathways to Change (ed. R. Yuretich). Amherst (in press). URL: http://marlin.bio.umass.edu/biology/kunkel/stemtec/final/final-rev.html