~ ii~ ~ ~

A Decade of PCR Cold Spring Harbor Laboratory and The Perkin-

Elmer Corporation celebrate 10 years of amplifica- tion with a videotape library in which Nobel prize winners Kary Mullis and James Watson and 19 other distinguished scientists review the applications and evolution of the amplification technique hailed as one of the century's most important scientific tools.

In 1995, the polymerase chain reaction will be 10 years old. The technique that began as a late-night in- spiration by an unrenowned scientist is now the bed- rock of DNA research, gene discovery, diagnostics development, forensic investigation and environmen- tal science. It has built an industry, provoked a court case, and spawned a dozen books, countless papers and a journal. Along the way, it earned its inventor, Kary Mullis, a Nobel prize~

To mark this anniversary, a conference sponsored by The Perkin-Elmer Corporation was held at Cold Spring Harbor Laboratory in September 1994. Begin- ning with perspectives from James Watson, famed for the discovery of the structure of DNA, and PCR- inventor Kary Mullis, outstanding scientists from a variety of fields reviewed the impact of the technique on their specialties, discussing the present and future applications of PCR technology.

A day and a half of wide-ranging, highly il- lustrated talks have been captured in this unique videotape library. The collection will appeal to

working scientists from the graduate student level up- wards who apply PCR to problems in human, animal and plant genetics, cell biology, diagnostics, forensic science and molecular evolution.

CONTENTS

PERSPECTIVES James D. Watson, Cold Spring Harbor Laboratory Kary Mullis, La Jolla AN IN-DEPTH LOOK AT PCR Primers, Oligos and Hybridization Wojciech Rychlik, National Biosciences, Inc. Biology of DNA Polymerases Tom Kunkel, National Institutes of Health PCR Automation and Genotyping Stanley Rose, The Perkin-Eimer Corporation APPLICATIONS OF PCR Human Genome Project Glen A. Evans, University of Texas Human Genetics Henry Erlich, Roche Molecular Systems Molecular Diagnostics Tom Caskey, Baylor College of Medicine Forensic Analysis Bruce Budowle, FBI Academy Gene Evolution/Ancient DNA Svante P~i~ibo, University of Munich Agriculture and the Third World Richard Jefferson, CAMBIA Gene Expression in Single Cells Jim Eberwine, University of Pennsylvania In Situ PCR Ashley Haase, University of Minnesota Combinational Libraries and Rapid Evolution Andrew Ellington, Indiana University THE FUTURE OF PCR Applications of Long Distance PCR Elise Rose, The Perkin-Elmer Corporation PCR Quantitation Francois Ferre, The Immune Response Corporation Analysis of PCR Products in Microchips Stephen Fodor, Affymetrix, Inc. RNA Differential Display Peng Liang, Dana Farber Cancer Institute Representational Difference Analysis Nikolai Lisitsyn, Cold Spring Harbor Laboratory Summary Maynard Olson, University of Washington

1994, 7 edited videotapes (10 hours 15 minutes in total) VHS: ISBN 0-87969-473-4; PAL: ISBN 0-87969-474-2 Price: $300

L EA [ lk] l NG M RY

E d i t o r

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M a n a g i n g E d i t o r

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Copyright �9 1997 by Cold Spring Harbor Laboratory Press

Volume 3 Number 6

March/April 1997 Pages 445-600

Editorial . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vi

R e v i e w

The Cerebellum, LTD, and Memory: Alternative Views . . . . . . . . . . . . . . . . . . . . . . . . . . . . 445 Rodolfo Llin~s, Eric J. Lang, and John P. Welsh

Research pape r s

Preserved Performance by Cerebellar Patients on Tests of Word Generation, Discrimination Learning, and Attention . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 456 Laura L. Helmuth, Richard B. Ivry, and Naomi Shimizu

A Neural Model of Cerebellar Learning for Arm M o v e m e n t Control: Cortico-Spino-Cerebellar Dynamics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 475 Jose L. Contreras-Vidal, Stephen Grossberg, and Daniel Bullock

Multiple Subclasses of Purkinje Cells in the Primate Floccular Complex Provide Similar Signals to Guide Learning in the Vestibulo-Ocular Reflex . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 503 Jennifer L. Raymond and Stephen G. Lisberger

The Effects of Reversible Inactivation of the Red Nucleus on Learning-Related and Auditory-Evoked Unit Activity in the Pontine Nuclei of Classically Condit ioned Rabbits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 519 M. Claire Cartford, Elizabeth B. Gohl, Maria Singson, and David G. Lavond

The Learning-Related Activity That Develops in the Pontine Nuclei During Classical Eye-Blink Conditioning Is D e p e n d e n t On the lnterpositus Nucleus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 532 Robert E. Clark, Elizabeth B. Gohl, and David G. Lavond

Reversible Inactivation of the Cerebellar lnterpositus Nucleus Complete ly Prevents Acquisition of the Classically Condit ioned Eye-Blink Response . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 545 David J. Krupa and Richard F. Thompson

Acquisition of a New-Latency Conditioned Nictitating Membrane Response~Major , but Not Complete, Dependence on the Ipsilateral Cerebellum . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 557 Christopher H. Yeo, Dominic H. Lobo, and Mison Baum

Persistent Phosphorylation Parallels Long-Term Desensitization of Cerebellar Purkinje Cell AMPA-Type Glutamate Receptors . . . . . . . . . . . . . . . . . . . . . 578 Kazutoshi Nakazawa, Sumiko Mikawa, and Masao Ito

Author Index, Volume 2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 592

Subject Index, Volume 2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 593

Author Index, Volume 3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 595

Subject Index, Volume 3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 596

Cover Dissociation of cerebellar attention (yellow and blue) and motor (green and red) activation (yellow and green - overlap in activation of 3 or more subjects; blue and red = overlap of any 2 subjects). Three- dimensional volume rendering of the cerebellum and brain stem demonstrates that during an attention task, the most common site of activation was in the left superior posterior cerebellum, while during a motor task, the most common site was in the right anterior cerebellum. (For details, see Courchesne and Mien, volume 4, p. 1; image rendered using VoxelView 2.5.)

Coming next m o n t h in Learning & Memory, vol. 4, n u m b e r 1, May/June 1997, the second special issue devoted to the cerebel lum

Review

Prediction and Preparation, Fundamental Functions of the Cerebellum Eric Courchesne and Greg Allen

Research Papers

Impaired Capacity of Cerebellar Patients to Perceive and Learn Two-Dimensional Shapes Based on Kinesthetic Cues Yury Shimansky, Marian Saling, David A. Wunderlich, Vlastislav Bracha, George E. Stelmach, and James R. Bloedel

Lateral Cerebellar Hemispheres Actively Support Sensory Acquisition and Discrimination Rather Than Motor Control Lawrence M. Parsons, James M. Bower, Jia-Hong Gao, Jinhu Xiong, Jinqi Li, and Peter T. Fox

Cerebellar Guidance of Premotor Network Development and Sensorimotor Learning Sherwin E. Hua and James C. Houk

Role of Cerebellum in Adaptive Modification of Reflex Blinks John J. Pellegrini and Craig Evinger

Single-Unit Evidence for Eye-Blink Conditioning in Cerebellar Cortex is Altered, but Not Eliminated, by Interpositus Nucleus Lesions Donald B. Katz and Joseph E. Steinmetz

Effect of Varying the Intensity and Train Frequency of Forelimb and Cerebellar Mossy Fiber Conditioned Stimuli on the Latency of Conditioned Eye-Blink Responses in Decerebrate Ferrets P~ir Svensson, Magnus Ivarsson, and Germund Hesslow

Conditioned Response Timing and Integration in the Cerebellum John W. Moore and June-Seek Choi

A Model of Pavlovian Eyelid Conditioning Based on the Synaptic Organization of the Cerebellum Michael D. Mauk and Nelson H. Donegan

Local Dendritic Ca z+ Signaling Induces Cerebellar LTD Jens Eilers, Hajime Takechi, Elizabeth A. Finch, George J. Augustine, and Arthur Konnerth

Absence of Cerebeilar Long-Term Depression in Mice Lacking Neuronal Nitric Oxide Synthase Varda Lev-Ram, Zuryash Nebyelul, Mark H. Ellisman, Paul Huang, and Roger Y. Tsien

Editorial

The cerebellum has intrigued neuroscientists for more than a century. This fascination is in part due to its unique macroscopic appearance and the fact that it occupies a significant percentage of the brain. At the microscopic level, the beauty of its cellular architecture is universally appreciated. The limited number of cell types and the apparent modularity of its cellular organization have raised the possibility that the cerebellum will be the first brain structure to be understood completely. The structure is intrinsically appealing, but what is its function and importance? Based on evolutionary considerations alone, it appears to be quite impor- tant. For example, during the past 10 million years, the size of the cerebellum has increased in greater proportion to other brain structures, including the cerebral cortex. The question of its function is less clear. Historically, the cerebellum was considered to be important for motor function. Based on the comparison of the sizes of cerebella in different species having different postural demands, the pre- vailing view in the early part of this century was that the cerebellum was critical for the control of posture. In reviewing these considerations, Cajal, in 1914, concurred with this view. Moreover, he stated "It should be obvious already that the cer- ebellum has nothing to do with consciousness or other higher functions."

However, over the past 20 years, research has seriously challenged the traditional views of the cerebellum's exclusive role in postural control. Be- ginning with the theoretical work of Marr, and the empirical work of Ito, Thach, and their colleagues, it became clear that the domain of the cerebellum extends beyond postural control. It is involved in motor learning, as well. Moreover, a specific form of synaptic plasticity, long-term depression (LTD) at the parallel fiber-Purkinje cell connection, has been implicated in the motor learning mediated by the cerebellum. But studies of patients with cer- ebellar dysfunction, as well as recent positron

& vi

emission tomography (PET) and functional mag- netic resonance imagery (fMRI) studies, raise the possibility of a greatly expanded range of cerebel- lar function. These include attention, associative learning, practice-related learning, procedural learning, declarative memory, working memory, semantic association, conditioned anxiety, mental exploration, and complex reasoning and problem solving, as well as sensory, motor, and motor skill acquisition.

Despite the tremendous advances in cerebel- lum research at the anatomical, biophysical, and systems levels, there is still no general theory that encompasses and explains the functional role played by the cerebellum in these diverse motor and nonmotor domains. Indeed, there is significant debate whether the cerebellum does in fact sub- serve all of these diverse functions. Even the role of the cerebellum in motor learning itself has been questioned. Because of the intense interest in the cerebellum in general, and its role in motor learn- ing in particular, Learning & Memory has devoted two special issues to this topic. We cover the cer- ebellum's role from the molecular to systems ap- proaches, with techniques ranging from cell cul- ture to brain imaging. The approaches of math- ematical modeling and computer simulations are also represented, because these raise the possibil- ity of generating testable models of cerebellar func- tion and tests of theories of the computations that are performed by the cerebellum. These special issues certainly advance the understanding of the cerebellum, but as these issues illustrate, the field has not yet matured to the point of a general frame- work of cerebellar function. Learning & Memory will serve as a vehicle to foster the continued ex- change of information and debate in those areas.

John H. Byrne Editor

L E A R N / N G M E M 0 R Y