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CURRICULUM
VITAE - Dr. David Earnest
I. Personal Data Name: David J. Earnest Address:
IV. Professional Affiliations National Scientific Society Memberships American Association for the Advancement of Science Society for Neuroscience Society for Research on Biological Rhythms
V. Areas of Research Interest Multidisciplinary approaches to study of the neurobiology of circadian rhythms and their regulation by light-dark signals -elucidation of the roles of specific neurotransmitter systems, immediate-early genes and neurotrophins in the signal transduction pathway underlying the photoentrainment of circadian rhythms -application of immortalized cell lines and other in vitro models to identify core cellular and molecular components of the pacemaker mechanism within the suprachiasmatic nucleus -identification of the molecular and cellular changes in the suprachiasmatic nucleus that mediate the deterioration of circadian pacemaker function during aging
VI. Current Projects Functional Characterization of Immortalized Suprachiasmatic Cells Identification of Clock and Clock-Controlled Genes in the Suprachiasmatic Circadian Pacemaker Role of Neurotrophins in the Regulation of Circadian Rhythms and Their Relation to the Deterioration of Suprachiasmatic Pacemaker Function during Aging
University
of Rochester School of Medicine, Rochester, New York
Texas
A&M University Health Sciences Center, College of Medicine,
College Station, Texas
IX. Academic Committee Service
X. Honors and Awards Undergraduate Honors Year(s) University
of Michigan Regents-Alumni Scholar 1972-76
Predoctoral and Postdoctoral Awards Year(s) Northwestern
University Fellowship 1978-79
XI. Current Research Grants NINDS
P01 NS39546-01 "Coordination of Circadian 2000-2005 Physiology
of Diverse Species" (V. Cassone, PI) Project
2: "Analysis of Clock and Clock- Controlled Genes
in
Immortalized SCN Cells" (D. Earnest, Project Leader) NIH R01 MH60147-01A2 "Role of BDNF in 2001-2004 the Photic Control of Circadian Rhythms" ($450,000 direct total costs) D.J.Earnest, P.I., 30% effort NIH R01 AA13242-01 "Developmental 2001-2006 Alcohol and Circadian Clock Function" ($750,000 direct total costs) D.J.Earnest, P.I., 20% effort Licensing Agreements with with Pfizer, Hoffmann-La Roche, Hoechst Marion Roussel Inc. and Chiron for Use of Immortalized SCN Cells XII. Pending Research Grants None XIII. Past Research Grants NIH
First Award NS24661 "Neural Control of Suprachiasmatic
Function"
($344,234 direct costs) Air Force Office of Scientific Research " Photic Regulation of Gene Expression and Cellular Activity in the SCN" ($180,000 total costs) D.J. Earnest, P.I., 25% effort 1989-92 NIH Shannon Award NS30939 "Light Signal Transduction in the SCN Circadian Pacemaker" ($80,000 total direct costs) D.J. Earnest, P.I., 25% effort 1992-94 Air Force Office of Scientific Research " Transplantation and Cloning of an Immortal Cell Line from Rat SCN" ($300,000 total costs) D.J. Earnest, P.I., 25% effort 1993-94
XIII. Past Research Grants (cont.) NSF SGER Award "Functional Characterization of a Rat Suprachiasmatic Nucleus-Derived Cell Line" ($41,616 total direct costs) D.J. Earnest, P.I., 10% effort 1993-94 NSF Grant, IBN-9511238 "Functional Characterization of Immortalized Suprachiasmatic Cells" ($273,620 total costs) D.J. Earnest, P.I., 17% effort 1995-99 DHHS-PHS-NIH "Localization of N-Acetyl Transferase" ($65,972 total costs) D.J. Earnest, Co-Investigator 1999-2000
XIV. Supervision of Personnel
XV. Masters and Ph.D. Committees
XVI. SURF and Medical Student Summer Research Training
XVII. Physical Facilities
Techniques: Explant and dissociated cell cultures Establishment of immortalized neural cell lines Gene transfer using retroviral vectors RT-PCR and in situ hybridization analysis of mRNA expression in brain tissue Immunocytochemistry and radioimmunoassay for neuropeptides Neural transplantation of fetal tissue and immortalized cells Analysis of rodent wheel-running behavior
Research Facilities: Facilities and equipment necessary for performing cell culture, immunocytochemical, immunoassay and molecular methodologies;Computer-based data acquisition system for monitoring locomotor behavior, body temperature and heart rate.
XVIII. Publications Manuscripts Earnest, D.J. and Turek, F.W. (1982) Splitting of the circadian rhythm of activity in hamsters: Effects of exposure to constant darkness and subsequent re-exposure to constant light. J. Comp. Physiol. 145:405-411. Earnest, D.J. and Turek, F.W. (1983) Role for acetylcholine in mediating effects of light on reproduction. Science 219:77-79. Earnest, D.J. and Turek, F.W. (1983) Effect of one-second light pulses on testicular function and locomotor activity in the golden hamster. Biol. Reprod. 28:557-565. Earnest, D.J. and Turek, F.W. (1984) Periodic exposure to a brief light signal stimulates neuroendocrine-gonadal activity in golden hamsters. J. Andrology 5:64-69. Earnest, D.J. and Turek, F.W. (1985) Neurochemical basis for the photic control of circadian rhythms and seasonal reproductive cycles: Role for acetylcholine. Proc. Natl. Acad. Sci. U.S.A. 82:4277-4281. Earnest, D.J. and Turek, F.W. (1986) Responses of the split activity components in the hamster to a brief perturbation of the constant lighting environment. Naturwissenschaften 73:565-566. Earnest, D,J, and Sladek, C.D. (1986) Circadian rhythms of vasopressin release from individual rat suprachiasmatic explants in vitro. Brain Res. 382:129-133. Keefe, D.L., Earnest, D.J., Nelson, D., Takahashi, J.S. and Turek, F.W. (1987) A cholinergic antagonist, mecamylamine, blocks the phase-shifting effects of light on the circadian rhythm of locomotor activity in the golden hamster. Brain Res. 403:308-312. Earnest, D.J. and Sladek, C.D. (1987) Circadian rhythms of vasopressin release from perifused rat suprachiasmatic explants in vitro: effects of acute stimulation. Brain Res. 422:398-402. Earnest, D.J., Sladek, C.D., Gash, D.M. and Wiegand, S.J. (1989) Specificity of circadian function in transplants of the fetal suprachiasmatic nucleus. J. Neuroscience 9:2671-2677. Earnest, D.J., Iadarola, M., Yeh, H.H. and Olschowka, J.A. (1990) Photic regulation of c-fos expression in neural components governing the entrainment of circadian rhythms. Exp. Neurol. 109: 353-361. Earnest, D.J., DiGiorgio, S.M. and Sladek, C.D. (1991) Effects of tetrodotoxin on the circadian pacemaker mechanism in suprachiasmatic explants in vitro. Brain Res. Bull. 26: 677-682. Earnest, D.J., Ouyang, S. and Olschowka, J.A. (1992) Rhythmic expression of Fos-related proteins in the rat suprachiasmatic nucleus during constant retinal illumination. Neuroscience Lett. 140: 19-24. Earnest, D.J. and Olschowka, J.A. (1993) Circadian regulation of c-fos gene expression in the SCN pacemaker by light. J. Biol. Rhythms 8: S65-71. Earnest, D.J., DiGiorgio, S.M. and Olschowka, J.A. (1993) Light induces expression of Fos-related proteins within gastrin-releasing peptide neurons in the rat suprachiasmatic nucleus. Brain Res. 627: 205-209. Liang, F.-Q., Walline, R. and Earnest, D. (1998) Circadian rhythm of brain-derived neurotrophic factor expression in the rat suprachiasmatic nucleus. Neuroscience Letters 242: 89-92. Liang, F.-Q., Sohrabji, F., Miranda, R., Earnest, B. and Earnest, D. (1998) Expression of brain-derived neurotrophic factor and its cognate receptor, TrkB, in the rat suprachiasmatic nucleus. Experimental Neurology 151: 184-193. Earnest, D.J., Liang, F.-Q., Ratcliff, M. and Cassone, V.M. (1999) Immortal time: Circadian clock properties of cell lines derived from the rat suprachiasmatic nucleus. Science 283: 693-695. Earnest, D.J., Liang, F.-Q., DiGiorgio, S.M., Gallagher, M.J., Harvey, B., Earnest, B.J. and Seigel, G.M. (1999) Establishment and characterization of adenoviral E1A immortalized cell lines derived from the rat suprachiasmatic nucleus. J. Neurobiology 39: 1-13. Wade, S., Oommen, P., Conner, W.C., Earnest, D.J. and Miranda, D.J. (1999) Overlapping and divergent actions of estrogen and the neurotrophins on cell fate and p53-dependent signal transduction in conditionally immortalized cerebral cortical neuroblasts. J. Neuroscience 19: 6994-7006. Liang, F.-Q., Allen, G. and Earnest, D.J. (2000) Role of brain-derived neurotrophic factor in the circadian regulation of the suprachiasmatic pacemaker by light. J. Neuroscience 20: 2978-2987. Allen,
G., Rappe, J., Earnest, D.J. and Cassone, V.M. (2001) Oscillating
on borrowed time: diffusible signals from immortalized SCN cells
are necessary for circadian rhythmicity in cultured fibroblasts?
J. Neuroscience 21: 7937-7943. Hurst, W.J., Earnest, D.J. and Gillette, M.U. The SCN 2.2 Cell Line Expresses Characteristics of the Suprachiasmatic Nucleus. (Submitted) Earnest, D.J., West, J.R. and Chen, W.-J. Effects of alcohol exposure during rapid brain development on the rat suprachiasmatic nucleus and circadian behavior. (In preparation) Book Chapters Turek, F.W., Earnest,D.J. and Swann, J. (1982) Splitting of the circadian rhythm of activity in hamsters. In: Vertebrate Circadian Systems: Structure and Physiology. (J. Aschoff, S. Daan and G. Groos, eds.), Springer-Verlag, Berlin, pp. 203-214. Turek, F.W., Swann, J. and Earnest, D.J. (1984) Role of the circadian system in reproductive phenomena. In: Recent Progress in Hormone Research. (R.O. Greep, ed.), vol. 40, Acad. Press, New York, pp. 143-184. Earnest,D.J., Lehman, M., Roenneberg, T., van den Pol, T., Zatz, M. and Silver, R. Clocks at the cellular level. In: Understanding the Biological Clock. (J.C. Dunlap and J. Loros, eds.), American Physiol. Soc., in press.
Technical Reports Earnest, D.J., Miranda, R. and Sohrabji, F. (1998) Parallel analysis of BDNF and NT-3 in rat brain. Promega Neural Notes IV: 16-18.
Abstracts Earnest, D.J. and Turek, F.W. (1980) Effects of constant darkness and re-exposure to constant light on the circadian rhythm of locomotor activity after "splitting" has occurred. Soc. for Neuroscience Abstr. 6:833. Earnest, D.J. (1981) Effect of a 1-second skeleton photoperiod on testicular function and entrainment of the activity rhythm in golden hamsters. Biol. Reprod. 24 Suppl. 1:102A. Earnest, D.J. (1982) Maintenance of testicular function in hamsters exposed to short days by intraventricular injections of carbachol during the night. Biol. Reprod. 26 Suppl. 1:47A. Earnest, D.J. and Turek, F.W. (1983) Differential responses of the split activity components in the hamster. Fed. Proc. 42:1253. Earnest, D.J. (1983) Circadian injections of carbachol mimic the effects of light on the activity rhythm and reproduction in the hamster. Gordon Research Conference on Chronobiology, New London, New Hampshire. Earnest, D.J. and Turek, F.W. (1983) Phase shifting and entrainment of the hamster circadian system: role for acetylcholine in mediating the effects of light. Soc. for Neuroscience Abstr. 9:626. Earnest, D.J. and Turek, F.W. (1984) Neurochemical basis for the photoperiodic control of seasonal reproductive cycles: role for acetylcholine. Biol. Reprod. 30 Suppl. 1:72. Earnest, D.J. and Sladek, C.D. (1984) Circadian rhythm in vasopressin release from rat suprachiasmatic explants in vitro. Soc. for Neuroscience Abstr. 10:500. Earnest, D.J. and Sladek, C.D. (1985) Culture systems for studying circadian rhythms of vasopressin release from rat suprachiasmatic explants. Gordon Research Conference on Chronobiology, Plymouth, New Hampshire. Earnest, D.J. and Sladek, C.D. (1985) Circadian rhythms of vasopressin release from perifused suprachiasmatic explants in vitro. Soc. for Neuroscience Abstr. 11:385. Earnest, D.J. and Sladek, C.D. (1986) Mechanisms underlying the acute and circadian release of vasopressin from perifused rat suprachiasmatic explants in vitro. Soc. for Neuroscience Abstr. 12:845. Earnest, D.J., Sladek, C.D., Gash, D.M. and Wiegand, S.J. (1987) Transplantation of subtypes of vasopressinergic neurons into Brattleboro rats: suprachiasmatic grafts generate circadian vasopressin rhythms in the cerebrospinal fluid. Soc. for Neuroscience Abstr. 13:212. Earnest, D.J. and Sladek, C.D. (1988) Phase-shifting responses of suprachiasmatic explants to ionic manipulation in vitro. Soc. for Neuroscience Abstr. 14:385. DiGiorgio, S.M., Sladek, C.D. and Earnest, D.J. (1989) Effects of tetrodotoxin on the ciracdian pacemaker in suprachiasmatic explants in vitro. Soc. for Neuroscience Abstr. 15:1060. Earnest, D.J., Trojanczyk, L.A., van Lare, J., Yeh, H.H. and Olschowka, J.A. (1989) C-fos proto-oncogene expression in the suprachiasmatic nucleus: immunocytochemical characterization. Soc. for Neuroscience Abstr. 15:1058. Ouyang, S., Trojanczyk, L.A., Olschowka, J.A. and Earnest, D.J. (1990) Time-based changes in the photoinduction of Fos expression in the suprachiasmatic nucleus. Soc. for Neuroscience Abstr. 16:771. Earnest, D.J., Gallagher, M., Yagil, C. and Sladek, C.D. (1990) Effect of optic nerve stimulation on peptide release and mRNA levels expressed by cultured suprachiasmatic explants. Soc. for Neuroscience Abstr. 16:771. Earnest, D.J., DiGiorgio, S.M., Trojanczyk, L.A. and Olschowka, J.A. (1991) Light induces proto-oncogene expression in gastrin releasing peptide neurons within the suprachiasmatic nucleus. Soc. for Neuroscience Abstr. 17:24. Earnest, D.J. (1992) Circadian effect of optic nerve stimulation on c-fos mRNA expression in SCN explants in vitro. Soc. for Res. on Biol. Rhythms Abstr. 3:19. Bellinger, D.L., Earnest, D.J., Gallagher, M. and Felten, D.L. (1992) Presence and availability of VIP in primary and secondary lymphoid organs. Soc. for Neuroscience Abstr. 18:1009. Earnest, D.J., DiGiorgio, S., Gallagher, M.J., Gannon, R.L. and Rea, M.A. (1993) Establishment and characterization of an immortal cell line derived from the rat SCN. Soc. for Neuroscience Abstr. 19:1056. Earnest, D.J., Gallagher, M.J. and Harvey, B. (1994) Expression of AMPA-type glutamate receptors in immortal cell lines derived from the rat SCN. Soc. for Neuroscience Abstr. 20: 450. Liang, F.-Q., Sohrabji, F., Miranda, R., Earnest, B. and Earnest, D. (1995) Expression of brain-derived neurotrophic factor mRNA in SCN cells in vivo and in vitro. Soc. for Neuroscience Abstr. 21: 453. Earnest, D., Earnest, B. and Sohrabji, F. (1995) Age-related changes in the expression of brain-derived neurotrophic factor mRNA in the rat SCN. Soc. for Neuroscience Abstr. 21: 1235. Earnest, D., Liang, F.-Q., Earnest, B., Miranda, R. and Sohrabji, F. (1996) Brain-derived neurotrophic factor and trkB tyrosine kinase receptors in the rat SCN: Distribution and influence of aging. Soc. for Res. Biol. Rhythms Abstr. 5: 123. Hurst, W.J., Earnest, D. and Gillette, M.U. (1996) Antigenic characterization of SCN 2.2 cell line during IBMX/forskolin-induced differentiation: Comparisons with rat SCN. Soc. for Res. Biol. Rhythms Abstr. 5: 64. Liang, F.-Q., Miranda, R., Sohrabji, F., Brooks, C. and Earnest, D. (1996) Brain-derived neurotrophic factor and trkB tyrosine kinase receptors in the rat SCN: Distribution and functional implications. Soc. for Neuroscience Abstr. 22: 1141. Liang, F.-Q., Ratcliff, M., Cassone, V.M. and Earnest, D.J. (1997) Immortal time: circadian clock properties of cell lines derived from the rat SCN. Soc. Neuroscience Abstr. 23: 20. Earnest, D.J., Mahoney, J.C., Chen, W.-J.A., Sohrabji, F. and West, J.R. (1997) Alcohol exposure during rapid brain development mimics the effects of aging on BDNF expression in the SCN and circadian timekeeping. Soc. Neuroscience Abstr. 23: 20. Jezierski, M.K., Stuth, N.R., Choudhri, M., Earnest, D.J. and Sohrabji, F. (1997) Differential effects of olfactory bulb lesions on basal forebrain BDNF levels in ovariectomized and intact animals. Soc. Neuroscience Abstr. 23: 48. Liang, F.-Q., Allen, G., Walline, R. and Earnest, D. (1998). Role of brain-derived neurotrophic factor (BDNF) in the circadian regulation of the suprachiasmatic pacemaker by light. Soc. for Res. Biol. Rhythms Abstr. 6: 45. Earnest, D., Liang, F.-Q., Ratcliff, M., Walline, R. and Cassone, V. (1998). Immortal clock properties: circadian metabolic and neurotrophin rhythms in cell lines derived from the rat SCN. Soc. for Res. Biol. Rhythms Abstr. 6: 45. Liang, F.-Q., Allen, G., Walline, R. and Earnest, D. (1998). Role of brain-derived neurotrophic factor (BDNF) in the circadian sensitivity of the suprachiasmatic pacemaker to light. Soc. Neuroscience Abstr. 24: 15. Allen, G., Walline, R. and Earnest, D. (1998). Real time monitoring of gene expression in immortalized SCN cells. Soc. Neuroscience Abstr. 24: 1690. Allen, G., Walline, R. and Earnest, D. (1999). Luciferase reported expression of clock and clock-controlled genes within immortalized SCN cells. Soc. Neuroscience Abstr. 25: 1131. Allen, G., Rappe J., Cassone, V. and Earnest, D. (2000). Time passes on: immortalized scn2.2 cells confer circadian rhythmicity on NIH-3T3 fibroblasts. Soc. for Res. Biol. Rhythms Abstr. 7: 55. Earnest, D., Walline, R. and Allen, G. (2000). Illumination of molecular clockwork in the SCN: reporter gene analysis of clock and clock-controlled genes within immortalized cells. Soc. for Res. Biol. Rhythms Abstr. 7: 151. Allen,
G., Rappe J., Cassone, V. and Earnest, D. (2000). Oscillating
on borrowed time:cellular outputs from SCN2.2 cells are necessary
for rhythmicity in NIH-3T3 fibroblasts. Soc. Neuroscience Abstr.
26.
XVIV. Invited Seminars and Symposia
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