Neural - Tissue Tables

SpeciesSpecies DetailCellsEnzyme(s)Medium Reference
Bovine Bovine
(also rat)
Heart Adrenal chromaffin Paraneurons Trypsin: 0.06%
25mM HEPES buffered Locke's solution, CMF Monolayer Co-Culture of Rat Heart Cells and Bovine Adrenal Chromaffin Paraneurons , In Vitro Cell Dev Biol 26, 335, 1990
Bovine Bovine Microvascular endothelial Collagenase/Dispase: 0.1%
MEM Primary Culture of Microvascular Endothelial Cells From Bovine Retina , In Vitro 18 (7), 626, 1982
Bovine Calf
(also lamb)
Oligodendroglia Neural Trypsin: 0.1%
(see reference) Metabolic Properties of Maintained Oligodendroglia Purified from Brain , J Biol Chem 253, 1592, 1978
Chicken Chicken, White Leghorn, embryos, 17-21 day Neurons Papain: 40 u/ml
HEPES The Kinetics of the Response to Glutamate and Kainte in Neurons of the Avian and Cochlear Nucleus , Neuron 9, 173, 1992
Chicken Chick, embryo, 10-14 day Flat, retina Trypsin: 0.1%
Tyrode's solution, CMF Emergence of Flat Cells From Glia in Stationary Cultures of Embryonic Chick Neural Retina , In Vitro Cell Dev Biol 26, 1073, 1990
Chicken Chick, White Leghorn, embryo
(also rat, SD, embryo)
Spinal cord Trypsin: 0.05%
Phosphate buffer
(see reference)
Separation of cell types from embryonic chicken and rat spinal cord: characterization of motoneuron-enriched fractions. , J Neurosci 1, 204, 1981
Chicken Chick Dorsal root ganglion neurons Spinal cord Trypsin: 0.1%
Puck's saline, CMF GABA Conductance of Chick Spinal Cord and Dorsal Root Ganglion Neurons in Cell Culture , J Neurophysiol 45, 605, 1981
Chicken Chick, White Leghorn embryo, 8 day Neurons, ganglia Trypsin: 0.25%
HBSS, CMF Selective Survival of Neurons from Chick Embryo Sensory Ganglionic Dissociates Utilizing Serum-Free Supplemented Medium , Exp Cell Res 125, 183, 1980
Chicken Chicken, White Leghorn, fertile eggs Ciliary ganglion neurons Trypsin: 0.25%
Eagle's MEM Long-Term Survival and Development of Dissociated Parasympathetic Neurons in Culture , Brain Res 183, 161, 1980
Chicken Chicken, embryos, 9-10 days old Dorsal root ganglia neurons Collagenase: 0.01%
Eagle's MEM Enkephalin Inhibits Release of Substance P From Sensory Neurons in Culture and Decreases Action Potential Duration , Proc Natl Acad Sci U S A 76 (1), 526, 1979
Chicken Chick embryos Ganglion chains, sympathetic ganglia Trypsin: 0.25%
Krebs Phosphosaline Preparation of Pure Neuronal and Non-Neuronal Cultures From Embryonic Chick Sympathetic Ganglia: A New Method Based on Both Differential Cell Adhesiveness and the Formation of Homotypic Neuronal Aggregates , Brain Res 114, 391, 1976
Fish Black ghose knige fish, adult (Apteronotus albifrons) Neurons, spinal cord Trypsin: 0.4%
PBS, CMF Differences in Growth of Neurons from Normal and Regenerated Teleost Spinal Cord in vitro , In Vitro Cell Dev Biol 29A, 145, 1993
Frog Xenopus, embryonic Neuron Collagenase Type 1: 0.1%
Steinberg's solution Structure and Physiology of Developing Neuromuscular Synapses in Culture , J Neurosci 7, 473, 1987
Guinea-Pig Guinea-pig, newborn Neuron, enteric Trypsin: 0.125%
Medium 199 Tissue Culture of Mammalian Enteric Neurons , Brain Res 152, 573, 1978
Hamster hamster, male vomeronasal organ neurons Collagenase Type 1: 0.02%
Trypsin: 0.02%
PBS Regulation by voltage and adenine nucleotides of a Ca2+-activated cation channel from hamster vomeronasal sensory neurons. , J Physiol 548, 777, 2003
Human human, 9 month neurons Collagenase Type 4: 1.33%
Papain: 0.07 u/ml
Neutral Protease: 1 mg/ml
DMEM/F12 EIF2B5 mutations compromise GFAP+ astrocyte generation in vanishing white matter leukodystrophy , Nat Med 11, 277, 2005
Human human, 26 week neural progenitor cells Papain: 2.5 u/ml
Deoxyribonuclease I: 250 u/ml
Neutral Protease: 1 u/ml
DMEM/F-12 Asymmetric localization of LGN but not AGS3, two homologs of Drosophila pins, in dividing human neural progenitor cells. , J Neurosci Res 75, 782, 2004
Human human, adult ventricular epithelial Papain: 11.4 u/ml
Deoxyribonuclease I: 10 u/ml
DMEM/F12 Promoter-targeted selection and isolation of neural progenitor cells from the adult human ventricular zone. , J Neurosci Res 59, 321, 2000
Human Human, 5-65 years Retinal pigment epithelial (RPE) Trypsin: 0.25%
HBSS Maintenance of Retinoid Metabolism in Human Retinal Pigment Epithelium Cell Culture , Exp Eye Res 69, 97, 1999
Human human dorsal root ganglion neurons Collagenase Type 1: 0.2%
Dispase: 0.5%
DMEM/F-12 Two tetrodotoxin-resistant sodium channels in human dorsal root ganglion neurons. , FEBS Lett 462, 117, 1999
Mouse mouse, 6 mo neurons, neurospheres Papain: 0.2%
Hibernate Isolation and culture of adult neurons and neurospheres. , Nat Protoc Vol. 2,, , 1490-8, 2007
Mouse Mouse, C57BL/6, 1 year Neurons Papain: 0.2%
DMEM Culture of Adult Mouse Neurons , Biotechniques 38(1), 99-104, 2005
Mouse mouse, adult brain and spinal cord cells Trypsin: 0.25%
PBS Expression of a dominant negative IFN-gammareceptor on mouse oligodendrocytes. , Glia 51, 22-34, 2005
Mouse mouse, neonatal neurons PDS kit:
EBSS Developmental regulation of gonadotropin-releasing hormone neurons by fibroblast growth factor signaling. , Endocrinology 145, 3830, 2004
Mouse mouse, C57BL cerebellar granule cell precursors Papain: 16.5 u/ml
Deoxyribonuclease I: 0.008%
Dulbecco's PBS Cerebellar granule cell precursors can differentiate into astroglial cells , Proc Natl Acad Sci U S A 101, 1211, 2004
Mouse mouse, 1-3 day neurons, ganglia Papain: 20 u/ml
Deoxyribonuclease I: 100 u/ml
Collagenase: 0.3%
Trypsin: 0.05%
HBSS Cell surface trafficking of the antidepressant-sensitive norepinephrine transporter revealed with an ectodomain antibody. , Mol Cell Neurosci 24, 1131, 2003
Mouse Mouse, postnatal-day-1-old (P1) Neurons Trypsin: 0.25%
NGF-containing medium Caspase Inhibition Extends the Commitment to Neuronal Death Beyond Cytochrome c Release to the Point of Mitochondrial Depolarization , J Cell Biol 150 (1), 131, 2000
Mouse Mouse embryos Neurons, DRG and SCG Trypsin: 0.25%
L-15 medium P75-Eficient Embryonic Dorsal Root Sensory and Neonatal Sympathetic Neurons Display a Decreased Sensitivity to NGF , Development 120, 1027, 1994
Mouse 65 days neurons, neuronal precursors Trypsin: 0.1%
Deoxyribonuclease I: 0.001%
DMEM De Novo Generation of Neuronal Cells from the Adult Mouse Brain , Proc Natl Acad Sci U S A 89, 8591, 1992
Mouse Mouse, CD-1, neonate Neurons, dorsal root ganglion Trypsin: 0.25%
HBSS Enhanced Neuronal Regeneration by Retinoic Acid of Murine Dorsal Root Ganglia and of Fetal Murine and human Spinal Cord in vitro , In Vitro Cell Dev Biol 27, 55, 1991
Mouse Mouse, fetal Precursor Trypsin: 0.5%
PBS Isolation and Characterization Of Mouse Neural Precursor Cells in Primary Culture , In Vitro Cell Dev Biol 27, 615, 1991
Mouse Mouse (SWR or CF1), 1-3 months Papillae, taste receptor Pronase E: 0.15%
Carbonate-Phosphate buffer
(see reference)
A Method for Isolating and Patch-Clamping Single Mammalian Taste Receptor Cells , Brain Res 503, 326, 1989
Mouse Mouse, neonatal
(also chick)
PNS test neurons Trypsin: 0.08%
Eagle's Basal Medium (see reference) Low Molecular Weight Agents Support Survival of Cultured Neurons From the Central Nervous System , J Neurosci 4 (3), 654, 1984
Mouse Mouse (BALB/c), adult Neurons, spinal cord Collagenase Type 3: 0.25%
Hank's BSS, CMF Survival of Purified Motor Neurons In Vitro: Effects of Skeletal Muscle-Conditioned Medium , Neurosci Lett 38, 187, 1983
Mouse Mouse, 0-30 day Neural Trypsin NF 1:250: 50 0.25%
BSS Cultured Cell Systems and Methods for Neurobiology , Methods Enzymol 32, 765, 1974
Porcine porcine, adult, 60-100 kg superior cervical ganglia Papain: 2 u/ml
Collagenase: 0.12%
Dispase: 0.48%
HBSS Presynaptic alpha7-nicotinic acetylcholine receptors mediate nicotine-induced nitric oxidergic neurogenic vasodilation in porcine basilar arteries. , J Pharmacol Exp Ther 298, 122, 2001
Quail Quail Neural crest Trypsin: 0.05%
MEM, HBSS Clonal Analysis of Quail Neural Crest Cells: They are Pluripotent and Differentiate in Vitro in the Absence of Noncrest Cells , Dev Biol 80, 96, 1980
Rat Rat, Wistar, 4 day hippocampal neurons PDS kit:
Neurobasal A Cross-talk of vitamin D and glucocorticoids in hippocampal cells. , J Neurochem 96, 500-9, 2006
Rat rat, 2 month spinal cord progenitor cells Hybrid toxins PDS kit: see reference
Neurobasal A Analysis of Green Fluorescent Protein Expression in Transgenic Rats for Tracking Transplanted Neural Stem/Progenitor Cells , J Histochem Cytochem 53(10), 1215, 2005
Rat rat, SD, E19 dorsal root ganglia PDS kit:
MEM/Ham's F12 AMG 9810 [(E)-3-(4-t-butylphenyl)-N-(2,3-dihydrobenzo[b][1,4] dioxin-6-yl)acrylamide], a novel vanilloid receptor 1 (TRPV1) antagonist with antihyperalgesic properties. , J Pharmacol Exp Ther 313, 474, 2005
Rat rat, Wistar, E15 vomeronasal receptor neurons Collagenase/Dispase: 0.1%
Papain: 0.5 u/ml
DMEM/F12 Maturation of vomeronasal receptor neurons in vitro by coculture with accessory olfactory bulb neurons. , Chem Senses 30, 111, 2005
Rat rat, SD, 1-2 day cortical astrocytes Papain: see reference
DMEM Mechanical strain injury increases intracellular sodium and reverses Na+/Ca2+ exchange in cortical astrocytes. , Glia 51, 35-46, 2005
Rat rat, fetal brainstem and cortical neurons PDS kit:
DMEM Extrahypothalamic expression of the glucagon-like peptide-2 receptor is coupled to reduction of glutamate-induced cell death in cultured hippocampal cells. , Endocrinology 145, 3495, 2004
Rat rat, SD, male, 270-330 g spinal progenitor cells PDS kit: with modifications
Neurobasal medium Intrathecal spinal progenitor cell transplantation for the treatment of neuropathic pain , Cell Transplant 11, 17, 2002
Rat rat, Fisher, 8-9 week adult progenitor Papain: 2.5 u/ml
Deoxyribonuclease I: 250 u/ml
Dispase: 1 u/ml
DMEM/F-12 The adult substantia nigra contains progenitor cells with neurogenic potential , J Neurosci 22, 6639, 2002
Rat rat. 15 week neurons Papain: 0.2%
Hibernate A Characterization of mitotic neurons derived from adult rat hypothalamus and brain stem. , J Neurophysiol 87, 1076, 2002
Rat Rat Neurons, hippocampal Papain: 15 - 20 u/ml
Eagle's MEM
(see reference)
B-Amyloid Peptide Blocks the Response of a7-Containing Nicotinic Receptors on Hippocampal Neurons , PNAS 98 (8), 4734, 2001
Rat Rat, newborn, 7 days old Neurons Trypsin: 0.25%
MEM10 A Sensory Neuron Subpopulation with Unique Sequential Survival Dependence on Nerve Growth Factor and Basic Fibroblast Growth Factor during Development , J Neurosci 21 (22), 8873, 2001
Rat Rat, embryonic 18-19 day hippocampal neurons PDS kit: see reference
DMEM Phosphorylation of cAMP response element-binding protein in hippocampal neurons as a protective response after exposure to glutamate in vitro and ischemia in vivo. , J Neurosci 21, 9204-13, 2001
Rat Rat, embryonic, day 18 Cortical Papain:
Neurobasal medium and DMEM Immobilization of Neural Cells in Three-Dimensional Matrices for Biosensor Applications , Biosensors & Bioelectronics 14, 871, 2000
Rat Rat, Wister, postnatal, P1-3 days Neurons, hippocampal Papain: 20 u/ml
EBSS Activity-Dependent Formation of Perforated Synapses in Cultured Hippocampal Neurons , Eur J Neurosci 11, 4241, 1999
Rat Rat, SD, female, pregnant Sciatic nerves Trypsin: 0.025%
L-15 medium
(see reference)
Prospective Identification, Isolation by Flow Cytometry, and In Vivo Self-Renewal of Multipotent Mammalian Neural Crest Stem Cells , Cell 96, 737, 1999
Rat Rat
(also mice)
Spinal cord Trypsin: 0.133%
HBSS and PIPES Identification of a Neural Stem Cell in the Adult Mammalian Central Nervous System , Cell 96, 25, 1999
Rat Rat, (Long Evans), 2-5 day old Neurons, hippocampal Papain: 20 u/ml
EBSS Activation and Desensitization of Hippocampal Kainate Receptors , J Neurosci 17 (8), 2713, 1997
Rat Rat, Wister and SD, newborn, 0-21 days, either sex Myenteric ganglia Trypsin: 0.05%
MEM-HEPES A New Method for the Isolation of Myenteric Plexus from the Newborn Rat Gastrointestinal Tract , Brain Res Proto 1, 109, 1997
Rat Rat, pups, 24-48 h old Neurons, hippocampal Papain: 20 u/ml
Harvest buffer Differential Surface Expression and Phosphorylation of the N-Methyl-D-Aspartate Receptor NR1 and NR2 in Cultured Hippocampal Neurons , J Biol Chem 272 (7), 4135, 1997
Rat Rat, SD, adult, 250-300g, P8 Neurons, DRG Dispase: 0.5%
L-15 w/ CO2 Regeneration of Adult Axons in White Matter Tracts of the Central Nervous System , Nature 390, 680, 1997
Rat Rat, 1-4 day old Neurons, hippocampal Papain: 20 u/ml
MEM Cannabinoids Inhibit N- and P/Q-Type Calcium Channels in Cultured Rat Hippocampal Neurons , J Neurophysiol 78, 43, 1997
Rat SD, Fisher hippocampal neurons Papain: 0.2%
HibernateA/B27 Isolation and culture of adult rat hippocampal neurons of any age. , J Neurosci Methods 71, 143, 1997
Rat rat, E18 hippocampal neurons Papain: 20 u/ml
Deoxyribonuclease I: 0.01%
MEM Upregulation of GABAA current by astrocytes in cultured embryonic rat hippocampal neurons. , J Neurosci 16, 2912-23, 1996
Rat Rat, pups, 14 day old Neurons, sympathetic Dispase: 0.24%
HBSS Extrinsic Factors Influence the Expression of Voltage-Gated K Currents on Neonatal Rat Sympathetic Neurons , J Neurosci 13 (6), 2591, 1993
Rat Rats, SD, 11-14 day old, 29-32 g Basal forebrain neurons Trypsin: 0.125%
Gey's BSS The Whole-Cell Calcium Current in Acutely Dissociated Magnocellular Cholinergic Basal Forebrain Neurones of the Rat , J Physiol 460, 91, 1993
Rat Rat, Wistar-Hanover, 7-12 day Postnatal dopamine neurons Trypsin: 0.035%
(see reference) Identified Postnatal Mesolimbic Dopamine Neurons in Culture: Morphology and Electrophysiology , J Neurosci 12 (11), 4264, 1992
Rat Rat, SD, 18-day-old, fetus Hippocampal Trypsin: 0.2%
HBSS IGF-I and IGF-II Protect Cultured Hippocampal and Septal Neurons Against Calcium-Mediated Hypoglycemic Damage , J Neurosci 12 (4), 1558, 1992
Rat Rat Stem, neural crest Collagenase: 0.075%
Ringer's solution Isolation of a Stem Cell for Neurons and Glia from the Mammalian Neural Crest , Cell 71, 973, 1992
Rat Rat, SD, pregnant, Charles River Pyramidal neurons Nonpyramidal neurons Trypsin: 0.027%
HEPES Electrophysiological Comparison of Pyramidal and Stellate Nonpyramidal Neurons in Dissociated Cell Culture of Rat Hippocampus , Brain Res Bull 26, 333, 1991
Rat Rat, SD, embryos, 19-21 days gestation Schwann, dorsal root ganglia Trypsin: 0.25%
HBSS, CMF Effects of Ethanol on Rat Schwann Cell Proliferation and Myelination in Culture , In Vitro Cell Dev Biol 26, 129, 1990
Rat Rat
(also bovine)
Heart Adrenal chromaffin Paraneurons Trypsin: 0.06%
25mM HEPES buffered Locke's solution, CMF Monolayer Co-Culture of Rat Heart Cells and Bovine Adrenal Chromaffin Paraneurons , In Vitro Cell Dev Biol 26, 335, 1990
Rat Rat, SD, 8 day old pups Cerebellar neurons Trypsin: 0.025%
Eagle's MEM Glutamate Becomes Neurotoxic Via the N-methyl-D-aspartate Receptor When Intracellular Energy Levels are Reduced , Brain Res 451, 205, 1988
Rat Rat, fetuses, 18 day old Hippocampal neurons Trypsin: 0.2%
Eagle's MEM Isolated Hippocampal Neurons in Cryopreserved Long-Term Cultures: Development of Neuroarchitecture and Sensitivity to Nmda , Int J Dev Neurosci 6 (5), 439, 1988
Rat Rat, postnatal Septal neurons Papain: 0.05%
PBS, CMF Septal Cholinergic Neurons From Postnatal Rat Can Survive In The Dissociate Culture Conditions In The Presence Of Nerve Growth Factor , Neurosci Lett 79, 85, 1987
Rat Rat, Long Evans, 1-15 days Neurons, visual cortex Papain: 20 u/ml
BSS see reference (see reference) Primary Culture of Identified Neurons From the Visual Cortex of Postnatal Rats , J Neurosci 6, 3044, 1986
Rat Rat, SD, female, 4 month old CNS cells Trypsin: 0.25%
EBSS Evidence That Axons are Mitogenic for Oligodendrocytes Isolated From Adult Animals , Nature 320, 756, 1986
Rat Rat, embryo, 15 day Dorsal horn neurons Spinal Trypsin: 0.025%
Ham's F-12 Synaptic Transmission between Dorsal Root Ganglion and Dorsal Horn Neurons in Culture: Antagonism of Monosynaptic Excitatory Postsynaptic Potentials and Glutamate Excitation by Kynurenate , J Neurosci 5, 2281, 1985
Rat Rat, fetus, 18-20 day Hippocampal neurons Trypsin: 0.25%
HBSS, CMF An Electron Microscopic Study of the Development of Axons and Dendrites by Hippocampal Neurons in Culture. I. Cells Which Develop Without Intercellular Contacts , J Neurosci 4, 1944, 1984
Rat Rat, postnatal Ganglion, retina Papain: 12.5 u/ml
HBSS w/5 mM HEPES Monoclonal Antibody to Thy-1 Enhances Regeneration of Processes by Rat Retinal Ganglion Cells in Culture , Science 224, 303, 1984
Rat Rat, SD, neonatal Retinal ganglion Trypsin: 0.25%
CMF medium /Ham's F-10 with FCS Retrograde Labeling, Enrichment, and Characterization of Retinal Ganglion Cells from the Neonatal Rat , J Neurosci 3, 2532, 1983
Rat Rat, SD, pups Retina Trypsin: 0.25%
Ham's F-12 Retrograde Labeling, Enrichment, and Characerization of Retinal Ganglion Cells from the Neonatal Rat , J Neurosci 3 (12), 2532, 1983
Rat Rat, W/FU, 5-8 day Neurons and glial Trypsin: 0.25%
MEM
(see reference)
Cell-Type-Specific Markers for Distinguishing and Studying Neurons And the Major Classes of Glial Cells in Culture , Brain Res 174, 283, 1979
Rat Rat, fetus Neurons, sympathetic Trypsin: 0.25%
L-15 or HBSS, CMF Postnatal Rat Sympathetic Neurons In Culture. 1. A Comparison With Embryonic Neurons , J Neurophysiol 42, 1410, 1979
Rat Rat, Wistar/Furth, newborn
(also bovine)
Schwann Trypsin: 0.25%
DMEM Studies on Cultured Rat Schwann Cells. I. Establishment of Purified Populations From Cultures of Peripheral Nerve , Brain Res 165, 105, 1979
Rat Rat, embryo Neurons, cortical Trypsin: 0.027%
MEM Rat Cortical Neurons in Cell Culture: Culture Methods, Cell Morphology, Electrophysiology, and Synapse Formation , Brain Res 149, 279, 1978
Rat Rat, newborn Neurons, sympathetic Collagenase Type 1: 0.01%
Hank's solution, CF Neurotransmitter Synthesis and Uptake by Isolated Sympathetic Neurones in Microcultures , Nature 270, 147, 1977
Rat Rat, neonatal Neurons, superior cervical ganglia Trypsin: 0.1%
Basal L-15 medium Primary Cultures of Dissociated Sympathetic Neurons I. Establishment of Long-Term Growth in Culture and Studies of Differentiated Properties , J Cell Biol 59, 329, 1973
Salamander Salamander, 18-25 cm Retina Papain: 14 u/ml
Saline Rod Cells Dissociated from Mature Salamander Retina: Ultrastruture and Uptake of Horseradish Peroxidase , J Cell Biol 100, 175, 1985
Salamander Salamander (A.tigrinum) Photoreceptors, retina Papain: 0.05%
(see reference) Responses to Light of Solitary Rod Photoreceptors Isolated From Tiger Salamander Retina , Proc Natl Acad Sci U S A 75, 3507, 1978
Sheep Lamb
(also calf)
Oligodendroglia Neural Trypsin: 0.1%
(see reference) Metabolic Properties of Maintained Oligodendroglia Purified from Brain , J Biol Chem 253, 1592, 1978
Shellfish Snails (Helisoma trivolis), albino, adult Buccal ganglia; SLT muscle Trypsin: 0.2%
DMEM Target Contact Regulates the Calcium Responsiveness of the Secretory Machinery During Synaptogenesis , Neuron 6, 145, 1991
Shellfish Pond snail (Helisoma) albino, adult Somata, buccal ganglia Trypsin: 0.2%
Antibiotic saline, Leibowitz 50% The Formation of Chemical Synapses Between Cell-Cultured Neuronal Somata , J Neurosci 8, 1032, 1988
Shellfish Helisoma trivovlis Buccal ganglia Trypsin: 0.2%
Liebowitz L-15 medium Single Channel Activity Differs in Growing and Nongrowing Growth Cones of Isolated Identified Neurons of Helisoma , J Neurosci Res 13, 285, 1985
Shellfish Mollusc, juvenile 1-2 gm or adult 50-100 gm Neurons LUQ cells Protease: 1%
L15 medium Neurite Regeneration by Aplysia Neurons in Dissociated Cell Culture: Modulation by Aplysia Hemolymph and the Presence of the Initial Axonal Segment , J Neurosci 3 (12), 2403, 1983
Shellfish Aplysia californica LUQ cells RUQ cells Protease: 1%
L15 medium Identified Aplysia Neurons Form Specific Chemical Synapses in Culture , J Neurosci 3 (12), 2614, 1983
Turtle Turtle (Pseudemys scripta elegans) Retinal Papain: 0.1% (13.5 u/mg)
Kreb's Ringer Biosynthesis of Acetylcholine in Turtle Photoreceptors , Proc Natl Acad Sci U S A 69, 1987, 1972