Showing papers by "Rockefeller University published in 1973"

Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution.

Abstract: A novel cell type has been identified in adherent cell populations prepared from mouse peripheral lymphoid organs (spleen, lymph node, Peyer's patch). Though present in small numbers (0.1–1.6% of the total nucleated cells) the cells have distinct morphological features. The nucleus is large, retractile, contorted in shape, and contains small nucleoli (usually two). The abundant cytoplasm is arranged in processes of varying length and width and contains many large spherical mitochondria. In the living state, the cells undergo characteristic movements, and unlike macrophages, do not appear to engage in active endocytosis. The term, dendritic cell, is proposed for this novel cell type.

Leukocyte locomotion and chemotaxis. New methods for evaluation, and demonstration of a cell-derived chemotactic factor.

Abstract: Polymorphonuclear leukocyte (PMN) locomotion and chemotaxis have been evaluated by direct microscopic observation of individual cells in thin slide-cover slip preparations, and also by observations on populations of cells migrating into a Millipore filter. The direct microscopic method used the polarity of the locomoting PMNs (broad, advancing lamellipodium and knoblike constriction at the rear) to record the direction of movement. The Boyden chamber Millipore assay was made more reliable by following the front of cells advancing into the filter, rather than counting the number of cells on the lower filter surface. Special modifications of the Millipore assay were necessary in order to distinguish between influences on rate of locomotion and true chemotaxis. In both systems the results indicate that under certain conditions leukocytes, and in particular PMNs, release into the medium a factor stimulating locomotion and exerting chemotactic action on PMNs in the vicinity. This cell-derived factor appears not to require serum factors for its release or action.

Atlas of estradiol-concentrating cells in the central nervous system of the female rat.

Abstract: Two hours following intraperitoneal injection, estradiol-H3 is concentrated by cells in a system of limbic and hypothalamic structures. Preoptic-hypothalamic nuclei containing estrogen-concentrating cells include the medial preoptic area, medial anterior hypothalamus, ventromedial nucleus, arcuate nucleus and ventral premammillary nucleus. Limbic structures include the medial and cortical nuclei of the amygdala, lateral septum, bed nucleus of the stria terminalis, diagonal band of Broca, olfactory tubercle, ventral hippocampus, and prepiriform and entorhinal cortex. Labelled cells were also found in the lateral and ventrolateral portions of the mesencephalic central grey. Compared to these regions, most other regions of the nervous system, including the spinal cord, have very small numbers of labelled cells, which are relatively weakly labelled, and are not found in regular, specific locations. The distribution of estrogen-concentrating cells determined with the present autoradiographic method agrees with previous autoradiographic conclusions and with biochemical results from cell fractionation experiments. The locations of estrogen-concentrating cells coincide in several brain regions with locations of estrogen-dependent neuroendocrine control mechanisms, as determined by brain implants, lesions, electrical stimulation, and electrophysiological recording. Moreover, experimental neuroanatomical studies have provided evidence for several pathways connecting regions which concentrate radioactive estradiol. Taken together, the evidence suggests a limbic-hypothalamic system of estrogen-concentrating neurons which participate in the control of mating behavior and of gonadotrophin release from the pituitary.

Turnover of transmitter and synaptic vesicles at the frog neuromuscular junction

Abstract: Curarized cutaneous pectoris nerve-muscle preparations from frogs were stimulated at 10/s or at 2/s for periods ranging from 20 min to 4 h. End plate potential were recorded intracellularly and used to estimate the quantity of transmitter secreted during the period of stimulation. At the ends of the periods of stimulation the preparations were either fixed for electron microscopy or treated with black widow spider venom to determine the quantities of transmitter remainind in the terminal. Horseradish peroxidase or dextran was added to the bathing solution and used as a tracer to detect the formation of vesicles from the axolemma. During 4 h of stimulation at 2/s many new vesicles were formed from the axolemma and the quantity of transmitter secreted was several times greater than the quantity in the initial store. After this period of stimulation, the terminals were severely depleted of transmitter, but not of vesicles, and their general morphological organization was normal. During 20 min of stimulation at 10/s the nerve terminals swelled and were severely depleted both of vesicles and of transmitter. During a subsequent hour of rest the changes in morphology were largely reversed, many new vesicles were formed from the axolemma and the stores of transmitter were partially replenished. These results suggest (a) that synaptic vesicles fuse with, and re-form from, the membrane of the nerve terminal during and after stimulation and (b), that the re-formed vesicles can store and release transmitter.

An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. I. Chick embryo fibroblast cultures transformed by avian RNA tumor viruses.

Abstract: Chick embryo fibroblast cultures develop fibrinolytic activity after transformation by Rous sarcoma virus (RSV). This fibrinolytic activity is not present in normal cultures, and it does not appear after infection with either nontransforming strains of avian leukosis viruses or cytocidal RNA and DNA viruses. In cultures infected with a temperature sensitive mutant of RSV the onset of fibrinolysis appears after exposure to permissive temperatures and precedes by a short interval the appearance of morphological evidence of transformation. See PDF for Structure The rate of fibrinolysis in transformed cultures depends on the nature of the serum that is present in the growth medium: some sera (e.g., monkey or chicken serum) promote high enzymatic activity, while others (calf, fetal bovine) do not. Some sera contain inhibitors of the fibrinolysin. Based on the effect of a small number of known inhibitors, at least one step of the fibrinolytic process shows specificity resembling that of trypsin. The sera of sarcoma-bearing chickens contain an inhibitor of the fibrinolysin, whereas normal chicken sera do not. For general discussion, conclusions, and summary see the accompanying paper, part II, ( J. Exp. Med . 137 :112).

Ascending central gustatory pathways.

Abstract: The central gustatory pathways of the albino rat have been traced using a combined electrophysiological-neuroanatomical technique. Gustatory responses in the medulla were recorded in the region of the solitary nucleus which receives the seventh nerve primary afferents. Fibers traced from lesions of these recording sites did not cross as expected into the medial lemniscus, but instead travelled rostrally to terminate ipsilaterally in a small celled area dorsal and ventral to the brachium conjunctivum as it enters the pons. Since gustatory responses could be recorded in this region it represents a previously undescribed secondary “pontine taste area.” Lesions of PTA result in degeneration of a bilateral ascending pathway travelling in the dorsomedial tegmentum to terminate in the classical gustatory nuclei of the thalamus. Other fibers in this pathway continue rostrally and distribute in the subthalamus, dorsolateral hypothalamus and subpallidal gray in the ventral forebrain. These findings in a mammal resemble those established nearly three quarters of a century ago by Herrick ('05) in the carp, and confirm his prediction that “broad lines of similarity [would be found] between both the peripheral and central gustatory paths in all vertebrates”.

Luteinizing hormone-releasing factor potentiates lordosis behavior in hypophysectomized ovariectomized female rats.

Abstract: Subcutaneous injection of luteinizing hormone-releasing factor (LRF) in estrogen-primed hypophysectomized, ovariectomized female rats facilitates the appearance of the lordosis response. The LRF effect on lordosis was seen 90, 180, and 360 minutes after injection. This effect could help to synchronize the female9s mating behavior with the ovulatory discharge of luteinizing hormone.

Golgi fractions prepared from rat liver homogenates. I. Isolation procedure and morphological characterization.

Abstract: In devising a new procedure for the isolation of Golgi fractions from rat liver homogenates, we have taken advantage of the overloading with very low density lipoprotein (VLDL) particles that occurs in the Golgi elements of hepatocytes approximately 90 min after ethanol is administered (0.6 g/100 g body weight) by stomach tube to the animals. The VLDLs act as morphological markers as well as density modifiers of these elements. The starting preparation is a total microsomal fraction prepared from liver homogenized (1:5) in 0.25 M sucrose. This fraction is resuspended in 1.15 M sucrose and loaded at the bottom of a discontinuous sucrose density gradient. Centrifugation at approximately 13 x 10(6)g.min yields by flotation three Golgi fractions of density >1.041 and <1.173. The light and intermediate fractions consist essentially of VLDL-loaded Golgi vacuoles and cisternae. Nearly empty, often collapsed, Golgi cisternae are the main component of the heavy fraction. A procedure which subjects the Golgi fractions to hypotonic shock and shearing in a French press at pH 8.5 allows the extraction of the content of the Golgi elements and the subsequent isolation of their membranes by differential centrifugation.

Fibrinolysis associated with oncogenic transformation : requirement of plasminogen for correlated changes in cellular morphology, colony formation in agar, and cell migration

Abstract: Fetal bovine and dog serum were selectively freed of plasminogen by affinity chromatography. The resulting serum as well as native and reconstituted serum (obtained by the addition of purified plasminogen to the plasminogen-depleted serum) were used to examine the role of plasminogen in (a) growth of normal and SV-40-transformed hamster embryo fibroblasts in liquid medium, (b) growth of SV-40-transformed hamster embryo fibroblasts in soft agar, (c) aggregation — a characteristic morphological change of SV-40-transformed hamster cells, and (d) migration of SV-40-transformed and control 3T3 cells from a monolayer into a "wound." The results demonstrated that exponential growth of both normal and transformed cells in liquid medium proceeded at the same rate in the presence or absence of plasminogen. In contrast, removal of plasminogen markedly depressed the plating efficiency of transformed cells in soft agar, eliminated their characteristic aggregation, and substantially reduced the extent of migration. The role of plasminogen and its activation in oncogenic transformation is discussed.

An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. II. Mammalian fibroblast cultures transformed by DNA and RNA tumor viruses.

Abstract: Chick, hamster, mouse, and rat embryo fibroblast cultures, transformed by either DNA or RNA viruses, show fibrinolytic activity under suitable conditions of growth and in appropriate media; normal counterpart cultures do not. The fibrinolysin is produced by the interaction of two protein factors: one of these, a cell factor, is released by transformed cells and accumulates in the medium when cultures are incubated in the absence of scrum. The second factor, the serum factor, is a specific protein that is present in sera of many avian and mammalian species, including man. Not all sera yield fibrinolysin on interaction with any given transformed cell factor, and the spectrum of activating sera is distinctive for each cell factor. This pattern appears to be determined by the cell type, rather than by the transforming virus. An important role for the fibrinolysin in oncogenic transformation is suggested by the following correlations. (a) The initial appearance of fibrinolysin precedes the morphological change after the transfer to permissive temperatures of chick fibroblast cultures infected with a temperature-sensitive mutant of RSV. (b) The initiation of fibrinolysis and of morphological change both require the synthesis of new protein, but not the synthesis of either DNA or rRNA. (c) The activity of the fibrinolysin is correlated with the retention of abnormal morphology in hamster cells transformed by SV-40. (d) The sera of normal chicks effectively activate fibrinolysis with the cell factor from transformed chick cells. In contrast the sera of chicks with RSV tumors do not; these contain an inhibitor of the fibrinolytic activity.

Identification of human B and T lymphocytes by scanning electron microscopy.

Abstract: In this study a variety of human lymphocytes of known B or T cell type, obtained from multiple sources, were prepared for scanning electron microscopy (SEM) by the critical point drying method. Distinction between normal B and T lymphocytes was relatively easy in most instances, on the basis of their surface architecture. Using immunological methods, between 20 and 30% of normal peripheral blood lymphocytes (PBL) were identified as B cells and from 69 to 82% as T cells. SEM results showed that 20% of the PBL had a complex villous surface and approximately 80% of cells were smaller and had a relatively smooth surface. Comparison of the above data and enrichment of B cells from PBL, by centrifugation after T cell rosettes had formed, indicated that the "villous" cells were B lymphocytes and the "relatively smooth" cells were T lymphocytes. T cells obtained from two human thymuses were also of the generally smooth cell type. Further evidence for the distinction of B and T lymphocytes, on the basis of surface morphology, was obtained from the examination of cultured lymphoid cell lines of known B or T cell derivation. Cells from cases of chronic lymphocytic leukemia also provided support for the above interpretations. Five of six untreated cases were clearly of B cell type by immunologic and SEM criteria. One unusual case showed the presence of T and B lymphocytes in almost equal numbers by SEM and a mixture of B and T cells by immunologic markers. An additional case that had received chemotherapy showed numerous atypical cells that were difficult to classify by SEM. Detailed examination of the smoother T cells showed that at least half of them had a moderate number of surface digitations and a small proportion had an intermediate surface morphology with a relatively large number of surface digitations. The latter presented difficulties in classification and may correspond to different stages of differentiation and represent subpopulations of lymphocytes. The distinction between human B and T lymphocytes on the basis of their surface architecture can be made by SEM of critical point dried samples, with relative ease in most but not all instances. The effects of stimulation, cell cycle, differentiation, intercellular contact, and density of cell population, on the surface architecture of lymphoid cells, remain to be determined.

RIBOSOME-MEMBRANE INTERACTION : Nondestructive Disassembly of Rat Liver Rough Microsomes into Ribosomal and Membranous Components

Abstract: In a medium of high ionic strength, rat liver rough microsomes can be nondestructively disassembled into ribosomes and stripped membranes if nascent polypeptides are discharged from the bound ribosomes by reaction with puromycin. At 750 mM KCl, 5 mM MgCl2, 50 mM Tris·HCl, pH 7 5, up to 85% of all bound ribosomes are released from the membranes after incubation at room temperature with 1 mM puromycin. The ribosomes are released as subunits which are active in peptide synthesis if programmed with polyuridylic acid. The ribosome-denuded, or stripped, rough microsomes (RM) can be recovered as intact, essentially unaltered membranous vesicles Judging from the incorporation of [3H]puromycin into hot acid-insoluble material and from the release of [3H]leucine-labeled nascent polypeptide chains from bound ribosomes, puromycin coupling occurs almost as well at low (25–100 mM) as at high (500–1000 mM) KCl concentrations. Since puromycin-dependent ribosome release only occurs at high ionic strength, it appears that ribosomes are bound to membranes via two types of interactions: a direct one between the membrane and the large ribosomal subunit (labile at high KCl concentration) and an indirect one in which the nascent chain anchors the ribosome to the membrane (puromycin labile). The nascent chains of ribosomes specifically released by puromycin remain tightly associated with the stripped membranes. Some membrane-bound ribosomes (up to 40%) can be nondestructively released in high ionic strength media without puromycin; these appear to consist of a mixture of inactive ribosomes and ribosomes containing relatively short nascent chains. A fraction (∼15%) of the bound ribosomes can only be released from membranes by exposure of RM to ionic conditions which cause extensive unfolding of ribosomal subunits, the nature and significance of these ribosomes is not clear.

Human immunoglobulins: classes, subclasses, genetic variants, and idiotypes.

Abstract: Publisher Summary The immunoglobulin classes and subclasses represent a group of structurally related proteins, and in all instances consist of two pairs of polypeptide chains held together by disulfide bridges and noncovalent forces. The immunoglobulins can be classified according to three different parameters related to different antigenic and biochemical structures: (1) heavy-chain C regions constitute the basis for the different classes and subclasses; (2.) light-chain C regions specify the types or subtypes; and (3.) variable regions of both heavy and light chains can be divided into different groups and subgroups. Immunoglobulin G is the major immunoglobulin class making up about 80% of serum immunoglobulin. There are four subclasses of immunoglobulin G that were first distinguished by antigenic differences. These also show distinct peptide and amino acid differences and differences in biological activities that are discussed in details in the chapter. Immunoglobulin A is the dominant immunoglobulin class in secretions and may be present in serum as a monomer or polymer. The antigenic properties of the immunoglobulins can be divided into three main categories: (1) antigens present in all normal sera are called “isotypes;” (2) antigens present in some but not in all normal sera and segregate as if controlled by allelic genes are called “azlotypes;” (3) antigenic properties related to one particular antibody population are called “individual specificities” or “idiotypes.”

Isolation and Purification of the Envelope Proteins of Newcastle Disease Virus

Abstract: A procedure has been developed for the isolation of Newcastle disease virus (NDV) envelope proteins The two surface glycoproteins and the non-glycosylated membrane protein were solubilized with 2% Triton X-100 and 1 m KCl Removal of the KCl by dialysis yielded by precipitation a pure preparation of the non-glycosylated membrane protein, which is insoluble in solutions of low ionic strength The soluble fraction consisting of the two glycoproteins possessed full neuraminidase and hemagglutinating activities The two glycoproteins could be separated by rate zonal sedimentation in a sucrose gradient containing 1% Triton X-100 and 1 m KCl Under these conditions, the sedimentation coefficient of the larger glycoprotein, virus protein 1, was 93s, and that of the smaller, virus protein 2, was 61s Both hemagglutinating and neuraminidase activities were associated with virus protein 1; virus protein 2 had neither activity The results suggest that both activities reside on a single NDV glycoprotein Similar results were obtained previously with another paramyxovirus, simian virus 5 These findings suggest that the association of hemagglutinating and neuraminidase activities with one glycoprotein is a general property of the paramyxovirus group

GOLGI FRACTIONS PREPARED FROM RAT LIVER HOMOGENATES : II. Biochemical Characterization

Abstract: The three Golgi fractions isolated from rat liver homogenates by the procedure given in the companion paper account for 6-7% of the protein of the total microsomal fraction used as starting preparation. The lightest, most homogeneous Golgi fraction (GF(1)) lacks typical "microsomal" activities, e.g., glucose-6-phosphatase, NADPH-cytochrome c-reductase, and cytochrome P-450. The heaviest, most heterogeneous fraction (GF(3)) is contaminated by endoplasmic reticulum membranes to the extent of approximately 15% of its protein. The three fractions taken together account for nearly all the UDP-galactose: N-acetyl-glucosamine galactosyltransferase of the parent microsomal fraction, and for approximately 70% of the activity of the original homogenate. Omission of the ethanol treatment of the animals reduces the recovery by half. The transferase activity is associated with the membranes of the Golgi elements, not with their content. Galactose is transferred not only to N-acetyl-glucosamine but also to an unidentified lipid-soluble component.

An Analysis of Hamster Afferent Taste Nerve Response Functions

Abstract: Sensitivities to moderately intense stimuli representing four taste qualities to man were determined for 79 hamster chorda tympani fibers. Some fibers were very sensitive to sucrose, sodium chloride, or hydrochloric acid, but none were very sensitive to quinine. These sensitivities were not randomly distributed among fibers: the sucrose sensitivity was separated from and negatively correlated with the other sensitivities which were associated and positively correlated with each other. Moreover, there were a limited number of sensitivity patterns: (a) fibers responding best to sucrose responded second-best to salt, less to acid, not to quinine; (b) fibers responding best to salt either responded second-best to sucrose and not to acid or quinine; or second-best to acid, less to quinine, and not to sucrose; and (c) fibers responding best to acid responded second-best to salt, more to quinine, and less to sucrose than other fibers. Therefore, if four stimuli of different taste qualities are ordered from acceptable to unacceptable, neural response functions of most hamster chorda tympani taste fibers peak at one point. Sensitivities to nine other moderately intense stimuli which vary in quality to man were also determined for 46-49 of the fibers. Sensitivities to sweet stimuli were always associated with each other and separated from sensitivities to nonsweet stimuli. Sensitivities to nonsweet stimuli were all associated with each other; however, the strongest correlations were between sensitivities to stimuli of like quality, e.g., the three acids or the two sodium salts.

Hormone secretion by cells dissociated from rat anterior pituitaries

Abstract: A new procedure has been developed for dissociating anterior pituitary tissue and producing a viable suspension of single cells. The procedure involves incubation of small tissue blocks in 1 mg/ml trypsin (15 min), followed by incubation in 8 µg/ml neuraminidase and 1 mM EDTA (15 min), followed by mechanical dispersion. Cell yields are ∼55%, based on recovered DNA. By electron microscopy five types of secretory cells (somatotrophs, mammotrophs, thyrotrophs, gonadotrophs, and corticotrophs) plus endothelial and follicular cells can be identified and are morphologically well preserved up to 20 h after dissociation. Throughout this period, the cells incorporate linearly [3H]leucine into protein for up to 4 h at a rate 90% greater than hemipituitaries, and they synthesize, transport intracellularly, and release the two major pituitary secretory products, growth hormone and prolactin. Immediately after dissociation the cells' ability to respond to secretogogues (high K+ and dibutyryl cyclic AMP) is impaired, but after a 6–12-h culture period, the cells apparently recover and discharge 24% and 52%, respectively, of their content of prelabeled growth hormone over a 3-h period in response to these two secretogogues. This represents a stimulation of 109% and 470% over that released by cells incubated in control medium. The results demonstrate that function and morphologic integrity are preserved in this cell system. Therefore it is suitable for the study of various aspects of pituitary secretion and its control.

Genesis of Cardiac Arrhythmias

Abstract: Activity of Automatic Cells The normal rhythm of the mammalian heart results from spontaneous excitation of cells in the sinoatrial node. These cells possess the property of automaticity.' The transmembrane potential of working muscle fibers in the atria or ventricles demonstrates a rapid depolarization on excitation (phase 0), a period of variable duration during which the cell repolarizes (phases 1, 2, and 3), and then a stable resting potential (phase 4), which persists until the next propagated impulse arrives and causes excitation. In contrast, in cells of the sinoatrial node, repolarization is not followed by a period during which the transmembrane potential is stable. Instead, immediately after the end of repolarization the membrane potential begins to decrease slowly. This slow depolarization during phase 4 lowers the transmembrane potential toward the threshold potential, the value of transmembrane potential at which excitation occurs. If the slow depolarization attains the threshold potential, excitation occurs and the cell develops an action potential which then propagates to excite adjacent cells and, normally, the rest of the heart.2 All cells which demonstrate this slow diastolic depolarization are said to be automatic. This mechanism for spontaneous firing has been called the normal automatic mechanism to differentiate it from other

Permeability of muscle capillaries to exogenous myoglobin

Abstract: Whale skeletal muscle myoglobin (mol wt 17,800; molecular dimensions 25 x 34 x 42 A) was used as a probe molecule for the pore systems of muscle capillaries. Diaphragms of Wistar-Furth rats were fixed in situ at intervals up to 4 h after the intravenous injection of the tracer, and myoglobin was localized in the tissue by a peroxidase reaction. Gel filtration of plasma samples proved that myoglobin molecules remained in circulation in native monomeric form. At 30–35 s postinjection, the tracer marked ∼75% of the plasmalemmal vesicles on the blood front of the endothelium, 15% of those located inside and none of those on the tissue front. At 45 s, the labeling of vesicles in the inner group reached 60% but remained nil for those on the tissue front. Marked vesicles appeared on the latter past 45 s and their frequency increased to ∼80% by 60–75 s, concomitantly with the appearance of myoglobin in the pericapillary spaces. Significant regional heterogeneity in initial labeling was found in the different segments of the endothelium (i.e., perinuclear cytoplasm, organelle region, cell periphery, and parajunctional zone). Up to 60 s, the intercellular junctions and spaces of the endothelium were free of myoglobin reaction product; thereafter, the latter was detected in the distal part of the intercellular spaces in concentration generally equal to or lower than that prevailing in the adjacent pericapillary space. The findings indicate that myoglobin molecules cross the endothelium of muscle capillaries primarily via plasmalemmal vesicles. Since a molecule of this size is supposed to exit through both pore systems, our results confirm the earlier conclusion that the plasmalemmal vesicles represent the large pore system; in addition, they suggest that the same structures are, at least in part, the structural equivalent of the small pore system of this type of capillaries.

The synthesis and turnover of rat liver peroxisomes. V. Intracellular pathway of catalase synthesis.

Abstract: The subcellular distribution of the biosynthetic intermediates of catalase was studied in the livers of rats receiving a mixture of [3H]leucine and [14C]δ-aminolevulinic acid by intraportal injection. Postnuclear supernates were fractionated by a one-step gradient centrifugation technique that separates the main subcellular organelles, partly on the basis of size, and partly on the basis of density. Labeled catalase and its biosynthetic intermediates were separated from the gradient fractions by immunoprecipitation, and the distributions of radioactivity were compared with those of marker enzymes. The results show that catalase protein is synthesized outside the peroxisomes, but rapidly appears in these particles, mostly still in the form of the first hemeless biosynthetic intermediate. Addition of heme and completion of the catalase molecule take place within the peroxisomes. During the first 15 min after [3H]leucine administration, more than half of the newly formed first intermediate was recovered in the supernatant fraction, where it was found to exist as an aposubunit of about 60,000 molecular weight.

Cervical effects on abducens motoneurons and their interaction with vestibulo-ocular reflex

Abstract: The effect of neck afferents on abducens motoneurons and their interaction with the vestibulo-abducens reflex were examined in chloraloseanesthetized or unanesthetized, decerebrate cats. The test reflex elicited in the abducens nerve by stimulation of the contralateral vestibular nerve was inhibited by contralateral and facilitated by ipsilateral cervical dorsal root or neck joint stimulation. These reciprocal effects were obtained by stimulation at the level of C2 and C3, but not from C5 or lower. Contralateral and ipsilateral cervical stimulation induced IPSPs and EPSPs, respectively, in abducens motoneurons. The latencies were 2.8–6.0 msec for the IPSP and 2.8–5.3 msec for the EPSP after stimulation of the dorsal root. The labyrinthine-induced disynaptic IPSP or EPSP was facilitated by conditioning stimulation of the contralateral and ipsilateral cervical dorsal root, respectively. It is thus postulated that the cervico-abducens and vestibuloabducens reflex pathways converge upon common inhibitory or excitatory interneurons in the vestibular nuclie. Labyrinthine- and cervical-induced responses of the presumed interneurons in the vestibular nuclei or those of their axons recorded in the abducens nuclie were consistent with the above view. Lesion experiments in the brain stem indicated that afferent volleys from the neck joint ascend ipsilaterally in the spinal cord, cross to the contralateral side in the brain stem, and eventually project to the vestibular nuclei, thus interacting with the vestibulo-ocular reflex activity. A possible functional role of the cervical effects on the ocular motoneuron was briefly discussed.