scholarly journals Electrophysiological properties of gap junctions between dissociated pairs of rat hepatocytes.

1986 ◽  
Vol 103 (1) ◽  
pp. 135-144 ◽  
Author(s):  
D C Spray ◽  
R D Ginzberg ◽  
E A Morales ◽  
Z Gatmaitan ◽  
I M Arias

Physiological properties of isolated pairs of rat hepatocytes were examined within 5 h after dissociation. These cells become round when separated, but cell pairs still display membrane specializations. Most notably, canaliculi are often present at appositional membranes which are flanked by abundant gap and tight junctions. These cell pairs are strongly dye-coupled; Lucifer Yellow CH injected into one cell rapidly diffuses to the other. Pairs of hepatocytes are closely coupled electrically. Conductance of the junctional membrane is not voltage sensitive: voltage clamp studies demonstrate that gj is constant in response to long (5 s) transjunctional voltage steps of either polarity (to greater than +/- 40 mV from rest). Junctional conductance (gj) between hepatocyte pairs is reduced by exposure to octanol (0.1 mM) and by intracellular acidification. Normal intracellular pH (pHi), measured with a liquid ion exchange microelectrode, was generally 7.1-7.4, and superfusion with saline equilibrated with 100% CO2 reduced pHi to 6.0-6.5. In the pHi range 7.5-6.6, gj was constant. Below pH 6.6, gj steeply decreased and at 6.1 coupling was undetectable. pHi recovered when cells were rinsed with normal saline; in most cases gj recovered in parallel so that gj values were similar for pHs obtained during acidification or recovery. The low apparent pK and very steep pHi-gj relation of the liver gap junction contrast with higher pKs and more gradually rising curves in other tissues. If H+ ions act directly on the junctional molecules, the channels that are presumably homologous in different tissues must differ with respect to reactive sites or their environment.

1989 ◽  
Vol 61 (6) ◽  
pp. 1142-1152 ◽  
Author(s):  
R. O. Brown ◽  
S. M. Pulst ◽  
E. Mayeri

1. The generation of egg-laying behavior in the marine mollusk Aplysia involves a prolonged burst discharge in the neuroendocrine bag cells, which secrete neuropeptides derived from the egg-laying hormone/bag cell peptide (ELH/BCP) precursor protein. 2. Besides the bag cells, which are located in the abdominal ganglion, small clusters of neurons in the cerebral and pleural ganglia also express the ELH/BCP neuropeptides. We made intracellular recordings from 32 of these ELH/BCP cells in right pleural ganglia, in 18 preparations, to characterize their physiological properties and their functional relationship to the bag cells. 3. The identification of these ELH/BCP cells was confirmed by pressure injection of Lucifer yellow and subsequent immunocytochemical processing for alpha-BCP immunoreactivity. 4. The basic electrophysiological properties of the pleural ELH/BCP cells were similar to those of the bag cells. These pleural cells were directly demonstrated to be electrically coupled, and direct intracellular stimulation of individual pleural ELH/BCP cells initiated prolonged, synchronous burst discharges in the entire cluster through a positive feedback mechanism. 5. Burst discharges elicited in the pleural ELH/BCP cells consistently initiated burst discharges in the bag cells. Bag cell burst discharges were less effective in initiating burst discharges in the pleural ELH/BCP cells, indicating that there were reciprocal but asymmetrical connections. 6. The results show that the pleural ELH/BCP cells are functionally coupled to the bag cells. They support the hypothesis that the pleural ELH/BCP cells are part of the descending pathway that initiates bag cell activity and egg-laying behavior, in vivo.


2013 ◽  
Vol 109 (7) ◽  
pp. 1876-1889 ◽  
Author(s):  
Caiping Hu ◽  
DiJon D. Hill ◽  
Kwoon Y. Wong

In the mammalian retina, some ganglion cells express the photopigment melanopsin and function as photoreceptors. Five morphological types of these intrinsically photosensitive retinal ganglion cells (ipRGCs), M1–M5, have been identified in mice. Whereas M1 specializes in non-image-forming visual functions and drives such behaviors as the pupillary light reflex and circadian photoentrainment, the other types appear to contribute to image-forming as well as non-image-forming vision. Recent work has begun to reveal physiological diversity among some of the ipRGC types, including differences in photosensitivity, firing rate, and membrane resistance. To gain further insights into these neurons' functional differences, we conducted a comprehensive survey of the electrophysiological properties of all five morphological types. Compared with the other types, M1 had the highest membrane resistance, longest membrane time constant, lowest spike frequencies, widest action potentials, most positive spike thresholds, smallest hyperpolarization-activated inwardly-rectifying current-induced “sagging” responses to hyperpolarizing currents, and the largest effects of voltage-gated K+ currents on membrane potentials. M4 and M5 were at the other end of the spectrum for most of these measures, while M2 and M3 tended to be in the middle of this spectrum. Additionally, M1 and M2 cells generated more diverse voltage-gated Ca2+ currents than M3–M5. In conclusion, M1 cells are significantly different from all other ipRGCs in most respects, possibly reflecting the unique physiological requirements of non-image-forming vision. Furthermore, the non-M1 ipRGCs are electrophysiologically heterogeneous, implicating these cells' diverse functional roles in both non-image-forming vision and pattern vision.


2021 ◽  
pp. 1-5
Author(s):  
David O. Carpenter ◽  
N Hori ◽  
Z Xu ◽  
N Akaike ◽  
Y Tan ◽  
...  

The effects of age on the physiological properties of cervical motor neurons were examined in slices made from an excised spinal cord graft of ICR mice from the second day after birth to age 350 days. The membrane potential of post-natal day 2 (PD2) to PD350 was about -65 mV and did not change greatly with age, although it was slightly higher at PD2. However, there were significant changes in membrane resistance, which increased with age from about 15 to 30 MΩ. The depolarization induced by the excitatory amino acid agonists, kainic acid, NMDA and AMPA, decreased with aging in spite of the increase in membrane resistance. The motor neurons of the aged mice showed delayed recovery from excitation caused by excitatory amino acid agonists. By injecting Lucifer yellow CH into motor neurons, it was observed that the dendrite trees become thin, and some of the dendrite branches were missing in older animals.


1985 ◽  
Vol 54 (4) ◽  
pp. 782-806 ◽  
Author(s):  
D. A. McCormick ◽  
B. W. Connors ◽  
J. W. Lighthall ◽  
D. A. Prince

Slices of sensorimotor and anterior cingulate cortex from guinea pigs were maintained in vitro and bathed in a normal physiological medium. Electrophysiological properties of neurons were assessed with intracellular recording techniques. Some neurons were identified morphologically by intracellular injection of the fluorescent dye Lucifer yellow CH. Three distinct neuronal classes of electrophysiological behavior were observed; these were termed regular spiking, bursting, and fast spiking. The physiological properties of neurons from sensorimotor and anterior cingulate areas did not differ significantly. Regular-spiking cells were characterized by action potentials with a mean duration of 0.80 ms at one-half amplitude, a ratio of maximum rate of spike rise to maximum rate of fall of 4.12, and a prominent afterhyperpolarization following a train of spikes. The primary slope of initial spike frequency versus injected current intensity was 241 Hz/nA. During prolonged suprathreshold current pulses the frequency of firing adapted strongly. When local synaptic pathways were activated, all cells were transiently excited and then strongly inhibited. Bursting cells were distinguished by their ability to generate endogenous, all-or-none bursts of three to five action potentials. Their properties were otherwise very similar to regular-spiking cells. The ability to generate a burst was eliminated when the membrane was depolarized to near the firing threshold with tonic current. By contrast, hyperpolarization of regular-spiking (i.e., nonbursting) cells did not uncover latent bursting tendencies. The action potentials of fast-spiking cells were much briefer (mean of 0.32 ms) than those of the other cell types.(ABSTRACT TRUNCATED AT 250 WORDS)


1982 ◽  
Vol 30 (2) ◽  
pp. 189-191 ◽  
Author(s):  
R L Michaels

Lucifer Yellow CH may be injected into pancreatic islet cells and visualized in Epon sections of the embedded tissue both before and after plastic removal and immunocyto-chemical staining. The dye retains its fluorescence, clearly marking the injected cell and adjacent dye-coupled cells, but does not interfere with the indirect immunofluorescent staining patterns that are characteristic of the islet cells


1983 ◽  
Vol 218 (1210) ◽  
pp. 95-110 ◽  

(i) Following previous work on the morphological and physiological properties of the two distal joints (J2, J3) of the atenna of the rock lobster Palinurus vulgaris , the mechanical, muscular and proprioceptive organization of the two proximal joints between the antennal segments S1 and S2 (J1) and between S1 and the cephalothorax (J0) have now been studied. (ii) Articulated by two classical condyles, J1 moves in a mediolateral plane. One external rotator muscle (ER) and three internal rotator muscles (IR1, IR2, IR3) subserve its movements. J0 is articulated by two different systems: a classical ventrolateral condyle and a complex sliding system constituted by special cuticular structures on the dorsomedial side of the S1 segment and on the rostrum between the two antennae. J0 moves in the dorsoventral plane by means of a levator muscle (Lm) and a depressor muscle (Dm). A third muscle, the lateral tractor muscle (LTm), associated with J0 and lying obliquely across S1, may modulate the level of friction between the S1 segment and the rostrum. (iii) Proprioception in J1 is achieved by a muscle receptor organ AMCO-J1 (antennal myochordotonal organ for the J1 joint) associating a small accessory muscle (S1.am) located in the proximal part of the S1 segment and a chordotonal organ inserted proximally on the S1.am muscle and distally on the S2 segment. J0 proprioception is ensured by a simple chordotonal organ (CO-J0) located in the anterior part of the cephalothorax. (iv) The S1.am muscle is innervated by three motoneurons characterized by their very small diameters and inducing respectively tonic excitatory postsynaptic potentials, phasic excitatory postsynaptic potentials and inhibitory postsynaptic potentials. Anatomical and physiological observations suggest functional correlation between S1.am and IR1 motor innervation. (v) Mechanical and muscular organization of J0 and J1 are compared with that of the other joints of the antenna. The properties of the AMCO-J1 proprioceptor are discussed in relation to the other muscle receptor organs described in crustaceans.


2001 ◽  
Vol 281 (4) ◽  
pp. G1004-G1013 ◽  
Author(s):  
Takashi Kojima ◽  
Alfredo Fort ◽  
Mingyuan Tao ◽  
Masao Yamamoto ◽  
David C. Spray

Primary cultures of adult mouse hepatocytes are shown here to reexpress differentiated hepatocyte features following treatment with 2% DMSO and 10−7 M glucagon. To examine the roles of gap junctional communication during hepatocyte growth and differentiation, we have compared treated and untreated hepatocytes from connexin (Cx)32-deficient [Cx32 knockout (KO)] and wild-type mice. In untreated cultures, DNA replication of Cx32 KO hepatocytes was markedly higher than of wild types. Although Cx26 mRNA levels remained high at all time points in wild-type and Cx32 KO hepatocytes, Cx32 mRNA and protein in wild-type hepatocytes underwent a marked decline, which recovered in 10-day treated cultures. Increased levels of Cx26 protein and junctional conductance were observed in Cx32 KO hepatocytes at 96 h in culture, a time when cell growth rate was high. Treatment with DMSO/glucagon highly reinduced Cx26 expression in Cx32 KO hepatocytes, and such treatment reinduced expression of both Cx32 and Cx26 expression in wild types. Dye transfer was not observed following Lucifer yellow injection into DMSO/glucagon-treated Cx32 KO hepatocytes, whereas the spread was extensive in wild types. Nevertheless, high junctional conductance values were observed in treated cells from both genotypes. These studies provide a method by which the differentiated phenotype can be obtained in cultured mouse hepatocytes and provide in vitro evidence that expression of gap junctions formed of Cx32 are involved in the regulation of growth of mouse hepatocytes.


1990 ◽  
Vol 36 (11) ◽  
pp. 1906-1910 ◽  
Author(s):  
J Osada ◽  
T Gea ◽  
C Sanz ◽  
I Millan ◽  
J Botella

Abstract A group of substances of molecular masses between 300 and 1500 Da have been found to be toxic metabolites in patients with uremia. We determined the concentration in serum of these molecules in the following groups of patients: two hemodialyzed groups (one with cuprophane and the other with polyacrylonitrile dialyzers), one group treated with continuous ambulatory peritoneal dialysis, one group of nondialyzed azotemic patients, and one control group of healthy persons. Ultrafiltrates of the subjects' sera were fractionated on Sephadex G-15 followed by ion-exchange chromatography. Eluates were monitored by absorbance at 254 and 206 nm. Partially characterized peaks P1 and P2, obtained by gel filtration, correlated with the concentration of creatinine in serum; their concentrations were significantly (P less than 0.01) larger in hemodialyzed groups than in peritoneal dialyzed or in nondialyzed azotemic patients. After ion-exchange chromatography, two peaks (P'5 and P'6) correlated with serum creatinine and also were larger in hemodialyzed patients than in the other groups. Apparently, adequate discrimination is obtained by gel-filtration analysis and further analysis by ion-exchange chromatography does not provide additional information in most of the affected patients.


Radiocarbon ◽  
2018 ◽  
Vol 60 (6) ◽  
pp. 1649-1656
Author(s):  
Simon Norris ◽  
Manuel Capouet

ABSTRACTThe European Commission CAST project (CArbon-14 Source Term) aimed to develop understanding of the potential release mechanisms of carbon-14 (radiocarbon, 14C) from radioactive waste materials under conditions relevant to waste packaging and disposal to underground geological disposal facilities. The project focused on the release of carbon-14 as dissolved and gaseous species from irradiated metals (steels, Zircaloys), from spent ion-exchange materials and from irradiated graphites. This paper provides an overview of the CAST project and its output. It also acts as an introduction and scene-setter to the other papers in this special edition of Radiocarbon.


1993 ◽  
Vol 40 (4) ◽  
pp. 559-562 ◽  
Author(s):  
P Widłak ◽  
J Rzeszowska-Wolny

The binding of [14C]benzo[a]pyrene (B[a]P) to DNA and proteins in total nuclei and subnuclear fractions of cultured rat hepatocytes was compared. The main targets of B[a]P were non-histone high molecular weight proteins of the nuclear matrix and DNA sequences attached to this structure. Following 24 h exposure to B[a]P the amounts of adducts in the nuclear matrix DNA and proteins were twice as high as in total nuclei. After withdrawal of the carcinogen containing medium the level of B[a]P-induced adducts gradually decreased but always remained the highest in the nuclear matrix proteins. Removal of adducts from the nuclear matrix DNA was more efficient than from the other DNA fractions, and 72 h after exposure to the carcinogen the level of DNA adducts in this fraction was similar to that in total nuclei.


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