Electrophysiological properties of neurons in submucosal ganglia of guinea pig distal colon

1991 ◽  
Vol 260 (6) ◽  
pp. G835-G841 ◽  
Author(s):  
T. Frieling ◽  
H. J. Cooke ◽  
J. D. Wood
Keyword(s):  
Guinea Pig ◽  
Ganglion Cells ◽  
Distal Colon ◽  
Submucosal Plexus ◽  
Electrophysiological Properties ◽  
Small Intestinal ◽  
Type 3

Intracellular recording methods were used in vitro to study the electrophysiological behavior of neurons in ganglia of the submucosal plexus in the distal colon of the guinea pig. The results revealed subpopulations of submucosal ganglion cells that corresponded to the AH/type 2, S/type 1, type 3, and type 4 subpopulations found elsewhere in the intestine. Electrical behavior of colonic submucosal neurons differed from the myenteric plexus of the colon, rectum, and stomach and the small intestinal submucosal plexus mainly in the relative proportions of the different subpopulations. Regional differences in this respect may be a reflection of functional specialization in the diverse regions of the alimentary canal.

Synaptic transmission in submucosal ganglia of guinea pig distal colon

1991 ◽  
Vol 260 (6) ◽  
pp. G842-G849 ◽  
Author(s):  
T. Frieling ◽  
H. J. Cooke ◽  
J. D. Wood
Keyword(s):  
Guinea Pig ◽  
Ganglion Cells ◽  
Input Resistance ◽  
Distal Colon ◽  
Presynaptic Muscarinic Receptors ◽  
The Neural Networks ◽  
Type 3 ◽  
Prolonged Response

Intracellular electrical recording was used to investigate synaptic behavior of ganglion cells in the neural networks of the submucosal plexus of the guinea pig distal colon. Fast excitatory postsynaptic potentials (EPSPs), mediated by nicotinic receptors, were found in all S/type 1 neurons, 70% of AH/type 2, 75% of type 3, and 95% of type 4 neurons. Slow EPSPs were characterized by membrane depolarization, increased input resistance, enhanced action potential discharge, and suppression of hyperpolarizing afterpotentials in 64% of the S/type 1 neurons, 74% of AH/type 2, 31% of type 3, and 70% of type 4 neurons. Micropressure application of acetylcholine evoked a two-component depolarizing response consisting of an initial transient with decreased input resistance followed by a prolonged depolarization associated with increased input resistance. The transient response was suppressed by nicotinic-blocking drugs. Muscarinic antagonists suppressed the prolonged response. Acetylcholine acted also at presynaptic muscarinic receptors to suppress stimulus-evoked fast EPSPs. No stimulus-evoked inhibitory synaptic potentials were observed. Norepinephrine, applied by microejection, acted at alpha 2-adrenoceptors to hyperpolarize the membrane potential in association with decreased neuronal input resistance.

Electrical behavior of myenteric neurons in guinea pig distal colon

1988 ◽  
Vol 254 (4) ◽  
pp. G522-G530 ◽  
Author(s):  
P. R. Wade ◽  
J. D. Wood
Keyword(s):  
Guinea Pig ◽  
Action Potentials ◽  
Distal Colon ◽  
Electrical Behavior ◽  
Myenteric Neurons ◽  
Single Action ◽  
Current Pulses

Intracellular recording was used in vitro to analyze electrophysiological properties of neurons in myenteric ganglia of guinea pig distal colon. The neurons were classified into six types based on their electrical behavior. Type 1 colonic neurons discharged action potentials throughout depolarizing current pulses and were otherwise similar to S/type 1 neurons found in the guinea pig small bowel. The second type had passive and active electrical properties similar to those of AH/type 2 myenteric neurons of the small intestine. These cells discharged only a single spike at the onset of depolarizing current pulses, and the spikes were followed by long-lasting hyperpolarizing afterpotentials. Excitability of the type 2 neurons was enhanced in the presence of elevated Mg2+ and reduced Ca2+, and the spikes were unaffected by tetrodotoxin. Type 3 colonic neurons showed fast synaptic potentials but did not generate action potentials. The majority of neurons were referred to as type 2 colonic neurons. Type 4 neurons discharged single action potentials only at the onset of depolarizing current pulses, and the spikes were not followed by prolonged hyperpolarizing afterpotentials. Unlike type 2 neurons, excitability remained unchanged in the presence of reduced extracellular Ca2+ and elevated Mg2+. Action potentials of type 4 neurons were suppressed or abolished by tetrodotoxin. A group of spontaneously active neurons was classified as type 5 colonic neurons. Type 6 cells were inexcitable and assumed to be glial cells.

Elevation of cAMP facilitates noradrenergic transmission in submucous neurons of guinea pig ileum

1993 ◽  
Vol 264 (3) ◽  
pp. G442-G446 ◽  
Author(s):  
D. H. Zafirov ◽  
H. J. Cooke ◽  
J. D. Wood
Keyword(s):  
Guinea Pig ◽  
Ganglion Cells ◽  
Phosphodiesterase Inhibitor ◽  
Input Resistance ◽  
Guinea Pig Ileum ◽  
Synaptic Excitation ◽  
Before And After ◽  
Intracellular Microelectrodes

Slow synaptic excitation and inhibition were studied with intracellular microelectrodes in submucous ganglion cells of the guinea pig ileum. Elevation of adenosine 3',5'-cyclic monophosphate (cAMP) after application of forskolin or the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) resulted in slowly activating depolarization of the membrane potential. The depolarization was associated with increased input resistance, enhanced excitability, and suppression of hyperpolarizing afterpotentials. This occurred in AH/type 2 but not S/type 1 neurons. The action of forskolin or IBMX mimicked slow synaptic excitation in the same neurons. Focal electrical stimulation also evoked slow inhibitory postsynaptic potentials (IPSPs). The amplitude and duration of the IPSPs were increased by forskolin or a membrane-permeant analogue of cAMP. Treatment with phentolamine, yohimbine or idazoxan suppressed the IPSPs before and after potentiation by forskolin, suggesting that the IPSPs were mediated by release of norepinephrine acting at alpha 2-adrenoceptors. Application of adenosine or selective adenosinergic A1 agonists suppressed or abolished the IPSPs. The results suggest that elevation of cAMP facilitates the release of norepinephrine at alpha 2-synapses on submucous neurons of guinea pig small bowel.

Partial resialylation of human asialotransferrin types 1 and 2 in the rat

1984 ◽  
Vol 62 (9) ◽  
pp. 853-858 ◽  
Author(s):  
Erwin Regoeczi ◽  
Paul A. Chindemi ◽  
Maria T. Debanne
Keyword(s):  
Sialic Acid ◽  
Electrophoretic Mobility ◽  
Acid Content ◽  
Sialic Acid Content ◽  
Small Doses ◽  
Type 3 ◽  
Time Type

125I-labeled asialotransferrin types 1 and 2 were administered in small doses to rats. The protein still in the plasma after 1–12 h was partially repurified and electrophoresed at pH 8.1, together with a transferrin standard that is composed of all six forms of the protein with respect to sialic acid content. The electrophoretic mobility of both asialotransferrins increased with time, type 2 being affected sooner than type 1. The changed mobility was due to increased electronegativity that was fully reversible by treatment of the samples with neuraminidase, thus identifying the underlying cause as partial resialylation. Asialotransferrin incubated in vitro with serum, plasma, or whole blood for 16 h exhibited no change in electrophoretic mobility. In conjunction with an earlier study on asialotransferrin type 3, it was found that the apparent speeds of resialylation of the three asialotransferrins were in the same order as their affinities for the asialoglycoprotein-binding hepatic lectin. This suggests the involvement of an endo- rather than of an ecto-transferase. Transfer of 59Fe from asialotransferrins to the liver was used to monitor the frequency of hepatocyte–asialotransferrin interactions. Iron deposition in the liver took place much more rapidly than the appearance of detectable quantities of partially resialylated asialotransferrin molecules in the circulation. It is concluded that each asialotransferrin molecule probably undergoes several passages through the hepatocyte before its glycans become modified.

43 EFFECT OF VITRIFICATION AT GERMINAL VESICLE STAGE ON THE MITOCHONDRIAL AND CYTOSKELETAL INTEGRITY IN BOVINE OOCYTES

2012 ◽  
Vol 24 (1) ◽  
pp. 134
Author(s):  
Y. Abe ◽  
K. Takakura ◽  
K. Kaito ◽  
T. Ogawa ◽  
M. Yokoo ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
Germinal Vesicle ◽  
Nylon Mesh ◽  
Developmental Rates ◽  
Type 3

In the previous report, we demonstrated that bovine germinal vesicle (GV) stage oocytes vitrified using a nylon mesh holder showed an in vitro maturation rate to the metaphase II (MII) stage similar to that of fresh ones. However, cleavage and developmental rates of vitrified oocytes were low. Because mitochondria and the cytoskeleton are thought to have a central role in energy supply and cellular division in mammalian embryogenesis, it seems possible that alternation in their function in vitrified GV oocytes may contribute to subsequent lower cleavage and developmental rates. The oxygen consumption rate reflects the mitochondrial activity and its measurement may be an effective way for non-invasive evaluation of oocyte quality. In this study, to ascertain that altered mitochondrial functions and cytoskeleton may contribute to reduce the quality of oocytes after vitrification, we evaluated the distribution of active mitochondria and the cytoskeleton in vitrified oocytes. We also examined the relationship between oxygen consumption rate and the distribution of active mitochondria in vitrified oocytes. Bovine GV oocytes connected with cumulus cells were exposed to the cryoprotectant (EFS40) in a stepwise way and transferred onto a nylon mesh holder, followed by plunging them directly into liquid nitrogen. After warming, vitrified oocytes were allowed in vitro maturation. After denuding, matured oocytes were stained with a mitochondria-specific probe, rhodamine-123 and then oxygen consumption rate using an embryo respirometer (HV-403; Research Institute for Functional Peptides, Yamagata, Japan) was measured in each oocyte. According to morphological distribution of mitochondria, oocytes were classified as follows: type 1, uniform distribution; type 2, spotted distribution; and type 3, a weak fluorescence. The oxygen consumption rate of the fresh oocytes at the MII stage was significantly (P < 0.05) higher than that of vitrified oocytes (5.24 and 4 × 1015 mol–1 s–1, respectively), although there was no difference between the fresh and vitrified groups at the GV stage (5.02 and 5.06 × 1015 mol–1 s–1, respectively). The oxygen consumption rates of type 1 oocytes in fresh and vitrified groups at the MII stage tended to be higher than those of type 2 and 3 oocytes (type 1, 5.29 and 5.27; type 2, 4.99 and 4.52; type 3, 4.77 and 4.48 × 1015 mol–1 s–1, respectively). In addition, the percentage of type 1 oocytes in the fresh group was significantly (P < 0.05) higher than that in the vitrified group (59.4 and 34.3%, respectively). The matured oocytes also were stained with α-tubulin monoclonal antibody or F-phalloidin independently to examine the morphological status of microtubules or microfilament. The rates of oocytes with abnormal microtubules and microfilament in the vitrified group were 29.7 and 43.5%, respectively, showing higher rates compared with corresponding fresh oocytes (9.8%; P < 0.05 and 25.0%; P = 0.21, respectively). These results suggested that the reduction of quality and subsequent developmental competence in vitrified oocytes might be related to damages of mitochondria and cytoskeleton.

New Fine Structure within the Adult Hematopoietic Stem Cell Compartment Revealed by Longterm Analysis of Mice Repopulated with Single Cells or Their In Vitro Generated Clonal Progeny.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1720-1720
Author(s):  
Brad Dykstra ◽  
David Kent ◽  
Lindsay McCaffrey ◽  
Kristin Lyons ◽  
Merete Kristiansen ◽  
...  
Keyword(s):  
Stem Cell ◽  
Hematopoietic Stem Cell ◽  
Single Cells ◽  
Myeloid Cells ◽  
Hematopoietic Stem ◽  
Type 3

Abstract Assessments of hematopoietic stem cell (HSC) repopulating activity in vivo have historically relied on calculated average longterm (12–16 wk) progeny outputs using non-purified transplants, thereby precluding definitive clonal assignments of donor-derived cells. Viral marking circumvents this problem, but has not been used for large scale surveys. Heterogeneity observed in the repopulation patterns has generally been inferred to reflect stochastic processes. We now report the in vivo repopulation kinetics of 89 individual longterm repopulating cells (LTRCs) before (n=49) and after (n=40) 4 days of clonal growth in vitro. LTRCs were defined here as cells whose WBC progeny could be detected at levels of ≥1% for at least 16 wk in sublethally irradiated Ly5-congenic W41/W41 hosts. Recipients were transplanted with either freshly isolated, single lin−Rho−SP LTRCs or 4-day clones generated from similar cells in serum-free cultures (+ 300 ng/ml SF, 20 ng/ml IL-11 & 1ng/ml Flt3-L). 4, 8, 12, 16, and 24 wk post-transplant, blood samples were stained for donor-derived B, T, and myeloid cells using a procedure that identifies donor/recipient doublets and Ly6g/Mac1low cells (which have features of lymphoid rather than myeloid WBCs) to exclude false-positive myeloid events. Four distinct patterns of repopulation were revealed. Type 1 showed a delayed production of predominantly myeloid WBCs (low or undetectable before 12 wk) that increased progressively (reaching 0.4–15% of all WBCs by 16 wk). Type 2 showed a robust multilineage repopulation that remained stable or increased over time (6–84% of all WBCs at 16 wk). Type 3 also showed an initially robust pattern of multilineage repopulation (peak numbers of WBCs at 8–12 wk and 1–51% at 16 wk), but the contribution of donor-derived myeloid cells was transient (&lt;0.5% by 16 wk). Type 4 showed a lymphoid-restricted pattern (myeloid contribution &lt;0.5% at all time points), with repopulation levels peaking at 8 wk and decreasing thereafter (1–22% at 16 wk). Persisting granulopoiesis, indicated by a high proportion of donor-derived cells in the Ly6g/Mac1+SSChi population at 16–24 wk, clearly distinguished the type 1 and 2 patterns from types 3 and 4 which showed near or complete cessation of donor-derived granulopoiesis beyond 12 wk. Preliminary secondary transplant experiments show that donor-derived LTRCs (with and without longterm granulopoietic activity) were exclusively generated in primary recipients with type 1 and 2 repopulation patterns. Amongst the freshly isolated LTRCs, 18% (9/49) were type 1, 41% (20/49) were type 2, 22% (11/49) were type 3, and 18% (9/49) were type 4. In contrast, 4-day clones derived from cells of the same phenotype and containing LTRC activity showed a marked decrease in type 1 and type 2 activity with a corresponding increase in type 3 and type 4 activity: type 1 = 5% (2/41), type 2 = 18% (7/40), type 3 = 28% (11/40) and type 4 = 50% (20/40). Collectively, these data identify a new hierarchy of four biologically discrete states within the compartment of cells currently defined as LTRCs. Proliferation of LTRCs either in vitro or in vivo appears to induce an irreversible transition from one state to another (from Type 1 to 2 to 3 to 4), suggesting the existence of intrinsic molecular correlates for each of these states and specific mechanisms that underlie their sequential appearance.

scholarly journals A mathematical model of rat ascending Henle limb. III. Tubular function

2010 ◽  
Vol 298 (3) ◽  
pp. F543-F556 ◽  
Author(s):  
Alan M. Weinstein
Keyword(s):  
Mathematical Model ◽  
Tubular Function ◽  
Proton Secretion ◽  
Luminal Membrane ◽  
Catalytic Role ◽  
Type 3

K+ plays a catalytic role in AHL Na+ reabsorption via Na+-K+-2Cl− cotransporter (NKCC2), recycling across luminal K+ channels, so that luminal K+ is not depleted. Based on models of the ascending Henle limb (AHL) epithelium, it has been hypothesized that NH4+ may also catalyze luminal Na+ uptake. This hypothesis requires that luminal NH4+ not be depleted, implying replenishment via either direct secretion of NH4+, or NH3 in parallel with a proton. In the present work, epithelial models of rat medullary and cortical AHL (Weinstein AM, Krahn TA. Am J Physiol Renal Physiol 298: F000–F000, 2009) are configured as tubules and examined in simulations of function in vitro and in vivo to assess the feasibility of a catalytic role for NH4+ in Na+ reabsorption. Modulation of Na+ transport is also examined by peritubular K+ concentration and by Bartter-type transport defects in NKCC2 (type 1), in luminal membrane K+ channels (type 2), and in peritubular Cl− channels (type 3). It is found that a catalytic role for NH4+, which is significant in magnitude (relative to K+), is quantitatively realistic, in terms of uptake via NKCC2, and in terms of luminal membrane ammonia backflux. Simulation of a 90% NKCC2 defect is predicted to double distal Na+ delivery; it is also predicted to increase distal acid delivery (principally as NH4+). With doubling of medullary K+, the model predicts a 30% increase in distal Na+ delivery, but in this case there is a decrease in AHL acidification. This effect of peritubular K+ on proton secretion appears to be akin to type 3 Bartter's pathophysiology, in which there is decreased peritubular HCO3− exit, cytosolic alkalinization, and a consequent decrease in luminal proton secretion by NHE3. One consequence of overlapping and redundant roles for K+ and NH4+, is a blunted impact of luminal membrane K+ permeability on overall Na+ reabsorption, so that type 2 Bartter pathophysiology is not well captured by the model.

scholarly journals Predatory activity of chlamydospores of the fungusPochonia chlamydosporia on Toxocara caniseggs under laboratory conditions

2013 ◽  
Vol 22 (1) ◽  
pp. 171-174 ◽  
Author(s):  
Juliana Milani Araujo ◽  
Jackson Victor de Araújo ◽  
Fabio Ribeiro Braga ◽  
Sebastião Rodrigo Ferreira ◽  
Alexandre de Oliveira Tavela
Keyword(s):  
Agar Medium ◽  
Toxocara Canis ◽  
Control Group ◽  
Vitro Assay ◽  
Petri Dishes ◽  
Egg Destruction ◽  
Type 3

The objective of this study was to use chlamydospores of the fungusPochonia chlamydosporia (isolates VC1 and VC4) against Toxocara canis eggs in a 15-day in vitro assay. One thousand T. canis eggs were placed in Petri dishes containing 2% water agar medium with different concentrations of chlamydospores (1,000, 10,000 or 100,000) of each fungal isolate of P. chlamydosporia (treated groups) and 1,000 eggs in Petri dishes without fungus (control group). Egg counts were performed to determine the ovicidal activity, which was classified as three effect levels: type 1, type 2 and type 3. Significant differences (P < 0.01) in egg destruction were found in comparison with the control group. The highest percentage of egg destruction was found in plates containing 100,000 chlamydospores (68.5% for VC1 and 70.5% for VC4). Chlamydospores of P. chlamydosporiawere effective in destroying T. canis eggs and may contribute in the future towards combating the eggs of this parasite.

Morphology of electrophysiologically identified myenteric neurons in the guinea pig rectum

1992 ◽  
Vol 262 (3) ◽  
pp. G545-G552 ◽  
Author(s):  
K. Tamura
Keyword(s):  
Guinea Pig ◽  
Circular Muscle ◽  
Type I ◽  
Type Ii ◽  
Fiber Tracts ◽  
Single Spike ◽  
Type 3 ◽  
Dogiel Type Ii

Dye-filled microelectrodes were used to investigate relations between morphology and electrophysiological behavior of neurons in the myenteric plexus of the guinea pig rectum. The neurons were divided into two general classes on the basis of morphology. The first class had smooth ovoid somas with multiple long processes that exited the ganglion in several different fiber tracts and fit the description of Dogiel type II neurons. Neurons of the second class had a single long process that exited the ganglion in a fiber tract and sometimes projected to the circular muscle. Neurons of the second class were subdivided into three groups. One of these had short club-shaped dendrites like Dogiel type I neurons. Another had short tapering filamentous processes. Neurons with a single long neurite that could not be classified made up the third group. AH/type 2 electrophysiological behavior was found in both general classes of neurons. S/type 1 electrophysiological behavior occurred only in neurons of the general class with a single long neurite. Type 3 electrical behavior, characterized by robust fast synaptic input in inexcitable neurons, was found only in the cells defined by a single long neurite. Single-spike neurons behaved electrophysiologically like AH/type 2 neurons without long-lasting hyperpolarizing afterpotentials. This behavior was limited to the class of neurons characterized morphologically by a single long neurite.

scholarly journals Toxoplasma gondii ADSL Knockout Provides Excellent Immune Protection against a Variety of Strains

Vaccines ◽  
2020 ◽  
Vol 8 (1) ◽  
pp. 16 ◽  
Author(s):  
Luyao Wang ◽  
Ding Tang ◽  
Chenghang Yang ◽  
Jing Yang ◽  
Rui Fang
Keyword(s):  
Toxoplasma Gondii ◽  
Protozoan Parasite ◽  
Growth Conditions ◽  
Economic Losses ◽  
Immune Protection ◽  
Cellular Immune ◽  
Type 3

Toxoplasma gondii is a protozoan parasite, occurring worldwide, endangers human health and causes enormous economic losses to the Ministry of Agriculture. A safe and effective vaccination is needed to handle these problems. In addition, ideal vaccine production is a challenge in the future. In this study, we knocked out the adenylosuccinate lyase (ADSL) gene and found that the gene reduces the growth rate of T. gondii tachyzoites in vitro under standard growth conditions by plaque or replication experiments. Furthermore, mice that were immunized with tachyzoites of the ME49ΔADSL strain induced 100% protection efficacy against challenge with the type 1 strain RH, type 2 strain ME49 and type 3 strain VEG. All mice that were immunized with ME49ΔADSL had a survival rate of 100% when they were reinfected with wild-type strains, either 30 days or 70 days after immunization, and immunization was also protective against homologous infection with 50 T. gondii ME49 tissue cysts. In addition, the level of Toxoplasma-specific IgG was significantly elevated at 30 and 70 days after immunization. ME49ΔADSL induced high levels of Th1 cytokines (interferon gamma (IFN-γ), interleukin (IL)-12) at 4 weeks after immunization and spleen cell cultures from mice vaccinated for 150 days were able to produce robust INF-γ and IL-12 levels in the supernatant. The results of the present study showed that ΔADSL vaccination induced a T. gondii-specific cellular immune response against further infections. These results suggest that the ADSL-deficient vaccine can induce anti-Toxoplasma gondii humoral and cellular immune responses and has 100% immune protection against post-challenge by the type 1 strain RH, type 2 strain ME49 and type 3 strain VEG. It will be used as an excellent candidate for live vaccines and may contribute in a positive meaning to control human toxoplasmosis.

Sign in / Sign up

Export Citation Format

Share Document