Sodium dependence of luminal HCO3− transport in rat cecum in vitro

Under in vitro conditions the rat cecum transported HCO3- from the serosal to an unbuffered solution in contact with the mucosal side [Js----m = 7.12 +/- 0.18 mumol.cm-2.h-1 (n = 149)]. With reversed tissues, a significantly lower flux was obtained [Jm----s = 2.47 +/- 0.11 mumol.cm-2.h-1 (n = 42)]. Both fluxes were stable for several hours. Increasing the H+ gradient across the tissue for 60 min did not change either flux. Anoxia for 45 min reversibly reduced Js----m by 65 +/- 3% (n = 20) but had no effect on Jm----s. Both fluxes were linearly related to HCO3- concentration on the buffered side, but the slope for Js----m was 3.5 times that for Jm----s. When tissues were initially set up in HEPES buffer rather than HCO3-, Js----m was 0.12 +/- 0.05 mumol.cm-2.h-1 (n = 6), which is not significantly different from zero. Replacement of Na+ by choline reduced Js----m by 40 +/- 3% (n = 11) and ouabain (1 mM) by 24 +/- 3% (n = 5). Replacement of Cl- with isethionate or K+ with Na+ for 60 min did not alter Js----m. Serosal application of DIDS (0.5 mM) reduced Js----m by 24 +/- 6% (n = 6), but SITS (0.5 mM), furosemide (1 mM), acetazolamide (0.1 mM), amiloride (1 mM), and a proton pump inhibitor (Sch 28080, 50 microM) had no effect. Mucosal application of DIDS, furosemide, and amiloride had no effect on Js----m. Serosal tetrodotoxin (1 microM) and indomethacin (28 microM) were also without effect.(ABSTRACT TRUNCATED AT 250 WORDS)

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Mucosal calcium uptake by rat cecum: identity with transcellular calcium absorption

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1983.244.6.g618 ◽

1983 ◽

Vol 244 (6) ◽

pp. G618-G622

Author(s):

H. N. Nellans ◽

R. S. Goldsmith

Keyword(s):

Calcium Binding ◽

Calcium Uptake ◽

Calcium Absorption ◽

Calcium Influx ◽

Maximal Velocity ◽

Similar Reduction ◽

High Calcium ◽

Wide Range ◽

Rat Cecum

Unidirectional intestinal calcium uptake (JCame) at the mucosal surface of rat cecum was investigated in vitro with intact tissue. Uptake is linear for 2–3 min with no indication of rapid calcium binding. Kinetic parameters reveal a maximal velocity of 333 nmol . cm-2 . h-1 with a half-maximal concentration of 0.98 mM. High-calcium diet decreased JCame by more than 60% with respect to both control and low-calcium diets; 1 mM N-ethylmaleimide caused a similar reduction. The activation energy of JCame is significantly less than that of transepithelial mucosal-to-serosal calcium absorption. Mucosal uptake was compared with transepithelial calcium fluxes in rat cecum and revealed a 1:1 correlation over a wide range of transport rates. These results are interpreted to implicate a feedback control system between basolateral calcium efflux and brush-border calcium influx.

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Water and ion handling in the rat cecum

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1990.259.5.g786 ◽

1990 ◽

Vol 259 (5) ◽

pp. G786-G791 ◽

Cited By ~ 3

Author(s):

E. Escobar ◽

C. Ibarra ◽

E. Todisco ◽

M. Parisi

Keyword(s):

Permeability Coefficient ◽

Driving Force ◽

Water Movement ◽

Short Circuit ◽

Short Circuit Current ◽

Chemical Gradient ◽

Osmotic Permeability ◽

Rat Cecum ◽

Good Agreement

The minute-by-minute net water movement (Jw) in the rat cecum was correlated with the transepithelial potential difference (PD), short-circuit current (Isc), and the unidirectional Na+, Cl-, and Rb+ fluxes, with the following results. 1) Jw was a linear function of the applied hydrostatic or osmotic transepithelial gradients (hydrostatic permeability coefficiency = 0.164 +/- 0.018 cm/s, n = 13; osmotic permeability coefficient = 0.0014 +/- 0.0002 cm/s, n = 6). 2) A fraction of this absorptive Jw (0.17 +/- 0.03 microliter.min-1.cm-2, n = 13) was independent of the presence of any osmotic, hydrostatic, or chemical gradient. 3) This fraction was Na+ dependent, associated with an amiloride-insensitive PD and net Na+ (2.37 +/- 0.68 mu eq.h-1.cm-2, n = 6) and Cl- influxes (3.45 +/- 1.46 mu eq.h-1.cm-2, n = 6), measured under short-circuit conditions. No net Rb+ movement was detected. 4) The absorptive Jw increased when HCO3- was replaced by tris(hydroxymethyl)aminomethane (Tris+) buffer or Cl- by SO4(2-). A good agreement between the observed and the expected Jw (assuming isosmotic reabsorption) was observed in the absence of HCO3-. 5) The presence of an osmotic but not a hydrostatic transepithelial gradient generated a transepithelial PD. These results show that water movement across the rat cecum in vitro is the result of a combination of hydrostatic-, osmotic-, and transport-associated transfers. Concerning this last driving force, the observed results indicate that the transport-related Jw results from the addition of an absorptive Jw, coupled to a nonelectrogenic NaCl entry, plus a secretory Jw probably coupled to HCO3- secretion.

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Transepithelial calcium transport by rat cecum: high-efficiency absorptive site

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1981.240.6.g424 ◽

1981 ◽

Vol 240 (6) ◽

pp. G424-G431 ◽

Cited By ~ 3

Author(s):

H. N. Nellans ◽

R. S. Goldsmith

Keyword(s):

Calcium Transport ◽

High Efficiency ◽

Calcium Absorption ◽

Short Circuit ◽

Calcium Fluxes ◽

Calcium Diet ◽

Low Calcium Diet ◽

High Calcium ◽

Rat Cecum

Transepithelial calcium transport has been investigated in rat cecum under in vitro voltage-clamp conditions. Under short-circuit conditions, the cecum behaves as a relatively tight epithelium for calcium fluxes, where mucosal-to-serosal (JCam leads to s) flux exceeds the reverse flux by at least 15-fold. JCanet is abolished in the presence of 1 mM N-ethylmaleimide, is inhibited by 40% with 1 mM ouabain, and is decreased by at least 60% when medium sodium is replaced by choline. Voltage-clamping experiments suggest that both electroneutral- and electrogenic-mediated calcium fluxes traverse the cell in the mucosal-to-serosal direction. Serosal-to-mucosal flux is purely diffusional and probably constrained to the paracellular pathway. In rats weighing less than 175 g, a low-calcium diet has no significant stimulatory effect on JCam leads to s, but a high-calcium diet markedly reduces this flux. These results suggest that the cecum possesses the highest density of calcium transport sites in the rat intestine and is ideally suited for bulk calcium absorption, which may be “down regulated” in response to an increased calcium load in growing animals.

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Ultrastructural Investigations of the Contractile Filaments of Amoebae

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100050950 ◽

1974 ◽

Vol 32 ◽

pp. 188-189

Author(s):

P.L. Moore

Keyword(s):

Cytoplasmic Streaming ◽

Three Dimensional ◽

Freeze Fracture ◽

Amoeboid Movement ◽

Different Types ◽

Chemical Conditions ◽

Complete Interpretation

Previous freeze fracture results on the intact giant, amoeba Chaos carolinensis indicated the presence of a fibrillar arrangement of filaments within the cytoplasm. A complete interpretation of the three dimensional ultrastructure of these structures, and their possible role in amoeboid movement was not possible, since comparable results could not be obtained with conventional fixation of intact amoebae. Progress in interpreting the freeze fracture images of amoebae required a more thorough understanding of the different types of filaments present in amoebae, and of the ways in which they could be organized while remaining functional.The recent development of a calcium sensitive, demembranated, amoeboid model of Chaos carolinensis has made it possible to achieve a better understanding of such functional arrangements of amoeboid filaments. In these models the motility of demembranated cytoplasm can be controlled in vitro, and the chemical conditions necessary for contractility, and cytoplasmic streaming can be investigated. It is clear from these studies that “fibrils” exist in amoeboid models, and that they are capable of contracting along their length under conditions similar to those which cause contraction in vertebrate muscles.

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In Vitro Induction of Crystals of MT Protein by Vinblastine-Colcemid in Mouse Spermatids

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100050676 ◽

1974 ◽

Vol 32 ◽

pp. 132-133

Author(s):

John J. Wolosewick ◽

John H. D. Bryan

Keyword(s):

Endoplasmic Reticulum ◽

Smooth Endoplasmic Reticulum ◽

Nuclear Ring ◽

Rough Er ◽

Distal Direction ◽

In Vitro Induction

Early in spermiogenesis the manchette is rapidly assembled in a distal direction from the nuclear-ring-densities. The association of vesicles of smooth endoplasmic reticulum (SER) and the manchette microtubules (MTS) has been reported. In the mouse, osmophilic densities at the distal ends of the manchette are the organizing centers (MTOCS), and are associated with the SER. Rapid MT assembly and the lack of rough ER suggests that there is an existing pool of MT protein. Colcemid potentiates the reaction of vinblastine with tubulin and was used in this investigation to detect this protein.

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Induction and Differentiation of Dental Epithelium in Vivo and in Vitro

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100053425 ◽

1982 ◽

Vol 40 ◽

pp. 142-145

Author(s):

E. J. Kollar

Keyword(s):

Connective Tissue ◽

Oral Mucosa ◽

Basal Lamina ◽

Functional Derivatives ◽

Specific Source ◽

Dental Epithelium ◽

Connective Tissue Stroma

The differentiation and maintenance of many specialized epithelial structures are dependent on the underlying connective tissue stroma and on an intact basal lamina. These requirements are especially stringent in the development and maintenance of the skin and oral mucosa. The keratinization patterns of thin or thick cornified layers as well as the appearance of specialized functional derivatives such as hair and teeth can be correlated with the specific source of stroma which supports these differentiated expressions.

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Immunoenzymatic demonstration of the simultaneous presence of transferrin, hemopexin and albumin in a same hepatocyte and their ultrastructural localization

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081309 ◽

1978 ◽

Vol 36 (2) ◽

pp. 88-89

Author(s):

M. Kraemer ◽

J. Foucrier ◽

J. Vassy ◽

M.T. Chalumeau

Keyword(s):

Plasma Proteins ◽

Rat Hepatocytes ◽

Ultrastructural Localization ◽

Carrier Proteins ◽

Normal Cells ◽

Adult Rat ◽

Adult Rat Hepatocytes ◽

Monospecific Antisera ◽

Different Levels

Some authors using immunofluorescent techniques had already suggested that some hepatocytes are able to synthetize several plasma proteins. In vitro studies on normal cells or on cells issued of murine hepatomas raise the same conclusion. These works could be indications of an hepatocyte functionnal non-specialization, meanwhile the authors never give direct topographic proofs suitable with this hypothesis.The use of immunoenzymatic techniques after obtention of monospecific antisera had seemed to us useful to bring forward a better knowledge of this problem. We have studied three carrier proteins (transferrin = Tf, hemopexin = Hx, albumin = Alb) operating at different levels in iron metabolism by demonstrating and localizing the adult rat hepatocytes involved in their synthesis.Immunological, histological and ultrastructural methods have been described in a previous work.

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