Role of sodium on H+ excretion in the integument of the leopard frog Rana pipiens

1. The northern leopard frog, Rana pipiens, pipiens, in contrast to the southern leopard frog, Rana pipiens, berlandieri, did not demonstrate any significant H+ excretion across its integument even during a challenge of chronic metabolic acidosis. Likewise, no increase in the number of H+ secreting mitochondria-rich cells were observed in the northern frogs. 2. Under normal acid-base conditions in the southern frogs, H+ excretion was found to be dependent on mucosal sodium concentrations, whereas during chronic metabolic acidosis, H+ excretion was independent of mucosal sodium concentrations, but was amiloride sensitive. 3. High salinity adapted southern frogs, under normal and acidotic conditions, had enhanced H+ excretion rates as compared to the control non-salt adapted frogs. 4. Blood analyses demonstrated that significant acid-base changes were the result of systemic acidosis and not due to salt adaptations. Blood Na+ and K+ concentrations were also efficiently maintained during salt adaptations or chronic metabolic acidosis. 5. The results suggest that H+ excretion in epithelia can be influenced by the sodium transport state of the cell and the systemic acid-base profile. Models are proposed explaining these relationships.     

Properties of the oscillatory cAMP binding component of Dictyostelium discoideum cells and isolated plasma membranes.

The cAMP receptor on the surface of aggregation competent Dictyostelium discoideum cells specifically binds [3H]cAMP in an oscillatory manner with a periodicity of 2 min. The oscillatory cAMP-binding component is developmentallly regulated and has the nucleotide specificity expected for recognition of chemotactic signals. The concentration dependence of the peak amplitudes of cAMP binding exhibit an apparent threshold at 10(-8) M cAMP. The threshold concentration for cAMP binding that we measure is consistent with the concentration dependence of signal relay (cAMP secretion) and the chemotactic response. The kinetic data of binding and dissociation are very rapid, consistent with the time course of oscillations in receptor capacity (affinity). Specific binding oscillations are destroyed by heat or chymotrypsin but are insensitive to trypsin or glycosidase. A plasma membrane localization of receptor is supported by enrichment of cAMP binding in a plasma membrane preparation from differentiated cells. Receptor oscillations with a 2-min period are preserved in the membrane preparations, and the peak amplitudes are increased about 10-fold consistent with the enrichment of other plasma membrane markers. The alternating change in the receptor's binding capacity for cAMP may be the basis of the relay refractory period as well as the primary oscillator involved in the generation of postreceptor events such as stimulation of adenylate cyclase, cAMP secretion, and cellular movement, all of which have been previously shown to oscillate.     

Hemodynamic effects of anti-G suit inflation in a 1-G environment

This study evaluated effects of various anti-G inflation pressures on cardiac volumes and the relationship of these volume changes to mean arterial pressure changes. Ventricular volumes were calculated using two-dimensional echocardiography. An anti-G suit was inflated to 2, 4, and 6 psi in the standing and supine positions for 10 male subjects. In the supine position, mean arterial pressure increased from base line for all three inflation pressures (P = 0.05). The end-diastolic volume increased after 2-psi inflation (P = 0.03). Cardiac output or stroke volume did not change. After standing, mean arterial pressure (P = 0.002), end-diastolic volume (P = 0.002), and stroke volume (P = 0.05) fell after suit deflation. Peripheral vascular resistance fell in the 2- and 4-psi inflation profiles. In the standing protocol, mean arterial pressure, end-diastolic volume, stroke volume, and cardiac output rose with all three inflation pressures (P less than 0.05). After reclining, heart rate increased (P = 0.02) and mean arterial pressure fell (P less than 0.05) in the 4- and 6-psi inflation profiles after suit deflation. Increases in mean arterial pressure are caused by increases in cardiac preload and cardiac output after inflation of the anti-G suit while subjects were standing. Increased cardiac preload was not consistently seen after inflation while subjects were supine. Changes in end-diastolic volume and mean arterial pressure were dependent on the pressure used to inflate the anti-G suit.     

Characterization of two molecular weight classes of cadmium binding proteins from the mussel, Mytilus edulis (L.)

Inducible cadmium binding proteins (Cd-BP) in the mussel, Mytilus edulis, were resolved into two molecular weight components by gel permeation chromatography on Sephadex G-75. Each of these two molecular weight components was further resolved into four subcomponents by DEAE ion exchange chromatography. All eight subcomponents bound cadmium and exhibited significant u.v. absorption at 254 and little absorption at 280 nm. Based on amino acid composition analysis two classes of proteins were identified, one having higher cysteine (approximately 25 mole %) and lower serine and glutamic acid contents compared to the other class.     

Diaphragm afferent modulation of phrenic motor drive

Experiments were performed in eight lightly anesthetized thiopental sodium (Pentothal) cats to examine whether diaphragmatic afferents can significantly alter the neural drive to the diaphragm when the animal is exposed to lower body negative pressure. Moving-time-averaged diaphragmatic electromyograms (EMGma) were recorded and compared before and during exposure to lower body negative pressure in each of three consecutive conditions: C7 spinalization, bilateral vagotomy, and cervical dorsal rhizotomy. Application of lower body negative pressure in C7-spinalized animals resulted in a decrease in inspiratory time and peak diaphragmatic activity compared with control levels. After bilateral vagotomy, EMGma activity was prolonged with the application of lower body negative pressure. However, there was no increase in peak EMGma activity. After transection of the cervical dorsal roots subserving the phrenic nerve, the prolongation of diaphragmatic activity negative was eliminated. Therefore, we conclude that the significant increase in duration of inspiration in response to application of lower body negative pressure in the C7-spinalized, bilaterally vagotomized cat is mediated by phrenic nerve afferents.     

Differential expression of metastasis-associated cell surface glycoproteins and mRNA in a murine large cell lymphoma

A metastatic variant cell subline of the Abelson virus-transformed murine large lymphoma/lymphosarcoma RAW117 has been selected in vivo ten times for liver colonization. Highly metastatic subline RAW117-H10 forms greater than 200 times as many gross surface liver tumor nodules as the parental line RAW117-P. Analysis of cellular proteins and glycoproteins indicates reduced expression of murine Moloney leukemia virus-associated p15, p30, and gp70, and increased expression of a sialoglycoprotein, gp150, in the highly metastatic H10 cells. Northern analyses of oncogene expression suggested that mRNA of various oncogenes was expressed equally or not expressed in the RAW117 cells of differing metastatic potential. Differential gene expression was examined using a cDNA library of 17,600 clones established from poly A+ mRNA isolated from H10 cells. The cDNA library was screened by the colony hybridization technique using probes made from both RAW117-P and -H10 cells. Approximately 99.5% of these cDNA clones were expressed identically in P and H10 cells. Of the few differentially expressed cDNA clones (approx. 150/17,600), one-half of these were identified as Moloney leukemia virus sequences in a separate probing with a radiolabeled Moloney leukemia virus probe. The remainder of the differentially expressed mRNA detected by colony hybridization of the cDNA library were expressed at higher levels (approx. 1/6) or lower levels (approx. 1/3) in the highly metastatic H10 cells.     

Involvement of the fastigial nuclei in vagally mediated respiratory responses

Xu, Fadi, and Donald T. Frazier. Involvement of thefastigial nuclei in vagally mediated respiratory responses.J. Appl. Physiol. 82(6):1853-1861, 1997.Previous studies have demonstrated that thecerebellum, especially the fastigial nucleus (FN), is capable ofmodulating respiratory responses to chemical and mechanical stimuli.Because there is evidence to show projections from vagal afferents tothe FN, the goal of this study was to determine the role of the FN inthe respiratory reflexes elicited by activation of vagal afferents.Experiments were performed in anesthetized (chloralose), paralyzed, andartificially ventilated cats with an occipital exposure of thecerebellum. Administration of capsaicin (Cap; 5-10 µg/kg) viathe right external jugular vein at the end of inspiration andapplication of lung inflation (LI; 10 cmH₂O) during inspiration werecarried out to stimulate nonmyelinated and myelinated vagal afferents,respectively. The phrenic neurogram was recorded as anindex of the respiratory motor output. Control cardiorespiratoryvariables [expiratory duration(TE), arterial bloodpressure] and their immediate responses to stimuli were comparedbefore and after bilateral lesions of the FN. The results showed thefollowing. 1) Capinjection and LI resulted in a dramatic increase inTE (apnea).2) FN lesions did not significantlyalter the control TE; however,the apneic duration induced by Cap injection was prolonged.3) Neither FN lesions norcerebellectomy affected the apneic duration that resulted fromapplication of LI. 4) Cold blockadeof the vagi (6-8°C) eliminated the respiratory responses elicited by LI but not Cap injection; vagotomy abolished the responses to both stimuli. 5) FN lesions didnot change the control ABP or its responses to either LI or Capinjection. It is concluded that the FN is involved in vagally mediatedrespiratory reflexes elicited by activation of nonmyelinated (C-fiber)vagal afferents.

     

Cellular changes in the toad urinary bladder in response to metabolic acidosis

Summary The urinary bladder ofBufo marinus excretes H⁺ and NH ₄ ⁺ , and the H⁺ excretion is increased when the animal is placed in metabolic acidosis. The mitochondriarich (MR) cells mediate the H⁺ excretion by the bladder. The purpose of this study was to determine if there is a change in MR cells of the bladder during metabolic acidosis. Bladders from normal toads and from toads that had been placed in metabolic acidosis were used. The bladders were mounted between plastic chambers and H⁺ excretion measured. The bladder was then fixed and prepared for scanning (SEM) and transmission (TEM) electron micrograph studies. SEM's at low magnification were used to count the various cell types and the TEM's were used to confirm the different cell types. Fields were randomly selected and a total of 2500 cells counted in each group. The bladders from toads in metabolic acidosis had a consistently higher ratio of MR cells to granular cells than did the normal bladders. These results indicate that during metabolic acidosis there is an increased number of MR cells in the bladder, and this increases the bladder's capacity to excrete H⁺.