Amphibian Lens Research Articles

Immunofluorescence localization of the crystallins in amphibian lens development, with special reference to the γ-crystallins

The ontogeny and localization of the crystallins in the developing lens of R. pipiens has been investigated by means of the immunofluorescence technique. Antibodies to adult R. pipiens total-lens proteins and adult R. pipiens γ-crystallins—whose antigen, tissue, and developmental specificity had been demonstrated—were used in both the direct and indirect (or “sandwich”) immunofluorescence techniques; both methods gave comparable results. These antibody-dye conjugates were applied to sections of embryonic R. pipiens; a series of lens developmental stages was recognized and defined by us to aid in identification of those stages subjected to immunofluorescence analysis. A positive reaction for the anti-total-lens-protein immunofluorescence reagent was first noted at Lens Developmental Stage VI, in a number of prospective primary lens fiber cells, and a weak, irregular reaction was also noted in the external layer. The intensity of immunofluorescence with this reagent increased in the fiber area and external layer until at Lens Developmental Stage X all cells of the lens were positive; the primary and secondary lens fiber cells and lens epithelium exhibited immunofluorescence in order of decreasing intensity. The first positive reaction with the anti-γ-crystallin immunofluorescence reagent could also be observed at Lens Developmental Stage VI. However, the immunofluorescence was present only in the region of the prospective primary lens fiber cells and was absent in the external layer. The intensity of the immunofluorescence increased in the fiber area until Lens Developmental Stage X, at which time it was greater in the primary fiber cells than that in the secondary fiber cells. No immunofluorescence could be detected in the external layer or in the lens epithelium with this reagent until Stage X. At this time, after the lens is well differentiated and has assumed the basic adult configuration, a weak immunofluorescence reaction was noted in the lens epithelium in a small percentage of cases. At no time, with either immunofluorescence reagent, could an immunofluorescence reaction be observed outside the lens tissue. Such results indicate that the γ-crystallins, previously implicated in bovine and regenerating newt lens fiber differentiation, are also indicative of lens fiber differentiation in the normal, developing lens of the anuran amphibian, R. pipiens. In addition, the relatively late appearance in lens development (i.e., after formation of a lens vesicle) of any immunofluorescence reaction for the crystallins suggest that they are not necessary for lens induction and subsequent lens placode formation.

Reversal by potassium of an effect of barium on the frog gastric mucosa.

ARTICLESReversal by potassium of an effect of barium on the frog gastric mucosaAD Pacifico, M Schwartz, TN MacKrell, SG Spangler, SS Sanders, and WS RehmAD Pacifico, M Schwartz, TN MacKrell, SG Spangler, SS Sanders, and WS RehmPublished Online:01 Mar 1969https://doi.org/10.1152/ajplegacy.1969.216.3.536MoreSectionsPDF (1 MB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat Previous Back to Top Next Download PDF FiguresReferencesRelatedInformation Cited ByAmmonia and gastric acid secretion: a key to understanding activity and regulation of the H+,K+-ATPaseInflammoPharmacology, Vol. 10, No. 4-6NH4Cl inhibition of acid secretion: possible involvement of an apical K+ channel in bullfrog oxyntic cellsSusan J. Hagen, Hui Wu, and Sarah W. Morrison1 August 2000 | American Journal of Physiology-Gastrointestinal and Liver Physiology, Vol. 279, No. 2Effect of melittin on PD, resistance and short-circuit current in the frog gastric mucosaBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 1369, No. 2Potential difference responses to nutrient K+, Na+ and Cl− changes with varying nutrient HCO3− in resting frog stomachBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 979, No. 3[31] Direct current electrical measurement in epithelia: Steady-state and transient analysisInhibition of acid secretion of fundus of Rana pipiens with a high concentration of potassium on the secretory sideBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 899, No. 1Inhibition of Ca2+-activated K+ channels in pig pancreatic acinar cells by Ba2+, Ca2+, quinine and quinidineBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 819, No. 2Effect of Ba2+ on the K+ conductance pathways in the frog corneaBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 819, No. 1Barium modifies the concentration dependence of active potassium transport by insect midgutThe Journal of Membrane Biology, Vol. 86, No. 2Effects of antidiuretic hormone on cellular conductive pathways in mouse medullary thick ascending limbs of Henle: I. ADH increases transcellular conductance pathwaysThe Journal of Membrane Biology, Vol. 80, No. 3Channels across Epithelial Cell LayersMicroelectrode study of K+ accumulation by tight epithelia: I. Baseline values of split frog skin and toad urinary bladderThe Journal of Membrane Biology, Vol. 72, No. 3The role of divalent cations in controlling amphibian lens membrane permeability; The mechanisms of toxic cataractsExperimental Eye Research, Vol. 36, No. 4Normal and anomalous potential responses due to K+ changes in bullfrog antrumBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 727, No. 1Anomalous potential response and (Na+ + K+)-ATPase in in vitro frog gastric mucosaBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 649, No. 2Mechanisms of cation permeation across apical cell membrane ofNecturus gallbladder: Effects of luminal pH and divalent cations on K+ and Na+ permeabilityThe Journal of Membrane Biology, Vol. 59, No. 3The effects of barium on the electrical properties of the basolateral membrane in proximal tubuleNaunyn-Schmiedeberg's Archives of Pharmacology, Vol. 318, No. 2K+-permeability of the outer border of the frog skin (R. temporaria)The Journal of Membrane Biology, Vol. 52, No. 2Relative roles of Ca2+, Sr2+, Ba2+ and Mg2+ in controlling lens permeability characteristicsCell Calcium, Vol. 1, No. 1Inhibition of potassium conductance by barium in frog skin epitheliumBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 552, No. 2Mechanism of the Inhibitory Action of Electric Current on H+ Secretion by Frog Gastric MucosaGastroenterology, Vol. 76, No. 4Gastric SecretionIon and Water Transport in Gastric MucosaElectrical Events During Activation and Inhibition of Gastric HCI SecretionGastroenterology, Vol. 73, No. 4Effects of barium on some stimulus evoked inhibitory post-synaptic potentials in the brain of Helix pomatiaComparative Biochemistry and Physiology Part A: Physiology, Vol. 58, No. 2Barium inhibition of sodium ion transport in toad bladderBiochimica et Biophysica Acta (BBA) - Biomembranes, Vol. 436, No. 3IMPLICATIONS OF THE NEUTRAL CARRIER Cl − -HCO 3− EXCHANGE MECHANISM IN GASTRIC MUCOSA fn116 December 2006 | Annals of the New York Academy of Sciences, Vol. 264, No. 1Secretory Events in Gastric MucosaConductance of epithelial tissues with particular reference to the frog's cornea and gastric mucosaExperimental Eye Research, Vol. 15, No. 5Influence of Fixation on Physiological Properties of Frog Gastric MucosaActa Physiologica Scandinavica, Vol. 88, No. 1Proton TransportUnstirred-layer model for the long time-constant transient voltage response to current in epithelial tissueJournal of Theoretical Biology, Vol. 32, No. 1 More from this issue > Volume 216Issue 3March 1969Pages 536-541 Copyright & PermissionsCopyright © 1969 by American Physiological Societyhttps://doi.org/10.1152/ajplegacy.1969.216.3.536PubMed5765605History Published online 1 March 1969 Published in print 1 March 1969 Metrics

Synthesis of macromolecules in epithelial cells of the cultured amphibian lens. IV. Requirement of DNA replication for the synthesis of RNA and protein during the "G2" period of the cell cycle.

Synthesis of macromolecules in epithelial cells of the cultured amphibian lens. IV. Requirement of DNA replication for the synthesis of RNA and protein during the "G2" period of the cell cycle.

Synthesis of macromolecules in epithelial cells of the cultured amphibian lens. 3. Involvement of protein synthesis in mitotic activation.

Synthesis of macromolecules in epithelial cells of the cultured amphibian lens. 3. Involvement of protein synthesis in mitotic activation.

Synthesis of macromolecules in epithelial cells of the cultured amphibian lens: II. Effects of puromycin and actinomycin D on protein synthesis

Synthesis of macromolecules in epithelial cells of the cultured amphibian lens: II. Effects of puromycin and actinomycin D on protein synthesis

Induction and competence in morphogenesis: amphibian lens development from non-lens ectoderm, influenced by mercaptoethanol.

Induction and competence in morphogenesis: amphibian lens development from non-lens ectoderm, influenced by mercaptoethanol.

Synthesis of macromolecules in epithelial cells of the cultured amphibian lens: I. DNA and RNA

1. 1. When bullfrog lenses are introduced into a culture system, the epithelial cells undergo bursts of DNA synthesis and mitosis which reach peaks at 52 and 87 hr post-isolation, respectively. 2. 2. These events are preceded by an augmentation of RNA synthesis which begins within 10 hr of isolation and attains peaks at 36 and 72 hr. 3. 3. If the increment in RNA synthesis is blocked by treatment with 10 −1 μg/ml actinomycin D the waves of DNA synthesis and mitosis are suppressed even though uridine incorporation in “blocked” lenses is still significantly higher than it is in freshly isolated material. 4. 4. It was shown by autoradiographic means that when the actinomycin D was supplied in tritiated form it was incorporated almost exclusively into the nucleus. 5. 5. The foregoing results were discussed with reference to the mechanisms which control proliferation.

In vitro culture of amphibian lenses.

BECAUSE the lens is an organ which lacks both blood and nerve supplies it has long been felt that it could be cultured with relative ease. Consequently, many attempts have been made in this direction, primarily with material of mammalian origin. Much of this work is reviewed by Schwartz1 and Bito2. Harding, Rothstein and Newman3 have demonstrated that the epithelial cells of rabbit lenses maintained in systems perfused with combinations of various tissue culture media are caused to undergo DNA synthesis and cell division at varying periods of time following isolation, the specific interval depending on the composition of the perfusate. This ‘activation’ of DNA synthesis is of interest inasmuch as large numbers of cells are synchronized in a stage of the nuclear cycle the characteristics of which one would like to be able to elucidate. The fact that the activation is occurring in vitro naturally makes the whole system much more amenable to biochemical dissection than would ordinarily be the case.

STUDIES OF AN INJURY-INDUCED GROWTH IN THE FROG LENS.

AbstractA tumor‐like growth in the lens of Rana pipiens occurs following mechanical injury to the lens. The frequency of occurrence and size of the growth are roughly dependent on the extent of the injury inflicted. The growth first appears four days after wounding, grows actively for three weeks and usually begins to regress after a month. The cells of the growth appear to originate by proliferation of the lens epithelialcells surrounding the wound which grow into and actively invade the lens fibers. The lens growth was not transplantable either homologously into the anterior eye chamber after dissociation, nor isologously in a subcutaneous site. Protein, LDH, MDH, and G‐6‐PD patterns obtained after electrophoretic separation in starch gels were similar for normal and the abnormal lenses. Whether a true tumor or hyperplasia, the froglens exhibits an unique reaction to the injury stimulus which is not found in mammals (man, rabbit, mouse). The controversial tumor immunity of the mammalian lens appears to the related to an innate resistance of the epithelium to proliferation which is illustrated in part by the species difference in the reactivity of the mammalian lensand the amphibian lens to injury.