Retinoic acid resistance of the variant embryonal carcinoma cell line RAC65 is caused by expression of a truncated RARα

P19 embryonal carcinoma (EC) cells differentiate when treated with retinoic acid (RA). The P19 EC-derived mutant cell line RAC65 is resistant to the differentiation-inducing activity of RA. We show that these cells express a truncated retinoic acid receptor alpha(mRAR alpha-RAC65), probably due to the integration of a transposon-like element in the RAR alpha gene. This receptor lacks 71 C-terminal amino acids and terminates in the ligand-binding domain. In CAT assays in RAC65 cells, mRAR alpha-RAC65 fails to trans-activate the RAR beta promoter, which contains a RA-response element. In wild-type P19 EC cells mRAR alpha-RAC65 functions as a dominant-negative repressor of RA-induced RAR beta activation. Gel retardation assays demonstrate that mRAR alpha-RAC65 is still able to bind to the RA-response element of the RAR beta promoter, indicating that competition with functional RARs for the same binding site leads to the observed dominant-negative effect. In addition, in two RAC65 clones in which wild-type hRAR alpha was stably transfected RA-sensitivity was restored and in one RAR beta expression could be induced by RA. Taken together, these data show that the primary cause of RA-resistance of RAC65 cells is the expression of a defective RAR alpha, which prevents the trans-activation of RA-responsive genes and results in a loss of the ability to differentiate.     

Glycerol alters cytoskeleton and cell adhesion while inhibiting cell proliferation.

Our previous studies with various cell lines and human glioma cells showed that glycerol suppresses cell proliferation. In this study, we examined the effects of glycerol on the cytoskeleton, general morphology and attachment of Baby Hamster Kidney (BHK) cells. In glycerol-treated cells proliferation was suppressed, the microfilament network was extensively reorganized and the microtubule network was more clearly defined, while the cell thickness, as observed by scanning electron microscopy (SEM), was decreased. In addition, glycerol treatment resulted in an increase in the number of nucleoli per cell and in the appearance of non-mitotic rounded cells. Glycerol treatment increased the time needed to trypsinize BHK cells from the culture dishes and reduced the time required for cell reattachment. We suggest that glycerol treatment provides a non-toxic tool to examine mechanisms of proliferation arrest.     

Influence of calcium pretreatment on wheat germination on saline media

Summary Wheat grains (Triticum vulgare L. var. Lemhi 53) were pretreated for about 6 hours with various solutions. They were then rinsed and planted on filter paper moistened with NaCl solution in petri dishes. Percent germination was measured after 3 days. Germination of water pretreated seeds on 1% NaCl was 8 per cent, while pretreatment with 1% CaCl₂. 2H₂O resulted in 90 per cent germination on 1% NaCl. Pretreatment with sodium and potassium chlorides enhanced germination only slightly. The beneficial effects of calcium pretreatment could be duplicated only partially by increments of CaCl₂ to the NaCl germination medium. Preatreatment concentrations ranging from 1 to 5% CaCl₂. 2H₂O, and times ranging from 3 to 24 hours were about equally effective. The beneficial effect of pretreatment persisted even though the seeds were subsequently dried and planted several months later. Pretreatment with calcium resulted in about a 25 per cent reduction in Na²² uptake from the germination medium.United Nations F.A.O. André Meyer Post Doctoral Research Fellow, on leave from Sind University, Hyderabad, West Pakistan.     

Dispersion of viable pig liver cells with collagenase.

Viable suspended hepatocytes were prepared from surgical biopsy specimens of pig and human liver by digestion with collagenase. Initial perfusion of the tissue through cannulated blood vessels with 0.5 mM EGTA followed by 0.2% collagenase gave the best results. 20−870 × 10⁶ cells of which 60–95 % excluded trypan blue were obtained from 5–30 g pig liver pieces, while results with human liver specimens were usually less satisfactory. In some experiments, however, viable cells, as judged by vital stain exclusion and ability to synthesize lipids were obtained in sufficient yield. In the pig hepatocytes glycerolipid synthesis from [³H] glycerol and oxidation and esterification of [¹⁴C] oleic acid had the same characteristics as those observed earlier in rat hepatocytes.     

Male behavior through the breeding season in Saimiri boliviensis boliviensis

The Bolivian squirrel monkey (Saimiri boliviensis boliviensis) is a seasonal breeder. Male squirrel monkeys show distinct morphological and behavioral changes prior to and during the breeding season. A “fatting syndrome” includes increased body weight, increased levels of androgens, and in the Bolivian subspecies, an increasingly active role in the social organization of the group. In this study, the behavior of ten adult male Bolivian squirrel monkeys was analyzed over a 6-month period prior to, during, and after the breeding season. Each was housed as the only adult male in a breeding unit with six to ten adult females and one juvenile male. Employing a principle components method, 11 behavioral clusters were generated from 27 responses. Their activity clusters were identified as follows: sexual activity that showed a peak around the time of peak conceptions; excitatory activity that was initially high but decreased throughout the breeding season; and maintenance activity that did not change across the breeding season. The changing social behavior of the male squirrel monkey parallels physiological changes and is correlated with changing androgen levels.     

Effect of phenological development on radiophosphorus translocation from leaves in crested wheatgrass

Summary The effect of phenological development on the pattern of photosynthate translocation was studied in crested wheatgrass [Agropyron cristatum (L.) Gaertn.] plants grown in a nursery under semi-natural environmental conditions at Logan, Utah. Radiophosphorus was used to trace the photosynthate translocation from April 13, 1968, through December 2, 1968.In early spring photosynthates were translocated about equally to the roots and younger leaves. Translocation was chiefly upward during the flower stage but was reversed with raturation of the inflorescence. In late summer the plants appeared quiescent but substantial ³²P was transported to the underground portions of the plant. After quiescence was broken in the fall, shoot and root growth were resumed simultaneously with peak ³²P movement to the crown and roots.The concentration of total available carbohydrates in the roots and crowns reached a maximum level just prior to quiescence and decreased during shoot production in the fall. Presumably, the carbohydrates were used in growth and the accumulation of carbohydrates for the winter is apparently not critical in crested wheatgrass. Since the plants have many basal leaves throughout the winter, they may have the capacity to carry out photosynthesis on warm, sunny winter days and are capable of immediately initiating growth in the spring.Utah Agricultural Experiment Station Paper 1074.