Electron microscopic studies on liver 5-aminolaevulinate synthase

The structure of chick embryo liver 5-aminolaevulinate synthase has been examined by electron microscopic studies using negative staining. From the different projections of the enzyme particles observed in electron micrographs, a model for the enzyme molecule has been proposed. In this model, an enzyme molecule consists of two curved and identical subunits associated in opposite polarities. From the dimensions of an enzyme molecule subunit measured from electron micrographs, the relative molecular mass of each subunit is estimated to be 70 000.     

Initial steps in receptor-mediated endocytosis : The influence of temperature on the shape and distribution of plasma membrane clathrin-coated pits in cultured mammalian cells

We have examined the shape and distribution of clathrin-coated pits in Swiss 3T3 cells at 4 or 37 degrees C using electron microscopy with serial sections and immunofluorescence light microscopy. Both groups were fixed in glutaraldehyde and preserved further using a membrane contrast enhancement technique consisting of sequential osmium-ferrocyanide, thiocarbohydrazide and osmium-ferrocyanide treatment in situ. Concanavalin A-horseradish peroxidase (conA-HRP) was used to identify these structures participating in endocytosis. Two hundred twenty-two clathrin-coated structures were analysed; 126 from cells fixed at 4 degrees C, and 96 from cells fixed after a 3 min warm-up to 37 degrees C. All coated structures labeled with conA-HRP had demonstrable connections to the plasma membrane. These coated structures were morphologically classified into three categories: (a) flat pits; (b) curved pits; and (c) pits with narrow-neck connections to the plasma membrane. At 37 degrees C, 27% of coated pits had narrow neck connections to the plasma membrane whereas at 4 degrees C only 1% had such connections. Receptosomes (endosomes) labeled with conA-HRP were found only after incubation at 37 degrees C, indicating that active endocytosis was occurring in cells at 37 degrees C, but not at 4 degrees C. Immunofluorescence with anti-clathrin antibody was used to quantitate the number of clathrin-coated pits in Swiss 3T3 cells incubated at 4 and 37 degrees C prior to fixation. No difference was detected. There were 426 +/- 122 pits per cell at 37 degrees C and 441 +/- 106 at 4 degrees C. These results support the hypothesis that formation of a narrow neck connected a coated pit to the cell surface is an early step in the mechanism of receptor-mediated endocytosis.     

Localized Photosynthate Deposition in Citrus Fruit Segments Relative to Source-Leaf Position

Photosynthate translocation into fruit segments was examinedin ‘Pineapple’ sweet orange Citrus sinensis (L.)Osbeck to determine whether previously reported patterns ofdistribution [Koch (1984) HortScience 19: 260] would changeover time or with alterations in balance between source leavesand sink fruit. In control plants, ¹⁴CO₂ was supplied to a sourceleaf nearest the fruit for 1 h, followed by 5 h translocation.Over 89% of [¹⁴C]assimilates in the fruit were localized in4 segments directly aligned with the source and 73% of thesewere in the center 2 segments. Peel, pulp and seeds showed similarpatterns. Little or no lateral spreading of [¹⁴C]photosynthatesoccurred when an additional 7 days were allowed for translocation,but distribution was slightly broader when the source leaf was8 nodes farther from the fruit. Defoliation and girdling toreduce the source/sink ratio gave variable results if done 18h before experiments, but widened the area receiving [¹⁴C]assimilatesto approximately half the fruit if done 7 days earlier. Thisoccurred only when an entire fruit, was dependent upon a singlesource, leaving the opposite half fruit without an externalsupply of photosynthates. These data show an extreme degreeof preferential translocation and inflexibility which can occurin a transport path. ¹Supported by United States Department of Agriculture CompetitiveResearch Grant 59-2121-1-1-752-0, Regional Project NC-142, andthe Institute of Food and Agricultural Sciences, Universityof Florida. (Received January 12, 1984; Accepted May 14, 1984)     

X-inactivation patterns in lymphocytes and skin fibroblasts of three cases of X-autosome translocations with abnormal phenotypes

Summary X-inactivation patterns were studied by replication analyses both in lymphocytes and skin fibroblasts of two patients carrying balanced X-autosome translocations, t(X;10)-(pter;q11) and t(X;17)(q11;q11), and one patient with an unbalanced translocation t(X;22)(p21;q11). Preferential late replication of the normal X chromosome was found in lymphocytes of both patients carrying balanced translocations and in skin fibroblasts of the patient carrying the translocation t(X;17). However, skin fibroblasts of the patient with a translocation t(X;10) showed preferential late replication of the abnormal der(X) chromosome with no spreading of late replication to the autosomal segment. In the case of unbalanced translocation t(X;22) there was preferential late replication of the der(X) chromosome both in lymphocytes and skin fibroblasts. The abnormal phenotype of the patients is discussed in relation to the observed X-inactivation patterns and the variability of the patterns in different tissues.     

Proteinase-sensitive release of enzymes from pancreatic microsomal fraction.

Suspensions of rat pancreatic microsomal fraction release alpha-amylase and ribonuclease on incubation at 37 degrees C, but not at 2 degrees C. The release is abolished by proteolytic enzymes. Ribonuclease associated with the microsomal fraction is protected from subtilisin BPN' attack, but is sensitive after release.     

Isolation and initial characterization of human basement membrane collagens

Neutral solutions of pepsin extracted human collagens derived from glomeruli, kidney, aorta, lung, heart, bowel, spleen, skeletal muscle and skin were subjected to heat gelation at 37°C. Centrifugation of the gel provided two fractions: gelled pellet and non-gelled supernatant. Analysis of these two fractions by gel electrophoresis, molecular sieve and ion exchange chromatography, and amino acid and carbohydrate determinations indicated that the non-gelled supernatant contained a substantial enrichment of basement membrane like collagen. The initial characterization of lung derived basement membrane collagen indicated close similarities with those derived from glomeruli and whole kidney and differences with that obtained from the spleen.     

The influence of foraging benefits on association of cattle egrets (Bubulcus ibis) with cattle

Summary The degree of association between cattle egrets (Bubulcus ibis) and cattle was studied during one summer on Saint Catherines Island, Georgia, USA. Previous work by Grubb (1976) and others indicated that cattle egrets foraging with cattle require fewer steps and less time to catch prey than egrets foraging without cattle and single egrets catch prey at a higher rate than egrets foraging in groups of two or more with cattle. Accordingly, we predicted that when given a choice egrets should forage with cattle rather than alone, egrets should prefer to associate with standing rather than sitting cattle, and single egrets associated with cattle should be more common than expected by chance.In excess of two-thirds of the egrets accompanied cattle. Neither time of day nor month influenced the degree of association, but egrets in forest were more likely to be associated with cattle than egrets in pasture. Standing cattle were more likely to be accompanied by egrets than were sitting cattle. Single egrets occurred more frequently than expected by chance when accompanying standing cattle but not when associated with sitting cattle. Thus, cattle egrets usually distributed themselves among cattle in the way predicted by optimal foraging theory.We dedicate this paper to the memory of Michael D. Sabath     

Analysis of picogram quantities of indole-3-acetic acid by high performance liquid chromatography-fluorescence procedures

The use of spectrofluorimeter coupled to a reverse phase high performance liquid chromatography column permits selective detection of indole-3-acetic acid at the low picogram level. The value of the technique is demonstrated by the analysis of endogenous IAA in elongating shoots, xylem sap and callus of Douglas-fir. The data are also used to illustrate a procedure whereby the accuracy of chromatographic analyses can be verified within definable probability limits.Abbreviations GC-MS combined gas chromatography-mass spectrometry - HPLC high performance liquid chromatography - IAA indole-3-acetic acid - SEC steric exclusion chromatography - SICM selected ion current monitoring Technical Paper No. 5379 from the Oregon State University Agricultural Experiment Station