Molecular cloning of the cDNA for androgen-dependent sperm-coating glycoproteins secreted by the rat epididymis

cDNA clones coding for two closely related androgen-dependent sperm-coating glycoproteins secreted by the rat epididymis were selected by screening an epididymal cDNA library constructed in lambda gt 11 with affinity-purified antibody directed against the glycoproteins. The largest clone of 956 nucleotides provided coding information for a protein of 246 amino acids of which the first 19 residues comprise a putative signal peptide sequence which when cleaved would produce a mature protein of 227 residues and a molecular mass of 26 kDa. Confirmation of the identity of the clone was provided by a match between the amino acid sequence predicted from the cDNA sequence and the actual amino acid sequence determined for a tryptic peptide fragment of one of the pure glycoproteins. It is probable that the primary amino acid sequence of the two glycoproteins is identical. Northern blot and slot-blot analysis revealed that the mRNA for the glycoproteins is approximately 1250 nucleotides long and that the concentration of the mRNA in the epididymis is androgen-dependent. The glycoproteins and their mRNAs were unique to the epididymis as determined by Western and Northern blots, respectively, since signals were absent from skin, brain, liver, kidney, heart, skeletal muscle and testis. Cross-reacting proteins of slightly smaller apparent molecular mass were detected in extracts of mouse and guinea-pig epididymis, but not rabbit or bull epididymis. Comparison with existing protein data bases revealed that the epididymal glycoproteins display significant sequence homology with yeast carboxypeptidase Y.     

Lymphopoiesis in the nude fetal thymus following sympathectomy

This study was carried out to examine the innervation of the nude fetal thymus during ontogeny and to see if lymphopoietic activity would occur within these thymic lobes in the absence of sympathetic neuronal input. Fetal thymic rudiments from nu/nu mice were removed and examined for galoxylic acid-induced histofluorescence to detect the catecholaminergic nerves. Some of these lobes were organ cultured for 5 to 7 days in the presence of deoxyguanosine to eliminate any existing lymphoid cells within the rudiments. Such "nonlymphoid" thymic rudiments were implanted into the anterior eye chambers of syngenic BALB/c mice (heterozygous) from which cervical sympathetic ganglia and part of the sympathetic chain had been surgically removed (right side) one week earlier. The left side was only sham operated. The thymic implants were allowed to grow for up to 21 days on both sides; they were then removed and examined by histofluorescence, immunofluorescence, and light microscopy. The results indicate for the first time that the nude fetal thymus is innervated by sympathetic nerves and that following sympathectomy the nude thymus is able to sustain lymphopoietic activity and generate lymphoid cells which have characteristics present on thymocytes during in vivo development in normal mice, such as binding to peanut agglutinin and expression of Thy-1 antigen. The relationship between the presence of sympathetic inhibitory influence and the thymic atrophy seen in the nude mice during ontogeny, is being investigated.     

Root turnover and production by14C dilution: implications of carbon partitioning in plants

Summary Estimates of belowground net primary production (BNP) obtained by using traditional soil core harvest data are subject to a variety of potentially serious errors. In a controlled growth chamber experiment, we examined the aboveground-belowground, labile to structural tissue, and plant to soil dynamics of carbon to formulate a¹⁴C dilution technique for potential successful application in the field and to quantify sources of error in production estimates.Despite the fact that the majority of net¹⁴C movement between above- and belowground plant parts occurred between the initial labeling and day 5, significant quantities of¹⁴C were incorporated into cell-wall tissue throughout the growing period. The rate of this increase at late sampling dates was greater for roots than for shoots. Total loss of assimilated¹⁴C was 47% in wheat and 28% in blue grama. Exudation and sloughing in wheat and blue grama, respectively, was 15 and 6% of total uptake and 22 and 8% of total plant production.When root production estimates by¹⁴C dilution were corrected for the quantities of labile¹⁴C incorporated into structural carbon between two sampling dates, good agreement with actual production was found. The error associated with these estimates was ±2% compared with a range of –119 to –57% for the uncorrected estimates. Our results suggest that this technique has potential field application if sampling is performed the year after labelling.Sources of errors in harvest versus¹⁴C dilution estimates of BNP are discussed.     

Emergence of some caddisflies (Trichoptera) from a wooded stream in southern Ontario

Twenty-eight species of nine families of caddisflies (Trichoptera) were identified in 170 samples taken over an 8-month period from five emergence traps placed on a second-order, forested, cold-stenothermal stream on the Niagara Escarpment, Ontario, Canada.A mean of 980.9 caddisflies m^–2 of streambed was obtained over the entire sampling period. Eleven common species accounted for 92.8% of the total emergence with specific proportions ranging from 23.8% (Wormaldia moesta) to 0.11 % (Rhyacophila sp.). The use of various kinds of traps in other studies and their effects on the detection of species composition and abundance are discussed and compared with the present study.Although the distributions of all the common species were invariant over time, four species showed low to high degrees of patchiness in the streambed; the other seven common species were uniformly distributed. However, a large residual variance suggested a subtle mechanism of microhabitat selection by the larvae and (or) pupae, not detectable by even the small emergence traps used.Both sexes of 15 species, only males of 4 and only females of 9 species were collected. Eight of the eleven common species showed significant departures from a balanced sex ratio and five exhibited a protandry of from 1 to 3 weeks. Neither this study nor others have been able to establish a predictable pattern of sex ratios in Trichoptera.The emergence periods and patterns of the eleven common species are described and compared with other studies. Of these common species, one emerged in the spring, seven during the summer and three during the late summer or early fall. Ten species had a short emergence period with a distinct peak and a significantly skewed pattern. One species,Parapsyche apicalis, exhibited a prolonged emergence period, no distinct peak and a significantly platykurtotic² pattern. With the exception ofLepidostoma sp. A, the emergence patterns of the common species were unimodal.     

An autoradiographic study of the uptake of tritiated proline by osteoblasts during hibernation

Twenty-four LSH and LVG strain golden hamsters, Mesocricetus auratus, were used. Experimental animals were maintained at 5 C and allowed to hibernate. Control animals were kept at 27 C. Six animals (3 experimental, 3 control) were injected subcutaneously with 1 microCi of 3H-proline/gm body wt. (Spec. act. 3 Ci/mM) after hibernation lasting 12 hours, 1 day, 3 days, or 7 days. Animals were killed 1 hour after injection and autoradiographs were prepared from 5 microns thick decalcified sections of femurs. A greater number of endosteal cells were labeled than periosteal cells and also exhibited a greater magnitude of labeling throughout the study. Differences between endosteal and periosteal cells both in percentage of cells labeled and magnitude of labeling were maximum in control animals and progressively decreased with increasing periods of hibernation. A reduction in synthesis of matrix proteins during the early period of hibernation was seen and was attributed to a significant reduction both in average cell activity and in the number of active cells during hibernation. The latter phenomenon apparently made a large contribution to the reduced matrical synthesis. 3H-proline uptake by osteoblasts probably reflects the reduced requirements of matrical synthesis during hibernation.     

The structural gene for the mitochondrial aldehyde dehydrogenase maps to human chromosome 12

Summary A cloned 850 bp cDNA fragment corresponding to the 3-coding part of human ALDHI-mRNA was used as a probe for the chromosomal assignment of the ALDHI gene. Southern blot analysis of human-rodent somatic cell hybrids indicates that the human ALDHI gene resides on chromosome 12.Dedicated to Prof. Dr. H. Holzer on the occasion of his 65. birthday     

Bkm sequences are polymorphic in humans and are clustered in pericentric regions of various acrocentric chromosomes including the Y

Summary Probes of uncloned Bkm satellite DNA and a Drosophila clone 2(8), consisting mainly of GATA repeasts related to a major sequence component in Bkm, have been used to probe Southern blots of human male and female DNAs obtained from a Caucasian and an Australian aboriginal population and to human chromosomes in situ. Hybridization was observed to a distinct and an indistint series of bands against a smeared background. The same distinct bands are identified in the DNA samples with both probes, but are most readily detected using the uncloned Bkm probe. Most restriction bands are common to both populations and some are polymorphic. However, certain bands appear to be characteristic of the Australian aboriginal samples. There are no distinct sex-linked patterns. However all of the small acrocentric human chromosomes, including the Y chromosome show hybridization to uncloned Bkm in situ.     

Mode of uptake of insoluble solid substrates by microorganisms. II: Uptake of solid n-alkanes by yeast and bacterial species

Using two species of yeast and one of bacterium, evidence has ben obtained which indicates that the microbial uptake of solid alkane powders occurs primarily through a substrate solubilization mechanism. EDTA, a strong inhibitor of hydrocarbon solubilization by the cells, inhibited the growth of these organisms on alkane powder; the inhibition could be removed vai a supply of artificially solubilized alkane. One of the yeast strians, which was a mutant incapable of growing on solid alkane powder and liquid alkane, could grow very well on artifically solubilized alkanes. It was demonstrated that the solid alkane solubilization rate during microbial growth could satisfactorily account for the maximal alkane uptake rate actully observed during growth. The specificity of solubilization for the solid alkane used as the growth substrate was demonstrated.     

Haemolysis of rabbit erythrocytes by a lectin from the seeds of<Emphasis Type="Italic">Croton tiglium</Emphasis>

The kinetics of haemolysis of rabbit erythrocytes byCroton tiglium lectin was studied as a function of concentration of the lectin and erythrocytes. The length of the prelytic period decreased with increasing lectin concentrations, indicating that the secondary events at the membrane which follow the binding of the lectin to cell surface carbohydrate receptors are accelerated at higher surface concentrations of the lectin. The rate or extent of haemolysis was not affected by the inclusion of ions like K⁺, Ca²⁺ and Mg²⁺ in the medium or by the substitution of ionic medium by a non-ionic medium. The inhibition of haemagglutination and haemolysis of rabbit red cells byCroton tiglium lectin by antilectin rabbit serum was observed. A possible mechanism of haemolysis by the lectin is discussed.     

Plant lectins specific for N-acetyl-β-D-galactosamine

N-Acetyl-D-galactosamine in β-linkage being ubiquitous in cell surface glycoproteins, their interaction with lectins specific for this sugar moiety may be a significant event in cell adhesion phenomena. This article discusses the common β-N-acetyl galactosamine-specific lectins, with particular stress on the lectin from winged beans (Psophocarpus tetragonolobus).