Cytochemical Localization of Guanylyl Cyclase Activity in Rabbit Taste Bud Cells

Guanylyl cyclase activity was cytochemically demonstrated inrabbit foliate taste buds. The enzymatic activity was localizedin the apical portion (microvilli and neck) of taste bud cells.Especially strong activity was observed on the microvillousmembrane of type I (dark) cells and often on a blunt processof type III cells. The microvilli of type II (light) cells showedweak enzymatic activity. Considering that the apical portionof taste cells is a likely site of interaction between tastestimuli and the cells, the results support the idea that cyclicGMP is involved in taste transduction.     

Bombardment-mediated transformation of plant cells

Significance of bombardment-mediated transformation in plant research is discussed. Apart from general subjects related to this transformation study, a particular emphasis has been placed on the pollen and organelle transformation and on integration mechanism. Over 70 literatures related to this field are cited here.     

Influence of Axis Removal on the Expression of a Gene for a Cysteine Endopeptidase (EP-C1) in French Bean Cotyledons during Germination

Levels of a cysteine endopeptidase, EP-Cl, and its mRNA increasedslightly in cotyledons of French bean detached from embryonicaxes. A transient expression assay by a particle gun indicatedthat the downstream region from position –340 of the EP-Clgene was sufficient for gene expression in cotyledons of germinatingseeds. (Received August 5, 1994; Accepted February 17, 1995)     

House mouse Mus musculus dispersal in East Eurasia inferred from 98 newly determined complete mitochondrial genome sequences

The Eurasian house mouse Mus musculus is useful for tracing prehistorical human movement related to the spread of farming. We determined whole mitochondrial DNA (mtDNA) sequences (ca. 16,000 bp) of 98 wild-derived individuals of two subspecies, M. m. musculus (MUS) and M. m. castaneus (CAS). We revealed directional dispersals reaching as far as the Japanese Archipelago from their homelands. Our phylogenetic analysis indicated that the eastward movement of MUS was characterised by five step-wise regional extension events: (1) broad spatial expansion into eastern Europe and the western part of western China, (2) dispersal to the eastern part of western China, (3) dispersal to northern China, (4) dispersal to the Korean Peninsula and (5) colonisation and expansion in the Japanese Archipelago. These events were estimated to have occurred during the last 2000–18,000 years. The dispersal of CAS was characterised by three events: initial divergences (ca. 7000–9000 years ago) of haplogroups in northernmost China and the eastern coast of India, followed by two population expansion events that likely originated from the Yangtze River basin to broad areas of South and Southeast Asia, including Sri Lanka, Bangladesh and Indonesia (ca. 4000–6000 years ago) and to Yunnan, southern China and the Japanese Archipelago (ca. 2000–3500). This study provides a solid framework for the spatiotemporal movement of the human-associated organisms in Holocene Eastern Eurasia using whole mtDNA sequences, reliable evolutionary rates and accurate branching patterns. The information obtained here contributes to the analysis of a variety of animals and plants associated with prehistoric human migration.Subject terms: Evolution, Genetic variation     

High frequency stable transformation ofArabidopsis thaliana by particle bombardment

Root explants ofArabidopsis thaliana ecotype C24 were bombarded with the plasmid pCH harboring the hygromycin phosphotransferase gene (hpt). A selection condition with post-bombardment culture of 3 days followed by culture with 20 mgl⁻¹ hygromycin gave the highest yield of transformants. More than 44% of explant clumps formed transformant shoots.     

Fish otolith contains a unique structural protein, otolin-1.

A collagen-like protein was identified from the otoliths of the chum salmon, Oncorhynchus keta. The otolith, composed mainly of calcium carbonate with small amount of organic matrices, is formed in the inner ear and serves as a part of the hearing and balance systems. Although the organic matrices may play important roles in the growth of otolith, little is known about their chemical nature and physiological function. In this study, a major organic component of the otolith, designated otolin-1, which may serve as a template for calcification, was purified. The sequences of two tryptic peptides from otolin-1 revealed high homology with parts of a saccular collagen which had been described previously [Davis, J.G., Oberholtzer, J.C., Burns, F.R. & Greene, M.I. (1995) Science 267, 1031-1034]. Cloning of a cDNA coding for otolin-1 revealed that the deduced amino-acid sequence contained a collagenous domain in the central part of the protein. Although collagen is the most abundant structural protein in the animal body, otolin-1 mRNA was expressed specifically in the sacculus. Immunohistochemical studies showed that otolin-1 is synthesized in the transitional epithelium and transferred to the otolith and otolithic membrane. This is the first report concerning characterization of a structural protein containing many tandem repeats of the sequence, Gly-Xaa-Yaa, typical for collagen from the biomineral composed of calcium carbonate.     

Histochemical localization of adenylate cyclase and phosphodiesterase activities in the folliate papillae of the rabbit I. Light microscopic observations

Adenylate cyclase and cyclic AMP phosphodiesterase activitieson the foliate papillae of rabbit were studied by means of histochemistry.In unfixed papillae the reaction product for adenylate cyclaseactivity was localized in the apex of taste buds, lamina proprioand connective tissue core of the papillae, but in fixed papillaeit was limited to the apex of taste buds. The reaction productfor cyclic AMP phosphodiesterase activity was limited to theapex of taste buds in unfixed and fixed papillae. Neither anacceleratory nor an inhibitory effect of sweet and bitter substanceson the adenylate cyclase and cyclic AMP phosphodiesterase activitieswas demonstrated, but NaCl prevented the formation of reactionproduct for the adenylate cyclase activity at the apex of tastebuds.     

Molecular analysis of the gene encoding a protein component of the Eikenella corrodens adhesin complex that is close to the carbohydrate recognition domain

A monoclonal antibody against a lectin-like substance (LS) of Eikenella corrodens (Ec) was used for screening the Ec DNA library. Three positive clones that carried an identical 12-kb segment were obtained. A 25-kDa protein, which specifically binds to the antibody, was overproduced in all of the Escherichia coli clones. Deletion analysis showed that the gene encoding the 25-kDa protein was located within a 1.2-kb segment. The nucleotide (nt) sequence of this segment contained an open reading frame encoding a protein of 24 600 Da. We purified the 25-kDa protein from the cloned E. coli strain. The sequence of the first 10 amino acids (aa) from the N-terminus of the purified 25-kDa protein agreed with that deduced from the nt sequence. Since the monoclonal antibody used in this study inhibits the physiological activity of EcLS, we concluded that the 25-kDa protein is a component of the adhesin complex, which is located near the carbohydrate recognition domain of lectin in EcLS.