Histochemical studies on the morphology of the Golgi apparatus and on the distribution of hexokinase,phosphoglucomutase and some dehydrogenases in the dorsal vagal and hypoglossal nuclei of the rabbit

Summary Detailed histochemical studies have been made on the distribution of various enzymes such as thiamine pyrophosphatase (TPPase), hexokinase and glucokinase (HK), phosphoglucomutase (PG), glucose-6-phosphate dehydrogenase (G6PD) and succinate dehydrogenase (SDH) in every component of the dorsal vagal nucleus (X) and the hypoglossal nucleus (XII) of adult healthy male rabbits.The neurons of both nuclei were classified into the same six categories on the basis of the morphology of the Golgi apparatus (GA). Many intermediate forms were observed among these different categories. In general, the neurons of the X showed much more developed GA compared with those of the XII. The present results concerning TPPase may indicate that two different types of neurons are present in both nuclei and that they go through cyclic activity. The GA extensions in the dendrites may also change their forms depending on the functional state.The cytoplasm of the neuron and the neuropil reacted more strongly in the X than in the XII for the HK test. All the glial cells were very strongly positive for HK and they were closely arranged in the X. Both nuclei showed a low level of PG activity. These results indicate that glucose in the blood is the main energy source, that not only oligodendrocytes but also astrocytes work as energy donators to the neurons, and that the neurons of the X get more energy supply than those of the XII. Inverse relationship was found on the distribution of G6PD and SDH in the neurons of the X and XII: The cytoplasm of the neuron in the X was rich in G6PD and relatively poor in SDH whereas this, in the XII, was rich in SDH and poor in G6PD. These findings may provide the support to the conception of Exceptional Nuclei. The predominance of hexose monophosphate shunt in the neurons of the X may provide TPNH which is important for synthesis.The present author assumes that the neurons of the X have vigorous secretory activity on the basis of well-developed GA, rich energy supply and the predominance of hexose monophosphate shunt.     

Histochemical studies on the morphology of the golgi apparatus and on the distribution of hexokinase,L-gulonolactone oxidase,xylulose reductases,and ascorbic acid in the locus coeruleus of the rabbit

Summary Detailed histochemical studies have been conducted on the distribution of thiamine pyrophosphatase (TPPase), hexokinase and glucokinase (HK), L-gulonolactone oxidase (GO), D-xylulose reductase (DX), L-xylulose reductase (LX) and ascorbic acid (AC) in every component of the locus coeruleus (LC) of the healthy adult male rabbit.The LC consisted of medium-sized neurons and small neurons. Both types of neurons were classified into the same five categories on the basis of the morphology of the Golgi apparatus (GA). Many intermediate forms were observed between these different categories. The present results concerning TPPase may indicate that each type of neuron goes through cyclic activity.The GA of the small neurons showed little variation in its reactivity and volume in each category and no disintegration or budding-off. These neurons were mildly positive for the HK test, and negative for the GO, DX, LX and AC tests in contrast to the medium-sized neurons. These results may suggest that the small neurons are metabolically inactive, and that they have a different function from the medium-sized neurons.The morphology of the GA of the medium-sized neurons was basically similar to that described for motor neurons. It was considerably different from the morphology of the GA reported in the dorsal vagal nucleus (X) and hypothalamic neurosecretory nuclei (HMN) of the rabbit. These results suggest that the medium-sized neurons of the LC may be motor neurons, and that they may not have a neurosecretory function.The medium-sized neurons showed strong activity whereas the surrounding glial cells and neuropil exhibited mild activity in the HK test. These findings may suggest that these neurons get their energy source directly from the circulating blood.The medium-sized neurons were mildly to moderately positive for the DX and LX tests, and some of them were strongly positive for the GO test. Positive granules showed the tendency to accumulate in a proximal part of the main cell process and the part of perikaryon adjacent to it for the AC test. On the basis of these results, it is suggested that there is a strong possibility that at least some of the medium-sized neurons of the LC have the ability to synthesize vitamin C. This ability may be intimately related to the ontogenetical development of catecholamine.     

实验性氟中毒大鼠中缝背核酶细胞化学研究

以饮用高氟水（１００ｐｐｍ）的方法制造了雄性大鼠慢性氟中毒模型。对中缝背核神经元Ｍｇ２＋－腺苷三磷酸酶（Ｍｇ２＋－ＡＴＰａｓｅ）和硫胺素焦磷酸酶（ＴＰＰａｓｅ）进行了定性、定量分析。结果表明：氟中毒大鼠中缝背核神经元内Ｍｇ２＋－ＡＴＰａｓｅ、ＴＰＰａｓｅ染色均浅于对照组,定量分析Ｍｇ２＋－ＡＴＰａｓｅ、ＴＰＰａｓｅ含量均减少。电镜观察氟中毒大鼠神经元内ＴＰＰａｓｅ颗粒减少,同时出现超微结构改变。这些结果提示：高氟可抑制某些酶的活性,并对中枢神经系统有直接损害     

Electron microscopic investigation of synaptic organization of the trochlear nucleus in cat

Summary Two distinct types of neuron in the cat trochlear nucleus (one large, one small) are described, the - and -motoneurons, respectively. Four types of terminals are observed which establish axo-dendritic synapses. Two of them (Types I and II) perform axo-somatic synapses as well. Terminals en passant (Types I and II) are predominant. The Type I terminal is long and slender with a characteristic distribution of the axoplasmic organelles and the unique feature of a relative narrowing of the synaptic cleft as compared to the width of the neighboring extracellular space. Its vesicle population is pleomorphic and a conspicuous glial barrier surrounds the synaptic zones. The Type II terminal differs slightly from Type I, revealing a wider synaptic cleft and lacking a characteristic distribution of the axoplasmic organelles. The type III terminal is rarely observed performing axo-somatic synapses, but is a common finding in the neuropil. Post-junctional dense bodies are often present in its axodendritic synapses. The Type IV nerve terminal performs axo-dendritic synapses and is characterized by a rich content of large granulated vesicles. Axo-axonal synapses are observed only very rarely.The synaptic organization of the feline trochlear nucleus is compared with the synaptic morphology of the oculomotor nuclei of inframammalian species (Waxman and Pappas, 1971). In addition to certain similarities (e.g., richness of synapses en passant), significant differences are encountered: the present study provides no morphological evidence for electrotonic transmission in the trochlear nucleus of cat.Preliminary report made at the Annual German Jahresversammlung der Anatomischen Gesellschaft, Lausanne, Switzerland, 1973.The authors wish to thank Professor R. Hassler for his valuable discussion. — Dr. W. B. Choi is on leave from the Department of Anatomy, Catholic Medical School, Seoul, Korea.     

L-α-Aminoadipate Inhibits Kynurenate Synthesis in Rat Brain Hippocampus and Tissue Culture

Intracerebral administration of L--aminoadipic acid (L-AAA) at 500 mg/kg body weight to rats caused a complex behavioral change with sporadic wet-dog shakes. Animals developed severe limbic seizures between 1 and 6 h after L-AAA injection, characterized by generalized convulsions. Twenty days after L-AAA injection kynurenine aminotransferase (KAT) activity measured in hippocampal brain tissue slices prepared with a McIlwain chopper at 30 m showed a significant 43% decrease. Subcutaneous injection of kynurenine at 500 mg/kg showed a 63% increase in KAT activity twenty days later. This increase was offset by a concomitant administration of 500 mg/kg L-AAA stereotaxically on day one. In astrocyte culture kynurenic acid synthesis is inhibited by L-AAA and L-pipecolic acid. The possible involvement of kynurenic acid in the modulation of neuronal degeneration is discussed.     

Enzyme histochemistry of the mesenteric and dorsal root ganglion cells of cat and squirrel monkey

Summary A detailed histochemical study has been made on the mesenteric ganglia of the cat, and dorsal root ganglia of the squirrel monkey by the use of appropriate histochemical techniques accompanied by appropriate controls for phosphatases, esterases, and oxidative enzymes. The different neurons of a particular ganglion show varied amounts of enzyme activity at a particular time depending upon the functional state of the neurons. SDH, CYO and LDH reaction is prominent in the cytoplasm of the neurons, gliocytes and satellite cells, whereas the MAO preparations generally show a weak reaction. The AK is prominent in the neuropil, cell membranes and peripheral part of cytoplasm, whereas ATPase activity has been observed in blood vessels as well. In AC preparations the area of lipofuscin concentration shows more intense reaction than the rest of the cytoplasm. The activity of AChE and BChE varies from mild, to moderate to strong. The TPPase preparations show morphologically different types and amounts of TPPase positive Golgi material even in the adjoining cells. The relationship between the TPPase Golgi material and various oxidative and dephosphorylating enzymes has been briefly discussed.Abbreviations used AC Acid phosphatase - AChE Acetyl-cholinesterase (specific) - AK Alkaline phosphatase - AMPase Adenosine monophosphatase (5-nucleotidase) - ATPase Adenosine triphosphatase - BChE Butyryl-cholinesterase (nonspecific) - CYO Cytochrome oxidase - DPN-D DPN-diaphorase - G6P Glucose-6-phosphatase - LDH Lactic dehydrogenase - MAO Monoamine oxidase - MDH Malic dehydrogenase - NAD-D NAD-diaphorase - SDH Succinic dehydrogenase - SE Simple esterase - TPPase thiamine pyrophosphatase T. R. Shanthaveerappa in previous publications.     

Detection and cytoplasmic localization of two different microtubule motor proteins in basal epithelial cells of freshwater sponges

Summary Epithelial sponge cells (pinacocytes) contain a set of 50 to 60 microtubules radiating from the nuclear region to the cell periphery. Vacuoles of the endocytic pathway (endosomes, lysosomes) and mitochondria move along single microtubules in both directions; moreover, the ring-like arrangement of the Golgi apparatus around the nucleus and the net-like organization of the endoplasmic reticulum in the cytoplasmic matrix are also maintained, in an energy-dependent manner, by the microtubular system. Significant changes in the velocities of retrograde and anterograde transport as well as distinct differences in the sensitivity of organelle dynamics to ATPase inhibitors and ATP analogues indicate the existence of two microtubule-based motor proteins. Ion exchange chromatography of pinacocyte homogenates resulted in the enrichment of a 97 kDa kinesin-like protein (SKLP) with the ability to cross-react with antibodies against the kinesin heavy chain. Two other polypeptides, with molecular mass of 75 and 400 kDa, apparently belonging to a cytoplasmic dynein-like protein (SDLP) could be recognized in immunoblots with antibodies against the intermediate and heavy chains of cytoplasmic dynein. In addition, three MAP-like polypeptides (SMAPLPs), with molecular mass of 280, 250 and 70 kDa, obviously related to the MAP-2 and tau-family, have been identified. Immunocytochemical studies at the light and electron microscopical level localized SKLP, SDLP, and SMAPLPs at endocytic vacuoles and mitochondria, whereas the endoplasmic reticulum has SKLP and SMAPLPs, but the Golgi apparatus only SDLP.Abbreviations AMP-PNP 5-adenylylimidodiphosphate - ATP adenosinetriphosphate - DiOC₆ (3) 3,3-dihexyloxacarbocyanine iodide - DTT 1,4-dithiothreitol - EDTA ethylenediaminetetraacetic acid - EGTA ethylene glycol-bis(-aminoethyl ether) - EHNA erythro-9-(2-hydroxy-3-nonyl)-adenine - EM electron microscope - ER endoplasmic reticulum - FPLC fast protein liquid chromatography - GA Golgi apparatus - GTP guanosinetriphosphate - HC heavy chain - HSS high-speed supernatant - IC intermediate chain - LC light chain - MAP microtubule-associated protein - MT microtubule - PIPES 1,4-piperazine-N, N-bis-(2-ethanesulfonic) acid - PMSF phenylmethylsulfonyl fluoride - SDLP sponge dynein-like protein - SDS-PAGE sodium-dodecyl-sulfate polyacrylamide gel electrophoresis - SKLP sponge kinesin-like protein - SMAPLPs sponge MAP-like proteins - UTP uridinetriphosphate     

Histochemistry of implantation in the rabbit

Summary The distribution of glycogen, acid and neutral mucopolysaccharides, hydrolases alkaline phosphatases, and carbohydrate dehydrogenases is described in the rabbit blastocyst and its surroundings over the implantation period from 5 to 9 days.Glycogen is found mainly in the embryonic tissues, where peaks of concentration coincide with differentiation, some is also seen in the uterine epithelium and secretion, and large quantities accumulate in the developing decidua.Neutral mucopolysaccharide is found in the yolk-sac and embryonic endoderm, and as secretion droplets in the uterine epithelium and secretion.Acid mucopolysaccharide occurs in the embryonic coverings and uterine secretion.RNA is associated in the embryo with developing tissues, and accumulates in the developing decidual cells.Hydrolases (acid phosphatase, and B esterase) increase their activity in the trophoblast knobs, in developing syncytiotrophoblast, and in the embryonic endoderm. Degeneration of the uterine epithelium is associated with maximal hydrolase activity.Trophoblastic alkaline phosphatase activity (non-specific and specific) decreases from 5 to 7 days of gestation, then increases markedly in the developing syncytiotrophoblast. AMPase appears in the embryonic mesoderm. In the uterine epithelium intense brush border staining is seen, and TPPase and UDPase become visible for a short period in the Golgi region. Phosphatases increase their activity in the decidua to 8 days and then decrease.Carbohydrate dehydrogenases (except -glycero-phosphate and -hydroxy-butyrate dehydrogenases) increase their activity in embryonic tissues, particularly in the developing syncytiotrophoblast and endoderm. Symplasma formation in the uterine epithelium is also associated with increase in enzyme activity, and a similar increase, up to 8 days of gestation, is seen in the decidua with isocitrate, malate, glucose-6-phosphate, lactate, succinate, and furfuryl alcohol dehydrogenases.Some correlation is found between the histochemical findings and the phenomena of epithelial removal, uterine secretion, decidual formation and function, giant cell function, morphogenesis, and histiotrophic nutrition, and the results are compared with previous findings for the rat in which implantation is morphologically, and probably physiologically a very different process.     

CRF-containing neurons of the rat hypothalamus

Summary The immunoreactive CRF-neurons of the rat hypothalamus have been examined immunohistochemically employing anti-rat CRF serum. These neurons are confined to the paraventricular nucleus, dorsomedial-lateral hypothalamic area, and suprachiasmatic nucleus, and are, respectively, also immunoreactive to anti-Met-enk, -alpha-MSH, and -VIP sera. Intraventricular administration of colchicine (50 g/5 l/rat) induces a dramatic enhancement of the immunostainability of the cell somata, and also accelerates the development of immunoreactivity of other stored peptides, especially in the paraventricular nucleus.The CRF-neurons respond to adrenalectomy by showing increased immunoreactivity and an increase in the number of cell bodies; in the dorsomedial-lateral area and suprachiasmatic nucleus, there is also an enhanced immunoreactivity for alpha-MSH and VIP, respectively. CRF-cells in the paraventricular nucleus become markedly hypertrophied, but do not show any enhanced immunoreactivity for Met-enk. Since the axons of the paraventricular neurons run to the median eminence, it is probable that they are involved with the endocrine control of hypophysial ACTH release. It is concluded that the CRF-containing neurons in rat hypothalamus consist of three types which are functionally and morphologically different.     

A whole range of fine structural criteria for immunohistochemically identified LH cells in rats

Summary Pituitaries from normal, young and adult male rats were fixed either in sublimate-formalin or in glutaraldehyde-osmium. In adjacent Paraplast sections, almost all the gonadotrophs were immunostained with both LH and FSH antisera. The rat LH and FSH antisera used were shown to be highly specific by the absorption test and by double antibody radioimmunoassay. Thin and thick adjacent Epon sections were prepared for EM and immunohistochemical examination. Cells stained with the rat LH antiserum were identified by LM, and then observed in detail by EM. On the basis of these observations we suggest that the LH cells are arranged in a sequence of basophils, i.e., Types II/III, III, III/IV and IV: Type II/III basophils are elongate with a cytoplasmic process and less vesiculated. They have morphological features of Type II (classical thyrotrophs) and also of Type III basophils. Type III basophils are oval in shape and moderately vesiculated. Both Types II/III and III basophils can be divided into two classes of cell characterized mainly by the existence of only small secretory granules (150–220 nm in diameter) (Type A) or by the coexistence of small and large (350–500 nm) (Type B). Type III/IV basophils are cells intermediate between types III and IV basophils, and moderately vesiculated with an abundance of secretory granules (150–300 nm in diameter). Type IV basophils are large, spherical or oval cells whose RER cisternae are conspicuously dilated; they contain less numerous secretory granules (150–300 nm in diameter). It is concluded that LH cells are not a single cell type, but include a wide range of subtypes.     

Isolation,characterization, and numerical taxonomy of Simonsiella strains from the oral cavities of cats,dogs, sheep,and humans

Forty-nine strains of the gliding prokaryote Simonsiella were isolated from the oral cavities of cats (8), dogs (19), sheep (4), and humans (18) in Southern California by a direct isolation procedure using a complex serum-enriched medium. The numerical taxonomic analysis (unweighted pair-group method using arithmetic averages) of 57 differential traits for each strain was based on standard bacteriological diagnostic tests and included the molar guanine-pluscytosine contents of the DNA and the relative percentages of fatty acid contents reported earlier. The resulting phenogram clustered the strains of Simonsiella into groups that correlated with sources of origin. The study included the neotype strain of Simonsiella crassa (ATCC 27504, ICPB 3651, NCTC 10283) of Australian sheep origin. The strains isolated from dogs, sheep, and humans form clusters of organisms that appear to have become adapted to live in and possibly to have evolved with their respective hosts. In our judgment, these source-of-origin clusters represent different ecospecies.List of Abbreviations ATCC American Type Culture Collection - BSTSY bovine serum-tryptic soy-yeast extract - DNA deoxyribonucleic acid - ECL equivalent chain length of carbon atoms - ICPB International Collection of Phytopathogenic Bacteria - NCTC National Collection of Type Cultures To whom offprint requests should be sent.     

Inhibition of the L-dopa transport system in human epidermal Langerhans cells by omeprazole and its analogues

Summary. L-3,4-dihydroxyphenylalanine (L-dopa) transport into human Langerhans cells (LC) occurs by a saturable mediation. This plasma membrane agency is, due to its characteristics, distinguishable from systems transporting other neutral, cationic and anionic amino acids into other cells and serves to catalyze the flow of L-dopa, only, into LC. The uphill operation of this L-dopa transport system is believed to occur by down-gradient countermigration of H⁺. Due to the uniqueness of the L-dopa transport system, the widely used analogue inhibition approach was not applicable. Instead we studied omeprazole and its analogues in our search for suitable inhibitory candidates. Omeprazole and most of its analogues were indeed inhibitory in the concentration range 1–100mol/L. Conspicuously, the compounds with strongest polarity were least inhibitory. The inhibitory pattern displayed by omeprazole and the other analogues on L-dopa uptake in LC corresponded to some extent to what has been observed previously for purified H⁺,K⁺-ATPase from tubulovesicles of the stomach. No effects of the inhibitors were registered on energy charge and lactate production of epidermal biopsies, nor were any gross alterations of ultrastructure of LC noticed.     

Effects of extracellular osmolality on renin release and on the ultrastructure of the juxtaglomerular epithelioid cell granules

Summary Superfusion with hypoosmotic solutions stimulates renin release from rat epithelioid cells adherent to isolated glomeruli. This stimulatory effect may be related to the observed swelling of the secretory granules; the swelling may markedly increase the probability of pre-exocytotic fusions between the granule and cell membranes, and consequently increase the frequency of exocytotic events.These studies were supported by the Danish Medical Research Council King Christian X's Foundation and the German Research Foundation within the Forschergruppe Niere/Heidelberg     

Thiamine pyrophosphatase activity in secretory cells of the lateral prostate and seminal vesicle of normal and castrated guinea pigs and castrates treated with oestradiol

Summary Ultrastructural localization of thiamine pyrophosphatase (TPPase) activity was studied in secretory cells of the lateral prostate and seminal vesicle of normal and castrated guinea pigs and castrates treated with 17-oestradiol benzoate. The present study has demonstrated that TPPase reaction product is consistently localized in the three to four trans cisternae of Golgi complexes in both the lateral prostate and the seminal vesicle. The reaction was intense and the reaction product often filled the cisternae completely.After castration there was a decrease in TPPase activity in both glands as revealed by the reduction in the amount of the reaction product which was found mainly in one to two trans cisternae of the regressed Golgi complex. The reaction product changed from a dense to a more particulate or granular pattern or to discrete deposits of high electron-density.Administration of 17-oestradiol benzoate to the castrates caused changes in the localization and patterns of distribution of TPPase. In the lateral prostate there was an apparent increase in TPPase activity. The reaction product was found in two to four trans cisternae and occasionally in the trans-most cisternae of the dilated Golgi complex. The reaction product appeared as discrete, dense coarse precipitates. In the seminal vesicle TPPase reaction product was consistently found in one to two trans cisternae in cells with larger Golgi complexes. However, almost all cisternae of the smaller Golgi complexes were TPPase-positive. The cytochemicl results of the present study suggest that TPPase activity and possibly the process of glycosylation in secretory cells of the lateral prostate and seminal vesicle may have been affected after castration and after oestradiol administration.     

Immunocytochemical localization of GABA containing neurons in the ventrolateral medulla oblongata of the rat

Summary Localization of -aminobutyric acid (GABA) in the ventrolateral medulla oblongata of the rat was studied, using antisera directed against GABA molecule fixed to bovine serum albumin. Within the rostral portion of the ventrolateral medulla, GABA-like immunoreactive neurons were found in the lateral wing of the raphe magnus and in the region of the paragigantocellular reticular nucleus. In the caudal portion of the ventrolateral medulla, a lesser number of GABA-stained neurons were found in the region around the nucleus reticularis lateralis. GABA-like immunoreactive punctate structures were also found throughout the ventrolateral medulla. These results provide further evidence for the existence of GABAergic neurons in the ventrolateral medulla oblongata of the rat.     

Contractile effects of vanadate on aorta rings from virgin and pregnant rats

The present study was undertaken to characterize the contractile effects of vanadate on thoracic aorta rings from virgin and term-pregnant rats. Vanadate caused concentration-dependent contraction in rat aortic rings with an EC₅₀ (concentration producing 50% maximum response) of 0.10 mM. Contractions in response to vanadate were equivalent to the ones measured with 1 M phenylephrine. The effects of vanadate were not affected by indomethacin (up to 10 M), an inhibitor of prostanoid cyclooxygenase, but were blocked in a concentration-dependent manner by staurosporine (0.1–1.0 M), an inhibitor of protein kinase C. Vanadate exhibited a significant decrease of contractile responses in aorta of pregnant as compared to virgin rats. When aortic rings were bathed in presence of different concentrations of vanadate, the concentration-response curve to phenylephrine was shifted to the left, but maximum response was not affected. The potentiation of the contractions to phenylephrine by vanadate was significantly more prominent in aorta of virgin than of pregnant rats. These results suggest that the contractile effect of vanadate on rat aorta is independent of endogenous prostanoids and may be mediated by protein kinase C-dependent pathway. These results also show that the contractile response to vanadate on the rat aorta is impaired during pregnancy.     

Characterization of the la antigens involved in suppressor T cell generation in the rat

The WRC rat, an intra-class II recombinant strain (RT1.B ⁿ B ^a D _, ^a ), was used to study the relative roles of the two class II loci in mixed lymphocyte reaction (MLR) proliferation and suppressor T cell (Ts) generation. Both MLR proliferation and Ts generation were noted in cultures of WRC with DA (RTI ^{a) stimulator cells. In contrast, cultures of WRC with BN (RT1sun} ) stimulator cells proliferate but do not generate significant amounts of Ts. The data suggest that RT1.B incompatibility is important in the generation of Ts in the WRC rat. Suppressor cells generated in cultures of WF (RT1 ^u ) with WRC stimulator cells potently suppressed a WF+WRC_x test MLR, with less suppression when tested against either the WF+DA_x or WF+BN_x MLRs. The latter experiments suggest that Ts clones may be produced to either class II subregion, and therefore that MLR proliferation and Ts induction are not necessarily linked, but vary with particular genotypes. The current lack of other rat intra-class II recombinant strains precludes assignment of suppressor induction/activation to a single locus.     

Observations on the ultrastructure of nerve cells in the brain of the earthworm,Eisenia foetida,with special reference to neurosecretion

Summary The submicroscopic structure of nerve cells in the brain of the earthworm Eisenia was studied. Six types of neurons containing morphologically different inclusions are identified. Types 1, 2 and 3 contain vesicles filled with homogeneous materials of high electron density. These are essentially similar to elementary granules in neurosecretory systems of vertebrates and invertebrates. Type 4 shows dense-cored vesicles which resemble in size catechol-containing granules as described, for example, in chromaffin cells of the adrenal gland. Type 5 has clear vesicles with a mean diameter of 400 Å. Some of these vesicles have a dense osmium deposit. Type 6 contains electron lucent vesicles with diameters of 500–800 Å. Occasionally these have osmiophilic cores. Clear vesicles of types 5 and 6 are similar to classical synaptic vesicles, while granulated vesicles resemble in size and appearance those described in adrenergic nerve endings. All six vesicle types have the same mode of origin from Golgi membranes. All of these vesicles are considered to be discharged from the perikarya into the axons entering the neuropil.     

Beta-1,3-glucanase from unfertilized eggs of the sea urchin Strongylocentrotus intermedius. Comparison with beta-1,3-glucanases of marine and terrestrial mollusks

-1,3-Glucanase (Lu) was isolated from unfertilized eggs of the sea urchin Strongylocentrotus intermedius. A comparative study of some properties of -1,3-glucanase Lu and -1,3-glucanases with different action types—endo--1,3-glucanase from crystalline style of the marine mollusk Spisula sachalinensis (LIV) and exo--1,3-glucanase from the terrestrial snail Eulota maakii (LII)—was performed. It was found that -1,3-glucanase Lu hydrolyzes laminaran with a high yield of glucose in the reaction products. The enzyme hydrolyzes substrates with retention of the glycosidic bond configuration, is able to cleave modified substrates, and exhibits transglycosylating activity. All properties of -1,3-glucanase from S. intermedius were more similar to those of the endo--1,3-glucanase from the marine mollusk (LIV) than exo--1,3-glucanase LII from the terrestrial snail. The differences in the effect of LIV and Lu on laminaran are probably related to the functions of -1,3-glucanase Lu from sea urchin eggs (which, in contrast to LIV, is not a digestive enzyme).     

<Emphasis Type="Italic">In vitro</Emphasis> regeneration of <Emphasis Type="Italic">Perilla frutescens</Emphasis> from hypocotyl and cotyledon explants

Organogenetic buds were induced from hypocotyl and cotyledon explants of oil crop Perilla frutescens in Murashige and Skoog (MS) medium supplemented with 5.7 M indole-3-acetic acid (IAA) and 8.9 – 13.3 M 6-benzylaminopurine (BA). Shoots were rooted on MS medium with 2.9 M IAA and 1.4 M gibberellic acid (GA3) and the regenerated plants flowered and set seeds normally.