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1.
Hui Zeng Haihui Ye Shaojing Li Guizhong Wang Jingru Huang 《In vitro cellular & developmental biology. Animal》2010,46(5):431-437
Hepatopancreas is an important digestive and endocrine organ in crustacean. However, there are few reports on cell cultures
from crabs. Here, the cell cultures of hepatopancreas from Scylla paramamosain was studied in vitro. Both the primary cell culture and subculture were grown in Leibovitz’ L-15 medium, M199 medium, or
a specially designed medium for S. paramamosain (MSP). The results showed that hepatopancreas cells in vitro grew in compact clusters in 2–3 d. Four types of cells could
be identified. They were embryo cells, fibrillar cells, resorptive cells, and blister-like cells, respectively. Some of these
cells could be subcultured for three generations. The MSP supported the best survival of these hepatopancreas cells, while
M199 medium was the least effective of these three media. Fetal bovine serum and crab muscle extracts as supplements stimulated
growth, but the crab hemolymph inhibited cell growth. Taken together, MSP is an appropriate medium for hepatopancreas cell
cultures from S. paramamosain and can support cultures through several passages. 相似文献
2.
Haihui Ye Ping Song Jun Ma Huiyang Huang Guizhong Wang 《Marine and Freshwater Behaviour and Physiology》2013,45(1):25-35
Vertebrate-type steroids, such as progesterone, have been identified in crustaceans. The physiological activity of progesterone during vitellogenesis is still not well understood. In this study, progesterone levels in the female mud crab, Scylla paramamosain, were determined by enzyme-linked immunosorbent assay. Peak levels of progesterone were detected during the previtellogenic stage in the hemolymph, ovary, and hepatopancreas, whereas the progesterone level decreased significantly in vitellogenic stage I. During vitellogenic stage II, progesterone levels rose again in the hemolymph and ovary, but continued to decrease in the hepatopancreas. By using western blotting, progesterone receptor (PR), with an apparent molecular weight of 70 kDa, was identified in the ovary during both vitellogenic stages I and II. By means of immunohistochemistry, PR was detected mainly in the follicle cells during vitellogenic stage I and in the nuclei of oocytes in vitellogenic stage II. Our results strongly suggest that progesterone promotes vitellogenesis in the mud crab, S. paramamosain via a classical genomic mechanism. 相似文献
3.
Arun M. Khurad Min-Juan Zhang Chanchal G. Deshmukh Ravindra S. Bahekar Ashish D. Tiple Chuan-Xi Zhang 《In vitro cellular & developmental biology. Animal》2009,45(8):414-419
A new continuous cell line from ovarian tissue of commercial variety “Kolar Gold” of silkworm, Bombyx mori, was established and designated as DZNU-Bm-12. The tissue was grown in MGM-448 insect cell culture medium supplemented with
10% fetal bovine serum (FBS) and 3% heat-inactivated B. mori hemolymph at 25 ± 1°C. The migration of partially attached small round refractive cells from the fragments of ovarioles began
from the beginning of explantation. The cells multiplied partially attached in the primary culture initially, and some of
them become freely suspended after 20 passages. The cells were adapted to MGM-448 and TNM-FH media each with 10% FBS and the
population doubling time of cell line was about 36 and 24 hr, respectively. The chromosome number was near diploid at initial
passages and slightly increased at 176th passage, but a few tetraploids and hexaploids were also observed. DNA profiles using
simple sequence repeat loci established the differences between DZNU-Bm-12 and DZNU-Bm-1 and most widely used Bm-5 and BmN
cell lines. The cell line was found susceptible to B. mori nucleopolyhedrovirus (BmNPV) with 85–90% of the cells harboring BmNPV and having an average of 3–17 OBs/infected cell. We
suggest the usefulness of this cell line in BmNPV-based baculoviral expression system and also for studying in vitro virus
replication. 相似文献
4.
Huiyang Huang Shizhao Han Guizhong Wang 《Marine and Freshwater Behaviour and Physiology》2013,45(4):297-305
To elucidate the role of gonadotropins-like substances in mud crab Scylla paramamosain, hemolymph samples were measured for concentrations of follicle stimulating hormone (FSH), luteinizing hormone (LH) and steroid hormones by enzyme-linked immunosorbent assay (ELISA). Hormonal concentration data were analyzed in association with the stages of gonadal development. ELISA has shown that in the female crab, the level of FSH reaches its peak in the early stage of ovary development, while estradiol and LH peaked during the late maturing stage of the ovary. In the male crab, testosterone and FSH culminated during the spermatid stage, and the level of LH peaked during the sperm stage. These results indicated that substances resembling the vertebrate FSH and LH are present in the hemolymph of S. paramamosain, and they may be involved in the development of the gonad. 相似文献
5.
Singh AK Chaudhary P Macwan AS Diwedi UN Kumar A 《Applied microbiology and biotechnology》2007,76(4):895-901
The chlorinated insecticide γ-hexachlorocyclohexane (γ-HCH) is sequentially metabolized by the products of linA, linB, linC, linD, linE, and linF genes to β-ketoadipate, which is subsequently mineralized. Two or more copies of these genes are present in the bacterium
Pseudomonas aeruginosa ITRC-5 that was isolated earlier by selective enrichment on technical-HCH. At least one copy of linA, linB, linC, linD, and possibly linE is lost from ITRC-5 upon its growth on γ-HCH. All the lin genes, however, are lost when the bacterium was grown in Luria–Bertani (LB) medium. The loss of lin genes is accompanied with the loss/rearrangement of insertion sequence IS6100 genes. Concomitant to the loss of lin genes, the degradation of HCH-isomers by “γ-HCH grown cells” is slower, when compared with “technical-HCH grown cells”, and
is completely lost by “LB-grown cells”. The selective loss of lin genes during different growth conditions has not been reported before and is expected to help in understanding the dynamism
of degradative genes. 相似文献
6.
Dermal matrix proteins initiate re-epithelialization but are not sufficient for coordinated epidermal outgrowth in a new fish skin culture model 总被引:1,自引:0,他引:1
We have established a new culture system to study re-epithelialization during fish epidermal wound healing. In this culture
system, fetal bovine serum (FBS) stimulates the epidermal outgrowth of multi-cellular layers from scale skin mounted on a
coverslip, even when cell proliferation is blocked. The rate of outgrowth is about 0.4 mm/h, and at 3 h after incubation,
the area occupied by the epidermal sheet is nine times larger than the area of the original scale skin. Cells at the bottom
of the outgrowth show a migratory phenotype with lamellipodia, and “purse string”-like actin bundles have been found over
the leading-edge cells with polarized lamellipodia. In the superficial cells, re-development of adherens junctions and microridges
has been detected, together with the appearance and translocation of phosphorylated p38 MAPK into nuclear areas. Thus, this
culture system provides an excellent model to study the mechanisms of epidermal outgrowth accompanied by migration and re-differentiation.
We have also examined the role of extracellular matrix proteins in the outgrowth. Type I collagen or fibronectin stimulates
moderate outgrowth in the absence of FBS, but development of microridges and the distribution of phosphorylated p38 MAPK are
attenuated in the superficial cells. In addition, the leading-edge cells do not have apparent “purse string”-like actin bundles.
The outgrowth stimulated by FBS is inhibited by laminin. These results suggest that dermal substrates such as type I collagen
and fibronectin are able to initiate epidermal outgrowth but require other factors to enhance such outgrowth, together with
coordinated alterations in cellular phenotype.
This work was supported by grants from the Faculty of Science at Toho University and from Fujirebio to M.S. 相似文献
7.
Weizhe Li Yufeng Pan Zhipeng Wang Haiyun Gong Zhefeng Gong Li Liu 《Cell and tissue research》2009,336(3):509-519
The fan-shaped body is the largest substructure of the central complex in Drosophila melanogaster. Two groups of large-field neurons that innervate the fan-shaped body, viz., F1 and F5 neurons, have recently been found
to be involved in visual pattern memory for “contour orientation” and “elevation” in a rut-dependent manner. The F5 neurons have been found to be responsible for the parameter “elevation” in a for-dependent manner. We have shown here that the F1 neuron also affects visual memory for “contour orientation” in a for-dependent way. With the help of Gal4/UAS and FLP-out techniques, we have characterized the morphological features of these
two groups of neurons at single neuron resolution. We have observed that F1 or F5 neurons are groups of isomorphic individual
neurons. Single F1 neurons have three main arborization regions: one in the first layer of the fan-shaped body, one in the
ventral body, and another in the inferior medial protocerebrum. Single F5 neurons have two arborization regions: one in the
fifth layer of the fan-shaped body and the other in the superior medial protocerebrum. The polarity of the F1 and F5 neurons
has been studied with the Syt-GFP marker. Our results indicate the existence of presynaptic sites of both F1 and F5 neurons
located in the fan-shaped body and postsynaptic sites outside of the fan-shaped body.
This work was supported by the “973 Program” (2005CB522804 and 2009CB918702), the National Natural Sciences Foundation of
China (30621004, 30625022, and 30770682), and the Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-YW-R-28). 相似文献
8.
A. J. Grant R. J. O'Connell 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》2000,186(6):535-542
We recorded the electrophysiological responses of individual olfactory receptor neurons in sensilla trichodea on the antennae of adult arctiid moths, Utetheisa ornatrix, to stimulation with volatiles associated with both sexes. All trichoid sensilla contain at least two receptor neurons, each
with distinct action potential amplitudes and waveforms, that respond dichotomously to male and female odors. Although, neither
female neuron responds to extracts of coremata or the male-produced pheromone hydroxydanaidal, they do respond in a gender-specific
manner to the volatiles emanating from whole pupae, hemolymph, thoracic froth, and adult animals of several ages. Thoracic
froth, which contains pyrrolizidine alkaloids, is thought to play a role in defense. Froth from moths reared on diets, with
or without added pyrrolizidine alkaloids, were equally effective in eliciting gender-specific patterns of response. Male trichoid
receptor neurons respond to these same materials with similar patterns of activation. These receptor neurons provide information
about substances, which we have termed “gender odors,” that are persistently emitted by nearby animals. These substances do
not appear to be the same as those already known to be involved in defense or the sexual dialog between individuals.
Accepted: 16 March 2000 相似文献
9.
A cell line designated as ADG was established from an abalone digestive gland using ERDF medium supplemented with 8% fetal
bovine serum (FBS), 8% abalone hemolymph, and high concentrations of NaCl, KCl, MgCl2, MgSO4, and CaCl2. ADG cells proliferated better in protein-free medium than in FBS-supplemented medium. Among 9 kinds of media examined, ERDF
medium was shown to be optimal for cell growth. ADG cells secreted 13 different kinds of glycosidases in protein-free medium:
α-L-fucosidase, β-L-fucosidase, α-D-galactosidase, β-D-galactosidase, N-acetyl-α-D-galactosaminidase, N-acetyl-β-D-galactosaminidase,
α-D-glucosidase, β-D-glucosidase, N-acetyl-α-D-glucosaminidase, N-acetyl-β-D-glucosaminidase, α-D-mannosidase, β-D-mannosidase,
β-D-xylosidase, and 1-3 xylanase. When ADG cells were cultured in Grace’s insect cell medium, the activity of some secreted
glycosidases increased 25-fold to 65-fold per cell as compared with control cells cultured in ERDF medium. ADG - abalone digestive
gland; ERDF - enriched RDF; FBS - fetal bovine serum; L-15 - Leibovitz’s L-15 media; DME - Dulbecco’s modified Eagle medium;
F-12 - nutrient mixture (Ham); LDF - L-15; DME: F-12 = 10 : 7 : 3; MEM - minimum essential medium; RPMI - RPMI medium 1640;
199 - media 199; GIC - Grace’s insect cell medium; pNP -p -nitrophenol.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
10.
Arterial cells and CNS sheath cells from Aplysia californica produce factors that enhance neurite outgrowth in co-cultured neurons 总被引:2,自引:0,他引:2
Substrate-bound and soluble factors regulate neurite outgrowth and synapse formation during development, regeneration, and
learning and memory. We report that sheath cells from CNS connectives and arterial cells from the anterior aorta of the sea
slug, Aplysia californica, enhance neurite outgrowth from co-cultured Aplysia neurons. Sheath and arterial cell cultures contain several cell types, including fibrocytes, myocytes, and amoebocytes. When
compared to controls (neurons with defined growth medium alone), the percentage of neurons with growth and the average neurite
lengths are significantly enhanced by sheath and arterial cells at 48 h after plating of the neurons; these parameters are
comparable to those of neurons cultured in medium containing hemolymph. Our results indicate that sheath cells produce substrate-bound
factor(s) and arterial cells produce diffusible factor(s) that promote growth. These growth factors likely promote neuron
survival and neurite outgrowth during neural plasticity exhibited in the adult CNS.
Electronic Publication 相似文献
11.
Imjongjirak C Amparyup P Tassanakajon A Sittipraneed S 《Molecular biology reports》2009,36(5):841-850
Antimicrobial peptides (AMPs) are important components of the host innate immune response against microbial invasion. In the
present study, we report the identification and characterization of a crustin (CrusSp) from the hemocyte of mud crab, Scylla paramamosain using an expressed sequence tag (EST) and rapid amplification cDNA end (RACE) approaches. Analysis of the nucleotide sequence
revealed seven different variances of the CrusSp cDNA in mud crab. The open reading frame encodes a protein of 111 amino acids with 21 residues signal sequence. The predicted
molecular mass of the mature protein (90 amino acids) is 10.27 kDa with an estimated pI of 8.54. Analysis of the protein domain features indicated typical conserved cysteine residues containing a single whey acidic
protein (WAP) domain at the C-terminus. A neighbour-joining tree showed that S. paramamosain crustin is closely related to other crustin homologues, and displays the highest similarity to crustin antimicrobial peptide
in shore crab Carcinus maenas. Four exons and three introns were identified within the 999 bp genomic DNA sequence of CrusSp. Tissue distribution analysis showed that CrusSp was highly expressed in hemocytes, gills, intestines and muscle but it was not expressed in hepatopancreas and eyestalks.
To gain insight into the in vitro antimicrobial activities of CrusSp, the mature peptide coding region was cloned into E. coli for heterologous expression. The recombinant CrusSp could inhibit the growth of gram-positive bacteria but had no inhibition activity against gram-negative bacteria. These results
indicated the involvement of CrusSp in the innate immunity of S. paramamosain. 相似文献
12.
M. Ghirardi A. Casadio L. Santarelli P. G. Montarolo 《Invertebrate neuroscience : IN》1996,2(1):41-49
Hemolymph of adultAplysia californica significantly affects neurite outgrowth of identified neurons of the land snailHelix pomatia. The metacerebral giant cell (MGC) and the motoneuron C3 from the cerebral ganglion and the neuron B2 from the buccal ganglion
ofH. pomatia were isolated by enzymatic and mechanical dissociation and plated onto poly-l-lysine-coated dishes either containing culture medium conditioned byHelix ganglia, or pre-treated withAplysia hemolymph. To determine the extent of neuronal growth we measured the neurite elongation and the neuritic field of cultured
neurons at different time points.Aplysia hemolymph enhances the extent and rate of linear outgrowth and the branching domain ofHelix neurons. With the hemolymph treatment the MGC neuron more consistently forms specific chemical synapses with its follower
cell B2, and these connections are more effective than those established in the presence of the conditioned medium. 相似文献
13.
The mud crab Scylla paramamosain plays a significant role in fishery resources in China. In this study, we developed 18 polymorphic microsatellite markers
in this important crab by 5′ anchored PCR technique. A total of 125 alleles were detected in a single population of 32 individuals
of S. paramamosain. The number of alleles per locus ranged from five to nine, with the allele size ranging from 166 to 316 bp. The polymorphism
information content (PIC), observed and expected heterozygosity ranged from 0.39 to 0.88, from 0.33 to 0.92 and from 0.42
to 0.86, respectively. Three loci (Scypa13, Scypa14 and Scypa15) deviated significantly from Hardy–Weinberg equilibrium (HWE)
after Bonferroni correction (P < 0.0028), and no linkage disequilibrium was found between loci pairs. These polymorphic microsatellite markers will be useful
for the study of population genetic structure, construction of genetic linkage maps and mapping of economically quantitative
trait loci (QTL) in S. paramamosain. 相似文献
14.
W. -H. Tsai G. L. Holbrook C. Schal A. -S. Chiang 《In vitro cellular & developmental biology. Animal》1995,31(7):542-546
Summary An in vitro organ culture system was established to support growth of corpora allata from the cockroachDiploptera punctata. During a 1-wk incubation in L-15B medium supplemented with 10% fetal bovine serum (FBS) and 10% cockroach hemolymph, adult
male corpora allata exhibited a cycle of de novo DNA synthesis followed by cell division. The number of S-phase cells and
metaphase cells per corpus allatum were counted from whole-mount monolayers after labeling in vitro with 5′-bromo-2′-deoxyuridine
and exposure to colchicine, respectively. While both FBS and cockroach hemolymph were essential for proliferation of allatal
cells, the growth-promoting effect of insect hemolymph was not species-specific and adult female hemolymph was more potent
than hemolymph from adult males. Furthermore, DNA synthesis of corpus allatum cells was stimulated in vitro by 20-hydroxyecdysone.
This sensitive assay system will be of immense utility in the search for allatal growth factors. 相似文献
15.
Anatol Rapoport 《Bulletin of mathematical biology》1950,12(4):327-337
Input-output formulas are derived for a neuron upon which converge single axones of two other neurons, which are subjected
to a Poisson shower, where a number of different assumptions are made concerning the mechanism of inhibition.
In one assumption so-called “bilateral pre-inhibition” is considered. That is to say, both neuronsN
1 andN
2 may exciteN
3, but if the stimulus of one of them follows within a certain interval σ of the other, the second stimulus is not effective.
This model is essentially no different from that involving two excitatory neurons acting upon a neuron having a refractory
period.
Another mechanism considered involves so-called “pre-and-post” inhibition, in which if two stimuli fromN
1 andN
2 fall within σ,both are ineffective. This case being mathematically much more involved than the preceding, an approximation method is used for
deriving the input-output formula.
Previous papers of this series are denoted by I, II, and III in this paper. 相似文献
16.
The existence of progesterone receptors (PR) in the Scylla paramamosain (mud crab) was studied using immunological techniques. By Western blotting, PR with an apparent molecular weight of 70 kDa
is identified in both the brain and the thoracic ganglion. By immunohistochemistry, PR immunoreactive neurons are detected
mainly in the protocerebrum, the subesophageal ganglion and the leg ganglion. PR immunoreactivity is localized mainly in the
nuclei of these neurons, while only a few neurons show such activities in their cytoplasm. Our results provide evidence that
progesterone modulates the neuroendocrine system mainly via nucleus receptors. 相似文献
17.
The goal of this paper is to propose a model of the hippocampal system that reconciles the presence of neurons that look
like “place cells” with the implication of the hippocampus (Hs) in other cognitive tasks (e.g., complex conditioning acquisition
and memory tasks). In the proposed model, “place cells” or “view cells” are learned in the perirhinal and entorhinal cortex.
The role of the Hs is not fundamentally dedicated to navigation or map building, the Hs is used to learn, store, and predict
transitions between multimodal states. This transition prediction mechanism could be important for novelty detection but,
above all, it is crucial to merge planning and sensory–motor functions in a single and coherent system. A neural architecture
embedding this model has been successfully tested on an autonomous robot, during navigation and planning in an open environment.
Received: 28 June 1999 / Accepted in revised form: 26 April 2001 相似文献
18.
R. Solomonoff 《Bulletin of mathematical biology》1952,14(2):153-157
The problem of finding the “weak connectivity” of a random net is reduced to one involving a Markov process. This provides
a mathematically exact treatment of the problem which had previously been treated by an approximation, whose justification
was not rigorous. The exact method allows in principle not only the calculation of the “weak connectivity”, but also of the
“strong connectivity”, and, in general, the probability that from a randomly selected neuron in the net there exist paths
to a specified number of neurons. The computations become exceedingly involved for large nets. 相似文献
19.
Organization of the receptive fields (RFs) of neurons of the extrastriate associative region 21b of the cerebral cortex was
studied in cats. Most neurons under study (63%) were “monocular,” while 37% of the cells were “binocular” units. Among 178
neurons examined in detail, heterogeneous RF functional organization was typical of about 76% of the units; point-to-point
testing of the entire RF area by stationary stimuli resulted in the generation of various types of responses (on, off, or on-off). The rest of the neurons (24%) generated homogeneous responses. The dimension, form, and functional organization of RFs
of the neurons under study depended to a certain extent on the parameters of visual stimuli used for the measurements. Examination
of the influence of the visual space, which surrounded the RF, on responses of the neurons evoked by stimulation of the RF
per se showed that darkening of the visual space adjacent to the RF inhibited neuronal responses to moving stimuli; in some cases
the responses were totally suppressed. Analysis of spatial overlapping of the RF sequentially recorded in the course of each
insertion of the electrode showed that the density of distribution of the overlapping RF areas of neighboring neurons with
the RF of the examined neuron is irregular, and that the RF is of a mosaic nature. We hypothesize that the visual space surrounding
the RF plays a significant role in the formation of responses of visually sensitive neurons to presentation of moving stimuli.
Neirofiziologiya/Neurophysiology, Vol. 37, No. 3, pp. 223–234, May–June, 2005. 相似文献
20.
Sandra I. Pitta-Alvarez Fabricio Medina-Bolivar María A. Alvarez Agustina A. Scambatto Patricia L. Marconi 《In vitro cellular & developmental biology. Plant》2008,44(6):502-507
Berberis buxifolia Lam., known as “Calafate”, is a plant native to Argentina that exhibits antimicrobial activity. This biological activity
is attributed to the isoquinoline alkaloid berberine. The aim of this research was to test the antimicrobial properties of
different extracts of this species, taking berberine as the reference molecule, and to examine if the expression of bacterial
multidrug resistance (MDR) efflux pumps could be responsible for possible resistance mechanisms. To this end, a wild-type
and a mutant strain of Staphylococcus aureus with a defective MDR efflux pump were used and the minimum inhibitory concentrations of the extracts were determined. The
studies were carried out with infusions of in vivo shoots and “Calafate” commercial tea, as well as with the media derived from shoot cultures incubated with different plant
growth regulators (thidiazuron, picloram, and jasmonic acid). As far as antimicrobial activity is concerned, all the extracts
tested were significantly more effective than berberine standard. “Calafate” commercial tea and shoot tea had inhibitory concentrations
similar to the one observed for ampicillin standard. The media from the shoot cultures, however, were significantly more effective
than all the others, particularly the one derived from jasmonic acid, suggesting the presence of compounds that could be acting
synergistically with berberine. There were no differences in antimicrobial activity against the wild-type and the mutant S. aureus; no definite conclusions could be drawn concerning the relationship between MDR pumps and possible pathogen resistance to
extracts of B. buxifolia. 相似文献