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Z Gong

Publications and source records attributed to Z Gong.

At least 109 records · Page 6Linked to original sources

Several splicing variants of isl-1 like genes in the chinook salmon (Oncorhynchus tschawytscha) encode truncated transcription factors containing a complete LIM domain.

Several novel cDNA clones have been isolated from a chinook salmon pituitary cDNA library. Sequence analysis of these clones indicates that they are closely related to the rat LIM domain homeobox gene, isl-1. Due to differential splicing, several of the clones encode truncated polypeptides containing a complete copy of the LIM domain without the homeodomain and C-terminal activation domain. The roles of these truncated polypeptides are discussed.

Amino Acid Sequence↗

Presence of isl-1-related LIM domain homeobox genes in teleost and their similar patterns of expression in brain and spinal cord.

Three novel LIM domain homeobox cDNAs encoding proteins structurally related to the Isl-1 protein were isolated from a chinook salmon pituitary cDNA library. Southern blot analysis of genomic DNA indicate that they are derived from three distinct genes, designated as isl-2a, isl-2b, and isl-3 genes. Nucleotide sequence analysis of reverse transcriptase-polymerase chain reaction amplified products reveal that the isl gene family contains two members (a and b) each of both isl-1 and isl-2 genes, and one member of isl-3 gene in the two tetraploid salmonid species, chinook salmon and rainbow trout, and only one member each of isl-1, isl-2, and isl-3 genes in the diploid zebrafish. The expression of the three isl genes in the rainbow trout were studied by reverse transcriptase-polymerase chain reaction analysis of embryonic and adult RNAs, and by in situ hybridization analysis of 8-week-old hatchlings. The transcripts of all three genes could be detected as early as 4 weeks postfertilization (the eye stage) and increased dramatically in 5-week-old embryos. In the adult, the three isl mRNAs appear to be differentially distributed in various tissues. The level of isl-1 mRNA is generally higher than those of isl-2 and isl-3 mRNAs. In situ hybridization analysis indicates that the transcripts of all three genes are localized in subsets of neurons in the brain and spinal cord. In the retina, isl-1 mRNA could be found in both the ganglion and inner nuclear layers while isl-2 and isl-3 mRNAs could only be detected in the ganglion layer. High level of isl-1 mRNA could also be found in mid-gut and interrenal organ where endocrine cells are densely populated. Based on these observations, we speculate that the three structurally related isl genes may play similar roles in cell determination and differentiation in the developing nervous system.

Aging↗

The antifreeze protein genes of the winter flounder, Pleuronectus americanus, are differentially regulated in liver and non-liver tissues.

The synthesis of winter flounder (Pleuronectus americanus) antifreeze protein (AFP) mRNAs in the liver is seasonally regulated by the pituitary gland. With the recent discovery that AFP mRNAs are also present in several non-liver tissues, the aim of the present investigation was to compare the regulatory mechanisms of AFP genes in liver and non-liver tissues. Northern blot analyses indicate that the level of liver AFP mRNA undergoes a several hundred fold difference between the winter and summer months, while AFP mRNAs from gills and kidneys exhibit only a modest 5-10 fold seasonal variation. As expected, the liver AFP mRNA in the hypophysectomized fish was increased by over 40 fold. However, no significant increase was observed for the non-liver AFP mRNAs upon hypophysectomy. These investigations suggest that AFP mRNAs in liver and non-liver tissues are differentially regulated.

Animals↗

Molecular heterogeneity among primary motoneurons and within myotomes revealed by the differential mRNA expression of novel islet-1 homologs in embryonic zebrafish.

Zebrafish embryos have three or four identifiable primary motoneurons per hemisegment. We previously reported that, while several ventral cells initially express the zebrafish Islet-1 (Isl-1) gene, a member of the LIM/homeobox gene family, the expression of this gene becomes restricted to a single or a pair of cells slightly anterior to each segment border by 16 hr after fertilization. Double staining by in situ hybridization and immunohistochemistry strongly suggested that these cells were mainly rostral primary motoneurons. Here, we have isolated two novel zebrafish cDNA clones for more Isl-1 family genes, termed zfIsl-2 and zfIsl-3. zfIsl-2 mRNA starts to be expressed in the ventral midsegmental cells per hemisegment around 15 hr. Double labeling experiments have shown that these midsegmental cells are the caudal primary motoneuron (CaP) and its variant equivalence pair. Our results revealed the heterogeneity in the expressed genes among primary motoneurons before the fates of the primary motoneurons are irreversibly determined, and further suggest the involvement of the Isl-1 and zfIsl-2 genes in the determination of cellular identities by primary motoneurons in embryonic zebrafish. zfIsl-3 mRNA is not expressed in motoneurons but is expressed at 17 hr, mainly in the ventral myotomes. This suggests that zfIsl-3 may be involved in the regional specification of the myotome and also in target recognition by CaP. zfIsl-2 is also expressed throughout the developing eye and tectal region of the midbrain, the target for the retinal axons. In the ventral spinal cord of the spadetail mutant embryo, which has defects in the somites, the cells expressing zfIsl-2 mRNA significantly decreased in number in contrast to the increase in cells expressing Isl-1 mRNA, suggesting the influence of the somites on the expression of both genes.

Amino Acid Sequence↗

Expression of the antifreeze protein gene in transgenic goldfish (Carassius auratus) and its implication in cold adaptation.

Ocean pout (Macrozoarces americanus) antifreeze protein (AFP) genes (approximately 10(6) copies) were microinjected into the oocytes of goldfish (Carassius auratus). Out of a total of 303 oocytes injected, 235 (77.6%) were normally fertilized after in vitro maturation and insemination, and 136 (57.9%) of the fertilized eggs were hatched. Dot blot analysis of genomic DNA from 2-month-old fish indicated that 30 of the fish (22%) were positive for the AFP transgene. The numbers of integrated AFP genes ranged from a single copy to multiple copies per cell. Two P1 male founders, Y45 and GV16, were crossed separately with a control female, and the inheritance of the AFP transgene was analyzed. Results from Southern blotting and polymerase chain reaction were found to be 7% (5 out of 82) and 58.8% (20 out of 34), respectively, indicating that these two founders are mosaic. The percentage of positive F2 generated from Y45F1 crossed with a control female was 56%, consistent with a Mendelian inheritance. Mature AFP detected by immunoblotting was expressed in both F1 and F2 offspring. These studies indicate that the antifreeze protein gene is successfully transferred and expressed in goldfish. Transgenic goldfish are significantly more cold tolerant than controls when challenged with low temperatures. These studies suggest that the AFP gene may have application in providing cold tolerance in addition to freeze resistance for a variety of fish species.

Acclimatization↗

[Clinical research of muscular relaxation induced by alcuronium and pancuronium].

The clinical experiment was designed for observing effect of muscular relaxation induced by alcuronium and pancuronium. 40 patients were randomly divided into alcuronium group (A) and Pancuronium group (P). A group and P group were redivided into A1,A2 groups and P1,P2 groups according to differences of the initial doses of muscle relaxants. The results of neuromuscular monitoring indicate that onset time and main-tenance time of 0.3 mg/kg alcuronium were respectively 2.4 min and 63 min; 0.1 mg/kg pancuronium were 2.7 min and 70 min. The initial doses mentioned above are suitable to intubation of anesthesia and meet surgical need for muscular relaxation. The changes of MAP,HR,blood kalium and natrium were not evident in statistical significance before and after administration of relaxants.

Alcuronium↗

A comparison of the effects of the propofol versus midazolam during total intravenous anesthesia for gynecological surgery procedures.

The effects of propofol and midazolam as an intravenous anesthetic were compared in 40 ASA I-II patients undergoing gynecological surgery during total intravenous anesthesia (TIVA). They were divided into propofol group (P n = 20) and midazolam group (M n = 20) randomly. The anesthesia was designed for each group respectively. Here, we discuss the experimental method and the results, which indicate that propofol is not only an effective anesthetic but also has more rapid and head-clear recovery properties than midazolam.

Adult↗

Trimethyl lead neurotoxicity in the rat: changes in glial fibrillary acidic protein (GFAP).

The literature on the toxicology of lead provides little evidence of the neurotoxicity of organic lead compounds. Toxicant-induced changes in the concentration of glial fibrillary acidic protein (GFAP) in the brain may help clarify at which stage of neurotoxicity astrocytes are affected and whether GFAP may provide an index of toxicity. Male F344 rats (> 42 days old) were exposed to 0 (control), 8 or 16 ppm lead as trimethyl lead (TMPb) in drinking water for up to 14 days. Weight Gain was significantly reduced in both exposed groups. Control rats had the expected brain regional pattern of GFAP concentration with the highest in the hippocampus and cerebellum and lowest in the cerebral cortex. The hippocampus was the region very sensitive to TMPb, with increased GFAP in rats exposed to 8 and 16 ppm TMPb with decreases in GFAP in rats exposed to 8 and 16 ppm TMPb for 14 days. There was a significant time-response in rats exposed to 8 ppm TMPb with decreases in GFAP on day 7 and increases on day 14. A hypothesis concerning this biphasic change in GFAP concentrations is discussed. The results indicate that GFAP may be used to indicate the role of the astrocyte in the neurotoxicity of TMPb. GFAP concentration, as biomarker of TMPb effect, was as sensitive to TMPb as body weight and thus may provide a marker of neurotoxicity.

Animals↗

Zinc and DNA binding properties of a novel LIM homeodomain protein Isl-2.

LIM homeodomain proteins are a family of recently characterized proteins which contain, in addition to a homeodomain, two tandem repeats of conserved Cys-His motifs termed as LIM domains. We have recently isolated several clones from a chinook salmon pituitary cDNA library that encode two novel LIM homeodomain proteins, Isl-2 and Isl-3, which are structurally related to rat Isl-1. In the present study, we used the salmon Isl-2 to determine the role of LIM domains in DNA binding. Several glutathione S-transferase (GST) fusion proteins containing either full length Isl-2 or various portions of this protein were expressed in bacteria. Zinc blot analysis reveals that the LIM domains produced in bacteria are capable of binding zinc. Gel shift analysis indicates that all homeodomain-containing fusion proteins are able to bind to a TAAT target sequence while the fusion proteins containing only the LIM domain are not. In contrast to a previous observation that the LIM domains of rat Isl-1 have an inhibitory role in DNA binding, full length salmon Isl-2 containing both the LIM domains and a homeodomain can bind to a TAAT target sequence. To further examine the role of LIM domains in DNA binding, several GST fusion proteins were used to select specific target DNA sequences from a pool of randomly incorporated oligonucleotides. Specific target DNAs were selected by fusion proteins containing the homeodomain or the full length Isl-2, but not by LIM domain only fusion proteins, indicating that the LIM domain alone is not involved in DNA binding. The selected target DNAs were cloned and sequenced. They revealed two classes of consensus, C/TTAATG/TG/A and C/TTAAGTG, for both the homeodomain and full length Isl-2. The two classes of consensus competed with each other for binding to the homeodomain. The equilibrium dissociation constants for DNA binding, estimated by Scatchard analysis, were similar for the homeodomain and full length Isl-2.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

Bulk isolation and identification of fish genes by cDNA clone tagging.

The fish has long been a popular research model for various disciplines in the life sciences, but its molecular biology remains largely unexplored, and the number of fish genes identified at present is still limited. To increase the repertoire of fish genes to facilitate further research, we have embarked on cDNA clone tagging studies in several teleosts. The basic procedure is partial sequencing of randomly selected cDNA clones by a single sequencing reaction to obtain nucleotide sequences of about 200 to 300 bp for each clone. These partial DNA sequences can be used as tags to identify fish homologous genes by searching for sequence similarity in DNA databases. By this method, we have sequenced a total of 265 clones from pituitary and liver cDNA libraries from five species of fish and identified 83 clones encoding 55 different genes. These 55 genes are described for the first time in their species, and 37 of them are described for the first time in fish. These preliminary data indicate that cDNA clone tagging is useful for rapidly increasing the number and availability of genes in teleosts. Several applications of these tagged random clones are discussed.

Animals↗

Phylogenetic specificity of prolactin gene expression with conservation of Pit-1 function.

In mammals, the pituitary POU homeodomain protein, Pit-1, binds to proximal and distal 5'-flanking sequences of the PRL gene that dictate tissue-specific expression. These DNA sequences are highly conserved among mammals but are dramatically different from PRL 5' sequences in the teleost species, Oncorhynchus tschawytscha (chinook salmon). To analyze the molecular basis for pituitary-specific gene expression in a distantly related vertebrate, we transfected CAT reporter gene constructs containing 2.4 kilobases (kb) 5'-flanking sequence from the salmon PRL (sPRL) gene into various cell types. Expression of the sPRL gene was restricted to pituitary cells, but in rat pituitary GH4 cells levels of expression were at least 90-fold lower than those obtained with a -3 kb rat PRL (rPRL) construct. Conversely, in primary teleost pituitary cells, -2.4 kb sPRL/CAT was expressed at levels about 10-fold higher than -3 kb rPRL/CAT. To determine whether species-specific transactivation by Pit-1 was sufficient to explain these species differences in PRL gene expression, we isolated a cDNA clone encoding the salmon Pit-1 POU domain and constructed a rat Pit-1 expression vector that contained salmon Pit-1 POU domain sequences substituted in frame. The chimeric Pit-1 encoded 14 amino acids unique to salmon. Coexpression of rat Pit-1 with salmon or rat PRL/CAT in transfected HeLa cells resulted in specific and strikingly comparable levels of promoter activation. Moreover, the specificity and efficacy of the chimeric salmon/rat Pit-1 was similar to wild type rat Pit-1 in activating salmon and rat PRL/CAT.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Effect of low benzene exposure on neurobehavioral function, AChE in blood and brain and bone marrow picture in mice.

The purpose of this study was to examine the effect of low level benzene exposure on neurobehavioral functions, AChE in blood and brain, bone marrow picture in Kunming mice. Forty adult Kunming male mice were divided into 4 groups. They were exposed to 12.52, 3.13, 0.78 and 0 ppm benzene for 2 h.d-1 for 30 d. Central nervous system (CNS) function was inhibited by 12.52 ppm and excited by 0.78 ppm benzene exposure, but irregularly affected by 3.13 ppm. AChE in blood and brain was decreased in 12.52, 3.13 ppm group. The weight of liver to body weight ratios in 12.52 ppm group was higher than those of control group significantly. Bone marrow picture revealed inhibited proliferation of white and red cell systems, especially in 12.52 ppm group, consisting of decrease of percentage of myeloblast, premyelocytes, myelocytes, erythroblasts and megakaryocytes, especially in 12.52 ppm group.

Acetylcholinesterase↗

Functional analysis and temporal expression of promoter regions from fish antifreeze protein genes in transgenic Japanese medaka embryos.

Several series of sequences that are upstream of the transcriptional start site of different types of fish AFP genes were fused to the bacterial CAT gene, and their transcriptional role was examined in a transient expression assay after microinjection into Japanese medaka (Oryzias latipes) embryos at the 1-4 cell stage. Our studies demonstrated that the AFP genes have functional promoter regions containing positive as well as negative regulatory regions, indicating that these genes could be regulated at multiple sites. We also observed a promoter-specific pattern of temporal expression. Typically, the CAT expression was low in the first 4 days of embryonic development or before the stage of body pigmentation, followed by a sharp increase. The high level was maintained until hatching (11-13 days after fertilization), by which time the activity decreased to a very low level.

Animals↗