ACTA
ZOOLOGICA
SINICA
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ACTA ZOOLOGICA SINICA
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ACTA ZOOLOGICA SINICA
Vol. 47 No.3 June. 2001
ABSTRACT
Effect of imposed sterility and removal on reproductivity and population
size of ratlike hamster in enclosures
ZHANG Zhi-Bin* ZHANG Jian-Xu WANG Fu-Sheng WANG Yong-Qing CAO Xiao-Ping WANG Yu-Shan
( National Key Laboratory of Integrated Pest Management, Institute of Zoology,
Chinese Academy of Sciences, Beijing 100080, China )
Abstract The effect of imposed sterility and culling on the reproductivity and population size of Ratlike hamster( Cricetulus triton ) in enclosures was studied by using surgical operation. This study aims to testify two modeling predictions: fertility control can achieve similar result as simple culling, and sterility of both sexes can achieve an extra result in reducing reproductivity of hamsters. The key factors in population regulation of the hamster were also discussed. The experiment was composed of 4 treatments, 2 replicates for each treatment: Control (9 normal males + 9 normal females); Culling (3 normal males + 3 normal females); Female and malesterility (3 normal males + 3 normal females + 6 sterile males + 6 sterile females); Female sterility (9 normal males + 3 normal females + 6 sterile females). The results by using one-way ANOVA analysis indicated that the average reproductivity of control group and culling group were higher than that of female sterility group, Female and male sterility group, and the average reproductivity of female sterility tended to be larger than that of female and male sterility. There were no significant difference between normal and sterile adults, females and males adults, male and female juveniles. The sex ratio of female: male is 53:49 = 1. 0816:1, not significantly different from the ratio of 1: 1. The growth of body mass was well fit by logistic model. The relative growth rate of sterile female and the maximum body mass were larger than those of normal female. The relative growth rate of normal male was smaller than that of normal female. When food and water were abundant, the density-dependent social factor did not affect the reproductivity, which implied that food instead of social factor might be the key factor in determining the change of reproductivity of the hamster in field. In the enclosures, the survival rate of males was lower than that of females and the survival rate of juveniles was obviously lower than that of adults. The survival rate of confined adults was very similar to that of adults in field, but the survival rate of the confined juveniles was obviously lower than that of juveniles in field. This implied that the impact of density-dependent social pressure would affect the mortality, especially that of juveniles. [Acta Zoologica Sinica 47(3):241-248, 2001, in English]
Key words Ratlike hamster ( Cricetulus triton ), Reproductivity, Body mass, Growth, Population regulation,Fertility control, Rodent control
*E-mail: zhangzb@panda.ioz.ac.cn
Population growth and productivity of the ciliated protozoa community in
the enclosures of saline-alkaline ponds*
ZHAO Wen* DONG Shuang-Lin ZHANG Zhao-Qi LI De-Shang
( Ocean University of Oingdao, Oingdao 266003, Shandong, China )
Abstract We studied the population growth and productivity by the ciliated protozoa community in the enclosures with and without Silver carp (Hypophthalmichthys molitrix)of saline-alkaline ponds productivity, during April-September in 1998 .The experimental enclosure was a land-based enclosure with an area of 0.10 hm2 in ponds. The enclosure was 20 m2(4m×5m) in area, 2.0 m in height (1.5 m in water depth), and is made of plastic-coated polyethylene woven cloth with a frame composed of wood and bumboo poles. During the experiment, water temperature of experimental enclosures were 22～32℃, conductivity ranged from 1 050～2400 μs/cm, total alkalinity were 3.02～5.64 mmol/L. Population growth rate and generation times were obtained through in situ experiment. During the culture season, the generation time of major species of protozoa: Askenasia volvoac , Ciclidium citrullus , Strobilidium gyrans , Prorodon viridis and Strombidinopsis sp. was 6.93～23.10, 6.30 ～18.73, 9.90, 9.90～14.75 and 7.62 hr, respectively; population growth rates were 0.030～0.100, 0. 037～0.110, 0. 070, 0.047～0.070 and 0. 091/hr, respectively. The mean daily productivity of total planktonic ciliates in in situ experimental bottles of the enclosures with and without fish were 1070.25 and 944.25 mg/m3·d, respectively, during May-August, the productivity of the both enclosures at the same times were 1.79 and 9.72 mg/m3· d, and the ratio of daily P/B calculated by month were 2.50 and 2.03, respectively.
The species composition, biomasses, productivity of planktonic ciliated protozoa were influenced by Silver carp and other environmental factors. It is concluded that the biomass and productivity of ciliated protozoa in the enclosures without fish were lower than those of the enclosure with Silver carp. The population growth of ciliated protozoa was subjected to water temperature and trophic status. [Acta Zoologica Sinica 47(3):249-255, 2001]
Key words Protozoa, Population growth, Productivity, Saline-alkaline ponds
E-mail: zhaowen@mail.dlptt.in.cn
Effects of thermal and hydric environments on incubating eggs, hatching'success,
and hatchling traits in the Chinese skink (Eumeces chinensis )
JI Xiang* ZHANG Chao-Hua
(Department of Biology, Hangzhou Normal College, Hangzhou 310036, China)
Abstract Effects of thermal and hydric environments on incubating eggs, embryonic use of energy and nutrients, and hatchling traits were studied in the Chinese skink (Eumeces chinensis ) from a population in Lishui, Zhejiang, eastern China. The eggs were incubated at temperatures of 24, 26, 30, and 32℃ on substrates with water potentials of 0 and - 220 kPa using a 4 ×2 factorial design. All viable eggs increased in mass over the course of incubation due to absorption of water, and mass gain during incubation was dependent on initial egg mass, temperature, and substrate water potential. Variation in the wet mass of hatchlings among treatments stemmed mainly from variation in water content. Temperature profoundly affected duration of incubation, water uptake by eggs during the course of incubation, embryonic use of energy and nutrients, embryonic mobilization of inorganic material from the shell, and almost all hatchling traits examined. However, incubation temperature did not affect hatching success within the range from 24℃ to 32℃. Hatchlings from eggs incubated at 32℃ were less developed, as indicated by smaller carcasses and more unutilized yolks, and hence were smaller than their siblings from lower incubation temperatures. Moreover, hatchlings from eggs incubated at 32℃ did not perform as well as their siblings from the temperatures lower than 32℃ in the racetrack, even when the hatchling size (snout-vent length) was kept constant. This indicates that high incubation temperatures have an impaired effect on locomotor performance of E. chinensis hatchlings. Eggs incubated in wetter substrates produced larger hatchlings that contained smaller residual yolks than did eggs incubated in drier substrates. At the time of hatching, shells form eggs incubated at 24℃ were heavier in dry mass and contained more ash than those from eggs incubated at lower temperatures, suggesting that less inorganic material be withdrawn from the shell by embryos developing at the temperature. Incubation temperature subtly affected body shape and head size of E. chinensis hatchlings: hatchlings from eggs incubated at 30℃ had the longest tails, and hatchlings from eggs incubated at 32℃ had the smallest heads. On the contrary, substrate water potential was not an important source of variation for body shape and head size of hatchlings. The effects of temperature on hatchling traits were independent of the effects of water potential. Variation in size and mass induced by incubation thermal and hydric environments would be important to post-hatching survival and fitness of hatchlings. Although temperature did not significantly influence hatching success within the range of 24 to 32℃, 26℃ and 30℃ were more suitable for incubating E. chinensis eggs, as eggs incubated at temperatures within this range produced larger and well-performed hatchlings at a lower energy expenditure. [Acta Zoologica Sinica 47(3):256-265, 2001]
Key words Chinese skink (Eurneces chinensis), Egg, Incubation, Hatchling traits, Locomotor performance
*E-mail: xji@mail.hz.zj.cn
Mate choice and related characteristics of mandarin vole( Microtus mandarinus )
TAI Fa-Dao①* WANG Ting-Zheng① ZHAO Ya-Jun②
(① College of Life Sciences, Shanxi Normal University, Xi 'an 710062, China)
②Institute of Laboratory Animal Sciences, the Chinese Academy of Medical Sciences, Beijing 100021, China )
Abstract The microtine rodents are an ideal group of rodents for comparative studies because many of these species, in spite of their close taxonomic relationship, show profound differences in reproductive biology and social organization. In order to ascertain the prediction from field studies that Mandarin vole (Microtus mandarinus) is monogamous and to analyse its mechanism of evolution and function of adaptation, we did this study. Many behavioral experiments show that the mating systems are associated with mate choice among individuals. We studied the mate choice of Mandarin vole in a T-shaped tether box. two end compartments(30 cm×30 cm×30 cm) and a central area(45 cm×30 cm ×30 cm). The subject individuals were able to move freely within the three compartments, and the two object individuals were linked at the choice chambers respectively during 30 minute trial. The mate choices by Mandarin vole for partner versus strange heterosexual vole and for familiar versus strange heterosexual vole were done in the maze. Familiarity was established by housing male with an unrelated female in a plastic breeding cage until they were found to display sexual behavior. Partnership referred as the relationship between a pair of male and female having bred at least one litter of offspring. Behavioral comparison between the two stimulus voles was achieved by a Wilcoxon matched-pairs test. In these tether tests, the male and female Mandarin voles spent significantly more time on social- investigating, mounting, copulating and being amicable to the partner and familiar heterosexual voles than the strange voles and attacked the strange voles for longer time. Moreover, this species spent less time in the neutral area and displayed the monogamous characteristics. Further more, the parameters such as dimorphism based on ratio of male and female body mass, ratio of testes and body mass, mean litter size, age of last nipple attachment, distance of nipple clinging, duration of social contacting, parental behavior in cage and social organization in field were calculated and summarized. Through comparing these characteristics of Mandarin vole with two vole species known to be monogamous tendencies (M. ochrogaster and M. pinetorurn) and two known to be promiscuous under most conditions (M. pennsylvanicus and M. montanus), It was shown that the interspecific difference in individual behaviors and other characteristics is consistent with interspecific difference of mating system. Mandarin vole is generally similar to the monogamous praire vole and pine vole and dissimilar significantly to the polygynous meadow vole and montane vole. Based on these data, we suggested that of Mandarin voles might be monogamous. This result is important for studying of social behavior because monogamous species can provide models not only for studying social behavior of voles with diverse mating system but also for studying the neurobiological basis of social behavior such as parental care, and inbreeding avoidance so. [Acta Zoologica Sinica 47(3):266-273, 2001]
Key words Mandarin vole (Microtus mandarinus), Mate choice, Monogamy, Related characteristics
*E-mail: taifadao@snnu.edu.cn
Cloning and expression in prokaryote of grass carp IGF- I cDNA
HUA Yi-Min LIN Hao-Ran*
( Institute of Aquatic Economic Animal, School of Life Science,
Zhongshan University, Guangzhou 510275, China )
Abstract Fragment of open reading frame of Grass carp (Ctenopharyngoden idellus ) cDNA was cloned for the first time from liver by RT-PCR technique using a set of primers designed according to the closely related Common carp IGF-I cDNA. Sequence analysis showed that similar to Common carp IGF-I cDNA, the cloned Grass carp IGF-I cDNA was also Ea-2 subtype, and the homology of nucleotide sequence in open reading frame was up to 95 % compared with the counterpart of Common carp, only nine amino acid were different between the two pre-pro-IGF-I s. The homology of nucleotide sequence in ORF of Grass carp was 63 % compared with the counterpart of human. Grass carp and Common carp mature protein differed in only two amino acids, while Grass carp and human mature protein differed in 13 amino acids. The fragment of Grass carp IGF-I DNA which expressed mature gcIGF-I protein, was subcloned into a GST fusion expression vector, pGEX-4T-3, to construct a recombinant expression vector, pGEX-4T-3-gcIGF-I. The pGEX-4T-3-gclGF-I was then transformed into E. coli, BL21. When cultured and induced by IPTG, high amount of GST-gcIGF-I fusion protein was expressed in the transformed BL21. The recombinant IGF-I protein had immune activity showed by Western Blotting technique. [Acta Zoologica Sinica 47(3):274-279, 2001]
Key words Grass carp ( Ctenopharyngoden idellus), IGF- I , Cloning, Expression, Prokaryote
*E-mail: LS32@zsu.edu.cn
Karyotypes and G-banding patterns of three eared-pheasant (Crossoptilon) species
SHI Xing-Di* ZHANG Zheng-Wang LIU Ling-Yun
( Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, College of Life Sciences, Beijing Normal University, Beijing 100875, China )
Abstract The eared-pheasants ( Crossoptilon ) from a genus endemic to China, which consists of four species: white eared-pheasant ( C. crossoptilon ), Tibetan eared-pheasant ( C. harmani ), blue eared-pheasant ( C. auritum ) and brown eared-pheasant (C. mantchuricum ). These species are listed as the national protected species of China. However the phylogenetic relationship among C. crossoptilon, C. auritum and C. mantchuricum remains unclear. In this paper, we used cytogenetic method to study phylogenetion relationship of the three species.
Chromosome preparations were made from PHA-stimulated peripheral lymphocytes by standard techniques and chromosome G-banding patterns were obtained with the NaOH method. The karyotype analysis consisted of comparisons of diploid numbers and the locations of the centromeres on each chromosome. The results showed that the karyotypes of the three species were identical to each other. All three species had a diploid number of 82, with seven pairs of macrochromosomes including the sex-chromosomes and 34 pairs of microchromosomes. There was no sharply demarcated boundary between macro- and microchromosomes. Sex-chromosomes were heteromorphic, males are homogametic with the sex chromosome constitution ZZ, while females were heterogametic with ZW sex chromosomes. The Z-chromosome was the forth longest of all chromosomes. Ranked 6th or 7th in length, W-chromosome was much shorter than the Z, of these macrochromosomes, the first pair of chromosomes was metacentric and the Z-chromosome was submetacentric, whereas the others were centromere telocentric.
Analysis of G-banding patterns for the macrochromosomes of the three species indicated that the band patterns of Z-chromosomes were very similar while the other six pairs were quite different in the three species. The telocentric W-chromosomes was highly heterochromatic. After quantifying G-banding patterns, we calculated the similarity coefficients and made a cluster analysis of the three species. The data from this study indicated a closer evolutionary relationship of blue eared-pheasant to brown eared-pheasant than to white eared-pheasant, in contrast to the previous result based on studies of protein electrophoresis. [Acta Zoologica Sinica 47(3):280-284, 2001]
Key words Crossoptilon, Karyotype, G-banding pattern, Relationship, Cluster analysis
*E-mail: shixd@bnu.edu.cn
Effects of estradiol-17βon immunocompetence in rainbow trout(Oncorhynchus mykiss )
HOU Ya-Yi* HAN Xiao-Dong
(Medical School, Nanjing University, Nanjing 210093, China)
Abstract The oral administration of estradiol-17β(E2) elevated plasma E2 concentration in juvenile Rainbow trout. This elevation significantly reduced plasma IgM levels. E2 also reduced dose-dependently the number of IgM producing cells (IPC) in vitro lymphocytes from the peripheral blood, head kidney, and spleen in juvenile Rainbow trout. In addition, to evaluate the effects of E2 on non-specific cellular defense, we investigated the amount of reactive oxygen intermediates produced from the head kidney macrophages. The result showed that macrophage (φ) activity was clearly suppressed by E2. These results suggest that low immunocompetence with the gonadal maturation may be related to the elevation of plasma E2 level in female trouts. The susceptibility to infectious diseases in the fish during spawning season may result from the suppression of E2 on both specific and non-specific immune activities. [Acta Zoologica Sinica 47(3):285-291, 2001, in English]
Key words Rainbow trout (Oncorhynchus mykiss), Estradiol-17β, IgM, IgM-producing cell, Macrophage, Immunocompetence
E-mail: yayihou@nju.edu.cn
Hatching of resting eggs produced by Brachionus calyciflorus
cultured under different conditions
Xl Yi-Long* HUANG Xiang-Fei
(Institute of Hydrobiology, The Chinese Academy of Sciences, Wuhan 430072, Chiina)
Abstract In order to identify the optimal condition of resting egg formation, or to increase the hatching rate of resting eggs produced by freshwater rotifer Brachionus calyciflorus under certain culture conditions, we experiments were made to examined the hatching of resting eggs produced by B. calyciflorus cultured under different temperatures, alga diets, illumination periods and pHs, respectively. The results showed that the hatching duration of resting eggs under different conditions in which resting eggs were produced and hatched was the initial seven days of the experiment, and the hatching rate peak occurred during the initial three days. There were highly significant effects of temperatures at which resting eggs was produced and hatched independently and in interaction on the accumulative hatching rate of resting eggs. The accumulative hatching rate of resting eggs which was produced and hatched at 20℃ was 52.50 ± 6.89 %, and the highest among all the tested experimental combinations. Both the rotifer's feeding history and hatching temperature influenced very significantly on the accumulative hatching rate of resting eggs, respectively. The mean value of accumulative hatching rate of resting eggs produced by B. calyciflorus which fed on Chlorella pyrenoidosa was 27.92 %, and higher than that on Scenedesmus obliquus or mixed algae composed of C. Pyrenoidosa and S. obliquus, which was 12.29 % and 18.75 %, respectively. There were no significant effects of the illumination conditions in which resting eggs were produced and hatched on the accumulative hatching rate. The pHs at which resting eggs were produced influenced very significantly on the accumulative hatching rate of resting eggs. The accumulative hatching rate of resting eggs formed at pH6.5 and pH7.5 were 32.50 ± 7.07% and 38.75 ± 15.53%, respectively, and both of them were higher than that at pH8.5～10.5. From the above-stated and our other published results, one can state that all the hatching pattern of resting eggs produced and hatched under different conditions are synchronous. 20℃ and pH7.5 are the optimal temperature and pH value for resting egg formation of B. calyciflorus, respectively. [Acta Zoologica Sinica 47(3):292-297, 2001]
Key words Brachionus calyciflorus, Resting eggs, Culture conditions, Hatching pattern, Hatching rate
*E-mail: yilongxi@mail.ahwhptt.net.cn
Fatty acid composition of sharphead dogshark( Scoliodon sorrakowah)
liver oil during developmental stages
LIU Xiao-Chun*
( School of Life Science, Zhongshan University, Ouangzhou 510275, China )
QIU Shu-Yuan
(Department of Oceanography, Xiamen University, Xiamen 361005, Fujian , China )
Abstract Fatty acid composition of the liver oil from different developmental stages of Sharphead dogshark (Scoliodon sorrakowah) of the southern Fujian coastal waters, caught from May, 1993 to April, 1995, was determined with capillary gas chromatography. Saturated, monounsaturated and polyunsaturated fatty acids represented 15.3 %～34.6 % 13.8 % ～ 25.6 % and 38.8 %～ 68.1% of the total fatty acids in the liver oils of Sharphead dogshark, respectively. The major fatty acids in each sample were 16: 0 (7.9 %～20.9 % ), 18: 0 (4.5 % ～ 12.1% ), 18:1ω9(6.1%～14.3% ), 20:4ω6(2.8% ～13.9% ) and 22:6ω3(16.3 % ～56.2% ).
Characteristics of fatty acid composition of the liver oils in Sharphead dogshark was closely associated with the developmental stages of the animals examined. In mate sharphead dogshark, 16:0(36.0 % ～39.8 % ), 16:1 (17.1% ～19.2 % ) and 18:1 (ω9 +ω7) (21.1% ～23.1% ) were the major constituents of fatty acids in the liver oils of embryos, the levels of 22: 6ω3 (docosahexaenoic acids, DHA ) were very low (3.8 ± 1.34, n = 3). Con- centrations of 22: 6ω3 were much higher in the liver oils of immature males (27.1 ± 2.47, n = 3) than in that of embryos, and reached maximum values at prespawning stage (51.0 ± 7.42, n = 5) and then decreased significantly at regressed stage (34.4 ± 2.40, n = 3). Contents of 22: 6ω3 in the liver oils in female increased during the development of the fish and reached maximun at the pre-gestation stage (49.7 ± 4.04, n = 5), which were significantly higher than that at the immature stage (25.7 ± 0.64, n = 3) and at ovary developing stage (33.3±3.46, n = 4), respectively, and decreased remarkably at the end of gestation (25.6 ± 8.08, n = 3). It showed that DHA (22: 6ω3) might play an important role in growth and development of Sharphead dogshark embryos. [Acta Zoologica Sinica 47(3):298-303, 2001]
Key words Sharphead dogshark (Scoliodon sorrakowah ), Liver oil, Fatty acid composition, Developmental stages
*E-mail: ls32@zsu.edu.cn
Immunocytochemical localization of dopamine and norepinephrine in the nervous system
and gonads of amphioxus (Branchiostoma belcheri)
WENG You-Zhu FANG Yong-Qiang* HU Xiao-Xia ZHOU Jing
(Third Institute of Oceanography, SOA, Xiamen 361005, Fujian , China)
Abstract It is well documented that dopamine (DA) and norepinephrine (NE) can regulate gonadotropin secretion in fishes. The presence of DA and NE in the brain of mature Amphioxus (Branchiostoma belcheri Gray) has been demonstrated by high performance liquid chromatography (HPLC), however, whether these two neurotransmitters are involved in the regulation of the secreton of epithelial cell (primitive gonadotropic cell) of Hatschek's pit (homologous to pituitary in vertebrates) remains unknown. In the present study, the immunocytochemical localization of DA and NE in the nervous system and gonads of Amphioxus was investigated with (avidin-biotin complex ABC) method.
Amphioxus (18～52.4 mm in total length) were collected from the Tong'an-Qiongtou sea area near Xiamen). Each animal was divided into three segments, rostal (from tentaculum peristomial to the first pair of gonads), medial (from the first pair of gonads to the anus) and caudal. They were fixed in Bouin's fixative without acetic acid, and embedded in paraplast. Longitudinal or cross sections of 6 btm thickness were cut. Sections with the structure of the brain, neural tube and gonads were identified under a light microscope and selected for immunostaining.
Medium and small types of DA and NE immunopositive nerve cells were found in the posterior part of telencephalon, anterior and middle part of midbrain, and the positive substance occurred in the cytoplasm of nerve cells. No immunoreactivity for DA and NE in this study was detected in the posterior of midbrain and hindbrain of Amphioxus. We found for the first time that DA immunopositive nerve cells and fibers also located in the infundibular part of Amphioxus, a region extending from the middle region of midbrain to the Hatschek's pit and corresponding to the fish infundibular which connected hypothalamus with pituitary. Large type of NE immunopositive nerve cells were showed in the posterior part of telencephalon. NE immunopositive nerve cells and fiber connecting with another nerve cells was observed in the middle part of midbrain. On the other hand, DA
and NE immunopositive nerve cells were identified on the dorsal side and middle part of spinal cord, and the immunopositive nerve fibers could be found on the ventral side. The distribution pattern of DA and NE in the nervous system of adult Amphioxus was different from that in larvae. Additionally, we observed for the first time that the immunoreactivity of DA and NE also located in the ovary and testis of Amphioxus in various development stages except in the mature testis.
In conclusion, the results suggested a direct action of DA and an indirect action of NE on the secretion Hatschek's pit through the brain GnRH neuronal system in Amphioxus, as has been shown in fishes. [Acta Zoologica Sinica 47(3):304 -309, 2001]
Key words Amphioxus (Branchiostoma belcheri ), Dopamine, Norepinephrine, Nervous system, Gonads
*E-mail: fant98@ public.xm.fj.cn
Acrosome reaction in sperm from the mud crab (Scylla serrata)
WANG Yi-Lei①* ZHANG Zi-Ping② XIE Fang-Jing① LI Shao-Jing③
(①Institute of Aquaculture Biotechnology，Jimei University，Xiamen 361021， Fujian, China)
(②Department of Biology and Chemistry，City University of Hong Kong，Kowloon，Hong Kong, China)
(③Oceanography Department，Xiamen University，Xiamen 361005，Fujian, China)
Abstract The nonmotile sperm with a large and complex acrosome body is a well-known characterization of decapod crustaceans. The acrosome reaction of this kind of sperm is more complex than others. Although a lot of work focused on the structure of acrosome and the acrosome reaction of decapod crustaceans has been published, little information is available about how to artificially induce the acrosome reaction with a high rate of success.
The ultrastructure of mature sperm and the spermatogenesis of Scylla serrata, a commercially important species of crab, have been reported in our previous works. As a part of our ongoing research, we investigated the optimal conditions for artificial induction of the acrosome reaction of Scylla serrata and observed the process of the acrosome reaction in the present study.
The crabs were collected from the local market. Spermathecaes from five female crabs were cut open, and the contents of the spermatophore were transferred into Ca2+ free artificial seawater and pooled in a single tube. The spermatophore was gently homogenized in Ca2+ free artificial seawater by using a glass tissue grinder. The debris were removed using nylon mesh filter and the sperm, suspended in Ca2+ free artificial seawater, were pelleted by centrifugation at 400μg for 3～5 min and then resuspended in fresh Ca2+ free artificial seawater.
A stock solution of A23187 was dissolved in DMSO and diluted to a concentration of 2mg/ml in fresh Ca2+ free artificial seawater.
The optimised period for the acrosome reaction was assayed by ionophore-induction. Spermatozoa exposed in 0 (control group) and 50 μg/ml for 10, 0, 30, 40, 50, 60, and 120 min were fixed in 3% glutaraldehyde.
Three factors with five levels including a series of pH (6.0, 7.0, 8.0, 9.0 and 10.0), a series of different concentrations in A23187 (32, 40, 48, 56 and 64μg/ml) and Ca2+ (0.10%, 0.15%, 0.20%, 0.25% and 0.30%) were chosen to assay the optimal condition for the acrosome reaction in sperm of the mud crab Scylla serrata using an orthogonal experiment. After incubation in the various test solutions for 40 min, the spermatozoa were fixed by adding an equal volume of fresh Ca2+ free artificial seawater-buffered 6% glutaraldehyde, the acrosome reaction was scored by light microscopy. Each score involved random examination of 200 spermatozoa. Some samples were then post fixed in 1% OSO4, dehydrated in acetone, embedded in an epoxy resin, cut with glass knives, stained with uranyl acetate and lead citrate, and viewed under transmission electron-microscope (JEM-CXII) and scanning electron-microscope (S-520) electron-microscope to observe the fine structural events of the reaction.
The results showed that most of the acrosome reaction of Mud crab sperm were completed in 40 minutes (Table 1).
The orthogonal experiment results in acrosome reaction of Scylla serrata were shown in table1, the results were analysed in table 3. Comparing the R value of each column of table 3, the largest R value, 75.6 derived from pH, therefore, pH (A) is the most important effect factor, the second most important effect factor is the concentration of A23187 (C, R=31.8), the third most important effect factor is the concentration of Ca2+ (B, R=26.2). The highest rate of acrosome reaction is achieved when sperm are exposed in pH 9.0 solutions. Both the higher and lower pH suppressed the success of the acrosome reaction. According to the K value of table 3, the optimised combination for acrosome reaction is A4B4C5. In all 25 of the tested combinations, No. 17 combination (A4B2C5) induced the highest acrosome rate. The re-test proceeded using the combination of A4B2C5, A4B4C5, 200 sperm were counted in each treatment. 73.5% and 82.5% of acrosome reaction rate were achieved in A4B2C5 and A4B4C5 respectively. It concludes that the highest rate (82.5%) of the acrosome reaction in sperm can be achieved when sperm isolated from the spermathecae of the female crab are exposed to 64μg/ml ionophore A23187，0.25% Ca2+ at pH 9 in Ca2+ free artificial seawater for 40min.
As depicted in PlateⅠ:1a, 2a, un-reacted sperm taken from the spermathecae of female crabs appeared to have a uniformly top-like shape with several slender radiating nuclear arms and a larger spherical acrosome overlying the nuclear mass. The acrosomal vesicle includes an apical cap, a subcap zone, an inner layer and an outer layer. The apical cap can be divided into three thin layers (two dense layers and one lucent layer). Outside of the outer layer are 2～3 lamellar structures. Microtubules of the performatorium in the central cannal present a conical appearance (PlateⅠ:3).
The acrosome reaction in the sperm of Mud crab occurs in two stages, initially with swelling of the acrosomal vesicle followed by extrusion of its contents. In this first stage, the large part of the nuclear arms retracted into the nucleus and the apical cap is swollen (PlateⅠ:1b, 2b, 4), then the apical cap is broken to form an aperture (PlateⅠ:5). Accompanied by retraction of nuclear arms, the acrosome vesicle including an inner layer, an outer layer and a part of the lamellar structures were everted through the apical acrosomal cap (PlateⅠ: 1c, 2c, 6). Subsequently, in the second stage, the acrosomal tubule projected forward and passed through the everted vesicle. The reacted sperm perforatorium projected forward about 2.5μm. The ejected perforatorium becomes the leading edge of the reacted sperm. Data show that one function of the acrosome reaction of crab sperm is to generate forward movement of this immotile cell. The importance of this forward movement in sperm is to penetrate the chorion. [Acta Zoologica Sinica 47(3):310 - 316, 2001]
Key words Mud crab (Scylla serrata)， Sperm， Acrosome reaction， Artificial inducing， Structure, Ultrastructure
*E-mail: yileiwang@hotmail.com
Embryonic development of abalone( Haliotis diversicolor reeve)
LU Jun-Yi* CHEN Zhi-Sheng WU Jin-Ying ZENG Hua SU Guan-Hua
( School of Life Science, Zhongshan University, Guangzhou 510275, China)
Abstract The embryonic development in Abalone (Haliotis diversicolor Reeve), was divided into four periods, including 32-stages (Table 1). (1) Cleavage period containing 13-stages: artificial fertilization, the first polar body stage, the second polar body stage, early 2-cell stage, 2-cell stage, 4-cell stage, 8-cell stage, 16-cell stage, 32-cell stage, 64-cell stage, 128-cell stage, morula stage and blastula stage; (2) Embryonic development period including 4-stages: gastrula stage, early trochophore stage in egg membrane, late trochophore stage in egg membrane and at the moment of hatching stage; (3) Postembryonic development composed of 13-stages: trochophore, early veliger stage, mid-veliger stage, late veliger stage, early creeping larva stage, mid-creeping larva stage, late creeping larva stage, early peristomial shell larva stage, mid-peristomial shell larva stage, late peristomial shell larva stage, early differentiation epipodes stage, mid-differentiation epipodes stage and late differentiation epipodes stage; (4) Juvenile period made up of 2-stages. early emergence stage of the first breathing poreand emergence stage of the first breathing pore.
Although the fertilized eggs of the Abalone may were incubated under the range of temperature from 18℃ to 34℃, the instantaneous incubation rate and hatching time are veried. The incubative duration of fertilized egg needed eleven hours and thirty-two minutes after the artificial fertilization under the temperature of 18℃, the instantaneous incubation rate was 2.5‰, the average incubation period was twenty-six hours and fifteen minutes. The speed of hatching rate was 0.0381, the finite rate of increase was 1.0010 per hour. But under the water temperature of 22℃, the beginning of hatching time was seven hours and thirty minutes, the average incubation period was twenty-three and five minutes, the speed of hatching rate was 0. 0415, the finite rate of increase was 1.0202 per hour.
Water temperature showed a marked influence upon the incubation period of the fertilization egg. Within the range of 20～28℃, the higher the water temperature, the shorter the incubation time and the higher the incubation rate (Table 2). The time that an embryonic egg developed into a juvenile (emergence of the 1st breathing pore) was 23～ 24 days under water temperature 23.7～28. 5℃ and salinity 32.8‰～34.5‰ (specific gravity 1.0223～1.0229). [Acta Zoologica Sinica 47(3):317 - 323, 2001]
Key words Abalone (Haliotis diversicolor Reeve), Embryonic development, Water temperature
E-mail: ls61@zsu.edu.cn
Structure and function of mucous epithelium of the intestine in Haliotis discus hannai
CUI Long-Bo* LIU Chuan-Lin LIU Xun LU Yao-Hua
(Department of Biochemistry, Yantai University, Yantai 264005, Shandong , China)
Abstract Disk ablone (Haliotis discus hannai Ino) is one of the important marine cultivated species in North China. Brown algae and red algae are the chief foods of disk abalone. They can also swallow small animals. Adopting to their food habit, they have a very long digestive tract. It's about 3 times longer than their bodies. In previous years, studies on the digestion of disk abalone were mostly concentrated to the roles of the digestive glands and those of the digestive tract has been neglected. In this paper the intestine and the rectum of Haliotis discus hannai Ino were studied with TEM and SEM observations, histochemical methods and tests for some enzymatic activities. The intestine can be evenly divided into anterior, meddle and posteriov. The epithelia of the intestine and the rectum are composed of 5 types of cells: (1) The cells with microvilli were the main cell type in the mid and prointestine. Their free borders were closely aggregated of microvilli. They were 2.0 μm long and 0.1 μm in diameter. Under the cell membrane there were many pinocytic vesicles. On the top of the cell there were a lot of mitochondria and multivesicular bodies. In the middle of the cell some secondary lysosomes and many lipid drops of different size were found. In the postintestine at the basal part of the cells, the cell membrane went inward, deeply into the cell to form some finger-like protrusions. Among them, there were many mitoehondria. Some nerves could be seen between the cell membrane and the basal membrane. (2) Ciliated cells' structures were similar to those of the microvilli cells. Instead of microvilli, the free border of the cell was aggregated of cilia. The basal bodies and the ciliary rootlets could be seen clearly. There were a lot of mitochondria among the rootlets. Ciliated cell was the main cell type in the prointestine, typhlosoles of intestine groove and rectum. (3) Gland cells I were distributed in the intestine. Its amount increased from the beginning to the end. There was only a small amount in rectum. The cells were narrow at top and there were some short microvilli on the free border. The striking character of these cells was that a large amount of secreting granules surrounded by membrane dispersed in the cells. At the basal part of the cells, tens or hundreds of granules were surrounded together by membrane again to form bulks which squeezed the neighbor cells and made them narrow in shape. Histochemical studies showed that the granules contained proteinase, nonspecific esterase and lipase, and there were 4 and 3 kinds of polysaccharide enzymes in intestine and rectum respectively. On the apices of many cells, there were swellings containing enzymatic secretion. (4) Gland cells II were mainly distributed in the rectum and typhlosoles. The top and the middle part of the cells were full of secretary granules, the diameter of the granule was about 1.1 μm. The content looked like sand and dense in electron density. The results of the analyses of proteinase, nonspecific esterase and lipase were negative. (5) The goblet cells were distributed mainly in rectum. The histochemical test showed that the goblet cells secreted mucopolysaccharide, and a large amount of mucoprotein filled in the cells. The results showed that the intestine and the rectum of the Haliotis discus hannai Ino were not only tracts for passing food and feces, they also played important roles in digestion. The cells with microvilli exhibited the ultrastructure feature of absorptive cells. They can absorb nutrition and water from the lumen of the intestine. The activity of the alkaline phosphatase at the top region of the cells showed that transportation through cell membrane were carrying on. The gland cells I may secrete digestive enzymes by the apocrine, so extracellular digestion could take place in the lumen. The secretion of the gland cells II may be used to consolidate feces. The vibration of the ciliated cells can facilitate the movement of the food and feces. The mucopolysaccharide secrated by the goblet cells can lubricate the lumen and cement feces. [Acta Zoologica Sinica 47(3):324 -338, 2001]
Key words Haliotis discus hannai Ino, Intestine, Rectum, Structure, Function, Enzyme
*E-mail: lbcui@163.net
Mapping，expression and RFLP of Quox-1 gene in human cell
ZHU Fan* MAO Xin LI Wen-Xin
(College of Life Science, Wuhan University,Wuhan 430072, P.R. China)
Abstract In order to study the relationship between the Quox-1 and the tumorigenesis and cell maligimization, cellular genomic DNA of normal human leukocyte and some tumor cells (LCE and HeLa cells) were extracted using proteinase K method and were restricted with EcoR I . After electrophoresis, and transferred to NC membrane, 32P-labeled Quox-1 cDNA was used as probe to detect the presence of Quox-1 homologous in genomic DNA of normal human, and cancer cells. Normal human leukocyte and some tumor cells (LCE and HeLa cells) were cultured respectively on glass slide with CO2 incubator. Using antibodies of Quox-1, the expression of Quox-1 protein in Normal human leukocyte, and some tumor cell was detected (LCE cell, HeLa cell). Results There was homologous sequence of Quox-1 in normal human leukocyte, and some tumor cells (LCE and HeLa cells), it appears Restriction Fragment Length Polymorphism (RFLP) and may undergo gene amplification. In LCE cells, the gene amplification was much greater than that in HeLa cells. The expression of Quox-1 protein could not be detected both in cell nucleus and cytoplasm of normal human leukocyte. But it was activated and expressed in LCE and HeLa cells. However, in the cell nucleus of LCE and Hela cells，Quox-1 gene had strong expression. There was weak expression of Quox-1 protein even in the cytoplasm. Conclusion These results suggest that the Quox-1 gene may be related with the tumorigenesis and cell aligimization. [Acta Zoologica Sinica 47(3):329-332, 2001]
Key words: Quox-1; Homoebox; RFLP; immunological histochemistry
*E-mail: pzhushzh@public.wh.hb.cn
Anatomical observation on the coronary arteries of Sichuan
golden monkey (rhinopithecus r. roxellanae)
YU Shi-Yuan*
(College of Life Science, Northwest Normal University, Lanzhou 730070, China)
GAO Hui-Xia
(Information center, Environment Protection Bureau of Gansu Province, Lanzhou 730030,China)
Abstract In order to investigate the blood supply features and coronary artery's branch characteristic of Rhinopithecus r. roxellanae heart, we observed the branches and distribution of the coronary arteries of Rhinopithecus r. roxellanae by arterial vascular casting and tissue transparance method.
The results showed that the nutrition of Rhinopithecus r. roxellanae heart is supplied by left and right coronary artery.
The left coronary artery of Rhinopithecus r. roxellanae divided into anterior descending branch and circumflex branch. Anterior descending branch gives off left fringe branch and interventricular septum branch in proper order. Left fringe branch is frequently the main branch of the anterior descending branch and gives off branches to lateral and posterior wall of the 1eft ventricle. The interventricular septum is supp1ied by a 1arge septal branch that originates from the anterior descending branch of the left coronary artery. The main trunk of the anterior descending branch distributes over anterior wall of the 1eft ventricle. Circumflex branch of left coronary artery gives off branches to left atrium and lateral and diaphragm wall of the 1eft ventricle.
The right coronary artery of Rhinopithecus r. roxellanae gives out arterial cone branch, anterior branch and posterior branch of right ventricle, and right fringe branch. Anterior interventricu1ar branch is an extension of arterial cone branch and gives off branches to part of anterior wall of the 1eft and right ventricle and cardiac apex et al. Anterior branch of right ventricle gives off branches to part of anterior wall of the right ventricle. Right fringe branch gives off branches to below fringe wall of right ventricle. Posterior branch of right ventricle distributes over the upper of diaphragm wall of right ventricle. Posterior interventricular branch is an extension of right coronary artery and branches to part of diaphragm wall of left and right ventricle and posterior 1/3 interventricular septum.
The distribution type of left and right coronary arteries on the diaphragmatic surface of Rhinopithecus r. roxellanae heart is the balance circulation type. [Acta Zoologica Sinica 47(3):333-337, 2001]
Key words Rhinopithecus r. roxellanae, Heart, Coronary artery, Casting
*E-mail: syyu@nwnu.edu.cn
Application of noninvasive sampling in conservation genetics *
LI Ming① WEI Fu-Wen① RAO Gang①② FANG Sheng-Guo②③ FENG Zuo-Jian①
(① Institute of Zoology, The Chinese Academy of Sciences, Beijing 100080, China)
(②The Key Laboratory of Conservation Genetics and Reproduction Biology for Endangered Wildlife
Ministry of Education, Hangzhou 371200, China )
(③Scholl of Life Sciences, Zhejiang University, Hangzhou 370012, China )
Abstract Noninvasive sampling technique can be used to address genetic questions of free-ranging animal by getting samples such as hairs, feathers, feces, urine etc. We reviewed different kinds of non-invasive sampling technique, its advantages and disadvantages in this paper. [Acta Zoologica Sinica 47(3):338-342, 2001]
Kev words Noninvasive sampling, Conservation genetics, Endangered species
Effects of hypergravity on the structures and functions
of vestibular and related system
SUN Jiu-Rong*
( College of Life Sciences, Peking University, Beijing, 100871, China)
Abstract Changes in behaviour relevant to the vestibular and related system were studied in long-evans rats conceived, born and housed in 2 acceleration of gravity for 4 months and thereafter exposed to 1 acceleration of gravity. Data from the hypergravity rats were compared respectively with those from the rotation group and the labyrinthectomized group. Static and locomotion modes of the hypergravity rats were changed, tension of extensor was enhanced and the abilities for locomotion equability, orientation in swimming and air-righting reflex were reduced. The adaptation process varied with different behaviours. The time for the recovery of the ability of orientating in swimming was the longest taking more than 1 month.
The adaptation mechanism related to the plasticity of otoconia, hair cell and neurons of vestibular sensory organ as well as evoked the change of releasing of some neurotransmitters and neuromodulators, development of muscles (especially the extensile muscles), expression of Fos in some brain regions. The Fos protein expression provides a useful tool for mapping brain functional activities after sensory stimulation, and showed a low basal level in normal and labyrinthectomized groups. The hypergravity rats, on the another hand, exhibited more Fos- positive cells in the superior colliculus, inferior colliculus, periaqueductal gray, raphe dorsal nucleus and solitary nucleus. In contrast, the inferior olivary nuclei, locus coeruleus and vestibular nuclei were not strongly labeled. These spatial patterns of Fos expression suggest that a reduction in gravity-inertial force may activate a neural pathway different from the vestibulo-olivar pathways activated by an increase in gravity-inertial force. [Acta Zoologica Sinica 47(3):343-346, 2001]
Key words Hypergravity, Vestibular system, Locomotor activity
*E-mail: sjr@pku.edu.cn
Characteristics of resting metabolic rate in little bunting
(Emberiza pusilla) and chestnut bunting (E. r utila)
LIU Jin-Song* ZHANG Zhi-Yan MA Hong HOU Zeng-Suan
( College of Life Science and Technology, Qiqihar University, Qiqihar 161006, Heilongjiang, China )
Abstract The resting metabolic rate (RMR) of birds correlates broadly with the climate of origin. In order to study the characteristics of thermoregulation in Little buntings and Chestnut buntings, are measured their resting metabolic rate (RMR) at ambient temperature (Ta) ranged from 5℃ to 30℃ and calculated thermal conductance. The results are mainly as follows.
1. Thermal neutral zone (TNZ) for Little buntings ranged from 20℃ to 25℃ and Chestnut buntings from 25℃ to 32. 5℃.
2. The minimum metabolic rate of Little buntings and Chestnut buntings were 4.19 ml O2/g·hr and 3.99ml O2/g·hr, respectively, which were 181% and 146 % of the expected values that calculated from their body mass according to Aschoff and Pohl (1970). Above the upper critical temperature, RMR and Tb increased.
3. The minimum thermal conductance (C) of Little buntings and Chestnut buntings were 0.26 ml O2/g·hr·℃ and 0.23 ml O2/g·hr·℃, respectively, which were 138% and 140% of the predicted values calculated according to Aschoff (1981).
In conclusion, the adaptation of Little buntings and Chestnut buntings to environmental climate were by the means of the slight higher level RMR and thermal conductance, and high intensity of chemical thermoregulation. [Acta Zoologica Sinica 47(3):347-350, 2001]
Key words Little buntings (Emberia pusilla ), Chestnut buntings ( E. rutila ) , Resting metabolic rate( RMR ) , Thermoregulation
E-mail: liujinsong@sina.com
Acute myocardial necrosis by an iodinated monoclonal antibody
of cardiac myosin heavy chins in rat
ZHU Xiao-Juan①③* YUAN Mao-Kun③ ZHAO Jian-Zeng① LI Rong-Wen①
GAO Feng-Tong③ HOU Li-Zhong① YANG Tong-Shu①
(① Institute of Biology Engineering, Jilin University, Changchun 130021, China )
(② Third Clinical Medical College, Jilin University, Changchun 130021, China )
(③Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China )
Abstract Monoclonal antibody (MAb) attained from hybridoma lA5 was against human cardiac myosin heavy chains (CMHC) and didn't react with smooth muscle. It was labeled with Na 125I and injected to the rats suffered from acute myocardial necrosis, which were treated by isoproylarterenol. After injection, the rats were studied from the 4th to the 48th hours by relative radioactivity in blood, thyroid gland, stomach, liver, lung, muscle and heart, respectively. The results indicated that l25I-CMHC-MAb could specifically reach the target necrotic organ in the 4th hours (2.03±0.60 ) and remained up to the 48th hours (2.55 ± 0.49) or longer, at which the 8th was higher (4.89 ± 0.44). Meanwhile 125I-CMHC-MAb showed much higher affinity to necrotic cardiac tissue than the normal (P<0.001). The same results were should in the blood from 4th (4.94 ± 0.45) to 48th (2.70 ± 0.12). Thus 125 I-CMHC-MAb can be considered as an effective tool for image diagnosis of acute myocardial [Acta Zoologica Sinica 47(3):351-354, 2001]
Key words Myosin, Myocardial necrosis, Monoclonal antibody
*E-mail: xingzhi@nenu.edu.cn
Patterns of lectin binding in the hemopoietic organs and blood cells of
Ctenopharyngodon idellus and Trionyx sinensis
GUO Qiong-Lin*
( Institute of hydrobiology, The Chinese Academy of Sciences, Wuhan 430072, China )
Abstract Lectin binding patterns in the hemopoietic organs and blood cells of Grass carp (Ctenopharyngodon idellus), Chinese soft-shelled turtle (Trionyx sinensis) were observed with two affinity histochemical methods ABC staining and fluorescence-bound lectin staining. Concanavalin (ConA), Phytohemagglutinin (PHA) Pokeweed agglutinin (PWM) and Lipopolysaccharide (LPS) binding sites were distributed mainly on the plasma membrane of lymphocytes. ConA, PHA binding sites were distributed in thymus, cordic structure of head kidney and scatterly spleenic tissue of C. idellus, also in thymus and periellipsoidal lymphatic sheaths and marginal region of T. sinensis. PWM, LPS binding sites were scatterly distributed in cordic structure of head kidney of C. idellus, spleenic tissue of C. idellus and T. sinensis, but positive cell groups were not obvious. ConA, PHA and PWM positive percents of lymphocytes in blood smears were repectively 36 %～41%, 33 %～37 %, 38 %～43 % in C. idellus and 37 % ～48 %, 36 % ～45 %, 44 % ～53 % in T. sinensis, but percents of positive cells were repectively 15%～18%, 14% ～17%, 9% ～12% in imprints of head kidney of C. idellus. [Acta Zoologica Sinica 47(3):355-359, 2001]
Key words Grass carp (Ctenopharyngodon idellus), Chinese soft-shelled turtle (Trionysc sinensis), Lymphocyte, Lectin binding pattern (receptor), Histochemistry
*E-mail: mxiao@wuhee.edu.cn
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