бб

ACTA

ZOOLOGICA

SINICA

бб

CHINESE

бб

ACTA ZOOLOGICA SINICA

бб

Vol. 47 No.6 Aug. 2001

бб

ABSTRACT

бб

Maximum metabolic rate in plateau pikas

(Ochotna curzoniae) and root voles (Microtus oeconomus)

WANG Yu-Shan WANG De-Hua WANG Zu-Wang*

( State Key Laboratory of Integrated Management of Pest Insects and Rodents,

Institute of Zoology, The Chinese Academy of Sciences, Beijing 100080, China)

Abstract The maximum metabolic rates (MMR) of mammals may influence their survival by putting the upper limit for sustained activity or thermogenesis, and affect rates of reproduction and distriibution. For the small mammal which inhabit on Qinghai-Tibetan Plateau, the thermogenesis may be most important to their survival. Plateau pikas (Ochotona curzoniae) and Root voles (Microtus oeconornus) are two dominant species in alpine meadow on Qinghai-Tibetan Plateau, the former is an epidemic species to the area, and the latter is a species with wide geographical range. We determined their cold-induced MMR in summer and winter. The metabolic rates were measured as oxygen consumption in closed-system Kalabukhov-Skvortsov respirometer under different temperatures (15бц, 10бц , 5бц , 0бц , - 5бц and - 10бц ). For each animal, the experiment was first carried out in normal atmosphere that initially filled the chamber, after 30бл40 minutes, the resting metabolism rate was got, then enough helium-oxygen mixture (79 % He and 21% O₂) was blown rapidly through the chamber replacing the air in it. The chamber was sealed at once again, and the metabolic rates were measured for not less than 15 minutes under same ambient temperature. The metabolic rates in helium-oxygen mixture were much larger than that in air. We got the MMR under 5бц and 0бц for Root voles and Plateau pikas respectively in summer, and in winter, 0бц and -5бц respectively (Fig.1 and Fig.2). Their maximum metabolic rates change from 5.93 (118 g) and 12.70 (27.8 g) ml O₂/gбдh in summer to 6.33 (115.8 g) and 18.29 ml O₂/gбдh (17.5 g) in winter for Plateau pikas and Root voles respectively (Table 1 and Table 2). The variations were not significant. We suggested that the two mammals lived under low temperature. Even in summer, the environmental temperature may be low, and it could decrease to 0бц around at night. This led to relatively large increase in MMR in summer. On the other hand, the mammals may use other mechanisms such as behavior for wintering, and huddling is very important for thermoregulation for them. These increased the MMR relatively in summer with comparison of that in winter. The mammals are stressed by low temperature which can increase MMR and low oxygen pressure which can decrease MMR, and this let the maximum metabolic rates similar to that of small mammals living in other habitats. The MMR of Root voles living on Qinghai-Tibetan Plateau and Red-backed vole (Clethrrionomys rutilus) from Alaska, both of them belonging to Microtine are higher than their sympatric species, Plateau pikas and Snowshoe hares (Lepus americanus), which belong to Lagomorphs (Table 3). This suggested that existed inter-species variation in MMR for mammals. The thermogenic capacity of small mammals inhabiting on Qinghai-Tibetan Plateau may have been evolved under the unique climate, just as Hayes and O' Connor suggested for high-altitude Deer mice. Further studies such as genetics, biochemistry and molecular biology on the mammals living in this area are needed. [Acta Zoologica Sinica 47(6): 601-608, 2001]

Key words Plateau pika (Ochotona curzoniae), Root vole (Microtus oeconornus), Maximum metabolic

Qinghai-Tibetan Plateau

*E-mail: wangzw@panda.ioz.ac.cn

E-mail: wangys@panda.ioz.ac.cn

бб

Analysis of helminth population and community composition

parasitic in striped mullet (Mugil cephalus L.)

LÜ Jun-Yi WU Jin-Ying YANG Da-Wei CHEN Zhi-Sheng ZENG Hua

(School of Life Science, Zhongshan University, Guangzhou 510275, China )

Abstract We reported ecological characters of the helminth community parasitic and relationship between them and Mugil cephalus L. (1 018 fish) along Yuedong area in Guangdong Province, China. We collected 2 133 indiulduals of helminthes from January to December in 1995. The results indicated that the hosts were infected by one to nine species of helminthes, including eight kinds of Digeneans and a classification of Acanthosentis.

The component communities of helminth parasitic in the hosts were dominated by digeneans. The total prevalence was 24.4 %, the range of intensity and mean intensity was 1бл81, and 8.8 respectively. The total mean abundance was 2.3.

The prevalances of Elonginus mugilus and Prohaplosplanchnus diorchis decreased with the increasing of body length of hosts (F > F_0.01, F-test), but the relationship between the body length of hosts and the prevalances of Haplosplanchnus cuneatus, H. elongatus, H. purii, Saccocoelioides huidongensis, Tandernorchis yuedongensis, Elliptobursa singlorchis and Neoechinorhynchus tylosuri was not obvious (F < F_0.05, F-test), and it was irregularly fluctuated. The results indicated that the intensity of infection of Haplosplanchus cuneatus, H. elongatus, Saccocoelioides huidongensis and Haplosplanchnus purii increased with the increasing of the body length of the hosts, the relationship between the intensity of other five species and the body length of the hosts was not apparent (P >0.05, F-test). The prevalence of helminth parasitic in host depended on the body length group of hosts, which was arranged as following 70бл90 mm, 90бл110 mm, 110бл130 mm, 50бл70 mm, 130бл150 mm, 150бл170 mm and >170 mm.

The prevalance and abundance in different month was different (F > F_0.01, P < 0.01, F-test), except in August and September, October and November (F< F_0.05, P >0.05, F-test). On the basis of the prevalence and abundance variety, Saccocoelioides hundongensis is the dominent population. The prevalence and population size of helminth parasitic in hosts arranged according to months of the year from the most to the least as follows: January, February, December, October, November, September, June, July, August, March, April and May. The helminth component community can be divided into 3 sub-component communities according to fuzzy clustering analysis, each having its own composition features. The results showed that the number of mean species, dominance index, infection index, species diversity, mean abundance and intensity of infection in the subcomponent community of helminths parasitic in host of less 70 mm body length were relatively low, and that the characteristic value of subcomponent community of helminths parasitic in host of more than 70 mm body length increased with the increasing of body size, except the number of mean species. [Acta Zoologica Sinica 47(6): 609-619, 2001]

Key words Helminth, Striped mullet (Mugil cephalus), Population and community composition

E-mail: Ls61@zsu.edu.cn

бб

Spatial distribution patterns of Chinese Gerbils

(Gerbilline) in relation to environmental factors

ZHOU Li-Zhiв┘в┌ MA Yongв┘* LI Di-Qiangв█

(в┘ Institute of Zoology, The Chinese Academy of Sciences, Beijing 100080, China )

(в┌ Biology Department, Huaibei Coal Industry Teachers' College, Huaibei, Anhui 235000, China)

(в█ Institute of Forest Ecological Environment and Protection,

The Chinese Academy of Forestry, Beijing 100091, China)

Abstract There are seven species of Gerbils belonging to three genera mainly in the arid regions of Mongolian-Xinjiang Plateau and its adjacent arid regions. Their distributional ranges include Xinjiang, Gansu, Ningxia, Qinghai, Inner Mongolia, Shannxi, Shanxi, Hebei, Jilin, Liaoning and Henan. They are important biodiversity indicator in the arid ecosystems.

An approach of geographical information systems (GIS) along with multivariate analysis was applied to study the spatial distribution patterns of the gerbil species and their correlation with environmental variables using resources and environment data of China on a scale of 1: 4 000 000. Geographic distribution data were collected from literatures and museum collections. All the available geographic distribution data were used in habitat analysis with MapInfo Professional Version 4.0, and then used to get digital species range maps. Environmental variables in the distribution area of gerbils collected with the 12 392.4 km2 equal-area grids were used for factor analysis to determine the principal factors that affected the distribution patterns of gerbils. We found that distribution of gerbils ranged from 34.4~to 50.2~N and from 74. l~to 124. I~E, with altitudes from -100 to 2 500m. The gerbils mainly inhabited desert and semi-desert with altitudes from 1 000 to 1 500 m. There were more species in west than in east part of the distribution area, where the richest species of gerbils were along north piedmont of Tianshan from the Alataw Shankou to Jiangjun Gobi, and in western Hexi Hallway and Ruo Shui Valley as well. The factor analysis on the environmental variables which affect the distribution of gerbil revealed that diversity of soil, vegetation and geomorphologic and altitude difference played a dominant role as a basic landscape factor, that total annual solar radiant energy and mean annual rainfall consisted of another principal factor, a factor of aridity and desertlization tendencies, and that'the third was temperature. The figure of R-analysis on the loadings of environmental variables performed by the three principal factors indicated that species number of gerbils in grids was close related to mean annual temperature and desertlization tendencies. The analysis of the scores in the gerbils' distribution grids revealed that higher the species richness, higher the probability of aridity and desertlization, higher temperature and more complex the habitats were. [Acta Zoologica Sinica 47(6): 616-624, 2001]

Key words Gerbils, GIS, Spatial distribution patterns, Species richness, Factor analysis

E-mail: hb196311@mail.ahbbptt.net.cn

бб

Genetic differentiation within Drosophila auraria

species complex revealed by random polymorphic DNA (RAPD)

ZHAO Zhong-Ming LU Jian DAI Zhuo-Hua*

(College of Life Sciences, Peking University, Beijing 100871, China)

Abstract The genetic polymorphisms of 12 geographic populations of Drosophila auraria species complex (D. auraria, D. biauraria, D. triauraria, D. quadraria and D. subauraria) were analyzed by PCR Random Amplified Polymorphic DNA (PCR-RAPD). Of the 40 random primers used in this analysis, 30 efficiently produced 161 RAPD bands, of which 129 were polymorphic. The dendrogram constructed using UPGMA algorithm based on the RAPD data showed that D. subauraria was the oldest species in the species complex, D. biauraira completed speciation next. Compared with D. subauraria and D. biauraria, D. auraria, D. quadraria were relatively younger. This result does not agree with conclusions of the previous studies that D. quadraria was the ancestral species of Drosophila auraria species complex. RAPD markers provided and valuable information for reconstructing speciation history of Drosophila auraria species complex. [Acta Zoologica Sinica 47(6): 625-631, 2001]

Key words Drosophila, Drosophila auraria species complex, RAPD, Phylogenetic relationship

*E-mail: dzhuoha@pku.edu.cn

бб

On the taxonomic status of Yangtze sturgeon, Asian and American

green sturgeon based on mitochondrial control region sequences

ZHANG Si-Mingв┘в┌ WU Qing-Jiangв█ ZHANG Ya-Pingв┘

(в┘Lab of Cellular and Molecular Evolution, Kunming Institute of Zoology,

The Chinese Academy of Sciences, Kunming 650223, China)

(в┌Yangtze River Institute of Fisheries, Chinese Academy of Fisheries Sciences, Jingzhou 434000, Hubei, China)

(в█Institute of Hydrobiology, The Chinese Academy of Sciences, Wuhan 430072, China)

Abstract It has been controversial whether Asian Green sturgeon and American Green sturgeon(Acipenser medirostris) are valid species, and the relationship among Yangtze sturgeon (A. dabryanus) and other species,especially Chinese sturgeon (A. sinensis),has also been confusing. The control region (D-loop) of mitochondrial DNA (mtDNA) was amplified and sequenced to address these problems. Our sequence data support the following hypotheses: (a) Yangtze sturgeon may be the most closely related to the Chinese sturgeon, and is likely to be a landlocked form of Chinese sturgeon; and (b) green sturgeon inhabiting North America and Far East Asia may belong to the same species, A. medirostris. However, more data are needed to test these ideas. [Acta Zoologica Sinica 47(6): 632-639, 2001 in English]

Key words Green sturgeon, Yangtze sturgeon (Acipenser dabrayanus), Chinese sturgeon (Acipenser sinensis), Systematics, Mitochondrial control region

E-mail: zhangsm@unm.edu

бб

Molecular taxonomy and phylogeny of four species of

Eriocheir (Decapoda: Brachyura: Grapsidae)

QIU Gao-Feng XU Qiao-Ting WANG Li-Qing FAN Zheng CHEN Xue-Jing

( Key Laboratory of Ecology and Physiology in Aquaculture, Ministry of Agriculture,

Fisheries College, Shanghai Fisheries University, Shanghai 200090, China )

Abstract The taxonomy and phylogenic relationships of four species of Eriocheir were investigated by mitochondrial DNA sequencing analysis. Parts of 16S ribosomal DNA (16S rDNA) were amplified with the polymerase chai reaction (PCR). A total of 537 bp nucleotide site was sequenced for 6 individuals from Eriocheir sinensis an E. leptognathus, which were collected from Chong-ming island and San-jla Port in Shanghai, respectively. 3 (8.78%) polymorphic nucleotide sites were detected. 28 (7.45 % ) sites were interspecific sequence variatio and the intraspecific genetic difference was small. Four haplotypes were found in both E. sinensis and E. leptognathus. Combined with the previously reported sequences (376 bp) from Pachygrapsus transversus (Sturm. bauer et al., 1996), Sesarma ayatum, Chasmagnathus granular (Schubart et al., 1998), E. japmmonicus and E. formosa (Cheng et al., 1998), phylogenetic trees were constructed with Maximum Parsimony and Neighbor-Joining method. The results indicated that E. sinensis to be more closely related to E. japonicus than to any other Eriocheir species. E. formosa formed a clade with E. sinensis and E. japonicus that was separated from the relatively independent clade formed by E. leptognathus. However, The two clades were clustered into the same group, although the evolutionary speed of E. leptognathus was faster than those of other Eriocheir species. This result was not consistent with Sakaiбпs and Guoбпs views (Sakai, 1983; Guo et al., 1997) that E. leptognathus and E. formosa were refered to a new genus, respectively. This research also indicated that the genetic variation was very low in the commercially important species E. sinensis because of the fast flow of gene caused by artificial transplant in aquaculture. [Acta Zoologica Sinica 47(6): 640-647, 2001]

Key words Eriocheir, 16S rDNA sequence, Molecular taxonomy, Phylogeny

E-mail: qiugf@online.sh.cn

бб

Modulation of GABAA receptors by pregnenolone

and pregnenolone sulfate in mouse brain during convulsion

ZHOU Xue-Rui

( Department of Biology, Huaiyin Teachers College, Huaiyin 223001, Jiangsu, China)

.LI Xiao-Yu ZHU Jian-Qin*

(Department of Biological Science and Technology, Nanjing University, Nanjing 210093, China)

Abstract Recent investigation shows the regulation of GABAA receptor by neurosteroids in mammalian brain. Though progesterone can be used as psychotropic drug via its interaction with GABAA receptor, the role of pregnenolone (Pe) and pregnenolone sulfate (Pes) remains unclear. To further elucidated neurosteroids on the modulation of GABAA receptor, we investigated the effect of Pe and Pes on the ³H-GABA binding to the GABAA receptors in hypothalamus, cerebral cortex, hippocampus and cerebellum in mice using a convulsion model induced by kainic acid. The results showed that Pe increased the binding of ³H-GABA to the GABAA receptors in the tested brain areas. The increase of binding in the hypothalamus, hippocampus and cerebellum was significant (P < 0.05 or P<0.001), but the increase of binding in cerebral cortex was not significant (P >0.05). The modulation effect of Pe on GABAA receptors was blocked by picrotoxin, and consequently the kainic acid-induced convulsion was inhibited. On the other hand, Pes markedly decreased the binding of the ³H-GABA to GABAA receptors (P <O.01 or P< 0.001) in all of the tested brain areas and enhanced the potency of kainic acid-induced convulsive. These results suggest that Pe had marked sedative and anti-convulsion effects, which were probably mediated by the GABAA receptors. [Acta Zoologica Sinica 47(6): 648-653, 2001]

Key words Mouse, GABAA receptor, Pregnenolone, Pregnenolone sulfate, Kainic acid, Picrotoxin

*E-mail: zjq@nju.edu.cn

E-mail: xueruizhou@263.net

бб

The expression of nov gene coincides with development and

differentiation of brain during the evolution from fish to mammals

SU Bing-Yin CAI Wen-Qin ZHANG Cheng-Gang*

( Department of Histology and Embryology, Third Military Medical University, Chongqing 400038)

Bernard Perbal

(Laboratoire dбпOncologie Virale et MoleculaireгмUFR de Biochimieгм

Universite Paris 7-DгоDiderotгм2 Place Jussieuгм75005гмParisгмFrance)

Abstract The nov gene is a founding member of the expanding CCN family of genes, which encode proteins reported to be involved in a variety fundamental biological functions. Our previous studies have confirmed that nov gene might be involved in the acquisition of specific functions during postnatal development of rat central nervous system. The aim of this study is to reveal the relationship between nov expression and development, differentiation of brain during the evolution from lower vertebrate----fish to higher vertebrate----mammals. Comparative distribution of nov mRNA-positive neurons in the brain of Silver carp, chicken, cattle, dog and cat was studied by using in situ hybridization histochemistry. The results were as follows: a few of nov mRNA-positive neurons were detected in dorsal motor nucleus of vagus, motor nucleus of the trigeminal nerve, anterior thalamic nucleus, posterior thalamic nucleus, dorsal hypothalamic nucleus and lateral hypothalamic nucleus of lower vertebrate Silver carp. In addition to the pons positive neurons were also found in cerebellum, most nucleus of thalamus and hypothalamus, and corpus striatum of chicken. Whereas a lot of nov mRNA-positive neurons were widely localized in the brain of mammals--cattle, dog and cat, strong positive signal was found in pons, cerebellum, thalamus, hypothalamus and cerebrum of these species. Our results indicated that nov gene is very conservative during the evolution of brain from lower vertebrate--fish to higher vertebrate--mammals, and the expression of nov gene coincided with the development and structure and function differentiation of brain. [Acta Zoologica Sinica 47(6): 654-658, 2001]

Key words Silver carp, Chicken, Mammal, Brain, Nephroblastoma overexpressed gene (nov) mRNA, In situ hybridization histochemistry, Comparative development

E-mail: bysu@mail.tmmu.com.cn

бб

Effects of the short interval successive partial

hepatectomy on rat survival and liver tissue structure

XU Cun-Shuan LI Yong-Hui DUAN Rui-Feng LU Ai-Ling XIA Min JI Ai-Ling

(College of Life Sciences, Henan Normal University, Xinxiang 453002, Henan, China)

Abstract In the paper, we analyzed 4 types of short interval successive partial hepatectomy (SISPH) models (E, G, K and M) were established after the possibility of that were analyzed with 12 types of SISPH models. The effects of SISPH on rat survival and its liver tissue structure were observed with histological method. According the results of A~D models, when the amount of liver once resected is up to 46 %, 75 % or 85 %, the survival rates of rat (Rattus norvegicus) were above 75%, but when the amount of liver once resected was up to 92.5%, the rats were all dead after 24 hours. Among the eight types of SISPH models (EблM), lobus exter-nalis sinister resected in first time, lobus centralis sinister and lobus centralis resected in second time, lobus dexter sinister resected in third time and lobus caudatus sinister resected in forth time, and E (3 times of liver resection, interval time between resections was 4, 4 and 4 hours, respectively in turn), G (3 times of liver resection, interval time between resections was 36, 4 and 4 hours, respectively in turn), K (4 times of liver resection, interval time between resections was 36, 36, 36 and 36 hours, respectively in turn) and M models (4 times of liver resection, interval time between resections was 4, 36, 36 and 36 hours respectively in turn) were selected out, according the survival rate and interval time, to proceed histological analysis. SISPH could affect the liver on the histological changes, and among that, the changes of hyperplasia of liver and mitosis of chymohepatocytes were, like in E model, most conspicuous. Almost all the hepatocytes of remanent liver lobes became hyperplasia, and the mitotic index was 15 times of the control. The sharp hyperplasia and quick proliferation gave rise to a series of changes of the liver structure, like obvious confusion of the structure of liver lobes and narrowing of the liver sinusoid. In the other hand, the percentages of binuclear hepatocytes of all the hepatoctes might also be affected by the SISPH, but there were much distinct among the 4 models. The results showed that the amount of the liver once resected might not exceed 85 % of the whole liver, and the sustain limit of rat to SISPH is dependent on the amount of liver resected and the interval time between successive partial hepatectomy. It was found that the models as represented by groups E, G, K and M, can be considered as useful models for studying the change of liver structure, the rate of rat survival in SISPH as well as the molecular mechanism of rat liver regeneration. [Acta Zoologica Sinica 47(6): 659-665, 2001]

Key words Rat, Model, Short interval successive partial hepatectomy (SISPH), Liver regeneration, Liver tissue structure

E-mail: xucs@henannu.edu.cn

бб

Immunohistochemical localization of the endocrine cells

in the digestive tract of Trimeresurus stejnegeri

ZHANG Zhi-Qiang ZHANG Sheng-Zhou WU Xiao-Bing*

(College of Life Science, Anhui Normal University, Wuhu 241000, Anhui , China)

Abstract At present, the gastrointestinal tract of reptiles has received a little attention from comparative endocrinologist and the available information being mainly limited to the crocodiles and lizards. To date there have been few studies of regulatory peptides in the gastrointestinal tract of snakes. However, Masini reported immunoreactivitl for gastrin \cholecystokinin (combined) and somatostatin in small intestinal endocrine cells of three species of snake, the Asp viper (Vipera aspis), the Grass snake (Natrix natrix) and the Viperine snake (N. maura), but found that immunoreactivity to pancreatic glucagon, secretin or motilin was absent. In addition, somato-statin-containing cells had been detected in the stomach and small intestine of the European adder (Viper aberus). There is, however, no systematic study of regulatory peptides of the entire length of the digestive tract in any snake. In view of the evolutionary position of reptiles, an investigation of the gastrointestinal regulatory peptide system was considered to be of particular interest. Trimeresurus stejnegeri, a member of the reptilia suborder serpents, was studied as an example of poisonous snakes. The purpose of this study was to conduct a detailed examination of the distribution of the endocrine cells in the digestive tract of this snake. The tunica mucosa of the entire length of the digestive tract was examined to determine the types, relative frequencies and distributions of endocrine cells found there by using an immunohistochemical method (Avidin- biotin- peroxidase complex method). The eight gastrointestinal peptides involved, which had been reported to occur in mammals, were 5-Hydroxytryptamine, somatostatin, giucagon, vasoactive intestinal polypeptide, gastrin, substance P, pancreatic polypeptide and insulin. The results showed that four of them were observed in the digestive tract of Trimeresurus stejnegeri, namely 5-Hydroxytryptamine, somatostatin, giucagon and vasoactive intestinal polypeptide. 5 - Hydroxytryptamine and somatostatin- immunoreactive cells distributed throughout the digestive tract, while 5-Hydroxytryptamine-immunoreactive cells were found with the highest density in the duodenum and ileum, the second highest in the jejunum, rectum and stomachus pyloricus, numberously in the stomachus corpus, occasionally in the oesophagus and stomachus cardiacus. Somatostatin-immunoreactive cells were mainly located in the duodenum, numberously in the stomach pyloric, stomachus corpus and jejunum, but rarely in any other regions. The glucagon-immunoreactive cells were only present in the duodenum and jejunum. The vasoactive intestinal polypeptide- immunoreactive cells were found moderately in the corpus stomachus and cardiacus, while sparsely in the oesophagus. Data are statistically different among these regions. No immunoreactivity was observed with the antisera to gastrin, substance P, pancreatic polypeptide and insulin. The evidence collected in this study indicates that the distributive mode of endocrine cells in the digestive tract of Trimeresurus stejnegeri differs from the other snakes. This may be related to its feeding and habitation. [Acta Zoologica Sinica 47(6): 666-671, 2001]

Key words Trimeresurus stejnegeri, Digestive tract, Endocrine cells, Immunohistochemistry

*E-mail: yxbwu@263.net

бб

Immunorecognition of estrogen and androgen receptors in the nervous

system and hatschek's pit of amphioxous (Branchiostoma belcheri)

WENG You-Zhu FANG Yong-Qiang* HU Xiao-Xia

(Third lnstitute of Oceanography , SOA , Xiarnen 361005, Fuiian , China)

Abstract Previous studies discovered that Amphioxus (Branchiostoma bekheri ) had primitive reproductive endocrin regulation axis (brain-Hatschekбпs pit-gonads) which synthesizes and secretes sex steriod hormones (estrogen androgen and progestergone). However, the feedback regulation of sex steroid hormones on brain am Hatschek's pit (homologous with the pituitary gland) of Amphioxus had not been reported so far. Thus, we used polyclonal antibodies against estrogen and androgen receptors to investigate the immuno-recognition of estrogen and androgen receptors in nervous system and Hatschekбпs pit of Amphioxus at different development stages.

Amphioxus were collected from the Tong'an-Qiongtou sea area near Xiamen. Each fish was divided into three segments, head (from tentaculum peristomial to the first pair of gonads), medial (from the first pair of gonads to the amus) and tail. Samples were fixed in Bouinбпs solution without acetic acid for 8--12 h, then embedded in paraplast. Sections with the structure of brain and Hatschekбпs pit were identified under a light microscope and selected for immunostaining with the SABC technique.

The results showed that the immuno-recognition of estrogen and androgen receptors existed in the telencephalon and the middle and posterior part of midbrain. Immunopositive substances mostly located in the nucleus of nerve cells, with a few in the cytoplasm. The dorsal side and middle region of nerve tube also showed immunopositive reaction, and the immunopositive substances of estrogen and androgen receptors distributed in the nucleus, cytoplasm and nerve fibers of nerve cells. But immunonegative reaction was showed in the hindbrain. On the other hand, immuno-recognition of estrogen and androgen receptors existed in the nucleus or the cytoplasm of epithelial cells (primitive gonadotrophic cells in Amphioxus) in the base part of Hatschekбпs pit at different stages of development. The other two cells in Hatschekбпs pit showed immunonegative reaction.

The results demonstrate for the first time that estrogen and androgen receptors exist in cephalochordata Amphioxus, and the immunorecognitive regions of the estrogen and androgen receptors in the brain and Hatschekбпs pit are similar to those of vertebrates. It suggests that sex steroid hormones exert feedback regulation on brain and Hatschekбпs pit, and stimulate the development and maturity of the reproductive endocrine regulation axis function in Amphioxus. These facts further demonstrate a system of reproductive endocrine regulation in Amphioxus similar to, or with close relationship with that in vertebrate. On the other hand, we suggest that the regulatory role of sex steroid hormones on gonads function and reproduction behavior may be direct or indirect through sex steroid hormones receptors in brain. [Acta Zoologica Sinica 47(6): 672-676, 2001]

Key words Amphioxus (Branchiostoma belcheri), Estrogen and androgen receptor, Immuocytochemistry, Nervous system, Hatschekбпs pit

*E-mail: fant98@public.xm.fj.cn

бб

In vitro induction and tissue and cell origin of

interferon in Grass carp (Ctenopharyngodon idellus)

SHAO Jian-Zhong XIANG Li-Xin

(College of Life science, Zhejiang University, Hangzhou 310012, China)

Abstract Interferon is an important cytokine that has been shown to have many functions such as anti-virus and immunomodulation. The tissue and cell origin of interferon had been well studied in humans and higher vertebrates, but less known in fishes. This paper reported the tissue and cell origin of Grass carp (Ctenopharyngodon idellus) interferon for the first time in order to investigate and understand the biological role of interferon in fish. The in vitro induction experiment of Grass carp interferon from nine tissues of peripheral blood, head kidney, kidney, thymus, spleen, liver, intestines, heart and muscle were undertaken. The results showed that the Grass carp interferon was mainly produced by the immune-related tissues of head kidney, spleen, peripheral blood and thymus which were full of leucocytes. Therefore, the leucocytes were further isolated from these tissues by Percoll gradient and density centrifugation and induced the interferon in vitro. The results confirmed that leucocytes were interferon-producing cells. Biochemical and biophysical characteristic studies indicated that the leucocyte interferon induced in vitro had same character with serum interferon induced in vivo. Furthermore, the antigenic characteristic analyses of neutralization test, dot-ELISA and Western blot were performed between two kinds of interferon, and the results also showed that the leucocyte interferon was the same as that of serum interferon. In order to find out what kind of cell in leucocytes was the interferon-producing cell, lymphocytes were purified from leucocytes by Ficoll-Urografin gradient and density centrifugation for the further induction of interferon, and the results showed that the lymphocytes were the interferon-producing cells. Finally, T and B lymphocyte was selectively separated by genetic incorporation of BudR and ultraviolet illumination inactivation technique to induce interferon respectively, the results indicated that the Grass carp interferon was produced by T lymphocyte. [Acta Zoologica Sinica 47(6): 677-683, 2001]

Key words Grass carp (Ctenopharyngodon idellus), Interferon, In vitro induction, Tissue and cell origin

E-mail: lscshaoj@mail.hz.zj.cn

бб

Effects of cycloheximide (CHX) on in vitro maturation of porcine oocytes

LI Guang-Pengв┘ MENG Qing-Gangв┘ WEI Pengв┘ YU Yuan-Songв┘

CHANG Zhong-Leв┌ TAN Jing-Heв┘в┌

(в┘Department of Biotechnology, Northeast Agricultural University, Harbin 150030, China)

(в┌ College of Animal and Veterinary Sciences, Shandong Agricultural University, Taian 271018, Shandong)

Abstract Since it was reported that protein synthesis and RNA transcription during early hours of culture is essential for germinal vesicle breakdown (GVBD) in in vitro maturing oocytes, a series of experiments have been designed to study the effects of cycloheximide (CHX), an inhibitor of protein synthesis, on GVBD, condensation of chromatin, oocyte maturation to MвЄstage and the cumulus expansion of porcine oocytes cultured in vitro.

Porcine cumulus-oocyte complexes (COCs) were aspirated with a syringe from the follicles of 2бл6 mm in diameter on the ovarian surface and cultured in mTCM-199 containing 10% FCS, 10 IU/ml PMSG and 10 % pFF, in an atmosphere of 5% CO2 in air at 38.5бцбл39бц. Five experiments were conducted in this study. In experiment 1, the nuclear changes during maturation of oocytes cultured under normal conditions were observed. In experiment 2, effects of CHX concentration on oocyte maturation were studied. In experiment 3, oocytes were first cultured in medium containing CHX for 24 hours and then transferred to and cultured in normal maturation medium to study the effect of protein synthesis during first hours of culture on porcine oocyte GVBD. In experiment 4, oocytes were first cultured in CHX-containing medium for 0, 6, 12, 24 and 48 hours before being transferred to and cultured in normal maturation medium up to 48 hours to study the effect of CHX during early hours of culture on oocyte maturation to M II stage. In experiment 5, at different hours of culture in normal maturation medium, oocytes were transferred to and cultured in CHX-containing medium up to 48 hours to study effects of CHX during later hours of culture on oocyte maturation to M II stage.

The results obtained were as follows: (1) GVBD was completely blocked when CHX was added at concentrations of 1, 10, or 50 ж╠g/ml. However, GVBD occurred in some oocytes when CHX was at 0.1 ж╠g/ml. The CHX inhibition of GVBD was fully reversible and once CHX was removed GVBD occurred very soon. When the COC was first cultured in the presence of 10 ж╠g/ml CHX for 24 h and then transferred to and cultured in normal medium (without CHX), GVBD occurred at 4бл8 h, 3 to 6 times earlier than that occurred under normal condition (20-24 h); (2) When the COC was first cultured in the presence of CHX for 0, 6, 12 and 24 h, and then transferred into normal medium and continued to culture up to 48 h, the M II maturation rates of oocytes were 84.1, 77.1, 48.9 and 27.8 %, respectively; (3) When the COC was cultured in normal medium for 0, 6, 12, 24, 36 and 48 h, then cultured in CHX media up to 48 h, the maturation rates were 0, 0, 0, 31.3%, 65.4% and 79.5%, respectively. In summary, results 2 and 3 indicated that with extension of exposure to CHX, whether during early or late hours of culture, oocyte maturation to M II stage was reduced markedly.; (4) CHX inhibited the expansion of cumulus cells. Based on these results, we conclude that the GVBD and maturation to MII stage of porcine oocytes needed de novo synthesis of proteins. [Acta Zoologica Sinica 47(6): 684-690, 2001]

Key words Pig, Oocytes, In vitro maturation, Cumulus expansion, Cycloheximide

E-mail: guangpengli@yahoo.com

бб

Cloning and sequence analysis of Yak (Poephagens grunnieus) ж┴-lactalbumin gene

FAN Bao-Liangв┘ Zhao Zhi-Huiв┌ LI Ningв█* WU Chang-Xinв▄

(в┘ Hebei Handan Agricultural College, Handan 057150, Hebei, China)

(в┌ The Quartermaster University of PLA , Changchun 130062, China)

(в█National Laboratory for Agrobiotechnology of China Agricurtal University, Beijing 100094 ,China )

(в▄ College of Animal Science and Technology of China Agricultural University, Beijing 100094, China )

Abstract Four pairs of primers (1, 2, 3 and B) overlapping each other were designed and synthesized according to the nucleotide sequence of bovine ж┴-lactalhumin gene. With these primers we had cloned and sequenced the ж┴-lactalbumin gene of Yak (Poephagens grunnieus) by PCR. The results demonstrated that the full length of this gene in Yak was 2 999 bp. The sequences covering the nucleotides from 1 to 674 and from 2 361 to 2 793 were the 5-flanking region and the 3'-flanking region respectively. The coding region of this gene covered more than 2 kb from nucleotide 675 to 2 793 and contained four exons (located in 675бл830 nt, 1 152-1 310 nt, 1 782бл1 857 nt, 2 361бл2 793 nt respectively) and three introns (located in 831бл1 151 nt, 1 311--1 781 nt, 1 858-2 360 nt respectively). It encoded a peptide containing 142 amino acids. The 19 amino acids in the N-terminus of this peptide constituted the signal peptide.

The result analyzed on the 5'-flanking sequence of this gene in Yak showed that it contained two potential TATA boxes, which were located in -25 бл- 33 and - 239 бл - 247 with the corresponding sequences of CAAATAAAA and TAAATAAA. The MGF recognition site was located in - 454бл- 467, with the sequence of CACTTCTTTTGTTT. The MPBF recognition site was located in - 267 бл - 279 with the sequence of GCTTCCTAGAACC. In addition a typical PMF recognition site was presented in - 623 бл- 635 with the sequence of TGATAGCATCTCA. The sequence б░AGTTCTб▒ located in - 583бл - 588 was a typical gtucocorticoid receptor binding site. The homology was about 85% between the sequences of б░AGGCTTGATGCCAб▒ located in - 55бл - 67 and that of the TGGCA banding protein recognition site in lysozyme gene. Perhaps this structure is related to the transcription regulation in this gene just as it was in lysozyme gene. All these show that the 5'-flanking sequence of Yak ж┴-lactalbumin gene we have cloned has most regulation element for high and specific expression in mammary gland. Compared with the corresponding sequence in cattle only the MGF recognition site showed difference and the sequence in Yak was in more accordance with the module sequence published by Groenen M. A. M. So it is possible that the 5'-flanking sequence of ж┴-lactalbumin gene in Yak is more suitable for preparing mammary gland bioreactors in transgenic animals.

Some mutations in the ж┴-lactalbumin gene of Yak resulted in the loose of some hairpin structure in the non-coding sequence of this gene. Perhaps this is one of the reasons why this gene expresses higher in Yak.

The high homology between Yak and Cattle in this gene shows that they have a very close phylogenic relationship. The present result is in accordance with the opinion that the Cattle and Yak is in the same subgenus--Bos subgenus. [Acta Zoologica Sinica 47(6): 691-698, 2001]

Key words Yak (Poephagens grunnieus), ж┴-lactalbumin, Gene, Sequence

*E-mail: ninglbau@public3.bta.net

E-mail: baoliangfan@hotmail.com

бб

Application of molecular scatology to animal ecology

WANG Rong-Jiang

( College of Life Sciences, Peking University, Beijing 100871, China)

Abstract Fecal DNA analysis is a new technique, by which animal DNA is extracted from the feces for involved researches. It is helpful for sampling problem in molecular ecology. In this paper, the methods, subjects, and advances of fecal DNA analysis were involved. Fecal DNA analysis is not only used in many areas of molecular ecology, but also might provide some ecological information, such as estimating population size and confirming the boundary of territory. Hence, fecal DNA analysis is an important complement of molecular ecology. [Acta Zoologica Sinica 47(6): 699-703, 2001]

Key words Fecal DNA analysis, Molecular ecology, Non-invasive sampling, Estimating population size

E-mail: rjwang@bio.pku.edu.cn

бб

Effect of water temperature on spawning of Dace (Leuciscus brandti) in the Suifen river

YIN Jia-Sheng SHEN Jun-Bao XU Wei WANG Wei-Kun

(Heilongjiang River Fishery Research Institute, the Chinese Academy of Fishery Sciences,

Harbin 150070, China)

Abstract The Dace (Leucisus brandti Dybowski) was a migration fish in Suifen river. The Dace can spawn at the water temperatures from 7бц to 25бц. Changing of water temperature is the leading factor cause the spawning population to ovulate. However, high or Iow current water temperature had no obvious effectuation on ovulating behavior of Dace. The ovulation of the Dace often occurred in the evening or next morning after the water temperature rapidly in creased. The fish spawning often occurred as the water temperature decreased at first, after a period of steady low water temperature, then increased. A massive ovulating action began at the time. If the rising of water temperature happens only after a period of steady level without any decreasing of the water temperature, the change can not stimulate ovulation of the Daces. Usually, every large-scale ovulate can sustain for 1 or 2 days. The peak time of ovulation Often appears in 48 hours after the water temperature increases. Once the spawning population began to ovulate, their actions will be very intense, so that even catching action of human

on them can not prevent the ovulation.

The test of artificial induced ovulation showed that only when variation of water temperature were more than 10бц ( increased degree + decreased degree), the Dace would ovulate. The Dace began to ovulate in 12-36 hours after the water temperature's change. The efficient time was shorter and the ovulation rate was higher the variation of water temperature were greater. At the range from 8бц to 24бц, as high or low water temperature had no effect on the efficient time stimulating ovulate, the Dace can not ovulate out of the temperature range. [Acta Zoologica Sinica 47(6): 704-708, 2001]

Key words Dace (Leuciscus brandti), Water temperature variation, Ovulation

бб

A primary report on inducing golden pheasant (Chrysolophus pictus)

reproduction in winter by artifical photo-periods

ZHANG Lu-Qiang YANG Zhen-Cai SUN Ru-Yong

(College of Life Science, Beijing Normal University, Beijing 100875, China)

(College of Life Science, Hebei Normal University, Shijiazhuang 050016, China)

Abstract Golden pheasant (Chrysolophus pictus) is a monotypic and species of China. In nature, it breeds once a year in spring-summer. From Oct. 1998 to Feb. 1999, we studied the effects of photo-periods on inducing the birds to reproduce in winter and succeeded. In the experiment, 36 adult females and 18 males were divided into three groups. Each group was divided into 6 replicates, each replicate included 2 females and 1 male, and fed in one cage. All the groups were exposed to short-day (8L:16D) for 8 weeks, and then transferred to long day (12L: 12D, 14L: 10D or 16L: 8D).

In result, photoperiods significantly affected Golden pheasant reproductive performance in winter. Considered of all of the reproductive performance indicators, the reproductive performance of the group exposure to 14L: 10D was the best, the 16L: 8D groupбпs was less and the 12L: 12D groupбпs was the worst. It may be that the birdsбп CDL (critical day-length) for inducing egg production was about 12 h, and the CDL for optimal egg production was 14L: 10D in winter.

The birds exposed to 12L: 12D had no fertilized egg. The other groups had fertilized eggs and the fertility was 15.05% (14L: 10D) and 20.41% (16L: 8D). The hatchability of fertilized egg were 76.47% (14L :10D) and 80.00% (16L: 8D).

The egg-laying pattern was affected by the size of photoperiod. The ratio of lay of the birds exposed to 14L: 10D, increased rapidely and sustained in a peak level for 5 weeks, and then fall down. But the other birds (group A and C) increased to the peak at the first week and then fall down quickly. [Acta Zoologica Sinica 47(6): 709-712, 2001]

Key words Golden pheasant (Chrysolophus pictus), Artifical photo-period, Reproduction, Winter

E-mail: zhangluq@yahoo.com.cn

бб

Construction of spider genomic DNA Cosmid and cloning of dragline silk gene

LI Min

( College of Biological Engineering, Fujian Normal University, Fuzhou 350007, China)

David L. Kaplan

( Biotechnology Center, Tufts University, Massachusetts 02155, USA)

Abstract The genomic DNA Cosmid library was constructed from Nephila clavipes spider muscle using SuperCos 1 Cosmid as a vector. The title of library was >5 б┴ 104 cfu/ж╠g ligated DNA. On the basis of published sequence from a partial cDNA sequence of the 3'end of the dragline silk gene, we designed and synthesized 3 oligonucleotides. Oligonucleotides were labeled with non-radioactive digoxigenin-dUTP and detected with chemilluminescent substrate. 56 positive recombinants were screen from the Cosmid library using DIG-Oligo 2 as a probe. DNA dot hybridization using DIG-Oligo 1 and DIG-Oligo 3 as the probes, respectively, 3 positive signals were identified from 56 colonies. They were appeared the same pattern when DNA from the colones digested by restriction enzymes. The spider dragline silk gene was confirmed again by Southern blot hybridization. [Acta Zoologica Sinica 47(6): 713-717, 2001]

Key words Spider, Genomic library, Dragline, Silk gene

E-mail: mli2@sina.com

бб

RT-PCR analysis on SOX gene of seven tissues from the

Chinese soft-shelled turtle (Pelodiscus sinensis)

NIE Liu-Wangв┘ SHAN Xiang-Nianв┌ WANG Mingв┘ GUO Chao-Wenв┘ LU Xiao-Xuanв┌

(в┘ Life Science College, Anhui Normal University, Wuhu 241000, Anhui , China)

( в┌ Life Science College, Nanjing Normal University, Nanjing 210009, China)

Abstract In this paper, RT-PCR analysis on SOX gene of seven tissues from the Chinese soft-shelled turtle (Pelodiscus sinensis) was studied, and SOX gene fragments of expression from the testicle, brain, heart, kidney and spleen were cloned using RT-PCR products. The results show that SOX genes has specific expression in the testicle, brain, spleen, cardiac muscle and kidney and isn't expression in muscle, liver and ovary of femel. The results of sequence reveal that the SOX genes of expression in the testicle are TSSOX1, TSSOX4 TSSOX5 and TSSOX9, and those are TSSOX2 and TSSOX4 in the brain, and this is TSSOX4 in the spleen and heart tissue, and those are TSSOX2 and TSSOX3 in the kidney tissue. This suggests that the SOX gene act important role not only on the sex determination, but also on the development of neural system, immunocyte system and the differentiation of male germ cell. [Acta Zoologica Sinica 47(6): 718-720, 2001]

Key words Chinese soft-shelled turtle (Pelodiscus sinensis), RT-PCR, Sequence analysis, Gene expression

E-mail: lwnie@mail.ahwhptt.net.cn

бб

Top

бб

Page Up

бб