Abstract   We observed male copulatory behavior in captive group of the Sichuan snub-nosed monkey comprised of 11 individuals, seven males and four females, with Focal-animal sampling technique, during May 2000¡«May 2001 at Shanghai Wild animal Park. The group consisted of two subunits: one was a one-male and multi-female unit (four females) and the other was an all-male unit (two adults, one subadult and one juvenile). During the whole study, four takeovers of dominant male took place, and all of the three adult males involved into the mating activities successively. Among about 1 630 sexual interactions, females were responsible for 94.2% of all sexual solicitations which suggested that it is females who were the main sexual solicitors in this species. About 47.9% of female sexual prostrations were responded by male mounting and only 38.9% of the total mounting interactions resulted in ejaculation by males. No successful female's sexual disturbance was observed for the reproductive reason. Although the dominant male monopolized all the females, solicitation by females still had dramatically effect on the copulatory frequencies among different mating pairs.

In this monkey species, a copulatory lock was absent, and the male was capable of ejaculating following a single mounting and may ejaculate more than once during 30-min observation session. The average mating duration was about 19.4¡À0.7 seconds. The percentage of mounting that were followed by ejaculation was about 26%. Copulatory patterns included No.11 (no lock, thrusting, multiple intromission, and single ejaculation) and No.12 (no lock, thrusting, single intromission, and single ejaculation) in Dewsbury's male-copulatory-patterned system, and No.12 (no lock, thrusting, single and brief intromission) in Dixson's classification. [Acta Zoologica Sinica  48(5):577-584, 2002]

Key words  Sichuan snub-nosed monkey  (Rhinopithecus roxellana), Copulatory patterns

*E-mail: zhangsy@a-1.net.cn

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Effects of sex and age on the behavior of captive giant pandas(Ailuropoda melanoleuca)

LIU Ding-Zhen^¢Ù* ZHANG Gui-Quan^¢Ú WEI Rong-Ping^¢Ú ZHANG He-Min^¢Ú FANG Ji-Ming^¢Ù SUN Ru-Yong^¢Ù

(¢Ù Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering & Institute of Ecology, Beijing Normal University, Beijing 100875, China)

(¢Ú China Conservation and Research Center for the Giant Panda, Wenchuan 623006, China)

Abstract   A behavioral observation was conducted from August of 1994 to January of 1995, and August to November 1995 in China Conservation and Research Center for the Giant Panda, Wolong, Sichuan Province. The focal-animal sampling and continuously recording methods were used throughout the observation. A significant difference in the frequencies of activity, rubbing anogenital area and investigating behavior were found between sexes, and with the aging of the panda, the frequencies of playing and investigating remarkably decreased. Additionally, males spent more time in rubbing anogenital area and sniffing, yet less time in resting than females, no differences were found in total duration of other behaviors. With the development of individuals, the time spent in playing  increased, while decreased in resting. The captive males show significantly more time in being active state (86.59¡À5.40%, n=11) than females (66.77¡À5.75%,n=13, P = 0.0500) in the daytime. These results suggest that: 1) The sexual difference of behaviors exists both in the wild and captivity; 2) The predominance of playing and investigating behavior by young pandas might indicate a process of behavioral learning and imitating, which may in return play an important role for the development of individual behaviors. The predominance of rubbing anogenital area and sniffing behaviors by males might be related to defending their territories and maintaining social relationships. [Acta Zoologica Sinica  48(5):585-590, 2002]

Key words  Giant panda(Ailuropoda melanoleuca), Sex, Age, Behavior, Behavioral frequency, Behavioral duration

*E-mail: dzliu@bnu.edu.cn

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Radiotelemetry of thermoregulation and thermal tolerance on Chinese cobras (Naja atra)ª«

overwintering in a laboratory enclosure

JI Xiang^¢Ù*  CHEN Hui-Li^¢Ù  DU Wei-Guo^¢Ù¡¡ZHU Bing-Quan^¢Ú

(¢ÙSchool of Life Sciences, Hangzhou Normal College, Hangzhou 310036, China)

(¢ÚSchool of Animal Sciences, Zhejiang University, Hangzhou 310029, China)

Abstract   We used commercially available electronic temperature recording devices, temperature- sensitive radiotransmitters (Holohil, SB-2T) and Hobo and Tinytalk dataloggers with an internal temperature sensor, to study thermoregulation and thermal tolerance in adult Chinese cobras (Naja atra) overwintering in an enclosure built in our laboratory. After being implanted with radiotransmitters according to procedures outlined by Reinert (1992), cobras were released into the enclosure. We received signals (pulses) from each radiotransmitter at 3-h intervals from 06:00 to 24:00 h, and visually checked the number of individuals thermoregulating outside the nest at daytime. Air and nest temperatures were automatically recorded at 1.2-h intervals over 24 h, using Hobo and Tinytalk dataloggers, respectively. The overall mean air temperature in the enclosure did not differ from the overall mean nest temperature, but air temperatures varied in a much wide range (-2.0 to 37.4 ¡ãC) than did nest temperatures (6.5 to 14.5 ¡ãC). Body temperatures varied in a range from 3.4 to 34.1 ¡ãC, which was wider than that of nest temperatures but narrower than that of air temperatures. The mean body temperatures differed among individuals, with the overall mean body temperature (15.0 ¡ãC) being higher than the overall mean air and nest temperatures. The mean lowest body temperature was 8.9 ¡ãC, which was similar to the empirically estimated lower limit (9 ¡ãC) of thermal tolerance for the species. Cobras could thermoregulate outside the nest using available warm spots in the enclosure at daytime when air temperatures were higher than 15 ¡ãC, and the recordings of body temperature for these individuals accounted for approximately 29% of the total recordings for all individuals under the same condition. Cobras thermoregulating outside the nest maintained higher body temperatures than did individuals inside the nest at night and daytime when air temperatures were either lower or higher than 15 ¡ãC. Cobras at daytime when air temperatures were lower than 15 ¡ãC maintained lower body temperatures than did those at night. Cobras that did not thermoregulate outside the nest when daytime air temperatures were higher than 15 ¡ãC maintained almost the same body temperatures as did those at night and daytime when air temperatures were lower than 15 ¡ãC. Body temperature was positively correlated with air temperature at night and daytime when temperatures were lower than 15 ¡ãC, but varied independently with air temperature at daytime when air temperatures were higher than 15 ¡ãC. The results imply that the species be a thermo-conformer at low temperatures without warm spots for thermoregulation but a thermo-regulator at relatively high temperatures with warm spots. A prolonged exposure of cobras to the temperatures lower than 9 ¡ãC was lethal, but a brief exposure of cobras to the temperatures lower than their lower limits of thermal tolerance does not necessarily increase mortality of animals. [Acta Zoologica Sinica  48(5):591-598, 2002]

Key words  Chinese cobra (Naja atra), Radiotelemetry, Thermoregulation, Thermal tolerance

*E-mail: xji@mail.hz.zj.cn

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Endemic bird species to China and their distribution

Lei Fu-Min ^¢Ù* Lu Jian-Li ^¢Ù,¢Ú Liu Yao^¢Ú Qu Yan-Hua^¢Ù Yin Zuo-Hua ^¢Ù

(¢Ù Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, CHINA)

(¢Ú Academy of Life Science, Hebei University, Baoding, Hebei, 071003,CHINA)

Abstract  The diversity and distribution patterns of endemic bird species in China were analysed with GIS software using a database of specimen collections and published references. There are 100 endemic bird species in China, belonging to 53 genera, 16 families and 7 orders, accounting for 8 % of China's avifauna.  Nine endemic genera are currently recognized including Tetraophasis, Chrysolophus, Crossoptilon, Ithaginis, Pseudopodoces, Rhopophilus, Kozlowia, Urocynchramus and Latoucheornis. The 2-dimensional distribution pattern shows that 61 species are found in Sichuan Province, 45 species in Gansu Province 41 species in Yunnan Province and 36 species in Tibet, with much fewer species in other provinces and only 3 species in Jilin Prov. In terms of recognised zoo-geographical subregions, 68 species are distributed in the "South-West Mountainous Subregion", 58 species in the "Western Mountainous Plateau Subregion" and 56 species in the "Qinghai-Zangnan Subregion" with much fewer species in other Subregions, and only 3 species in the "Tianshan Hilly Subregion" and 1 species in the "Da Hinggan Mt. Subregion".

Three centers of endemism were identified by overlapping distribution maps, namely the "Hengduanshan Mountain areas", "Mountain areas in north Sichuan Province and Qinling as well as south Gansu Province", and "Taiwan Island". These mainly are comprised of three habitat types: "High Mountain and Plateau", "Himalayan - Hengduanshan Mt." and "Island". The distribution patterns also show that most (45) endemic species are vertically distributed in areas of mid to high elevation ranging from 1 800m to 3 400m.  From 100 m to 1 800 m, species diversity increases with altitude, while from range 2 900 m to 5 500 m, the opposite occurs. [Acta Zoologica Sinica  48(5):599-610, 2002]

Key words: Chinese endemic birds, Center of species diversity, Distribution patterns, GIS

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

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Relationships between sexual masturbation behavior and serum testosterone

levels of a captive male Baiji

CHEN Dao-Quan  ZHAO Qing-Zhong  WANG Ke-Xiong  ZHANG Xian-Feng  WEI Zhuo

 KUANG Xin-An  GONG Wei-Ming  WANG Xiao-Qiang  LIU Ren-Jun  WANG Ding*

(Institute of Hydrobiology, The Chinese Academy of sciences, Wuhan 430072,China)

Abstract  The Baiji or Chinese River Dolphin(Lipotes vexillifer) are found only in the middle and lower reaches of the Yangtze River, China and are one of the most endangered cetacean species in the world. A male baiji was stranded in the Chenglingji section of the middle reaches of the Yangtze River on 11 January 1980 and removed to the Baiji Aquarium of the Institute of Hydrobiology of the Chinese Academy of Sciences. This animal, known as ¡°Qi Qi¡± has been successfully kept in captivity since that time.

In order to compare relationships between sexual masturbation behavior and serum testosterone levels in this male baiji, behavioral observation and analyse of serum testosterone levels from blood samples were carried out from March 1996 through December 1998.Blood samples were taken from the dorsal aspect of the tail fluke between 10:00 and 11:00 a.m. around the 10th of each month. Serum testosterone levels were determined using radio-immunoassay (RIA) techniques. Behavioral observation was conducted for one hour(from 09: 00 to 10: 00 a.m.)everyday in spring and one hour in every three days in other seasons using the focal-animal sampling method. Following a pilot study, the behaviors of ventrally contacting the wall of the pool and erection of the penis were defined as sexual masturbation behaviors and the frequency of both were recorded per hour.

Serum levels of testosterone showed two peaks in 1997.The first peak was from April to June and the second smaller peak was in October. The highest peak was 6.73 ng/ml, recorded in April. The lowest level was 1.24 ng/ml, recorded in January. Two peaks were once again observed in 1998.The first peak was from Februay to June and the second smaller peak was on October. The highest peak was 9.66 ng/ml, recorded in April and the lowest 1.79 ng/ml recorded in January. The frequency of penile erection on the day before blood sampling showed one peak in spring from March to June, and another smaller peak in September in 1997. However, in 1998, this occurred from January to October without any obvious seasonal pattern. The frequency of ventral contact with the wall one day before the blood sampling showed 3 peaks in 1997, one peak was from March to May, one was in October, with a smaller peak in August. In 1998, three main peaks were observed, in February, April and October, with a smaller peak on July. The frequency of penile erection to a length longer than two centimeters showed one peak in Spring with the highest occuring in May in 1997.In 1998, three peaks were apparent, in February, April and August respectively. The monthly average frequency of ventral contact with wall showed similar patterns in 1997 and 1998: two peaks each year, one from February to April, and the other from August to September. The monthly average frequency of penile erection showed only one large peak, from March to May in 1997, but in 1998 there were three small peaks in February, May to July and November respectively. It may be because the animal was seriously sick in the Spring of 1996, penile erection was only observed in July and August of this year, when the observations were conducted one day before blood sampling, and the length of of the erect penis was in all cases less than 2ª±0 cm. Ventral contact with the wall behavior during the same observation period only happened in April at a very low frequency. Monthly average occurrences of such behaviors in 1996 all happened at very low levels, compared with the 1997 and 1998 results. Meanwhile, serum testosterone levels in 1996 were also low, with a different seasonal pattern from that observed in 1997 and 1998.

Based on these results, our preliminarily conclusion is that both sexual masturbation behavior and serum testosterone levels of this male baiji show a periodic cycle and similar seasonal patterns: two peaks in the spring and autumn of each year with the spring peak the highest. We infer from this that the testes of the male baiji were active from February through October (T=2.48 to 9.66 ng/ml), with a higher peak (T=3.44 to 9.66 ng/ml) around April, and another smaller peak(T=3.33 to 4.25 ng/ml) in October. Testes were inactive (T=1.24 to 2.85 ng/ml) from November until January of the next year. The baiji is considered a seasonally reproductive animal species with the peak of mating occurring around April. [Acta Zoologica Sinica  48(5):611-617, 2002]

Key words  Chinese river dolphin(Lipotes vexillifer), Sexual masturbation behavior, Serum testosterone

*E-mail:wangd@ihb.ac.cn

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Effects of number of homosexual partners on copulating date in female captive masked palm civets (Paguma larvata), and fluctuation of urine estrogen during breeding season

JIA Zhiª²Yun DUAN En-Kui  JIANG Zhi-Gang^*ªªWANG Zu-Wang

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

Abstract  Previous studies demonstrated that female social stimuli from the individuals of the opposite sex can cause females to elevate estradiol levels in female insectivores, rodents and primates. Further more, the presence of the homosexual partners can increase the ability to reproduce in the species where males and females reproduce seasonally, by advancing the onset of the rutting season.  However, whether this is true of carnivores remains unclear. Previous studies also indicated that estradiol plays a role in the initiation of sexual behavior in primates. The masked palm civet, Parguma larvata, reproduces seasonally in variable social groups including multiple males and one female, multiple females and one male, or one male and one female. By studying captive masked palm civets, we aimed to determine if:  (1) female civets will come into estrous earlier when presented with homosexual partners, and (2) estradiol level become elevated during lactation. The target animals included three groups with one male and one female, five groups with two females and one male, and three groups with two males and one female. To increase statistical power, we combined groups with one female, since the results indicated that there was not difference in copulating date between the group with one female and two males, and that with one female and one male.  We identified the first day on which copulation occured by checking for the presence of the copulatory plug and mating behaviour, and determined the level of estradiol by using a radioimmunoassay kit. The results indicated that: (1) the five dominant females in the groups with one male and two females copulated 30.7 days earlier than the five subordinate females in the same groups, and 28.6 days earlier than the six females in the mono-female groups. However, there was no statistical difference in the date of first copulation between the subdominant females and the females in mono-female groups. The urine estradiol concentration of the dominant females was 1 909.7¡À336.7(4) pg/ml, significantly higher than that of the subordinate females (LSD test, P<0.05), which was 320.4¡À82.65 (5)pg/ml; (2) two days after the parturition, the estradiol concentration in five mother civets' urine rose peaking on the 5th day, and a second peak appeared 16.75¡À4.46 days later in four of the five mother civets.  The above results suggest that: (1) social influences only facilitate reproduction in dominant females, and (2) the elevation of estradiol in lactating females is the basis for female polyestry in this species. [Acta Zoologica Sinica  48(5):618-624, 2002]

Key words  Masked palm civet(Paguma larvata), Number of homosexual partner, Copulating date, Estradiol concentration, Urine

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

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Molecular evolution and variability in mitochondrial DNA in 10 species of Passeriformeª¬ª³

Li Qing-wei^¢Ù*  Li Shuang^¢Ù  Tian Chun-yu^¢Ù  Wang Yong-jun^¢Ú  Guo Yu-mei^¢Û

  (¢Ù College of Life Science, Liaoning Normal University, Dalian 116029, China )

        (¢ÚShenzhen Hongshulin National Research Administration, Shenzhen 518040, China )

    (¢ÛSchool of pharmacy University of Southern California Los Angeles. CA90089  USA)

Abstract  The mitochondrial RFLP analyzing method was used to study the phylogenetic relationships of ten species from three families of passeriformes,including Melanocorypha mongolica, Corvus corone, Pica pica, Carpodacus erythrinus, Caduelis flame, Coccothraustes coccohraustes, Loxia curvirostra, Pyrrhula griseiventris, Pyrrhula pyrrhula, and Emberiza elegans. Fourteen restriction enzymes were used in this study. Based on the electrophoretic patterns of mtDNA in ten species, phylogenetic trees were constructed using the computer program MEGA 2.1.

Mitochondrial genome polymorphism. MtDNA fragments from different species showed different restriction endonuclease maps. The length of the mitochondrial genome of Carduelis flamme, Melano corypha mongolica and other eight species was 23.13 kb, 20.3 kb and 16¡«17 kb, respectively. The MtDNA genome enlargement observed in this study has never been previously reported.It was initially thought that this result was due to the opposite replication of the non-coding region, but this requires more detailed sequence analysis. 

Interspecific heredity and mutation. Though the Alaudidae were represented by only one species (Melanocorypha mongolica ) in our study, Alaudidae ¢ñ was obviously different from the other two families in restriction endonuclease type: all species had 1-2 Kpn ¢ñ sites except Melanocorypha mongolica which had no Kpn I sites; Melanocorypha mongolica had 1 Xba ¢ñ site while other species had 2-3 Xba ¢ñ sites. From a morphological perspective, the scale of the tarsometatarsus in the Alaudidae was significantly different from other families. Furthermore, the karyotype of the Alaudidae also displayed some unique features. As far as Carduelis falmme was concerned, there were two apparent characters of restriction endonuclease type: one was that 57% of restriction sites belonged to a single site; the other was that genome length was longer (6¡«7 kb )than the other species in the Fringgillidae family. Consistent to a study of the karyotypes of 120 passeriforms species by other researchers, we found that the species demonstrating unique karyotypes showed the the same uniqueness in mtDNA RFLP.

Phylogenetic analysis. The mitochondrial gene tree constructed in this study suggests that the Alaudidae are a single branch separate from the other two families, and the Fringgillidae are more related to the Corvidae than the Alaudidae. It has been reported that mitochondrial DNA evolves at a rate of 2% per million years. Using this evolutionary rate, our results indicate that these taxa diverged 4 million years ago, which is identical to what is suggested by the fossil record. This suggests that mitochondrial DNA may provide an appropriate molecular clock for research on the evolutionary history of birds. [Acta Zoologica Sinica  48(5):625-632, 2002]

Key words  Passeriformes,  Mitochondrial DNA,  Molecular evolution

*E-mail:Liqw@yahoo.com.cn

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Distribution of the P-M System in Drosophila melanogaster Strains from Northeastern China and the Invasion Route of P-element

DENG Ling-Ling  ZENG Qing-Tao^* QIAN Yuan-Huai  LI Shou-Tao

      (Department of Biology, Hubei University, Wuhan, 430062, China)

Abstract   We reported the distribution of the P-M system in Drosophila melanogaste strains from northeastern China, and identified the origin of the initial P-element of China and the possible invading routes of P-element in China. By using two standard diagnostics (cross A and cross A*) as the standard method of assays, we measured P-element activity and cytotype regulatory ability of 108 isofemale lines of D.melanogaster strains from 15 local populations, which were collected in 1999 and 2000 in northeastern China. In terms of the P-M system, five local populations including Dalian, Yantai, Yanji, Changbai and Ha'erbin mainly from the coastal areas and the areas near Korea were of the Q type and the rest were all of the M¡ätype, but none was of the P type. We also found the geographical differentiation of the GD frequencies of the local strains in cross A*, which showed a gradational increase from southeast to northwest of northeast China and with the increase of latitude, on the other hand, the GD frequencies of the neighboring areas (i.e. Beijing and Huhehaote) was similar to that of the inland of northeastern China. In Korea and Japan, it was reported that the local strains were mainly Q type, and the strains from the areas along the coastal of Bo Sea and near Korea of China is Q type. From these facts, It was concluded that there are two routes for the P-elements initially invading China from Korea and Japan : one was from Korea and Japan to Dalian, Yantai by the sea, then spread to the northeast and south, and the other was from Korea to Changbai, Yanji, then to the northeastern areas and the extensive inland of China. The two routes were met in Ha'erbin area, which made the GD frequency in cross A* in this area much lower than that in the neighboring areas. [Acta Zoologica Sinica  48(5):633-640, 2002]

Key words  Drosophila melanogaster, P transposable element,hybrid dysgenesis,cytotype,GD frequency,the invasion of  P element

*E-mail: zengqit@hubu.edu.cn

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Localization of 13 neurochemical substances in the nervous system of the Chinese earthworm (Pheretima aspergillum):an immunocytochemical study

LUO Zhen-Guo^*ªªZHAO Yue  HAN Qing-Guo  ZHANG Xiao-Yun

(Department of Biotechnology, Shenzhen University,Shenzhen 518060, Guangdong,China)

Abstract  About 10 neurochemical substances have been found in the nervous system ofAnnelids, but no detailed research  in this area has been conducted on the Chinese earthworm,ª«P. aspergillum. This research aims to establish if neurochemical substances discovered in other Annelids also exist in P. aspergillum and also if there are any new neurochemical substances in this species. Chinese earthworms were fed with paper pulp and agar to cause them excrete previously ingested earth and sand. They were then deeply anesthetized with 10% ethanol and dissected. The samples were fixed in fixative containing 4% paraformaldehyde and 2% picric acid in 0.01 mol/L PBS (pH 7.2) at 4¡æ for 6 hours, the transferred to 30% sucrose-PBS until completely infiltrated. Samples were then embedded in OCT compound (USA), quick frozen in liquid nitrogen and sectioned into 20 ¦Ìm thicknesses using a LEICA CM 1100 cryostat (Pharmacia). The serial sections were stained with antibodies anti-adrenocorticotropin hormone (ACTH), anti-arginine vasopreesin (AVP), anti-calcitonin, anti-cholecystokinin (CCK), anti-¦Â-endorphin (¦Â-EP), anti-glutamate, anti-calbindin-D, anti-calcitonin gene-related peptide (CGRP), anti-neuropeptide Y (NPY), anti-oxytocin (OT), anti-somatostatin (SOM), anti-vasoactive intestinal polypeptide (VIP) and anti-¦Ã-amino-butyric acid (GABA) using immunocytochemistry techniques. The immunostained consecutive sections were observed and the positive cells and fibers were examined under a light microscope. It was found that ACTH positive neurons, AVP positive neurons, calcitonin positive neurons containing dense large granules, CCK positive neurons, NPY positive neuron and glutamate positive neurons were stained heavily and distributed densely,but the ¦Â-EP positive neurons were stained lightly. A few OT positive neurons were detected sparsely distributed in brain tissue. Calbindin-D, CGRP, GABA, SOM,VIP immunocytochemical reactions were egative. AVP positive neurons, glutamate positive neurons and NPY positive neurons also were detected in the subpharyngeal ganglion and ventral ganglion.¦Â-EP positive neurons, CGRP positive neurons, SOM positive neurons and VIP positive neurons were found in the enteric nervous system. Calcitonin positive epithelial cells, CGRP positive epithelial cells and VIP positive epithelial cells were distributed between negative epithelial cells.

The results indicate the presence of not only previously discovered neurochemical substances: ACTH, CCK, CGRP, NPY, VIP, ¦Â-endorphin also found in the nervous systems of other Annelids, but also new neurochemical substances: calcitonin, glutamate, AVP, OT,SOM. In adition, calcitonin, glutamate, VIP positive epithelial cells were distributed in the digestive system of this species. [Acta Zoologica Sinica  48(5):641-647, 2002]

Key words  Earthworm(Pheretima aspergillum), Neuropeptide, Immunocytochemistry

*E-mail: lqlzg@yahoo.com.cn

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Changes of HSF mRNA expression in rhesus monkey testes during germ cell apoptosis induced by heat stress or high dosage of testosterone undecanoate

 ZHOU Xin-Chang^¢Ù HU Zhao-Yuan^¢Ù ZUO Ru-Jin^¢Ú LIU Yi-Xun^¢Ú*ªª

(¢Ù    State Key Laboratory of Reproductive Biology,Institute of Zoology, ChineseAcademy of Sciences, Beijing100080,China)

(¢Ú Kunming Institute of Zoology,Chinese Academy of Sciences, Kunming 650200,China)

Abstract  In order to investigate the changes of HSF2 mRNA expression and its role in germ cell apoptosis induced by heat stress or a high of testosterone undecanoate, we established the operation-induced unilateral cryptorchid and TU-injected monkey models, and examined in situ analysis of testicular cell DNA fragmentation and the expression level of HSF2 mRNA in germ cells after treatment. The TUNEL results showed that the apoptotic signals after treatment increased and the strongest apoptotic signals appeared in the germ cells of the cryptorchid testes on day 5 or the TU-treated monkey testes on day 30,suggesting that the failure of spermatogenesis in the testes of monkeys after treatment may be due to the germ cell apoptosis induced by heat stress or high TU.The HSF2 mRNA decreased slightly at the early phase of the cell apoptosis just prior to the strongest signals of apoptosis,followed by dramatic decrease.This change was closely time-correlated to that of Hsp70-2 mRNA(unpublished observation). It,therefore,suggests that HSF2 may regulate the expression of Hsp70-2. Moreover,HSF2 may control the spermatogenesis and the germ cell apoptosis by other signal pathways. [Acta Zoologica Sinica  48(5):648-653, 2002]

Key words  Rhesus monkey,HSF2,Germ cell,Apoptosis, Heatstress, Testosterone undecnoate

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

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Effect of GnRH analogue on PKC activity in cultured rat stomach smooth muscle cells

CHEN Lei^¢Ù LI Dong-Hong^¢Ú YAO Yuan-Qing^¢Ú LIU Li-Hong^¢Û HUANGWei-Quan^¢Ù

(¢Ù     Department of Histology and Embryoloqy, the Fourth Millitary Medical University, Xi'an 710032, China)

(¢Ú     Department of Obsentrics and Gynocology of Tangdu Hospital, the Fourth Millitary Medical University,Xi'an 710038, China)

(¢ÛDcpartment of Pharmacology, the Fourth Millitary Medical University, Xi'an 710032, China)

 Abstract  Recently, Gonadotropin-releasing hormone(GnRH)and its receptor have been found in rat digestive system. Our previous studies demonstrated that GnRH receptor and its mRNA both existed in cultured rat stomach smooth muscle cells(SSMCs),and also, we found GnRH analogue could increase the intracellular £ÛCa²⁺£Ýof SSMCs. In order to investigate the effect of GnRH analogue on protein kinase C(PKC)activity in cultured rat stomach smooth muscle cells,we used Radioimmuoassay to determine the PKC activity.SSMCs which we used in this experiment were passage 3¡«5. After several days in culture medium, SSMCs were washed and treated with different agents in the following manners: (1) SSMCs were treated with Alarelin at 10^-5 mol/L for different time(1,2,3,6,8 and 10 min)to find the time-dependence. (2) The SSMCs were treated for 3 min with Alarelin at 10^-9, 10^-8,10^-7,10^-6,10^-5 mol/L respectively to identify the optimal dosage of Alarelin (3) SSMCs were preincubated with phorbol 12-myristate 13-acetate(PMA)for 10 min in Group A and Group B, and 12 hr in Group C respectively. Then Group B and Group C were treated with 10^-5 mol/L Alarelin to confirm the effects of PMA on PKC activity of SSMCs. (4) In order to find the calcium-dependence of GnRH analogueme diated PKC activity, the SSMCs from which extracellular calcium ion were removed the by the use of EGTA were treated with 10^-5 mol/L Alarelin. Then, total cells were scraped with a rubber policeman and transferred to plastic tubes containing the extraction PKC. Cell suspensions were centrifuged and cell pellets were resuspended and sonicated on ice. The homogenate was centrifuged at 100 000¡Ág for 30 min at 4¡æ.The supernatant was used as the ¡°cytosol¡± fraction, and the pellets were re-homogenized in 1% Triton X-100.The second homogenate was centrifuged at  100 000¡Ág for 30 min at 4¡æ, and the supernatant was used as the¡°membrane¡± fraction.PKC activity in both fractions were assayed by determining the transfer rate of ³²P from £Û¦Ã32P£ÝATP to myelin basic protein(MBP). Total PKC activity refers to PKC activity in ¡°cytosol¡± fraction plus the one in ¡°membrane¡±fraction.

The results show that Alarelin could induce activation of PKC, which acted in time-course manner, and the peak time of PKC activity was 3 minutes after treatment.When Alarelin was 10^-9,10^-8,10^-7,10^-6,10^-5 mol/L respectively, the PKC activity was increased gradually,and acted in dose-dependent way. The peak value reached 781.40¡À14.74 pmol/min/mg when Alarelin was 10^-5 mol/L. The smallest value was 103.12¡À11.04 pmol/min/mg. After cells were preincubated with 10 mmol/L PMA 10 min,then the 10^-5 mol/L Alarelin was administered(Group B), we found that PKC activity was not different from Group A or Group B(P>0.05). After PKC activity was depleted by 12 hr treatment of PMA in Group C,then incubated with 10^-5 mol/L Alarelin,we found PKC activity were significantly lower than that of Group A and Group B(^P<0.05).It was also observed that the alarelinstimulated PKC activity was dependent on the presence of extracellular calcium. These results indicated that PKC is an important signal molecule in the regulation of GnRH to cultured stomach smooth muscle cells and GnRH analogue could increase the PKC activity. [Acta Zoologica Sinica  48(5):654-659, 2002]

Key words  GnRH analogue, PKC activity, Radiochemicalassay, Stomach smooth muscle cells

*E-mail:huangwq@fmmu.edu.cn

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Fusion of Ehrlich's ascites cell plasma membrane with liposomes driven by transmembrane pronton efflux:effects of calcium ions and liposome phospholipid composition^*

JIAO Xuan-Mao  LI Qing-Huan  XU Mei  XIONG Jing-Wei  LI Lin-jiang  Shao Lan  LIU Shu-Sen*

(State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100080,China)

SHEN Zi-Wei

(State Key Laboratory of Biomembrane and Membrane Biotechnology, Department of Biological Sciences and Technology,Tsinghua University,  Beijing 100084, China)

Abstract  The Effects of Ca²⁺and the surface electric charge density of plasma membranes on membrane fusions driven by proton translocation were studied.It was shown that Ca²⁺ could promote the membrane fusion of Ehrlich's ascites cells with liposomes driven by proton translocation across plasma membranes, but the extent of the membrane fusion was not related to the concentration of calcium. The relationship between fusion extent and surface electric charge density of membranes was shown to be closely correlated with each other under various concentrations of Ca²⁺, and liposomes composed of different lipids Ca²⁺<6 mmol/L and PE¡ÃPC¡ÃCL=6¡Ã2¡Ã2, Ca²⁺>6 mmol/L as well as PC¡ÃPE¡ÃCL=6¡Ã2¡Ã2. No correlation was seen with Ca²⁺ and PE¡ÃCL=8¡Ã2, Ca²⁺=10 mmol/L and PE¡ÃCL=8¡Ã2. Besides, the effects of the ratio of PE to PC in the liposome on membrane fusion indicated that the extent of membrane fusion of Ehrlich's ascites cells with liposomes decreased with the decrement of PE and increment of PC. This implies that the ability to forme ¡®stalk' intermediates was one other factor to influence  membrane fusion of Ehrlich's ascites cells with liposomes. [Acta Zoologica Sinica  48(5):660-666, 2002]

Key words  Ehrlich's ascites cell, Membrane fusion, Calcium ions,PE/PC,Electric charge density, embrane surface, Proton translocation

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

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Growth of Selincuo schizothoracini (Gymnocypris selincuoensis) in Selincuo Lake,Tibeten Plateau

CHEN Yi-Feng^* HE De-Kui  CAO Wen-Xuan  DUAN Zhong-Hua

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

Abstract  Selincuo schizothoracini is an endemic Tibetan fish  that has been turned into the basis of an  economically important local fishery since 1992. However, there were no ecological studies on this species before.

This paper presents information on growth features of Selincuo schizothoracini, including growth functions, growth index and inflexion point. Specimens were captured between 1997¡«1998 using 3.0¡«6.0-cm-mesh nylon trifilament gill nets and throwing-nets during. The standard length and total weight of each specimen was measured after captured. Individuals were sexed on the basis of morphological sex characters and/or the gonads of sacrificed specimens. Both annulus and otolith radii were measured along the vertical line from the outboard ridge to the nucleil of sectioned sagittae using a micrometer and binocular microscope. A total of 252 sectioned otoliths were measured. The regression line between sectioned otolith radius and standard length was fitted for each sex in order to back-calculate standard length. Three growth models, the von Bertalanffy growth function, Gomperz growth function and decimal cubic function, were used to model the standard length growth of Selincuo schizothoracini using the weighted average of back-calculated standard lengths. The von Bertalanffy growth equations were L_t = 484.1906[1-e^{-0.06839(t-0.6028)}] (¡á) and L_t = 485.3285[1-e^{-0.0710(t-0.5679)}] (¡â), Gomperz growth equations were L_t = 382.5614e^{-exp[-0.1997(t-0.5.3719)]} (¡á) and L_t = 405.4271e^{-exp[-0.1889(t-5.7228)]} (¡â) and the decimal cubic equations were L_t=-25.7461+41.1394t-1.5622t²+0.0232t³(¡á)and L_t = -24.2198+40.1444t-1.3629t²+0.0174t³ (¡â), where t is age. Each of these growth functions provided a high goodness of fit and accurately described the growth of Selincuo schizothoracini well. Among them, the decimal cubic function had the highest goodness of fit. However, the von Bertalanffy growth function was extremely suited to model the growth of older animals (over 18 years), and the Gomperz growth function was especially suited to reveal the growth patterns of younger fish  (under 12 years). Because the adults mostly remain in the lake and juveniles live in its affiliated rivers, the growth characteristics of Selincuo schizothoracini should reflect their migratory (river-lake) life history.

The weight-length relationship could be described by the equations of W = 0.00023L^2.5303 (¡á) (n = 371) and W = 0.00046L^2.4072 (¡â) (n = 427), where W = total weight (g) and L = standard length (mm). The von Bertalanffy function was also used to model growth of total weight and could be described as W_t=1336.9932[1-e^{-0.06839(t-0.6028)}]^2.5303 (¡á) and W_t=1439.4753[1-e^{-0.0710(t-0.5679)}]^2.4072 (¡â).

The growth patterns of male and female Selincuo schizothoracini were significantly different in the Gomperz growth function and the decimal cubic function, and also in the relationship between  weight and length. The inflection points of total weight growth were 14.2 years (¡á) and 12.9 years (¡â). [Acta Zoologica Sinica  48(5):667-676, 2002]

Key words  Selincuo schizothoracini (Gymnocypris selincuoensis), Endemic fish, Growth, Functions, Weight length relationship, Tibet

*Email: chenyf@ihb.ac.cn

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Effects of cadmium on spermatogenesis in freshwater crab

(Sinopotamon yangtsekiense)

WANG Lan^¢Ù*ªªSUN Hai-Feng^¢Ú  LI Chun-Yuan^¢Û

(¢Ù College of Life Science and Technology, Shanxi University, Taiyuan 030006, Shanxi, China)

(¢Ú Collleg of Chemistry Engineering, Shanxi University, Taiyuan 030006,  Shanxi, China)

(¢Û China Institate for Radiation Protection, Taiyuan 030006, Shanxi, China)

Abstract  Reproduction of Crustaceana is a complex regulated process which can be affected in several ways by heavy metals and other pollutants. For the first time the effect of cadmium on the male germ cell of the freshwater crab (Sinopotamon yangtsekiense)  was studied by the means of body cavity injection (Cd²⁺ 1.5 ¦Ìg/g boby weight) in this paper. The germ cell at the different developmental stages in the treated and control individuals had been observed by transmission electron microscopy (TEM) in order to determine whether cadmium might have a morphological effect on them during spermatogenesis. The same experiment had been done twice in the period of March to May and October to December in 1999.

 In the experiment, male crabs were brought from aquatic product market in Taiyuan and raised in laboratory for more than a week before treatment. The mature male crabs in middle size were chosen and injected Cd²⁺ into the body cavity. The treated and control animals were dissected in different times (10 min, 0.5 h, 1 h, 2 h, 4 h, 24 h). The results showed that all kinds of observed cells had been changed ultrastructurally after 30 minutes of cadmium  treatment except for spermagone, and that the longer of cadmium treatment was, the more seriously the cells had been changed in morphology.

Spermagone had ultrastructural changes after two hours of cadmium treatment; and their cell membrane  had been broken partly and nuclear membrane distorted and most mitochondria vacuolated after four hours of treatment. In primary spermatocyte and secondary spermatocyte, ultrastructural changes had been observed after 30 minutes of Cd²⁺ treatment, and changes of organelle were distinct after one hour of Cd²⁺ treatment. The damaged order of organelle of spermatocyte was: mitochondria and endoplasmic reticula firstly (30 min), nuclear membrane and nucleoli secondly (2 h), cell membrane and Golgi complex and lysosome lastly (24 h). In secondary spermatocyte order, mitochondria had been damaged firstly (0.5 h) and cell membrane and nuclear membrane  and endoplasmic reticula secondly (1 h). Injected cadmium 30 minutes later, the cell membrane of spermoblast had been damaged and acrosome had been distorted and mitochondria had been shifted 4 hours later, i.e.injecting cadmium into the animals could cause all organoid of all sorts of male germ cells damaged shortly. Among five kinds of male germ cell, spermagone was the latest one to be damaged by cadmium. Golgi complex was the least to be damaged by cadmium, its ultrastructure had been observed normally in secondary spermatocyte ¢ò and had been damaged slightly in primary spermatocyte ¢ñ after 24 hours of treatment. Cell membrane and mitochondria were affected earlier than other organelle. The changes of cell membrane and mitochondria were including cell membrance swelled seriously (eg. spermoblast) and disintegrated partly, mitochondria distorted and vacluolated and broken, some mitochondria contracted and turned smaller than normal ones, some mitochondria cristae disappeared. At first nucleus had been distorted and nuclear membrane had been expanded slightly, chromatin of primary spermatocyte had been condensated to cluster and chromatin of secondary spermatocyte  had been turned free 4 hours later. Ribosome and nucleoli had been increased to three or five, and some of the latter had been shifted around nuclear membrane 2 hours later. Rough endoplasmica reticula (RER) had been vacuolated largely and turned into smooth endoplasmica reticula (SER) by disassemblying ribosome from RER, which led to the increase of SER and ribosome. There were more lysosome and vacuolus in cytoplasmic matrix and other organella had been damaged seriously twenty¨Cfour hours later.

The results showed that male germ cells of the experimented animals could be damaged by injecting cadmium into their body cavity. The organella with membrane structure were easily  damaged, such as mitochondria, nucleus, endoplasmica reticula and cell membrane. Among them mitochondria was damaged earliest, which suggested that mitochondria was a sensitive organelle to cadmium and could show damage degree led to by cadmium. [Acta Zoologica Sinica  48(5):677-684, 2002]

Key words  Freshwater crab (Sinopotamon yangtsekiense), Spermatogenesis, Cadmium, Ultrastructure, Crustacea

E-mail:lanwang@sxu.edu.cn

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An improved radioimmunoassay for plasma luteinizing hormone in Bactrian camel

(Camelus bactrianus)

LI Xi-Long^¢Ù¢Ú*ªªZHAO Xing-Xu^¢Ù SU Lei^¢Ú CHEN Bei-Heng^¢Ù

(¢ÙDepartment of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China)

(¢ÚKunming Institute of Zoology, CAS, Kunming 650223, China)

Abstract  A new radioimmunoassay (RIA) systems for plasma LH in Bactrian camel was developed by using CamLH, CamLH NZY 01, hLH or hCG as standard reference,¹²⁵I-hLH or ¹²⁵I-hCG as radioligand and anti-hLH or anti-hCG serum as antibody. After evaluating and comparing the specificity, sensitivity and accuracy of different combination, the best combination of radioimmunoassay which used CamLH as standard reference, ¹²⁵I-hLH as radioligand and anti-hLH serum as antibody was selected to measure plasma LH concentration of Bactrian camel. The validity of this combination was also testified and confirmed. [Acta Zoologica Sinica  48(5):685-688, 2002]

Key words  Bactrian Camel (Camelus bactrianus), Luteinizing Hormone, Radioimmunoassay

*E-mail:lixilong@hotmail.com

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 An overview of invertebrate ecology in China

XU Mu-Qi*  ZHANG Zhi-Bin

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

Abstract  This paper reviews the progress of the population and community ecological research on invertebrate in China during the past 30 years. On the basis of modern ecology's developing tendency and current situation of China, authors made several suggestions on the principal aspects of further strengthening and promoting ecological researches on the main groups of invertebrates in China. [Acta Zoologica Sinica  48(5):689-694, 2002]

Key words  Invertebrates, Population and community, Ecology

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

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Incubating behavior of the horned lark(Eremophila alpestris) and small skylark(Alauda gulgula)

 ZHAO Liang^*  ZHANG Xiao-Ai LI Lai-Xing

(Northwest Plateau Institute of  Biology, The Chinese Academy of Sicences, Xining 810001,China)

Abstract  The incubating behavior of the horned lark and small skylark was studied in alpine meadows. The key results are: ¢Ù There was no significant difference in the temperature outside the nests of horned larks and small skylarks (F_{1, 87}=3.286,P>0.05). ¢Ú If parents were off the nest, the temperature in the nest was not significantly different between the two species(F_1,87=3.009,P>0.05), however, if parent birds were on the nest, the temperature in horned lark nests was significantly higher than that in small lark nests (F_{1, 210}=7.555,P<0.01). ¢Û Temperature trends inside and outside the nests of the two species were similar, both following a low-high-low pattern, and showed a significantly positive correlation(horned lark:r=0.572, n=158, P<0.01;small skylark:r=0.511, n=141, P<0.01). ¢Ü During the hatching period, the temperature in nests was significantly higher than the outside temperature in horned larks(t=7.051, df=157,  P<0.01), however, there was no significant difference between the inside and outside in the small skylark(t=-1.558, df=149, P>0.05). ¢Ý The attendance time of horned larks and small skylarks showed three peaks;  before 09:00 h, between 12:00 and 15:00 and after 19:00 h, respectively. ¢Þ The attendance time and incubation behavior of the two species changed with variation in the environmental temperature. ¢ß There was no significant difference in the average attendance time per day between horned larks and small skylarks(P>0.05). ¢à The attendance frequency of horned larks was high (4.17 times/h), and visit duration short (524.32 s/period), wherase small larks showed the opposite pattern; low attendance frequency(1.79 times/h)with relatively long(1 510.00 s/ periods)visits.

Our results are consistent with the hypothesis that attendance time is a balance between the energy requirements of the parent birds and the temperature requirements of the developing embryos. It follows that a bird's choice of nest site should involve a trade-off between costs and benefits, e.g. microclimate versus proximity to food resources. [Acta Zoologica Sinica  48(5):695-699, 2002]

Key words  Horned lark (Eremophila alpestris), Small skylark (Alauda gulgula),Hatching behavior, Attendance time,Environmental temperature

*E-mail: lzhao@mail.nwipb.ac.cn

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Compensatory growth in the adult lacertid lizard (Eremias breuchleyi)ª«following starvation

XU Xue-Feng^*  WU Yi-Lian

(Department of Biochemistry, Chuzhou Normal College, Chuzhou 239012, Anhui, China)

Abstract  An experiment on compensatory growth in lacertid lizards from a population in Suzhou, Anhui, eastern China, after different periods of starvation, was conducted from May to June and September to October 2001. All lizards used in this study were adult males. The experiment lasted for 24 days for each test group. Group C, S3, S6, S9 were deprived of food for 0 (control), 3, 6 and 9 days, respectively. Then each group was fed for the rest of the experimental period. There were significant differences in food intake and mass change among the experimental groups during the course of weight recovery. S9 had higher food intake and mass change than S3 and group C. There were no obvious differences in the apparent digestive coefficient (ADC) or assimilation efficiency (AE) among experimental groups. The results indicate that there was a compensatory effect in weight recovery following starvation, and that the compensatory effect mainly resulted from significant increase in feeding intensity during weight recovery.

Key words  Lizard (Eremias breuchleyi),  Starvation,  Compensatory growth

*E-mail: xuefxu@cztc.edu.cn

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