Histone H1 variants as sperm-specific nuclear proteins of Rana catesbeiana,and their role in maintaining a unique condensed state of sperm chromatin

Amino acid analyses of nuclear basic proteins of an anuran amphibian, Rana catesbeiana, revealed that they are comprised of a full set of core histones and three types of lysine-rich, sperm-specific proteins. On the basis of their amino-acid compositions and partial amino-acid sequences of their trypsin-resistant cores, the sperm-specific proteins could be defined as members of the histone H1 family. Both micrococcal nuclease digestion and electron microscopy indicated that sperm chromatin consists of nucleosomal and fibrillar DNA structures which are irregularly interspersed with each other. When sperm nuclei were incubated with nucleoplasmin, nuclei decondensed to some extent, and the sperm-specific H1s were removed, but not completely. The residual sperm-specific histone H1 variants were also found in reconstituted male pronuclear chromatin, comprising regularly spaced nucleosomes. We conclude that sperm-specific histone H1 variants are essential for chromatin condensation in the sperm nuclei, but that their complete removal is not necessary for the remodeling into somatic chromatin that takes place after fertilization. Mol. Reprod. Dev. 47:181–190, 1997. © 1997 Wiley-Liss, Inc.     

A microsatellite‐based method for genotyping diploid and triploid water frogs of the Rana esculenta hybrid complex

Rana esculenta is a hybrid between Rana lessonae (LL) and Rana ridibunda (RR), and hybrids may be diploid (LR) or triploid (LLR or LRR). Genotypes can be roughly determined from erythrocyte size and morphometry in adult frogs, but accurate genotyping requires more labourious methods. Here I demonstrate that both the L and R genomes have specific microsatellite alleles, and that genotype and ploidy can be accurately inferred from the quantitative ratio of PCR‐amplified (polymerase chain reaction‐amplified) genome‐specific alleles. This method greatly facilitates genotyping in DNA studies of the R. esculenta complex and allows analysis of badly preserved samples and embryos.     

Behavioral responses of American toad and bullfrog tadpoles to the presence of cues from the invasive fish, Gambusia affinis

The introduction of non-native predators is thought to have important negative effects on native prey populations. The susceptibility of native prey to non-native or introduced predators may depend on their ability to respond appropriately to the presence of these non-native predators. We conducted a laboratory based behavioral experiment to examine the response of American toad (Bufo americanus) and bullfrog (Rana catesbeiana) tadpoles to the presence of cues from the introduced mosquitofish (Gambusia affinis), a potential tadpole predator. Neither the American toad tadpoles nor the bullfrog tadpoles responded behaviorally to the presence of mosquitofish cues. If tadpoles are unable to respond to the presence of mosquitofish cues appropriately, then their ability to avoid predation by mosquitofish may be compromised and this may contribute to the impacts of mosquitofish on some tadpole populations.     

Pool desiccation and developmental thresholds in the common frog, Rana temporaria

The developmental threshold is the minimum size or condition that a developing organism must have reached in order for a life-history transition to occur. Although developmental thresholds have been observed for many organisms, inter-population variation among natural populations has not been examined. Since isolated populations can be subjected to strong divergent selection, population divergence in developmental thresholds can be predicted if environmental conditions favour fast or slow developmental time in different populations. Amphibian metamorphosis is a well-studied life-history transition, and using a common garden approach we compared the development time and the developmental threshold of metamorphosis in four island populations of the common frog Rana temporaria: two populations originating from islands with only temporary breeding pools and two from islands with permanent pools. As predicted, tadpoles from time-constrained temporary pools had a genetically shorter development time than those from permanent pools. Furthermore, the variation in development time among females from temporary pools was low, consistent with the action of selection on rapid development in this environment. However, there were no clear differences in the developmental thresholds between the populations, indicating that the main response to life in a temporary pool is to shorten the development time.     

昆明地区滇蛙两性生殖器官的生长及发育同步性

月平均采集并解剖滇蛙14—16只(雌雄约各半),测量体长、体重、脂肪体重、精巢重、卵巢重、精巢体积和输卵管直径等。以体长为协变量做协方差分析,结果显示,精巢重、卵巢重、精巢体积、输卵管直径在年周期内差异显著,呈周年规律性变化。滇蛙的繁殖期为4—6月,5月下旬至6月上旬为繁殖高峰。异速生长分析和回归分析结果显示,性腺发育与体重和体长多呈正异速生长(b>1),个体增长对性腺的发育有显著影响。个体较大的滇蛙年生殖频次至少为2次,这是提高繁殖成功率的繁殖策略之一。性腺发育与脂肪体重呈负异速生长(b<1)。脂肪体为繁殖和冬眠提供能量储备,脂肪体重在繁殖高峰期达最小值。少数脂肪储存不足的个体通过延迟冬眠时间积累能量,以便安全越冬。相关性分析和交叉相关分析结果显示,两性生殖腺的发育具有显著的相关性,但生殖腺的发育并非完全同步。雄蛙精巢的发育具有一定的滞后性,因为卵的成熟比精子的成熟需要更长的时间。但精巢体积和输卵管直径发育具同步性,这恰是为授精做好了准备。     

Do rivers function as genetic barriers for the plateau wood frog at high elevations?

Using the plateau wood frog Rana kukunoris from the Hengduan Mountains as a model system, we tested whether rivers form significant genetic barriers (the riverine barrier hypothesis) to high elevation amphibians. Samples were collected from eight sites across three major river drainages, the Min, the Dadu and the Yalong Rivers, and the population genetic structure of these frogs was evaluated with data from eight microsatellite DNA loci. A large amount of genetic structure was found, and the pairwise F _ST ranged from 0.022 to 0.508 and a global F _ST was 0.215. Both analysis of molecular variance and isolation by distance analysis suggested that rivers, mountain ridges and geographic distances all contributed significantly to the population structure. However, no single landscape has prominent barrier effect to the plateau wood frog populations. An assignment analysis using the computer program Structure grouped the eight populations into four population clusters, and no single type of landscape can sufficiently explain the clustering. In conclusion, rivers do not appear to be the leading genetic barriers for the plateau wood frog. The strong population genetic structure is likely the consequence of attributes of the species, as opposed to environmental fragmentation, and the barrier effect of the landscapes is largely swamped by the large amount of intrinsic population structure.     

Temporal peculiarities of responses of auditory system neurons of the frog <Emphasis Type="Italic">Rana temporaria</Emphasis> to tone bursts with different modulation frequencies

Activity of medullar and midbrain auditory neurons at action of amplitude-modulated tone burst was recorded in immobilized common frogs Rana temporaria. Depth of modulation amounted to 10% or 80%, frequency of modulation varied from 5 to 150 Hz, and carrier intensity was in the range of 20–30 dB. Phasic neurons in medulla clearly reproduced the modulation frequency, but only at the 80% modulation depth. However, during presentation of signal with the 10% modulation depth, these neurons practically did not respond. Tonic neurons were able to reproduce the modulation frequency up to 10–150 Hz, but at the 10% modulation depth, the degree of reproduction of envelope was rather low, although for several first modulation periods it rose statistically significantly. In midbrain, the highest frequency of the reproduced modulation sharply fell. At greater modulation frequencies, the response of these neurons qualitatively reminds that of medullar neurons. At the low modulation frequencies, there is identified a group of midbrain neurons with a prominent increase of the signal modulation. This occurs in the frequency diapason up to 60 Hz; at an increase of the modulation frequency the time of achievement of maximal synchronization decreases. The optimal modulation frequency in many neurons of semicircular torus corresponds to parameters of the male nuptial call.     

Molecular characterization of major histocompatibility complex class II alleles in the common frog, Rana temporaria

Major histocompatibility complex (MHC) class II genes, which play a major role in the immune system response, are some of the most polymorphic genes in vertebrates. We developed polymerase chain reaction primers for part of the second exon of an expressed MHC class II gene in the common frog, Rana temporaria. We genotyped this locus in five frog populations in southeast England and detected eight alleles in 215 individuals. Five or six alleles were detected in each population with a maximum of two alleles per individual, indicating that only a single locus was amplified. We also inferred the possible existence of a null allele. There were 23 variable nucleotide sites (out of 136) and 13 variable amino acid sites (out of 44), many of which corresponded to amino acids involved in antigen recognition. We detected a significant excess of nonsynonymous substitutions at antigen binding sites, indicating that this gene is under positive selection. The level of variation we found was similar to that in other amphibian MHC class II loci, such as those in Bombina bombina, Xenopus laevis and Ambystoma tigrinum.     

镉对中国林蛙蝌蚪生长发育的毒性效应

将中国林蛙(Rana chensinensis)蝌蚪分别暴露于加Cd2 的自来水和去离子水中,统计得出24、48、72和96 h的半数致死浓度(LC50)、全致死浓度(LC100)、零致死浓度(LC0)和安全浓度(SC)。结果表明,随着Cd2 浓度的升高和染Cd2 时间的延长,蝌蚪的死亡率增高,Cd2 加入去离子水中对蝌蚪的毒性比在自来水中更强。另外,将26~27期林蛙蝌蚪饲养在含Cd2 0.05、0.1、0.2和0.4 mg.L-1的自来水中直至完全变态,通过对完全变态所需时间、平均体重和长度增长率的测定,研究在SC以下Cd2 对林蛙蝌蚪胚后发育的影响。随着Cd2 浓度的升高,蝌蚪的平均体重和长度的增长率降低,完全变态所需时间增长。说明Cd2 污染在SC以下仍可抑制中国林蛙蝌蚪的生长发育,延缓变态。