Morphometric analysis of interspecific and microgeographic variation of crayfish from a Mexican cave

Two undescribed species of Procambarus crayfish (Crustacea, Decapoda) inhabit the subterranean stream running in the recently explored Cueva de Los Camarones, Chiapas, Mexico. Individuals ranging from dark, robust, large-eyed forms with an epigean mien to pale, comparatively slender, microphthalmic ones resembling related troglobites were found, but only the extremes could be distinguished visually. Evolutionary relationships among individuals were investigated morphometrically by means of multivariate analysis'on 14 body dimensions. These analyses revealed a significant discontinuity between the two species, mainly determined by the shape of the rostrum, chelae and telson. The occurrence of a few aberrant phenotypes may indicate the existence of some gene flow between the two species.
Microgeographic variation was demonstrated within the less cavernicolous species. The characters involved were the same as those that discriminate between the two species. These results are compared with allozymic data presented in a parallel paper and the role of genie introgression and environmental induction are discussed.     

Light-evoked walking in crayfish: Behavioral and neuronal responses triggered by the caudal photoreceptor

The caudal photoreceptors (CPRs) of crayfish (Procambarus clarkii) can trigger walking and abdominal movements by their response to light.

The arthropod gap junction and pseudo-gap junction: Isolation and preliminary biochemical analysis

Summary The hepatopancreas of the crayfish, Procambarus clarkii, contains an unusual abundance of gap junctions, suggesting that this tissue might provide an ideal source from which to isolate the arthropod-type of gap junction. A membrane fraction obtained by subcellular fractionation of this organ contained smooth septate junctions, zonulae adhaerentes, gap junctions and pentalaminar membrane structures (pseudo-gap junctions) as determined by electron microscopy. A further enrichment of plasma membranes and gap junctions was achieved by the use of linear sucrose gradients and extraction with 5 mM NaOH. The enrichment of gap junctions correlated with the enrichment of a 31 Kd protein band on polyacrylamide gels. Extraction with 20 mM NaOH or 0.5% (w/v) Sarkosyl NL97 resulted in the disruption and/or solubilization of gap junctions. Negative staining revealed a uniform population of 9.6 nm diameter subunits within the gap junctions with an apparent sixfold symmetry. Using antisera to the major gap junctional protein of rat liver (32 Kd) and to the lens membrane protein (MP 26), we failed to detect any homologous antigenic components in the arthropod material by immunoblotting-enriched gap junction fractions or by immunofluorescence on tissue sections. The enrichment of another membrane structure (pseudo-gap junctions), closely resembling a gap junction, correlated with the enrichment of two protein bands, 17 and 16Kd, on polyacrylamide gels. These structures appeared to have originated from intracellular myelin-like figures in phagolysosomal structures. They could be distinguished from gap junctions on the basis of their thickness, detergent-alkali insolubility, and lack of association with other plasma membrane structures, such as the septate junction. Pseudo-gap junctions may be related to a class of pentalaminar contacts among membranes involved in intracellular fusion in many eukaryotic cell types. We conclude that pseudo-gap junctions and gap junctions are different cellular structures, and that gap junctions from this arthropod tissue are uniquely different from mammalian gap junctions of rat liver in their detergentalkali solubility, equilibrium density on sucrose gradients, and protein content (antigenic properties).     

Synaptic connections of the common inhibitory motoneurone within the fifth thoracic ganglion of crayfish

The common inhibitor (CI) has been studied morphologically and electrophysiologically in the fifth thoracic ganglion of crayfish (Procambarus clarkii). It has a large soma and possesses two separate dendritic fields arising from distinct integrative segments.In vitro preparations display motor outputs ranging from tonic activity to fictive locomotion. The CI's tonic firing frequency increases as more excitors are recruited, and displays two peaks of frequency during fictive locomotion, one during stance, the other during swing.Paired intracellular recordings have been used to demonstrate the different central synaptic connections received or made by the CI. At least 27% of the proximal excitors receive monosynaptic connections from the CI corresponding to post-synaptic depolarizations of small amplitude mediated by GABA. However as they do not change the overall activities of the excitors which receive them, they may be used for local inhibition within the dendrites. Besides, electrical synapses between several proximal excitors and the CI may synchronize their activity.The CI receives synaptic connections arising from interneurones. Some are direct either by inhibitory monosynaptic connections or by electrical couplings whereas others arise through polysynaptic pathways. All these connections are functionally significant in the control of the CI firing activity and in its motor coordinations.     

Amino acid sequence of the minor isomorph of the crustacean hyperglycemic hormone (CHH-II) of the Mexican crayfish Procambarus bouvieri (Ortmann): Presence of a d-amino acid

The primary structure of the neurohormone crustacean hyperglycemic hormone (CHH-II) was determined by means of enzymatic digestions, manual Edman degradation, and mass spectrometry. CHH-II is a 72 residue peptide (molecular mass 8388 Da), with six cysteines forming three disulfide bridges that connect residues 7–43, 23–39, and 26–52. The peptide has blocked N- and C-termini, and lacks tryptophan, histidine, and methionine. The CHH-I and CHH-II of Procambarus bouvieri have identical sequences and elicit levels of hyperglycemia that are not distinguishable. The difference between the two isomorphs consists in a posttranslational modification of a l-Phe in CHH-I to a d-Phe in CHH-II at the third position from the N-terminus.     

A narrow hybrid zone between two crayfish species from a Mexican cave

In Northern Chiapas (Mexico), two newly discovered species of Procambarus crayfish inhabit a subterranean stream. These species can be morphologically distinguished only by comparing extreme phenotypes (dark, thick-eyed, surface dwelling-like individuals vs light, elongated, microphtalmic, cave dwelling-like individuals). Individuals with intermediate phenotypes co-occur with those exhibiting extreme phenotypes. Crayfish were assayed electrophoretically and individual patterns at 23 gene loci were obtained. Unusually high levels of heterozygosity in both species and a clear discrimination between the two gene pools were revealed. The relationships between individuals were investigated by means of multivariate analysis on individual multilocus genotype profiles. Results showed the occurrence of individuals genetically intermediate between the two major clusters, which shared allozymic variants with both species. Due to the occurrence of alternative alleles in the two gene pools, we could quantify patterns of introgression, which revealed asymmetric gene flow between the two species. Moreover, differential levels of introgression in subsamples within the surface-like species were found: most introgressed individuals came from the inner section of the cave, where the two species were greatly mixed. These results are also discussed in reference to the morphometric results from a companion paper. A possible evolutionary pathway, leading to the situation in this cave, and possibly in neighbouring cave systems, is outlined. The hypothesis of a past history of allopatric divergence from a common ancestor and a subsequent secondary contact between these two Procambarus species is supported by geological studies. Crayfish sympatry and competitive exclusion are also discussed.     

SPONTANEOUS AND LIGHT-EVOKED DISCHARGE OF THE ISOLATED ABDOMINAL NERVE CORD OF CRAYFISH IN VITRO REVEALS CIRCADIAN OSCILLATIONS

We examined the well-known spontaneous discharge (SD) and lightevoked (PD) discharge of the crayfish caudal photoreceptor for the possible existence of a daily rhythm in spike frequency. To do this, we isolated the abdominal nerve cord in vitro and studied its discharge frequency in constant darkness. Single cosinor analysis revealed significant SD and PD circadian rhythms (P <. 05) with periods τ = 24.4h and 24.2h, respectively. These oscillations correspond to an endogenous circadian discharge of the caudal photoreceptor that is enhanced by light. The importance of this rhythm in the adaptive behavior of crayfish is discussed. (Chronobiology International, 18(5), 759–765, 2001)     

不同地位克氏原螯虾胜利者-失败者效应的稳定性

为探究不同地位的克氏原螯虾(Procambarus clakii)胜利者-失败者效应的稳定性。通过视频拍摄优势者和从属者在新的领域中各自与等重量级雄性螯虾连续遭遇战,统计胜负场数、格斗次数、时间、优势指数等多个参数。在与陌生对手的首次交战中,40个优势者中有23只螯虾取得了胜利,17只失败,胜负比例之间差异不显著;而从属者中获胜比例为18/40,与失败者的比例之间也没有差异,说明优势者和从属者之间原有的等级地位并不被陌生对手识别。在与陌生对手的第二次交战中,获胜的优势者中18/23的螯虾再次胜利,极显著高于失败的螯虾比例;获胜的从属者中,仅11/18的螯虾再次胜利,与失败者的比例之间不显著;而失败的优势者中,13/17的螯虾持续失败,失败的从属者中,17/22的螯虾持续失败,均显著高于获胜的比例,说明不同地位的螯虾胜利者-失败者效应稳定性不同,优势者的胜利者-失败者效应均比较稳定,而从属者的失败者效应稳定,胜利者效应并不稳定,一胜之后不能获得稳定、完全的二胜。对格斗次数、时间、优势指数等参数统计分析发现,胜利或失败的优势者以及从属者在格斗策略上有较大的差异。     

Colonization of freshwater habitats by an introduced crayfish, <Emphasis Type="Italic">Procambarus clarkii</Emphasis>, in Southwest Iberian Peninsula

The introduction of some crustacean species has produced alterations of freshwater environments and declines of native species worldwide. The red swamp crayfish, Procambarus clarkii Girard, was introduced in the Southwest Iberian Peninsula in the 1970’s, producing severe impacts on rice agriculture and on native biota such as macrophytes, gastropods, native crayfish and amphibians. We studied the distribution of P. clarkii in two areas of SW Iberian Peninsula: the Sado River basin (SW Portugal), an area colonized by this species around 1990, and the Parque Natural del Entorno de Doñana (SW Spain), colonized soon after its introduction in the Iberian Peninsula, in the 1970’s. Our main goal was to determine which factors limit crayfish distribution, which could help to identify the most effective management practices to contain its spread. Procambarus clarkii was found in most types of water bodies, including small and shallow ones. Distance to a crayfish source was the single predictor variable explaining crayfish occurrence in most types of habitats and in both areas. The only exception was for the Sado permanent stream points, where crayfish presence was negatively affected by an interaction between elevation and flow velocity. Other habitat characteristics have apparently little or no importance for its successful colonization. Moreover, this study indicated that overland dispersal is apparently a frequent phenomenon in this species. Our findings can be used to determine which habitats are most likely to be colonized by the crayfish and to develop practical measures which may limit its spread and minimize its impacts.     

Detoxification enzymes of freshwater crayfish Procambarus clarkii fed a diet enriched in selenium: Preliminary results

This study aimed at investigating the extent of accumulation of selenium in the hepatopancreas of the crayfish Procambarus clarkii and its effects on the enzymatic activities of catalase, glutathione peroxidase, and glutathione reductase. Crayfish were fed for 15 days either a diet enriched in Se, ED (1.21 mg kg^-1) or a standard diet, SD (0.30 mg kg^-1). Results showed that Se accumulation is higher in the sample subject to ED rather than to SD. A sexual difference was found for the response of the analyzed enzymes in the ED sample only. Catalase activity and glutathione peroxidase activity decreased in females and males, respectively, whereas glutathione reductase activity showed significant reductions in both sexes. These preliminary data might provide information about prooxidant effects of Se on P. clarkii when fed a Se enriched diet.