Chondrichthyes Chitinase: Molecular Cloning, Distribution, and Phylogenetic Analysis

We have previously reported the presence of three types of chitinase (acidic fish chitinase-1: AFCase-1, acidic fish chitinase-2: AFCase-2, fish chitinase-3: FCase-3) in Actinopterygii. In the present research, we report the identification of the novel chitinase genes HjChi (ORF: 1380 bp) and DkChi (ORF: 1440 bp) from the stomach of Chondrichthyes, Japanese bullhead shark (Heterodontus japonicas) and Kwangtung skate (Dipturus kwangtungensis), respectively. Organ-specific expression analysis identified the stomach-specific expression of HjChi, whereas DkChi was expressed widely in all organs. Chitinase activity was measured using pNP-(GlcNAc)n (n = 2, 3) as a substrate and β-N-acetylhexosaminidase (Hex) activity was measured using pNPGlcNAc. Relatively high values of chitinase activity were observed in the stomach, spleen, and gonads of the Japanese bullhead shark, H. japonicas , compared with that observed in the stomach of the Kwangtung skate D. kwangtungensis . However, Hex activity was detected throughout the body of both species. The optimal pH of chitinase in both the Japanese bullhead shark, H. japonicas, and the Kwangtung skate, D. kwangtungensis, were 3.5 - 5.5 and 3.5 - 4.0, respectively, and 4.0 for Hex in both species. Phylogenetic analysis revealed that Chondrichthyes chitinase forms a unique group (Chondrichthyes chitinase). These results suggested that the possibility of the formation of chitinase groups for each class in the phylogenetic analysis based on the observation of class-specific chitinase.

The complete mitochondrial genome of the grey bamboo shark (Chiloscyllium griseum) (Orectolobiformes: Hemiscylliidae): genomic characterization and phylogenetic application

The complete mitochondrial genome of the grey bamboo shark, Chiloscyllium griseum （Orectolobiformes： HemiscyUiidae） was obtained, with a total of 16 755 base pairs （bp）. The mitogenome encoded 13 protein- coding genes, two ribosomal RNAs, 22 transfer RNAs and a noncoding control region, a gene arrangement identical to the most common type found in vertebrates. All tRNA sequences were folded into typical clover- leaf secondary structure with the exception of tRNA-Ser2, in which the DHU arm stem was replaced with 12 unpaired nucleotides and formed a simple loop. In the control region, the putative termination-associated sequences （TAS） with hairpin-loop structure and the conserved sequence blocks （CSB） I-III were identified and considered to be associated with the replication and transcription of mtDNA. No repeat motifs were found in the mitogenome of C. griseum. On the basis of the mitogenomic data available in the Class Chon- drichthyes from both Genbank and this study, and the phylogenetic analyses using maximum likelihood （ML） and Bayesian inference （BI） methods, the results strongly support the basal division between batoids and sharks and the monophyly of the Superorder Galeomorphii, and confirm the phylogenetic position of C. griseum in the Subclass Elasmobranchii. This study suggests that the mitogenomic data are more robust for reconstructing phylogenetic relationships than individual genes in the Elasmobranchii.

Interaction between Spermatozoa and the Different Epithelia along the Female Reproductive Tract of Mustelus schmitti

The presence of sperm was observed in different organs of the reproductive tract of M. schmitti. These were the club, baffle and terminal zone of the oviducal gland, posterior portion of the uterus and in the cervix. This fact allowed for consideration not only its storage but its interaction with the female reproductive tract epithelia as well. SEM （microscopic observation studies） detected loose and sole spermatozoa in the club and baffle zones in a pregnant female. On the other hand, histochemical techniques showed sperm storage tubules of the terminal zone, which contain bundles of sperm, do not stain AB （alcian blue） or PAS （periodic acid schiff）. Disintegration of some of the male gametes was observed in the posterior part of the uterus with TEM （transmission electron microscopy） studies which would indicate a first uterine sperm selection. The uterine epithelial cells involved in this process showed an important level of secretion vacuoles suggesting an interaction with sperm cells. Sperm in the cervix was found without the matrix that constitutes the spermatozeugmata formed in the male tract. Some heads of the sperm were uncurled while they were interacting with the cervical cells.

Hox Gene Clusters of Early Vertebrates:Do They Serve as Reliable Markers for Genome Evolution?

Hox genes, responsible for regional specification along the anteroposterior axis in embryogenesis, are found as clusters in most eumetazoan genomes sequenced to date. Invertebrates possess a single Hox gene cluster with some exceptions of secondary cluster breakages, while osteichthyans （bony vertebrates） have multiple Hox clusters. In tetrapods, four Hox clusters, derived from the so-called two-round whole genome duplications （2R-WGDs）, are observed. Overall, the number of Hox gene clusters has been regarded as a reliable marker of ploidy levels in animal genomes. In fact, this scheme also fits the situations in teleost fishes that experienced an additional WGD. In this review, I focus on cyclostomes and cartilaginous fishes as lineages that would fill the gap between invertebrates and osteichthyans. A recent study highlighted a possible loss of the HoxC cluster in the galeomorph shark lineage, while other aspects of cartilaginous fish Hox clusters usually mark their conserved nature. In contrast, existing resources suggest that the cyclostomes exhibit a different mode of Hox cluster organization. For this group of species, whose genomes could have differently responded to the 2R-WGDs from jawed vertebrates, therefore the number of Hox clusters may not serve as a good indicator of their ploidy level.

Estimating the geographic range of a threatened shark in a data-poor region： Cetorhinus maximus in the South Atlantic Ocean

The distribution of the planktivorous basking shark Cetorhinus maximus is influenced by zooplankton abundance at small scales and temperature at medium scales in the North Atlantic. Here, we estimate the distribution of basking sharks on South Atlantic continental shelves, and the relative importance of chlorophyll concentration, as a proxy for zooplankton abun- dance, and temperature in determining habitat suitability for basking sharks at large scales. We used maximum entropy （MaxEnt） and maximum likelihood （MaxLike） species distribution modelling to test three hypotheses： the distribution of basking sharks is determined by （1） temperature, （2） chlorophyll concentration, or （3） both chlorophyll and temperature, while considering other factors, such as oxygen and salinity. Off South America, basking shark habitat included subtropical, temperate and cool-temperate waters between approximately 20°S and 55°S. Off Africa, basking shark habitat was limited to cool-temperate waters off Namibia and southern South Africa. MaxLike models bad a better fit than MaxEnt models. The best model included minimum chlorophyll concentration, dissolved oxygen concentration, and sea surface temperature range, supporting hypothesis 3. However, of all variables included in the best model, minimum chlorophyll concentration had the highest influence on basking shark distribution. Unlike the North Atlantic distribution, the South Atlantic distribution of basking sharks includes subtropical and cool-temperate waters. This difference is explained by high minimum chlorophyll concentration off southern Brazil as compared to North Atlantic subtropical areas. Observations in other regions of the world support this conclusion. The highest habitat suitability for basking sharks is located close to nearshore areas that experience high anthropogenic impact [Current Zoology 61 （5）： 811-826, 2015].