Transcriptome profiling of longissimus dorsi during different prenatal stages to identify genes involved in intramuscular fat deposition in lean and obese pig breeds.

BACKGROUND: There was significant difference in muscle development between fat-type and lean-type pig breeds. METHODS AND RESULTS: In current study, transcriptome analysis and bioinformatics analysis were used to compare the difference in longissimus dorsi (LD) muscle at three time-points (38 days post coitus (dpc), 58 dpc, and 78 dpc ) between Huainan (HN) and Large white (LW) pig breeds. A total of 24500 transcripts were obtained in 18 samples, and 2319, 2799, and 3713 differently expressed genes (DEGs) were identified between these two breeds at 38 dpc, 58 dpc, and 78 dpc, respectively. And the number and foldchange of DEGs were increased, the alternative splice also increased. The cluster analysis of DEGs indicated the embryonic development progress of LD muscle between these two breeds was different. There were 539 shared DEGs between HN and LW at three stages, and the top-shared DEGs were associated with muscle development and lipid deposition, such as KLF4, NR4A1, HSP70, ZBTB16 and so on. CONCLUSIONS: The results showed DEGs between Huainan (HN) and Large white (LW) pig breeds, and contributed to the understanding the muscle development difference between HN and LW, and provided basic materials for improvement of meat quality.

Real-time imaging and developmental biochemistry analysis during embryogenesis of Caridina pseudogracilirostris.

This paper reports on the real-time imaging and developmental biochemistry of the freshwater caridean shrimp, Caridina pseudogracilirostris. The complete time-lapse development of a single embryo was recorded in an artificial mold, developed in our lab, and imaged under a stereomicroscope. It took 8 days to complete the 5 stages of embryonic development (1 cleavage stage, 2 gastrulation stage, 3 nauplius stage, 4 prehatching embryo, and 5 zoea stages). As the decapod eggs are enriched with dense yolk, biochemical determination of the major components was made to evaluate the yolk utilization during embryogenesis. The concentration of protein, lipid, and carbohydrate declined drastically from Stage I (cleavage) to Stage IV (Zoea), reflecting sustained yolk utilization during embryogenesis. The increase in the size of the embryo correlates with changes in water content. Lipids, being the principal organic substrate, changes in the fatty acid (FA) composition of embryos during development were determined by GC. The FA composition was observed within the range of 25%-60.87% for saturated, 22.57%-56.45% for monounsaturated, and 5.64%-18.51% for total polyunsaturated FAs. The essential polyunsaturated fatty acid were higher in Stages I, IV, and V, suggesting a major role in embryogenesis. The cellular proliferation and organogenesis as visualized in the real-time imaging correlate well with the biochemical variations observed in C. pseudogracilirostris.

A developmental staging system and musculoskeletal development sequence of the common musk turtle (Sternotherus odoratus).

BACKGROUND: The extremely derived body plan of turtles has sparked a great interest in studying their developmental biology. Here, we describe the embryonic development of the Stinkpot, or common musk turtle (Sternotherus odoratus), a small aquatic turtle from the family Kinosternidae. RESULTS: We identify 20 distinct developmental stages, some comparable to stages described by previous studies on other turtles and some in between these, improving the resolution of the generalities of turtle development. We provide a detailed account of both the external morphology and skeletal development, as well as a general look at the early stages of muscular development until the attainment of the adult muscular anatomical pattern. CONCLUSIONS: Several potential skeletal and muscular apomorphies of turtles are identified or elaborated. The musk turtle, with its small size and hard-shelled egg, could become an important species for the study of turtle evolution and development, suitable for in ovo experimentation and late stage imaging of well-advanced anatomical features.

Intrauterine and post-ovipositional embryonic development of Amerotyphlops brongersmianus (Vanzolini, 1976) (Serpentes: Typhlopidae) from northeastern Argentina.

The reproductive biology and embryonic development of Typhlopidae have rarely been explored. This family of snakes includes mostly oviparous species with uterine egg retention, but the morphology and development of embryos remain unknown. This work aimed to describe the embryonic development of Amerotyphlops brongersmianus from the northeast of Argentina. For this purpose, embryos from intrauterine eggs of gravid females and eight post-ovipositional eggs incubated in the laboratory were analyzed. Embryonic stages, corresponding to the early, mid and advanced development, and a hatchling were described. The main organs and systems form during the period of intrauterine embryonic retention. Comparing to other snakes, differences in the development of cranial structures such as encephalic vesicles and mandibular and maxillary processes were identified. After oviposition the development and differentiation of the tissues and organs completes, the body scales develop, the characteristic pattern of pigmentation establishes and the embryo grows and consumes the yolk. On average, the incubation period lasts 55 days. Differences in the stage of development at oviposition among females of different populations were observed. Embryonic retention could extend up to advanced stages of development.

Development of the chondrocranium of two caiman species, Caiman latirostris and Caiman yacare.

Little is known about the embryonic development and variation of the chondrocranium in Crocodylia and there are no works on any Caiman species. Due to the importance of cranial features in the systematics of this clade, investigating the development of the skull in embryonic stages is essential. In this study, we present for the first time the development of the cartilaginous skull of two extant Caiman species. Anatomical descriptions of the embryonic chondrocranium of Caiman latirostris and Caiman yacare were made, paying special attention to their inter- and intraspecific variation. For this purpose, pre-hatching ontogenetic cranial series of these two caiman species were prepared with a double staining and diaphanization technique. The main differences with other crocodylian species were observed in the palatoquadrate, and interspecific variation within the genus was recorded in the hyobranchial apparatus and larynx. Some characters may be distinctive of Caiman (posterior and ventral surface of the otic process of the palatoquadrate articulated with the dorsal process of the columella auris, and otic process articulated with the lateral wall of the auditory capsule), Alligatoridae (presence of an epiphanial foramen) or C. latirostris and C. yacare (Corpus hyoidei with different number and position of foramina and different shapes of its anterior contour and anterior and posterior notch, different degrees of broadening of the distal end of the Cornu branchiale I, and presence/absence of a notch in the posteroventral surface of the cricoid). Homologies of the elements belonging to the hyobranchial apparatus could not be confirmed. As in other tetrapods the trachea consists of incomplete cartilaginous rings. Morphological changes and dissimilarities found in this study are useful as a context to start studying phylogenetic constraints. Moreover, in a heterochronic context, variations may be involved.

Molecular characterization and expression patterns of Nanog gene validating its involvement in the embryonic development and maintenance of spermatogonial stem cells of farmed carp, Labeo rohita.

BACKGROUND: The homeobox containing transcription factor Nanog plays crucial roles in embryonic development/proliferation and/or maintenance of spermatogonial stem cells (SSCs) via interacting with transcription factors such as Oct4 and Sox2 in mammals. However, knowledge of its exact mechanistic pathways remains unexploited. Very little is known about teleost Nanog. Information on the Nanog gene of farmed rohu carp (Labeo rohita) is lacking. We cloned and characterized the Nanog gene of rohu carp to understand the expression pattern in early developmental stages and also deduced the genomic organization including promoter elements. RESULTS: Rohu Nanog (LrNanog) cDNA comprised an open reading frame of 1,161 nucleotides bearing a structural homeodomain; whereas, the genomic structure contained four exons and three introns suggesting that it is homologous to mammalian counterparts. Phylogenetically, it was closely related to freshwater counterparts. Protein sequence (386 AA of 42.65 kDa) comparison revealed its low similarity with other vertebrate counterparts except that of the conserved homeodomain. Tissue distribution analysis revealed the existence of LrNanog transcripts only in adult gonads. The heightened abundances in the ovary and proliferating spermatogonia suggested its participations in maternal inheritance and male germ cell development. The potentiating abundances from fertilized egg onwards peaking at blastula stage vis- à-vis decreasing levels from gastrula stage onwards demonstrated its role in embryonic stem cell development. We also provided evidence of its presence in SSCs by western blotting analysis. Further, the promoter region was characterized, predicting a basal core promoter and other consensus elements. CONCLUSION: The molecular characterization of LrNanog and its documented expression profiling at transcript and protein levels are indicative of its functional linkage with embryonic/spermatogonial stem cell maintenance. This is the first report of LrNanog genomic organization including its promoter sequence information with predicted regulatory elements of a large-bodied carp species. This will be useful for elucidating its mechanism expression in future. Nanog could be used as a potential biomarker for proliferating carp SSCs.

RNA-sequencing of the sturgeon Acipenser baeri provides insights into expression dynamics of morphogenic differentiation and developmental regulatory genes in early versus late developmental stages.

BACKGROUND: Acipenser baeri, one of the critically endangered animals on the verge of extinction, is a key species for evolutionary, developmental, physiology and conservation studies and a standout amongst the most important food products worldwide. Though the transcriptome of the early development of A. baeri has been published recently, the transcriptome changes occurring in the transition from embryonic to late stages are still unknown. The aim of this work was to analyze the transcriptomes of embryonic and post-embryonic stages of A. baeri and identify differentially expressed genes (DEGs) and their expression patterns using mRNA collected from specimens at big yolk plug, wide neural plate and 64 day old sturgeon developmental stages for RNA-Seq. RESULTS: The paired-end sequencing of the transcriptome of samples of A. baeri collected at two early (big yolk plug (T1, 32 h after fertilization) and wide neural plate formation (T2, 45 h after fertilization)) and one late (T22, 64 day old sturgeon) developmental stages using Illumina Hiseq2000 platform generated 64039846, 64635214 and 75293762 clean paired-end reads for T1, T2 and T22, respectively. After quality control, the sequencing reads were de novo assembled to generate a set of 149,265 unigenes with N50 value of 1277 bp. Functional annotation indicated that a substantial number of these unigenes had significant similarity with proteins in public databases. Differential expression profiling allowed the identification of 2789, 12,819 and 10,824 DEGs from the respective T1 vs. T2, T1 vs. T22 and T2 vs. T22 comparisons. High correlation of DEGs' features was recorded among early stages while significant divergences were observed when comparing the late stage with early stages. GO and KEGG enrichment analyses revealed the biological processes, cellular component, molecular functions and metabolic pathways associated with identified DEGs. The qRT-PCR performed for candidate genes in specimens confirmed the validity of the RNA-seq data. CONCLUSIONS: This study presents, for the first time, an extensive overview of RNA-Seq based characterization of the early and post-embryonic developmental transcriptomes of A. baeri and provided 149,265 gene sequences that will be potentially valuable for future molecular and genetic studies in A. baeri.

Embryonic development of Columba livia(Aves: Columbiformes) from an altricial-precocial perspective / Desarrollo embrionario de Columba livia(Aves: Columbiformes) desde una perspectiva altricial-precoz / Desenvolvimento embrionário de Columba livia(Aves: Columbiformes) a partir de uma perspectiva altricial-precoce

Objective: this study characterized the morphogenesis of Columba livia and proposed a table with 43 embryonic stages. The ontogeny of this species was divided into three phases: early, middle and late. Methods and Results: the early phase (stages 1-26) includes the final period of segmentation, gastrulation, neurulation and somitogenesis, organization of the extraembryonic membranes, and the initial formation of the major organic systems. The middle phase (stages 27-36) was characterized mainly by the growth of limbs and the organization of the autopodium, the formation and growth of the peak, and the development of integumentary annexes such as feather germs and leg scales. The late phase (stages 37-43) shows the overall embryo growth, the final organization of the pterilosis pattern and growth of feather germs, as well as final consumption of the yolk, and hatching. Conclusions: these results allow making comparisons with the proposed developmental events of Gallus gallus domesticus, highlighting similarities and differences in the ontogenetic sequence of both species. Therefore we propose some hypotheses about possible heterochronic events related to altricial-precocial developmental models.

Objetivo: en el presente trabajo se caracteriza la morfogénesis de Columba livia y se propone una tabla con 43 estadios embrionarios. La ontogenia de dicha especie fue dividida en tres etapas: temprana, media y tardía. Métodos y Resultados: la etapa temprana (estadios 1-26) incluye el periodo final de la segmentación, la gastrulación, neurulación y somitogénesis, la organización de las membranas extraembrionarias y la formación de los esbozos de los principales sistemas de órganos. La etapa media (estadios 27-36) se caracteriza principalmente por el crecimiento de los miembros y la organización del autopodio, la formación y crecimiento del pico y el desarrollo de los anexos tegumentarios como plumones y escamas de las patas. En la etapa tardía (estadios 37-43) se evidencia el crecimiento general del embrión, la organización final del patrón de pterilosis y crecimiento de plumones, como así también el consumo final del vitelo y la eclosión. Conclusiones: en base a los resultados obtenidos se realizan algunas comparaciones con respecto a los eventos del desarrollo propuestos para Gallus gallus domesticus, destacando semejanzas y diferencias en la secuencia ontogénetica de ambas especies. A partir de esto se plantean algunas hipótesis acerca de posibles eventos heterocrónicos relacionados con los modelos de desarrollo altricial-precoz.

Objetivo: no presente trabalho caracteriza-se a morfogênese de Columba livia, mediante uma tabela com 43 estágios embrionários. A ontogenia desta espécie foi dividida em três etapas: inicial, intermediária e final. Métodos y Resultados: a etapa inicial (estágios 1-26) incluiu o período final de segmentação, gastrulação, neurulação e somitogênese, organização das membranas extraembrionárias e formação dos esboços dos principais sistemas de órgãos. A etapa intermediária (estágios 27-36) foi caracterizada principalmente pelo crescimento dos membros e organização do autopódio, formação e crescimento do bico e desenvolvimento de anexos tegumentários como as penas e as escamas das patas. Na etapa tardia (estágios 37-43) foi caracterizada pelo crescimento geral do embrião, a organização final do padrão de pterilose e o padrão de crescimento das penas, assim como o consumo final do vitelo e eclosão. Conclusões: com base nos resultados obtidos, foram realizadas algumas comparações com respeito aos eventos do desenvolvimento proposto para Gallus gallus domesticus, destacando semelhanças e diferenças durante a sequência ontogenética de ambas espécies. A partir disso, algumas hipóteses são propostas acerca dos possíveis eventos heterocrônicos relacionados com os modelos de desenvolvimento altricial e precoce.