Genome re-sequencing to identify single nucleotide polymorphism markers for muscle color traits in broiler chickens.

OBJECTIVE: Meat quality including muscle color in chickens is an important trait and continuous selective pressures for fast growth and high yield have negatively impacted this trait. This study was conducted to investigate genetic variations responsible for regulating muscle color. METHODS: Whole genome re-sequencing analysis using Illumina HiSeq paired end read method was performed with pooled DNA samples isolated from two broiler chicken lines divergently selected for muscle color (high muscle color [HMC] and low muscle color [LMC]) along with their random bred control line (RAN). Sequencing read data was aligned to the chicken reference genome sequence for Red Jungle Fowl (Galgal4) using reference based genome alignment with NGen program of the Lasergene software package. The potential causal single nucleotide polymorphisms (SNPs) showing non-synonymous changes in coding DNA sequence regions were chosen in each line. Bioinformatic analyses to interpret functions of genes retaining SNPs were performed using the ingenuity pathways analysis (IPA). RESULTS: Millions of SNPs were identified and totally 2,884 SNPs (1,307 for HMC and 1,577 for LMC) showing >75% SNP rates could induce non-synonymous mutations in amino acid sequences. Of those, SNPs showing over 10 read depths yielded 15 more reliable SNPs including 1 for HMC and 14 for LMC. The IPA analyses suggested that meat color in chickens appeared to be associated with chromosomal DNA stability, the functions of ubiquitylation (UBC) and quality and quantity of various subtypes of collagens. CONCLUSION: In this study, various potential genetic markers showing amino acid changes were identified in differential meat color lines, that can be used for further animal selection strategy.

Specialized synapse-associated structures within the calyx of Held.

The calyx of Held exhibits fast glutamatergic neurotransmission at high rates with low temporal jitter and has adapted specialized synaptic mechanisms to support its functional demands. We report the presence in calyces of an atypical arrangement of subcellular organelles, called the mitochondria-associated adherens complex (MAC). We demonstrate that MACs are located adjacent to synapses and contain membranous elements linked with coated and uncoated vesicles. Mitochondria that form MACs have more complex geometries than other mitochondria within the calyx and can extend between clusters of synapses. We estimate that the calyx contains 1600 MACs, 2400 synapses, and 6200 readily releasable vesicles. We also identify synaptic vesicle endocytotic regions close to MACs and synapses and hypothesize that calyces are composed of multiple activity modules, each containing machinery for vesicle release and recycling.

Ultrastructure of neurons and large synaptic terminals in the lateral nucleus of the trapezoid body of the cat.

Neurons of the lateral nucleus of the trapezoid body (LNTB), the most prominent periolivary nucleus of the cat superior olivary complex, form an important component of the descending auditory pathways and also innervate the medial superior olive. Cells forming the posteroventral subnucleus (pvLNTB), when investigated by light microscopy, exhibit morphological similarities with globular bushy cells of the cochlear nucleus and principal cells of the medial nucleus of the trapezoid body. These latter two cell types are integral components of brainstem circuitry mediating the early stages of sound localization. In this report, ultrastructural features of LNTB neurons are described. pvLNTB cell bodies are characterized by a round to oval shape, smooth nuclear membrane, and the relative paucity of stacks of rough endoplasmic reticulum. In addition, pvLNTB cell bodies and proximal dendrites are contacted by large synaptic terminals which contain round synaptic vesicles and form multiple asymmetric synaptic junctions. These ultrastructural characteristics are similar to those previously described for globular and principal cells and distinguish pvLNTB cells from cells of the main subnucleus. Large terminals contacting pvLNTB cells contain a specialized organelle assembly, including an adherens plaque associated by filamentous strands with a mitochondrion. We name this organelle assembly the mitochondria-associated adherens complex (MAC) and note its proximity to synaptic junctions. Because high activity rates are characteristic of large terminals in the lower auditory system, the MAC may play a specialized role in membrane stabilization at synapses which generate high rates of vesicle membrane turnover.

Hyperpolarization-activated channels in trigeminal ganglia innervating healthy and pulp-exposed teeth.

AIM: To use immunocytochemistry for determining the expression of HCN1, HCN2 and HCN3 (three subunits of the hyperpolarization-activated cyclic nucleotide-gated current channel) in rodent trigeminal ganglia (TG) that innervate healthy teeth and determine if expression of HCN subunits is increased in TG following pulp exposure. METHODOLOGY: Pulps were exposed in right maxillary incisors of male Sprague-Dawley rats. After fixation, TG were removed, cryostat sectioned, and immunocytochemistry was utilized to study the expression of HCN1-3 subunits. Immunoreactivity of individual neurons from the maxillary region of the TG was determined with ImageJ software. Differences in the number immunopositive neurons amongst groups were tested for statistical significance with either a Yates or Pearson's chi-square or Fisher's exact probability tests depending on neuron sample size. Differences in the intensity of immunoreactivity between groups were tested for statistical significance with a Student's t-test. RESULTS: The majority of TG neurons were immunopositive for HCN1-3. Moreover, statistically significant increases in the number of TG neurons immunopositive for HCN1 and the intensity of HCN1-3 immunoreactivity were observed within hours of exposing the tooth pulp. CONCLUSIONS: HCN1-3 expression, as determined by immunocytochemistry, is increased within hours after injury. Given that I(h) can facilitate neuronal excitability, results of the current study suggest that antagonists to HCN1-3 subunits could work as analgesics in the alleviation of orofacial pain.