Distinct types of dorsal root ganglion sensory neurons may have unique contributions to chronic pain. Identification of primate sensory neuron types is critical for understanding the cellular origin and heritability of chronic pain. However, molecular insights into the primate sensory neurons are missing. Here we classify non-human primate dorsal root ganglion sensory neurons based on their transcriptome and map human pain heritability to neuronal types. First, we identified cell correlates between two major datasets for mouse sensory neuron types. Machine learning exposes an overall cross-species conservation of somatosensory neurons between primate and mouse, although with differences at individual gene level, highlighting the importance of primate data for clinical translation. We map genomic loci associated with chronic pain in human onto primate sensory neuron types to identify the cellular origin of chronic pain. Genome-wide associations for chronic pain converge on two different neuronal types distributed between pain disorders that display different genetic susceptibilities, suggesting both unique and shared mechanisms between different pain conditions.

In the present study, the effects of seminal plasma (SP), cumulus-oocyte-complexes (COCs) conditioned medium (CCM) and hyaluronan (HA) on functional changes and in vitro fertilizing ability of porcine spermatozoa were examined. In in vitro fertilization (IVF) experiments, 10% (v/v) of exogenous SP in the fertilization medium prevented sperm penetration (using fresh-extended and frozen-thawed ejaculated spermatozoa). Analysis of frozen-thawed CCM revealed a HA content to levels of 30 ng/mL per incubated COC. Presence of frozen-thawed CCM did not, however, prove effective to increase (furthermore decreasing) oocyte penetration in vitro, and neither did supplementation with exogenous HA at the same concentration as that present in the CCM (secreted by COCs). Analysis of sperm capacitation using the chlortetracycline (CTC) assay showed that frozen-thawed CCM had no elevating effect on 'B-pattern' spermatozoa (implying capacitation-like changes) and that addition of 10% (v/v) SP held spermatozoa in the 'F-pattern' (intact) status. Dose of 500 microg/mL HA and freshly prepared CCM increased, however, the frequency of capacitated spermatozoa (B-pattern) without resulting in increased rates of 'AR-pattern' (acrosome-reacted) spermatozoa, compared with controls. The present results confirm the decapacitating effect of SP and suggest capacitating actions of HA (dose-related) and CCM (freshly prepared) on boar spermatozoa in vitro. The unclear effects of frozen-thawed CCM and a low dose of HA on penetration rates of boar spermatozoa call for further researches of their function in vivo.

Highlights d Subcutaneous and intramuscular injection target the skin and the muscle, respectively d The immunization route affects the anatomical LN cluster that is targeted d Priming of adaptive immune responses occurs in the local vaccine-draining LNs d Comparable innate and adaptive immune responses with both immunization routes

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An experimental series was carried out to examine the ability of exogenous hyaluronan (HA) to facilitate monospermic penetration of in vitro-matured (IVM) oocytes during conventional porcine in vitro fertilization (IVF). IVM oocytes were co-cultured in fertilization medium with frozen-thawed, ejaculated boar spermatozoa. Addition of 0.5 mg/mL HA to the fertilization medium increased the rates of monospermic penetration (p < 0.05) on cumulus-denuded oocytes, without detrimentally affecting penetration rates (rates of monospermy were 57% and 78% in 2 mM caffeine-containing fertilization medium without and with HA, respectively). Caffeine and cumulus cells affected the rate of monospermic fertilization (p < 0.02-0.03), and caffeine affected the rate at which oocytes were penetrated (p < 0.02). Moreover, exogenous HA (0.5 mg/mL) in the sperm pre-incubation medium promoted monospermic penetration rates without affecting overall penetration rates. The effects of exogenous HA on sperm penetration and monospermy were also compared among semen batches. After semen was pooled from three fertile boars and held in homologous seminal plasma (SP) for 3 (batch A), 10 (batch B) or 20 (batch C) h prior to being cooled, spermatozoa from each semen batch were co-cultured with IVM oocytes. Monospermy of cumulus-denuded oocytes was increased (p < 0.05) by the addition of exogenous HA within the same semen batch (batches B and C). Furthermore, co-culture with cumulus cells did not mask the effects of HA on batch-B semen. The concentration of exogenous HA (0, 0.05 and 0.5 mg/mL) during sperm-oocyte co-culture differentially affected the rates of penetration and monospermy of cumulus-intact oocytes, depending on the time spermatozoa were held in SP before being frozen. The highest penetration rate was obtained by exposing semen to SP for 20 h prior to being cooled (0.05 mg/mL HA). The results suggest that exogenous HA decreases polyspermy during conventional porcine IVF. Furthermore, it appears that the action of HA is dependent upon the priming action of seminal plasma.

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