Sperm contributes diverse RNAs to the zygote. While sperm small RNAs have been shown to impact offspring phenotypes, our knowledge of the sperm transcriptome, especially the composition of long RNAs, has been limited by the lack of sensitive, high-throughput experimental techniques that can distinguish intact RNAs from fragmented RNAs, known to abound in sperm. Here, we integrate single-molecule long-read sequencing with short-read sequencing to detect sperm intact RNAs (spiRNAs). We identify 3440 spiRNA species in mice and 4100 in humans. The spiRNA profile consists of both mRNAs and long non-coding RNAs, is evolutionarily conserved between mice and humans, and displays an enrichment in mRNAs encoding for ribosome. In sum, we characterize the landscape of intact long RNAs in sperm, paving the way for future studies on their biogenesis and functions. Our experimental and bioinformatics approaches can be applied to other tissues and organisms to detect intact transcripts.

This study provides the first description of the ultrastructural features of sperm storage tubules (SSTs) in the uterovaginal region of the oviduct of the Indian garden lizard, Calotes versicolor. Abundant spermatozoa along with copious secretory material were found in the lumen of the SSTs. These secretory granules appeared similar in electron density to those found in the epithelial cells lining the SSTs indicating their similar origin. The close physical proximity of sperm with these granules suggests an intimate association between the two. The present study is also the first report of recovery of motile sperm from the flushings of SSTs in C. versicolor. The density of sperm found in the flushings varied, being most abundant during the reproductive phase and minimum/absent during the regressive phase. Understanding the microenvironment of the SSTs, the nature of the secretory granules and their interaction with sperm can guide us in unraveling the biology of oviductal sperm storage. Anat Rec, 298:1932Rec, 298: -1937Rec, 298: , 2015. V C 2015 Wiley Periodicals, Inc.Key words: uterovaginal junction; sperm storage tubules; secretory materialAbbreviations used: PAS 5 periodic acid-Schiff; SST 5 sperm storage tubule; TEM 5 transmission electron microscopy This article includes an AR WOW Video, which can be viewed at

Female sperm storage is an intriguing adaptation exhibited by a wide array of both vertebrates and invertebrates. The mechanisms underlying female sperm storage have remained elusive. Using the Indian garden lizard Calotes versicolor as a model organism, we investigated the role of low and high molecular weight factors in this phenomenon. Previously, we demonstrated three distinct phases of the reproductive cycle in this animal with live, motile spermatozoa recovered from the uterovaginal region during the reproductive phase. In the present study, we analysed the uterovaginal contents using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified an abundant protein band corresponding to ~55 kDa regardless of the phase of the reproductive cycle. Analysis of the purified protein by liquid chromatography-tandem mass spectrometry suggested a unique protein without any homology to the National Center for Biotechnology Information database. Exogenous addition of this protein to washed spermatozoa derived from the epididymis reversibly inhibited sperm motility in a concentration- and time-dependent manner, suggesting it plays a key role in sperm storage. These studies are likely to offer new avenues to unravel the secrets of female sperm storage seen across the animal taxa and may have novel applications not only in reproductive biology, but also in general cell storage and preserving endangered animal species.

An optimal intranasal (IN) dose of human mesenchymal stem cell-derived extracellular vesicles (hMSC-EVs), 90 min post-traumatic brain injury (TBI), has been reported to prevent the evolution of acute neuroinflammation into chronic neuroinflammation resulting in the alleviation of long-term cognitive and mood impairments. Since hippocampal neurogenesis decline and synapse loss contribute to TBI-induced long-term cognitive and mood dysfunction, this study investigated whether hMSC-EV treatment after TBI can prevent hippocampal neurogenesis decline and synapse loss in the chronic phase of TBI. C57BL6 mice undergoing unilateral controlled cortical impact injury (CCI) received a single IN administration of different doses of EVs or the vehicle at 90 min post-TBI. Quantifying neurogenesis in the subgranular zone-granule cell layer (SGZ-GCL) through 5′-bromodeoxyuridine and neuron-specific nuclear antigen double labeling at ~2 months post-TBI revealed decreased neurogenesis in TBI mice receiving vehicle. However, in TBI mice receiving EVs (12.8 and 25.6 × 109 EVs), the extent of neurogenesis was matched to naive control levels. A similar trend of decreased neurogenesis was seen when doublecortin-positive newly generated neurons were quantified in the SGZ-GCL at ~3 months post-TBI. The above doses of EVs treatment after TBI also reduced the loss of pre-and post-synaptic marker proteins in the hippocampus and the somatosensory cortex. Moreover, at 48 h post-treatment, brain-derived neurotrophic factor (BDNF), phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2), and phosphorylated cyclic AMP response-element binding protein (p-CREB) levels were downregulated in TBI mice receiving the vehicle but were closer to naïve control levels in TBI mice receiving above doses of hMSC-EVs. Notably, improved BDNF concentration observed in TBI mice receiving hMSC-EVs in the acute phase was sustained in the chronic phase of TBI. Thus, a single IN dose of hMSC-EVs at 90 min post-TBI can ease TBI-induced declines in the BDNF-ERK-CREB signaling, hippocampal neurogenesis, and synapses.

Long‐term sperm storage by females in various regions of the oviduct is documented across many invertebrate and vertebrate species. Although, many reports emphasize on the histology, histochemistry and ultrastructural features of sperm storage, very little is known about the mechanisms underlying the sperm storage. The current review documents the occurrence of sperm storage by females in a wide array of invertebrate and vertebrate species. This review also provides an insight on the presence of various molecular factors of the sperm storage tubules presumably responsible for the prolonged sperm storage with an emphasis on a model reptile, the Indian garden lizard, Calotes versicolor which contains a unique approximately 55‐kDa protein in its utero‐vaginal lavage and found to inhibit washed epididymal sperm motility in a concentration and time‐dependent manner in a reversible fashion.