208 Sperm Retention, Storage and Release from the Oviduct: A Story of Sugars, Steroids, and Channels

Because mating is not always synchronized with ovulation, females from many species store sperm in the female reproductive tract until ovulation and fertilization. This may be done for short periods, a day or two for swine and cattle, or longer periods. Other mammals, such as some species of bats, store sperm for several months. Chickens and turkeys store sperm for 2–4 weeks and queens of some species of insects store sperm for over a decade in specialized structures. How sperm are retained, kept fertile for varying times and released is unclear. We have identified two specific carbohydrate motifs that are abundant in the porcine oviduct that bind and retain sperm in the isthmus. When immobilized, these two glycans lengthen sperm lifespan and suppress the normal increase in intracellular Ca2+ that normally accompanies capacitation. Porcine sperm can be released from oviduct cells and immobilized glycans by progesterone, perhaps of ovarian or cumulus-oocyte complex origin, which activates CatSper, a sperm-specific Ca2+ channel. Progesterone, as well as other compounds that stimulate hyperactivated motility, trigger sperm release, suggesting that hyperactivated motility is sufficient to release porcine sperm from oviduct glycans. We also have found that blocking proteasome-induced sperm protein lysis diminishes the number of sperm released from oviduct glycans. Finally, a transcriptomic approach has identified several groups of genes that are differentially regulated in both bovine and porcine oviducts from estrus animals that are storing sperm compared to oviducts from diestrus animals. This provides clues about how sperm lifespan is extended during storage.

An acute dose of intranasal oxytocin rapidly increases maternal communication and maintains maternal care in primiparous postpartum California mice

Maternal-offspring communication and care are essential for offspring survival. Oxytocin (OXT) is known for its role in initiation of maternal care, but whether OXT can rapidly influence maternal behavior or ultrasonic vocalizations (USVs; above 50 kHz) has not been examined. To test for rapid effects of OXT, California mouse mothers were administered an acute intranasal (IN) dose of OXT (0.8 IU/kg) or saline followed by a separation test with three phases: habituation with pups in a new testing chamber, separation via a wire mesh, and finally reunion with pups. We measured maternal care, maternal USVs, and pup USVs. In mothers, we primarily observed simple sweep USVs, a short downward sweeping call around 50 kHz, and in pups we only observed pup whines, a long call with multiple harmonics ranging from 20 kHz to 50 kHz. We found that IN OXT rapidly and selectively enhanced the normal increase in maternal simple sweep USVs when mothers had physical access to pups (habituation and reunion), but not when mothers were physically separated from pups. Frequency of mothers’ and pups’ USVs were correlated upon reunion, but IN OXT did not influence this correlation. Finally, mothers given IN OXT showed more efficient pup retrieval/carrying and greater total maternal care upon reunion. Behavioral changes were specific to maternal behaviors (e.g. retrievals) as mothers given IN OXT did not differ from controls in stress-related behaviors (e.g. freezing). Overall, these findings highlight the rapid effects and context-dependent effect a single treatment with IN OXT has on both maternal USV production and offspring care.

Leptin receptor neurons in the dorsomedial hypothalamus regulate diurnal patterns of feeding, locomotion, and metabolism

The brain plays an essential role in driving daily rhythms of behavior and metabolism in harmony with environmental light–dark cycles. Within the brain, the dorsomedial hypothalamic nucleus (DMH) has been implicated in the integrative circadian control of feeding and energy homeostasis, but the underlying cell types are unknown. Here, we identify a role for DMH leptin receptor-expressing (DMHLepR) neurons in this integrative control. Using a viral approach, we show that silencing neurotransmission in DMHLepR neurons in adult mice not only increases body weight and adiposity but also phase-advances diurnal rhythms of feeding and metabolism into the light cycle and abolishes the normal increase in dark-cycle locomotor activity characteristic of nocturnal rodents. Finally, DMHLepR-silenced mice fail to entrain to a restrictive change in food availability. Together, these findings identify DMHLepR neurons as critical determinants of the daily time of feeding and associated metabolic rhythms.

An acute dose of intranasal oxytocin rapidly increases maternal communication and maintains maternal care in primiparous postpartum California mice

Maternal-offspring communication and care are essential for offspring survival. Oxytocin (OXT) is known for its role in initiation of maternal care, but whether OXT can rapidly influence maternal behavior or ultrasonic vocalizations (USVs; above 50 kHz) has not been examined. To test for rapid effects of OXT, California mouse mothers were administered an acute intranasal (IN) dose of OXT (0.8 IU/kg) followed by a separation test with three phases: habituation with pups in a new testing chamber, separation via a wire mesh, and finally reunion with pups. We measured maternal care, maternal USVs, and pup USVs. In mothers, we primarily observed simple sweep USVs, a short downward sweeping call around 50 kHz, and in pups we only observed pup whines, a long call with multiple harmonics ranging from 20 kHz to 50 kHz. We found that IN OXT rapidly and selectively enhanced the normal increase in maternal simple sweep USVs when mothers had physical access to pups (habituation and reunion), but not when mothers were physically separated from pups. Maternal-pup USVs were correlated upon reunion, but IN OXT did not influence this correlation. Finally, mothers given IN OXT showed a more positive change in retrievals/carrying and greater total maternal care upon reunion. Behavioral changes were specific to maternal behaviors (e.g. retrievals) as mothers given IN OXT did not differ from controls in stress-related behaviors (e.g. freezing). Overall, these findings highlight the rapid effects and context-dependent effect a single dose of IN OXT has on both maternal USV production and offspring care.

Leptin-receptor neurons in the dorsomedial hypothalamus regulate the timing of circadian rhythms in feeding and metabolism in mice

Animal behavior and metabolism are tightly coordinated with sleep-wake cycles governed by the brain in harmony with environmental light:dark cycles. Within the brain, the dorsomedial hypothalamic nucleus (DMH) has been implicated in the integrative control of feeding, energy homeostasis, and circadian rhythms [1], but the underlying cell types are unknown. Here, we identify a role for DMH leptin receptor-expressing neurons (DMHLepR) in these effects. Using a viral approach, we show that silencing DMHLepR neurons in adult mice not only increases body weight and adiposity, but also shifts circadian rhythms in feeding and metabolism into the light-cycle. Moreover, DMHLepR silencing abolishes the normal increase in dark-cycle locomotor activity characteristic of nocturnal rodents. Furthermore, DMHLepR-silenced mice fail to entrain to a restrictive change in food availability. Together, these findings identify DMHLepR neurons as critical determinants of the daily time of feeding and associated metabolic rhythms.

Abiotic stress impact on aerosol mass spectra over a forest site in Lithuania

Volatile organic compounds (VOCs) emitted by trees in response to abiotic stress evoke high levels of secondary organic aerosol (SOA) compounds. Few techniques exist to provide chemically-resolved submicron (PM1) particle mass concentrations and source apportionment of stress-induced emissions from trees and SOA formation. The chemical composition of atmospheric aerosol particles was characterized using an aerosol chemical speciation monitor (ACSM) at a mixed-mature forest site – the Aukštaitija Integrated Monitoring Station in the eastern part of Lithuania. The organic fraction of PM1 consisted of SOA (76%) and of anthropogenic combustion related primary organic aerosol (POA) (24%). The analysis of tree trunk circumference revealed three shrinkage and three normal increase episodes. During the episodes of tree trunk circumference shrinkage, several m/z signal (m/z 42, 43, 45, 48, 50) intensities were found to be magnified together with the daily SOA concentration. The stress response analysis confirm that tree trunk circumference shrinkage may be observed through the enhancement of selected m/z signals and result in increased SOA levels.

Determinants of Speculative Demand of Wheat and Its Impact on Consumer Welfare Loss

A much-neglected concept of price speculation from the side of consumers plays an important role in the determination of prices and quantity traded in the market. Almost all the consumers speculate about future prices of commodities and at the first stage, whenever prices increase, most of the consumers believe and speculate that the prices will increase more in the future. The current study underhand is conducted to find the factors of speculative demand for wheat and its relationship with consumer’s welfare. The need for this study was to find the factors of speculative demand and its and its relationship with the welfare loss. For this purpose data were collected from one hundred wheat consumers who buy wheat monthly from rural and urban areas of District Mandi Baha ud Din, Pakistan at random by direct interviewing to analyze which factors compel them not to buy wheat at annual bases and how it is related to their income, family size, and monthly income. Results show a negative relationship between the speculative demand for wheat and income. There is also a positive relationship between speculation and family as well as between speculation and actual quantity demanded. The lower-income consumers do speculation about the prices of the wheat from September to March and cause prices to increase about 8.92% more than the normal prices which cost them to lose consumer welfare and surplus. It is concluded that if consumers avoid speculation, they can buy wheat at 8.92% lesser price and can enjoy more consumer welfare and surplus. An increase in speculative demand increases prices more than a normal increase. The addition of some portion of price increment merely is due to consumer’s psychological phenomenon of speculation.

The normal increase in insulin after a meal may be required to prevent postprandial renal sodium and volume losses

Despite the effects of insulinopenia in type 1 diabetes and evidence that insulin stimulates multiple renal sodium transporters, it is not known whether normal variation in plasma insulin regulates sodium homeostasis physiologically. This study tested whether the normal postprandial increase in plasma insulin significantly attenuates renal sodium and volume losses. Rats were instrumented with chronic artery and vein catheters, housed in metabolic cages, and connected to hydraulic swivels. Measurements of urine volume and sodium excretion (UNaV) over 24 h and the 4-h postprandial period were made in control (C) rats and insulin-clamped (IC) rats in which the postprandial increase in insulin was prevented. Twenty-four-hour urine volume (36 ± 3 vs. 15 ± 2 ml/day) and UNaV (3.0 ± 0.2 vs. 2.5 ± 0.2 mmol/day) were greater in the IC compared with C rats, respectively. Four hours after rats were given a gel meal, blood glucose and urine volume were greater in IC rats, but UNaV decreased. To simulate a meal while controlling blood glucose, C and IC rats received a glucose bolus that yielded peak increases in blood glucose that were not different between groups. Urine volume (9.7 ± 0.7 vs. 6.0 ± 0.8 ml/4 h) and UNaV (0.50 ± 0.08 vs. 0.20 ± 0.06 mmol/4 h) were greater in the IC vs. C rats, respectively, over the 4-h test. These data demonstrate that the normal increase in circulating insulin in response to hyperglycemia may be required to prevent excessive renal sodium and volume losses and suggest that insulin may be a physiological regulator of sodium balance.

The role of Nrf2 in skeletal muscle contractile and mitochondrial function

Nuclear factor erythroid 2-related factor 2 (Nrf2) is a transcription factor that confers cellular protection by upregulating antioxidant enzymes in response to oxidative stress. However, Nrf2 function within skeletal muscle remains to be further elucidated. We examined the role of Nrf2 in determining muscle phenotype using young (3 mo) and older (12 mo) Nrf2 wild-type (WT) and knockout (KO) mice. Basally, the absence of Nrf2 did not impact mitochondrial content. In intermyofibrillar mitochondria, lack of Nrf2 resulted in a 40% reduction in state 4 respiration, which coincided with a 68% increase in reactive oxygen species (ROS) emission. Nrf2 abrogation impaired in situ muscle performance, characterized by a 48% greater rate of fatigue and a 35% decrease in force within the first 5 min of stimulation. Acute treadmill exercise resulted in a 1.5-fold increase in Nrf2 activation via enhanced DNA binding in WT animals. In response to training, cytochrome- c oxidase activity increased by 20% in the WT animals; however, this response was attenuated in KO mice. Nrf2 protein was reduced 30% by training. Despite this, exercise training normalized respiration, ROS production, and muscle performance in KO mice. Our results suggest that Nrf2 transcriptional activity is increased by exercise and that Nrf2 is required for the maintenance of basal mitochondrial function as well as for the normal increase in specific mitochondrial proteins in response to training. Nonetheless, the decrements in mitochondrial function in Nrf2 KO muscle can be rescued by exercise training, suggesting that this restorative function operates via a pathway independent of Nrf2.

Acute changes in neurovascular reactivity after subarachnoid hemorrhage in vivo

Subarachnoid hemorrhage causes acute and long-lasting constrictions of pial arterioles. Whether these vessels dilate normally to neuronal activity is of great interest since a mismatch between delivery and consumption of glucose and oxygen may cause additional neuronal damage. Therefore, we investigated neurovascular reactivity of pial and parenchymal arterioles after experimental subarachnoid hemorrhage. C57BL/6 mice were subjected to subarachnoid hemorrhage by filament perforation or sham surgery. Neurovascular reactivity was assessed 3 h later by forepaw stimulation or inhalation of 10% CO2. Diameters of cerebral arterioles were assessed using two-photon microscopy. Neurovascular coupling and astrocytic endfoot Ca2+ were measured in brain slices using two-photon and infrared-differential interference contrast microscopy. Vessels of sham-operated mice dilated normally to CO2 and forepaw stimulation. Three hours after subarachnoid hemorrhage, CO2 reactivity was completely lost in both pial and parenchymal arterioles, while neurovascular coupling was not affected. Brain slices studies also showed normal neurovascular coupling and a normal increase in astrocytic endfoot Ca2+ acutely after subarachnoid hemorrhage. These findings suggest that communication between neurons, astrocytes, and parenchymal arterioles is not affected in the first few hours after subarachnoid hemorrhage, while CO2 reactivity, which is dependent on NO signaling, is completely lost.