Altricial Rodents Research Articles

Astrocyte extracellular matrix modulates neuronal dendritic development.

Major developmental events occurring in the hippocampus during the third trimester of human gestation and neonatally in altricial rodents include rapid and synchronized dendritic arborization and astrocyte proliferation and maturation. We tested the hypothesis that signals sent by developing astrocytes to developing neurons modulate dendritic development in vivo. We altered neuronal development by neonatal (third trimester-equivalent) ethanol exposure in mice; this treatment increased dendritic arborization in hippocampal pyramidal neurons. We next assessed concurrent changes in the mouse astrocyte translatome by translating ribosomal affinity purification (TRAP)-seq. We followed up on ethanol-inhibition of astrocyte Chpf2 and Chsy1 gene translation because these genes encode for biosynthetic enzymes of chondroitin sulfate glycosaminoglycan (CS-GAG) chains (extracellular matrix components that inhibit neuronal development and plasticity) and have not been explored before for their roles in dendritic arborization. We report that Chpf2 and Chsy1 are enriched in astrocytes and their translation is inhibited by ethanol, which also reduces the levels of CS-GAGs measured by Liquid Chromatography/Mass Spectrometry. Finally, astrocyte-conditioned medium derived from Chfp2-silenced astrocytes increased neurite branching of hippocampal neurons in vitro. These results demonstrate that CS-GAG biosynthetic enzymes in astrocytes regulates dendritic arborization in developing neurons.

Dual isolation of primary neurons and oligodendrocytes from guinea pig frontal cortex.

Primary cell culture is a technique that is widely used in neuroscience research to investigate mechanisms that underlie pathologies at a cellular level. Typically, mouse or rat tissue is used for this process; however, altricial rodent species have markedly different neurodevelopmental trajectories comparatively to humans. The use of guinea pig brain tissue presents a novel aspect to this routinely used cell culture method whilst also allowing for dual isolation of two major cell types from a physiologically relevant animal model for studying perinatal neurodevelopment. Primary neuronal and oligodendrocyte cell cultures were derived from fetal guinea pig's frontal cortex brain tissue collected at a gestational age of 62 days (GA62), which is a key time in the neuronal and oligodendrocyte development. The major advantage of this protocol is the ability to acquire both neuronal and oligodendrocyte cellular cultures from the frontal cortex of one fetal brain. Briefly, neuronal cells were grown in 12-well plates initially in a 24-h serum-rich medium to enhance neuronal survival before switching to a serum-free media formulation. Oligodendrocytes were first grown in cell culture flasks using a serum-rich medium that enabled the growth of oligodendrocyte progenitor cells (OPCs) on an astrocyte bed. Following confluency, the shake method of differential adhesion and separation was utilized via horizontally shaking the OPCs off the astrocyte bed overnight. Therefore, OPCs were plated in 12-well plates and were initially expanded in media supplemented with growth hormones, before switching to maturation media to progress the lineage to a mature phenotype. Reverse transcription-polymerase chain reaction (RT-PCR) was performed on both cell culture types to analyze key population markers, and the results were further validated using immunocytochemistry. Primary neurons displayed the mRNA expression of multiple neuronal markers, including those specific to GABAergic populations. These cells also positively stained for microtubule-associated protein 2 (MAP2; a dendritic marker specific to neurons) and NeuN (a marker of neuronal cell bodies). Primary oligodendrocytes expressed all investigated markers of the oligodendrocyte lineage, with a majority of the cells displaying an immature oligodendrocyte phenotype. This finding was further confirmed with positive oligodendrocyte transcription factor (OLIG2) staining, which serves as a marker for the overall oligodendrocyte population. This study demonstrates a novel method for isolating both neurons and oligodendrocytes from the guinea pig brain tissue. These isolated cells display key markers and gene expression that will allow for functional experiments to occur and may be particularly useful in studying neurodevelopmental conditions with perinatal origins.

Neocortex neurogenesis and maturation in the African greater cane rat

BackgroundNeocortex development has been extensively studied in altricial rodents such as mouse and rat. Identification of alternative animal models along the “altricial-precocial” spectrum in order to better model and understand neocortex development is warranted. The Greater cane rat (GCR, Thyronomys swinderianus) is an indigenous precocial African rodent. Although basic aspects of brain development in the GCR have been documented, detailed information on neocortex development including the occurrence and abundance of the distinct types of neural progenitor cells (NPCs) in the GCR are lacking.MethodsGCR embryos and fetuses were obtained from timed pregnant dams between gestation days 50–140 and their neocortex was analyzed by immunofluorescence staining using characteristic marker proteins for NPCs, neurons and glia cells. Data were compared with existing data on closely related precocial and altricial species, i.e. guinea pig and dwarf rabbit.ResultsThe primary sequence of neuro- and gliogenesis, and neuronal maturation is preserved in the prenatal GCR neocortex. We show that the GCR exhibits a relatively long period of cortical neurogenesis of 70 days. The subventricular zone becomes the major NPC pool during mid-end stages of neurogenesis with Pax6 + NPCs constituting the major basal progenitor subtype in the GCR neocortex. Whereas dendrite formation in the GCR cortical plate appears to initiate immediately after the onset of neurogenesis, major aspects of axon formation and maturation, and astrogenesis do not begin until mid-neurogenesis. Similar to the guinea pig, the GCR neocortex exhibits a high maturation status, containing neurons with well-developed dendrites and myelinated axons and astrocytes at birth, thus providing further evidence for the notion that a great proportion of neocortex growth and maturation in precocial mammals occurs before birth.ConclusionsTogether, this work has deepened our understanding of neocortex development of the GCR, of the timing and the cellular differences that regulate brain growth and development within the altricial–precocial spectrum and its suitability as a research model for neurodevelopmental studies. The timelines of brain development provided by this study may serve as empirical reference data and foundation in future studies in order to model and better understand neurodevelopment and associated alterations.

Growing underground: Development of thermogenesis in pups of the fossorial rodent Ctenomys talarum

In mammals, during the pup’s development and adult life, integrated requirements of all activities of the individual must conform to a sustained rate of metabolism. Thus, partitioning the available energy according to short-term priorities at a specific moment allows animals to survive and optimize long-term reproductive success. In altricial rodents, thermal balance is a key factor for survival. When no exogenous source of heat is present, altricial pups rapidly lose heat, reaching ambient temperature (Ta). Fossorial rodents showed a strong dependence on burrows, where Ta remains relatively stable within narrow ranges. Pups of the fossorial rodent Ctenomys talarum are altricial, making them an excellent model to evaluate the development of thermogenic capacity. In this study, the ontogeny of the thermogenic capacity of pups of C. talarum was evaluated. Using respirometry techniques, non-shivering thermogenesis (NST), total thermogenic capacity (cold-induced maximum metabolic rate, MMR), and resting metabolic rate (RMR) in pups until post-weaning age (day 60) were analyzed. No NST was present in pups until day 60 despite the presence of molecular markers for NST in brown adipose tissue deposits, which became functional in adults. Although pups are altricial at birth, they maintain their thermal balance behaviorally during lactation. Total thermogenic capacity became fixed at an early age, indicating an improvement in shivering thermogenesis (ST) efficiency after day 10, which might be related to the development of musculature related to digging. Before the aboveground dispersal period (~day 60), pups gradually reached adult Tb by improving ST and thermal isolation, allowing them to confront climatic fluctuations on the surface.

Early life parameters and personality affect oxidative status during adulthood in an altricial rodent

It is increasingly recognized that alterations of the cellular oxidative status might be an important cost underlying challenging early life conditions. For example, an increased litter size can impose challenges as the offspring will face increased competition for maternal resources. Within a litter, individuals with relatively higher starting mass typically show higher growth rates, which can lead to increased oxidative damage. We investigated the long‐term consequences of these early life parameters on the oxidative status in mature mound‐building mice (Mus spicilegus). Individual differences in the animals' exploration tendency were assessed by repeated open field and novel object tests. We predicted less exploratory phenotypes, which typically show a higher stress responsiveness, to be particularly susceptible to possible effects of these early life parameters on oxidative status. We quantified oxidative damage of DNA (8‐hydroxy‐2'‐deoxyguanosine levels, 8‐OHdG) and proteins (protein carbonyl content, PCC), and activities of the antioxidants catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD) in liver and skeletal muscle tissue. 8‐OHdG levels were positively associated with CAT and SOD in both tissues, indicating that increased oxidative DNA damage was associated with an upregulation of antioxidant production. Hepatic DNA damage after maturity was increased in animals from larger litters. In less exploratory animals, DNA damage and the activity of CAT and SOD in the muscle were increased, but only in individuals with higher relative starting mass (measured on postnatal day 9). This interaction may be explained by the typically higher adrenocortical activity in less exploratory phenotypes and by the higher growth in relatively heavier pups, two factors known to increase oxidative stress. These findings contribute to enlightening the complex interplay between early life conditions, personality, and oxidative status.

Nestling Altricial Rodents and Lagomorphs Are Not Ectotherms.

AbstractBased on the synthesis of data on metabolic rates of 15 species investigated at moderate ambient temperatures, nestlings of most studied species of altricial rodents and lagomorphs exhibit thermoregulatory control of thermogenesis within 3 d after birth, signifying that they express physiological thermoregulation for 86%-100% of their time as nestlings. Classifying nestlings as ectotherms (or poikilotherms) is thus inappropriate and fosters misconceptions regarding their body temperatures (Tb's), control of Tb-sensitive functions (e.g., cardiac output), and energetics of development. The fact that nestlings live as litters in nests means that their thermoregulatory capacities in their actual, natural lives often far exceed the capacities they exhibit as isolated individuals-pointing to a pressing need for improved understanding of the physiology of litters. Litters in nests are already known in two cases to exhibit true homeothermy at ages when individuals studied in isolation express only modest thermoregulatory abilities. Golden (Syrian) hamster nestlings are exceptional, requiring 2 wk to develop thermoregulatory thermogenesis. They are properly considered ectotherms at up to 2 wk of age, helping to clarify that most species-being dramatically different-are not.

Comparative Milestones in Rodent and Human Postnatal Central Nervous System Development.

Within the substantially different time scales characterizing human and rodent brain development, key developmental processes are remarkably preserved. Shared processes include neurogenesis, myelination, synaptogenesis, and neuronal and synaptic pruning. In general, altricial rodents experience greater central nervous system (CNS) immaturity at birth and accelerated postnatal development compared to humans, in which protracted development of certain processes such as neocortical myelination and synaptic maturation extend into adulthood. Within this generalization, differences in developmental rates of various structures must be understood to accurately model human neurodevelopmental toxicity in rodents. Examples include greater postnatal neurogenesis in rodents, particularly within the dentate gyrus of rats, ongoing generation of neurons in the rodent olfactory bulb, differing time lines of neurotransmitter maturation, and differing time lines of cerebellar development. Comparisons are made to the precocial guinea pig and the long-lived naked mole rat, which, like primates, experiences more advanced CNS development at birth, with more protracted postnatal development. Methods to study various developmental processes are summarized using examples of comparative postnatal injury in humans and rodents.

Developmental environment influences activity levels in a montane rodent, Phyllotis xanthopygus

Ambient temperature and thermal variability play a crucial role in diverse aspects of organisms’ biology. In the current context of climate change, it is critical to understand how temperature impacts traits that could affect fitness. In Phyllotis xanthopygus, a small altricial rodent inhabiting an altitudinal gradient in the Andes Mountains of Argentina, the behavioral response to temperature varies between populations from different altitudes. Animals from high altitude (cold environment) reduce their activity rate at high temperatures, in contrast to animals from low altitude (relatively warmer environment). The goal of this study was to unveil the mechanism underlying such intraspecific behavioral variability in P. xanthopygus. We characterized activity rates under different thermal treatments both for wild-reared and lab-reared animals. As we expected, the intraspecific variability shown by animals raised at different altitudes in the field disappeared in animals raised under homogenous conditions in the laboratory. Our results are indicative of ontogenetic plasticity in P. xanthopygus and suggest that the behavioral versatility of adult individuals to deal with thermal challenges is shaped by the range of environmental conditions experienced during their early life. This adds to the list of features that modulate the biological performance of individuals and could influence the relative vulnerability of populations inhabiting different elevations under the global disturbance of climate change.

Functional Postnatal Maturation of the Medial Olivocochlear Efferent-Outer Hair Cell Synapse.

The organ of Corti, the auditory mammalian sensory epithelium, contains two types of mechanotransducer cells, inner hair cells (IHCs) and outer hair cells (OHCs). IHCs are involved in conveying acoustic stimuli to the CNS, while OHCs are implicated in the fine tuning and amplification of sounds. OHCs are innervated by medial olivocochlear (MOC) cholinergic efferent fibers. The functional characteristics of the MOC-OHC synapse during maturation were assessed by electrophysiological and pharmacological methods in mouse organs of Corti at postnatal day 11 (P11)-P13, hearing onset in altricial rodents, and at P20-P22 when the OHCs are morphologically and functionally mature. Synaptic currents were recorded in whole-cell voltage-clamped OHCs while electrically stimulating the MOC fibers. A progressive increase in the number of functional MOC-OHC synapses, as well as in their strength and efficacy, was observed between P11-13 and P20-22. At hearing onset, the MOC-OHC synapse presented facilitation during MOC fibers high-frequency stimulation that disappeared at mature stages. In addition, important changes were found in the VGCC that are coupled to transmitter release. Ca2+ flowing in through L-type VGCCs contribute to trigger ACh release together with P/Q- and R-type VGCCs at P11-P13, but not at P20-P22. Interestingly, N-type VGCCs were found to be involved in this process at P20-P22, but not at hearing onset. Moreover, the degree of compartmentalization of calcium channels with respect to BK channels and presynaptic release components significantly increased from P11-P13 to P20-P22. These results suggest that the MOC-OHC synapse is immature at the onset of hearing.SIGNIFICANCE STATEMENT The functional expression of both VGCCs and BK channels, as well as their localization with respect to the presynaptic components involved in transmitter release, are key elements in determining synaptic efficacy. In this work, we show dynamic changes in the expression of VGCCs and Ca2+-dependent BK K+ channels coupled to ACh release at the MOC-OHC synapse and their shift in compartmentalization during postnatal maturation. These processes most likely set the short-term plasticity pattern and reliability of the MOC-OHC synapse on high-frequency activity.

Post-weaning infant-to-mother bonding in nutritionally independent female mice.

Infant-parent attachment is highly selective and continues beyond essential care in primates, most prominently in humans, and the quality of this attachment crucially determines cognitive and emotional development of the infant. Altricial rodent species such as mice (Mus musculus) display mutual recognition and communal nursing in wild and laboratory environments, but parental bonding beyond the nursing period has not been reported. We presently demonstrated that socially and nutritionally independent mice still prefer to interact selectively with their mother dam. Furthermore, we observed gender differences in the mother-infant relationship, and showed disruption of this relationship in haploinsufficient Nbea+/- mice, a putative autism model with neuroendocrine dysregulation. To our knowledge, this is the first observation of murine infant-to-mother bonding beyond the nursing period.

Mechanisms of intergenerational transmission of polycystic ovary syndrome.

Developmental origins of adult disease (DoHAD) refers to critical gestational ages during human fetal development and beyond when the endocrine metabolic status of the mother can permanently program the physiology and/or morphology of the fetus, modifying its susceptibility to disease after birth. The aim of this review is to address how DoHAD plays an important role in the phenotypic expression of polycystic ovary syndrome (PCOS), the most common endocrinopathy of women characterized by hyperandrogenism, oligo-anovulation and polycystic ovarian morphology. Clinical studies of PCOS women are integrated with findings from relevant animal models to show how intergenerational transmission of these central components of PCOS are programmed through an altered maternal endocrine-metabolic environment that adversely affects the female fetus and long-term offspring health. Prenatal testosterone treatment in monkeys and sheep have been particularly crucial in our understanding of developmental programming of PCOS because organ system differentiation in these species, as in humans, occurs during fetal life. These animal models, along with altricial rodents, produce permanent PCOS-like phenotypes variably characterized by LH hypersecretion from reduced steroid-negative feedback, hyperandrogenism, ovulatory dysfunction, increased adiposity, impaired glucose-insulin homeostasis and other metabolic abnormalities. The review concludes that DoHAD underlies the phenotypic expression of PCOS through an altered maternal endocrine-metabolic environment that can induce epigenetic modifications of fetal genetic susceptibility to PCOS after birth. It calls for improved maternal endocrine-metabolic health of PCOS women to lower their risks of pregnancy-related complications and to potentially reduce intergenerational susceptibility to PCOS and its metabolic derangements in offspring.

Hair cell maturation is differentially regulated along the tonotopic axis of the mammalian cochlea.

Key points Outer hair cells (OHCs) enhance the sensitivity and the frequency tuning of the mammalian cochlea.Similar to the primary sensory receptor, the inner hair cells (IHCs), the mature functional characteristics of OHCs are acquired before hearing onset.We found that OHCs, like IHCs, fire spontaneous Ca2+‐induced action potentials (APs) during immature stages of development, which are driven by CaV1.3 Ca2+ channels.We also showed that the development of low‐ and high‐frequency hair cells is differentially regulated during pre‐hearing stages, with the former cells being more strongly dependent on experience‐independent Ca2+ action potential activity. Sound amplification within the mammalian cochlea depends upon specialized hair cells, the outer hair cells (OHCs), which possess both sensory and motile capabilities. In various altricial rodents, OHCs become functionally competent from around postnatal day 7 (P7), before the primary sensory inner hair cells (IHCs), which become competent at about the onset of hearing (P12). The mechanisms responsible for the maturation of OHCs and their synaptic specialization remain poorly understood. We report that spontaneous Ca2+ activity in the immature cochlea, which is generated by CaV1.3 Ca2+ channels, differentially regulates the maturation of hair cells along the cochlea. Under near‐physiological recording conditions we found that, similar to IHCs, immature OHCs elicited spontaneous Ca2+ action potentials (APs), but only during the first few postnatal days. Genetic ablation of these APs in vivo, using CaV1.3−/− mice, prevented the normal developmental acquisition of mature‐like basolateral membrane currents in low‐frequency (apical) hair cells, such as I K,n (carried by KCNQ4 channels), I SK2 and I ACh (α9α10nAChRs) in OHCs and I K,n and I K,f (BK channels) in IHCs. Electromotility and prestin expression in OHCs were normal in CaV1.3−/− mice. The maturation of high‐frequency (basal) hair cells was also affected in CaV1.3−/− mice, but to a much lesser extent than apical cells. However, a characteristic feature in CaV1.3−/− mice was the reduced hair cell size irrespective of their cochlear location. We conclude that the development of low‐ and high‐frequency hair cells is differentially regulated during development, with apical cells being more strongly dependent on experience‐independent Ca2+ APs.

A Novel Continuously Recording Approach for Unraveling Ontogenetic Development of Sleep-Wake Cycle in Rats.

Sleep-wake development in postnatal rodent life could reflect the brain maturational stages. As the altricial rodents, rats are born in a very undeveloped state. Continuous sleep recording is necessary to study the sleep-wake cycle profiles. However, it is difficult to realize in infant rats since they rely on periodic feeding before weaning and constant warming and appropriate EEG electrodes. We developed a new approach including two types of EEG electrodes and milk-feeding system and temperature-controlled incubator to make continuously polysomnographic (PSG) recording possible. The results showed that there was no evident difference in weight gaining and behaviors between pups fed through the milk-feeding system and warmed with temperature-controlled incubator and those kept with their dam. Evolutional profiles of EEG and electromyogram (EMG) activities across sleep-wake states were achieved perfectly during dark and light period from postnatal day (P) 11 to P75 rats. The ontogenetic features of sleep-wake states displayed that the proportion of rapid eye movement (REM) was 57.0 ± 2.4% and 59.7 ± 1.7% and non-REM (NREM) sleep was 5.2 ± 0.8% and 4.9 ± 0.5% respectively, in dark and light phase at P11, and then REM sleep progressively decreased and NREM sleep increased with age. At P75, REM sleep in dark and light phase respectively, reduced to 6.3 ± 0.6% and 6.9 ± 0.5%, while NREM correspondingly increased to 37.5 ± 2.1% and 58.4 ± 1.7%. Wakefulness from P11 to P75 in dark phase increased from 37.8 ± 2.2% to 56.2 ± 2.6%, but the change in light phase was not obvious. P20 pups began to sleep more in light phase than in dark phase. The episode number of vigilance states progressively decreased with age, while the mean duration of that significantly increased. EEG power spectra in 0.5–4 Hz increased with age accompanied with prolonged duration of cortical slow wave activity. Results also indicated that the dramatic changes of sleep-wake cycle mainly occurred in the first month after birth. The novel approaches used in our study are reliable and valid for continuous PSG recording for infant rats and unravel the ontogenetic features of sleep-wake cycle.

Malnutrition vs calorie restriction: The effects on maternal nutrient reduction in cardiac fetal mitochondrial function

Despite the recognized beneficial effects of caloric restriction, poor maternal nutrition alters development, predisposing offspring to chronic diseases including cardiovascular disease. Mitochondrial bioenergetics are central to cardiac energy metabolism and growth. Most developmental programming studies are in polytocous altricial rodents, which show different trajectories of development and maternal nutrient burden from monotocous precocial mammals, including humans. To enable translation to human programming, we developed a primate baboon model of moderate maternal nutrient reduction (MNR) in which mothers receive 70% global nutrition eaten by controls during pregnancy and lactation and offspring often showed growth restriction. Our hypothesis is that MNR adversely impacts fetal cardiac mitochondrial metabolism and homeostasis. Our results show in the baboon model that MNR increased fetal mtDNA content and transcription of key mitochondrial genes. MNR fetal oxidative phosphorylation was decreased and mitochondria contained sparse disarranged cristae. This study provides the first association between MNR and cardiac mitochondrial remodeling in the primate fetus. Thus, disruption of cardiac mitochondrial fitness may contribute to later life cardiac dysfunction.

Effect of moderate, 30 percent global maternal nutrient reduction on fetal and postnatal baboon phenotype.

Most developmental programming studies on maternal nutrient reduction (MNR) are in altricial rodents whose maternal nutritional burden and offspring developmental trajectory differ from precocial non-human primates and humans. Control (CTR) baboon mothers ate ad libitum; MNR mothers ate 70% global control diet in pregnancy and lactation. We present offspring morphometry, blood cortisol, and adrenocorticotropin (ACTH) during second half of gestation (G) and first three postnatal years. Moderate MNR produced intrauterine growth restriction (IUGR). IUGR males (n=43) and females (n=28) were smaller than CTR males (n=50) and females (n=47) in many measurements at many ages. In CTR, fetal ACTH increased 228% and cortisol 48% between 0.65G and 0.9G. IUGR ACTH was elevated at 0.65G and cortisol at 0.9G. 0.9G maternal gestational weight gain, fetal weight, and placenta weight were correlated. Moderate IUGR decreased body weight and morphometric measurements at key time points and altered hypothalamo-pituitary-adrenal function.

Estrogen receptor alpha is required in GABAergic, but not glutamatergic, neurons to masculinize behavior

Masculinization of the altricial rodent brain is driven by estrogen signaling during a perinatal critical period. Genetic deletion of estrogen receptor alpha (Esr1/ERα) results in altered hypothalamic-pituitary-gonadal (HPG) axis signaling and a dramatic reduction of male sexual and territorial behaviors. However, the role of ERα in masculinizing distinct classes of neurons remains unexplored. We deleted ERα in excitatory or inhibitory neurons using either a Vglut2 or Vgat driver and assessed male behaviors. We find that Vglut2-Cre;Esr1lox/lox mutant males lack ERα in the ventrolateral region of the ventromedial hypothalamus (VMHvl) and posterior ventral portion of the medial amygdala (MePV). These mutants recapitulate the increased serum testosterone levels seen with constitutive ERα deletion, but have none of the behavioral deficits. In contrast, Vgat-Cre;Esr1lox/lox males with substantial ERα deletion in inhibitory neurons, including those of the principal nucleus of the bed nucleus of the stria terminalis (BNSTpr), posterior dorsal MeA (MePD), and medial preoptic area (MPOA) have normal testosterone levels, but display alterations in mating and territorial behaviors. These mutants also show dysmasculinized expression of androgen receptor (AR) and estrogen receptor beta (Esr2). Our results demonstrate that ERα masculinizes GABAergic neurons that gate the display of male-typical behaviors.

Early life experience drives short-term acclimation of metabolic and osmoregulatory traits in the leaf-eared mouse.

We studied the putative effect of early life experience on the physiological flexibility of metabolic and osmoregulatory traits in the leaf-eared mouse, Phyllotis darwini, an altricial rodent inhabiting seasonal Mediterranean environments. Adult individuals were collected in central Chile and maintained in breeding pairs. Pups were isolated after weaning and acclimated to different temperatures (cold or warm) and water availability (unrestricted and restricted) until adulthood. Subsequently, individuals were re-acclimated to the opposite treatment. Rodents reared in the warm and subjected to water restriction had lower basal metabolic rate (BMR), total evaporative water loss (TEWL) and body mass (Mb) compared with those developing in the cold treatment; nevertheless, individuals subjected to warm temperatures had greater relative medullary thickness (RMT) and urine concentrating ability (UCA). Cold-reared rodents re-acclimated to warm conditions exhibited physiological flexibility of metabolic traits; however, their osmoregulatory attributes did not vary. Conversely, warm-reared rodents re-acclimated to cold had reduced RMT and UCA, but the metabolic traits of these individuals did not change. These results suggest a trade-off between metabolic performance and renal capabilities that might hinder physiological acclimation. Our results support the hypothesis of ontogenetic dependence of short-term acclimation in osmoregulatory and metabolic traits in P. darwini.

Timing of cardiomyocyte growth, maturation, and attrition in perinatal sheep.

Studies in altricial rodents attribute dramatic changes in perinatal cardiomyocyte growth, maturation, and attrition to stimuli associated with birth. Our purpose was to determine whether birth is a critical trigger controlling perinatal cardiomyocyte growth, maturation and attrition in a precocial large mammal, sheep (Ovis aries). Hearts from 0-61 d postnatal lambs were dissected or enzymatically dissociated. Cardiomyocytes were measured by micromorphometry, cell cycle activity assessed by immunohistochemistry, and nuclear number counted after DNA staining. Integration of this new data with published fetal data from our laboratory demonstrate that a newly appreciated >30% decrease in myocyte number occurred in the last 10 d of gestation (P < 0.0005) concomitant with an increase in cleaved poly (ADP-ribose) polymerase 1 (P < 0.05), indicative of apoptosis. Bisegmental linear regressions show that most changes in myocyte growth kinetics occur before birth (median = 15.2 d; P < 0.05). Right ventricular but not left ventricular cell number increases in the neonate, by 68% between birth and 60 d postnatal (P = 0.028). We conclude that in sheep few developmental changes in cardiomyocytes result from birth, excepting the different postnatal degrees of free wall hypertrophy between the ventricles. Furthermore, myocyte number is reduced in both ventricles immediately before term, but proliferation increases myocyte number in the neonatal right ventricle.

Depolarizing chloride gradient in developing cochlear nucleus neurons: Underlying mechanism and implication for calcium signaling

Precise regulation of the chloride homeostasis crucially determines the action of inhibitory transmitters GABA and glycine and thereby endows neurons or even discrete neuronal compartments with distinct physiological responses to the same transmitters. In mammals, the signaling mediated by GABAA/glycine receptors shifts during early postnatal life from depolarization to hyperpolarization, due to delayed maturation of the chloride homeostasis system. While the activity of the secondary active, K+-Cl−-extruding cotransporter KCC2, renders GABA/glycine hyperpolarizing in auditory brainstem nuclei of altricial rodents, the mechanisms contributing to the initially depolarizing transmembrane gradient for Cl− in respective neurons remained unknown. Here we used gramicidin-perforated patch recordings, non-invasive Cl− and Ca2+ imaging, and immunohistochemistry to identify the Cl−-loading transporter that renders depolarizing effects of GABA/glycine in early postnatal life of spherical bushy cells in the cochlear nucleus of gerbil. Our data identify the 1Na+:1K+:2Cl− cotransporter 1 (NKCC1) as the major Cl−-loader responsible for depolarizing action of GABA/glycine at postnatal days 3–5 (P3–5). Extracellular GABA/muscimol elicited calcium signaling through R-, L-, and T-type channels, which was dependent on bumetanide- and [Na+]e-sensitive Cl− accumulation. The “adult like”, low intracellular Cl− concentration is established during the second postnatal week, through a mechanism engaging the NKCC1-down regulation between P5 and P15 and ongoing KCC2-mediated Cl−-extrusion.

Photoperiodic effects on reproductive development in male cavies (Cavia aperea)

Age at maturity is a particularly important life history parameter, as it predicts potential life time reproductive success in many small mammal species. Animals should therefore optimize the timing of maturation by reacting to environmental stimuli that predict future environmental conditions. Photoperiod often reliably predicts ecological conditions. Animals born into a photoperiod indicating favorable conditions (spring, summer) may mature earlier and at a lower weight than animals born into conditions indicating unfavorable conditions (autumn, winter). So far most work was done on small, altricial rodent species and we still lack knowledge about their precocial relatives. Precocial animals are born much further developed than altricials and might show less plasticity in their ontogenetic trajectory than the latter.We tested the influence of photoperiod simulated by increasing (spring) or decreasing (autumn) light in climate chambers on important life history parameters in a medium sized rodent, the highly precocial cavy (Cavia aperea). We wanted to determine whether photoperiod influences timing of maturation and early growth in male cavies and whether patterns of testosterone in blood are reflected by patterns of testosterone metabolites in feces.Males born into simulated spring grew faster and matured at an earlier age than males born into simulated autumn conditions. Patterns of testosterone in blood correlated with testosterone metabolites measured in feces. Male cavies strongly react to predictive photoperiod cues by adjusting growth and timing of maturation as we found previously for females, corroborating the importance of seasonal cues for adjustments of life history.